Dairy Processing

Jagrani
Minj
Aparna Sudhakaran V
Anuradha Kumari Editors
Dairy Processing:
Advanced
Research
to Applications
Dairy Processing: Advanced Research to
Applications
Jagrani Minj • Aparna Sudhakaran V •
Anuradha Kumari
Editors
Dairy Processing:
Advanced Research to
Applications
Editors
Jagrani Minj Aparna Sudhakaran V
Department of Food Science and Department of Dairy Microbiology
Technology, Nebraska Innovation College of Dairy Science and Technology,
Campus (NIC), University of Nebraska Kerala Veterinary and Animal Sciences
Lincoln, NE, USA University (KVASU)
Mannuthy, Thrissur, Kerala, India
Anuradha Kumari
Dairy Chemistry, Guru Angad Dev
Veterinary and Animal Sciences University
Ludhiana, Punjab, India
ISBN 978-981-15-2607-7 ISBN 978-981-15-2608-4 (eBook)

https://doi.org/10.1007/978-981-15-2608-4
© Springer Nature Singapore Pte Ltd. 2020

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Preface
Being in the field of dairy for more than 15 years, the idea of writing a book came
to our mind to cover especially the applications of research in dairy processing. We
mainly focused to collect the different aspects of dairy processing like Dairy
Engineering, Dairy Technology, Dairy Chemistry, Dairy Microbiology, Dairy
Extension, Management and the advances in these fields in a single place. The
research data in each and every field of dairy science and processing are increasing
day by day. So, updating all the new and advanced research and their applications
from the lab to land scale is an initial and small step in the creation of this book. The
collective information on the current trends in dairy processing, research and appli-
cation is the ultimate aim behind this work.
Dairy products are highly preferred by people of all age groups. So, knowing
more about milk and its processing will be a good idea for the consumers and
researchers as well. During the processing of milk and milk products, it undergoes
several processing treatments like pasteurization, sterilization, homogenization,
freezing, condensing and drying, which is intended to either increase its quality or
shelf life or ease of storage. Engineering is involved in each step of the dairy sector
whether it is milking, processing of milk, production of different dairy products,
quality control, packaging, storage and distribution, cleaning-sanitation or effluent
treatment. The twenty-first century has seen progress in the use of new processing
technologies, non-thermal technologies, value addition, quality improvement, bet-
ter novel analytical techniques, development of health-promoting foods, by-product
utilization, etc.
In order to gain a better understanding and to disseminate the latest development
in this expanding field, a humble effort has been made in the form of this book. The
16 chapters in this book have covered different aspects of dairy processing.
Advancement in research, technology, novel health-promoting dairy products,
advanced analytical methods, quality challenges, and extension activities are also
discussed in this book.
This book is an attempt to share collective information on the advanced research
of dairy science to the undergraduate and postgraduate students, research scholars
and the academic institutions. Research and innovations in biofunctional dairy
products, probiotics, functional beverages, etc., their processing technologies and
their analysis by such advanced and innovative tools and techniques are elaborated
in this book for easy access to our readers. This book will be of interest to a wide
v
vi Preface
range of readers including dairy scientists, dairy engineers, dairy technologists,

nutritionists and dairy laboratory personnel worldwide.
The writing of this book was impossible without the valuable authors who con-
tributed by writing about different subjects of dairy science.
We thank Springer publishers for giving us this opportunity to write a book titled
‘Dairy Processing: Advanced Research to Applications’. We also thank the whole
editorial and production staff for making this possible.
Lastly, we express our thanks to our families for their understanding and emo-
tional support during the writing and editing of this book.
Lincoln, NE, USA Jagrani Minj

Thrissur, India Aparna Sudhakaran V
Ludhiana, India Anuradha Kumari

Contents
1 Basic Facts About Dairy Processing and Technologies�� 1

Aparna Sudhakaran V and Jagrani Minj
2 Non-thermal Processing of Dairy Foods�� 25
K. G. Rashmi and Aswin S. Warrier
3 Dairy Engineering: A Keystone to the Dairy Industry�� 51
Kesha D. Vankar
4 Advances in Dairy Engineering Research Across the Globe�� 63
Aswin S. Warrier
5 Significance of Fortification of Beneficial Natural Ingredients
in Milk and Milk Products�� 87
Jagrani Minj and Sonam Dogra
6 Multifunctional Aspects of Probiotics and Prebiotics in Health
Management: An Overview�� 119
Kamna Saini and Jagrani Minj
7 Research-Based Biofunctional Aspects of Milk Protein-Derived
Bioactive Peptides�� 133
Suvartan Ranvir, Nancy Awasti, Pranali Nikam, and
Neelima Sharma
8 Whey: Importance and Techno-functional Applications�� 161
Rita Mehla, Anuradha Kumari, Jyotika Dhankhar, Mitul Bumbadiya,
and Anuj Tyagi
9 Overcoming the Quality Challenges Across the Supply Chain�� 181
C. H. Aysha and S. Athira
10 Application of Research Methodologies in Dairy Chemistry�� 197
Anuradha Kumari, Niraj Kumar Singh, Sonika Choudhary, Dev
Priya, and Arpna Sharma
11 Approaches for Detection of Dairy Microorganisms: An Update �� 217
Aparna Sudhakaran V and Santosh Anand
vii
viii Contents
12 The Role of Yeast and Molds in Dairy Industry: An Update �� 243
Nancy Awasti and Santosh Anand
13 Dairy Industry: Hurdles Ahead in an Economic Perspective�� 263
Denny Franco and Bulbul G. Nagrale
14 Novel Milk and Milk Products: Consumer Perceptions�� 283
Anuradha Kumari, Himanshi Solanki, and Aparna Sudhakaran V
15 Novel Dairy-Based Drinks: Changing Scenario�� 301
Swathi P. Anand and Nancy Awasti
16 Extending the Horizons of Dairying to the Common Man:
An Indian Perspective�� 327
Aiswarya S. Panicker, M. Misha Madhavan, and Himanshi Solanki
Editors and Contributors
About the Editors
Jagrani Minj, Ph.D. (Dairy Microbiology), is currently a Postdoctoral Research

Scholar at the Department of Food Science and Technology, University of Nebraska
Lincoln, NE, USA. She worked as a research associate and as a senior officer in the
dairy industry for 9 months and 10 months, respectively. She has published a number
of research papers, book chapters and technical articles. She has also received best
oral presentation awards at conference. She holds a B. Tech. (Dairy Technology), and
M. Tech. and Ph.D. (Dairy Microbiology). During her professional career, she has
received various prestigious fellowships, including an Indian Council of Agricultural
Research-Junior Research Fellowship (ICAR-JRF), University Grant Commission-
Rajiv Gandhi National Fellowship (UGC-RGNF), an institutional fellowship, and
currently has a National Overseas Scholarship for her postdoctoral research work.
Aparna Sudhakaran V, Ph.D. (Dairy Microbiology), is an Assistant Professor of

Dairy Microbiology at Kerala Veterinary and Animal Sciences University at the
College of Dairy Science and Technology, Kerala, India. She has 6 years of experi-
ence in teaching dairy microbiology at undergraduate and postgraduate levels. She is
the recipient of numerous academic awards, including Gold Medal for B.Tech.,
Divisional Topper for M.Tech. and Ph.D.; AIR-1 in the Entrance Examinations for PG
and Ph.D.; Fellowships like the DST-INSPIRE Fellowship, ICAR Senior Research
Fellowship, ICAR Junior Research Fellowship and KAU Merit Scholarship. She also
received the ICAR-sponsored ‘Best Teacher Award’ from the institution in 2016–2017,
as well as a ‘Young Investigator Award’ from the Probiotic Association of India
(2018), best paper award for an article published in Indian Dairyman (2012) and four
awards for best poster/oral paper presented at professional conferences.
Anuradha Kumari, Ph.D. (Dairy Chemistry), is an Assistant Professor of Dairy

Chemistry at Guru Angad Dev Veterinary and Animal Sciences University,
Ludhiana, Punjab, India. She holds a B. Tech (Dairy Technology), and M.Tech. and
Ph.D. (Dairy Chemistry). She worked as a Senior Research Fellow for a year and
worked in the dairy industry for 10 months. Her research interests include func-
tional foods, detection of artificial sweeteners, use of food by-products and product
ix
x Editors and Contributors
development. She has published more than 11 papers in leading national and inter-
national journals, as well as review articles, book chapters and popular-science
articles, and she has also attended 8 international and national conferences. She is
an editorial board member of various national and international research journals.
She received an award for the best poster at a national conference.
Contributors
Santosh Anand, Ph.D. (Dairy Microbiology) Dairy Microbiology Department,

PJM College of Dairy Technology, Parkheta, Jharkhand, India
Swathi P. Anand, M.Tech. (Dairy Microbiology) New Product Development
(Dairy), Britannia Industries Ltd., Bengaluru, India
S. Athira, Ph.D. (Dairy Chemistry) College of Food Technology, Kerala
Veterinary and Animal Sciences University, Thrissur, Kerala, India
Nancy Awasti, Ph.D. (Dairy Science) Department of Dairy and Food Science,
South Dakota State University, Brookings, SD, USA
C. H. Aysha, M.Tech. (Dairy Microbiology) College of Food Technology, Kerala
Mitul Bumbadiya, M.Tech. (Dairy Chemistry) ICAR-National Dairy Research
Institute, Karnal, Haryana, India
Sonika Choudhary, Ph.D. (Dairy Chemistry) National Dairy Development
Board Dairy Services, New Delhi, India
Jyotika Dhankar, Ph.D. (Dairy Chemistry) Food Science and Technology,
MDU, Rohtak, Haryana, India
Sonam Dogra, Ph.D. (Dairy Microbiology) Punjab Pollution Control Board,
Zonal Office Laboratory Ludhiana, Ludhiana, Pujnab, India
Denny Franco, Ph.D. (Agriculture Economics) WTC, ICAR-IARI, New
Delhi, India
Anuradha Kumari, Ph.D. (Dairy Chemistry) Department of Dairy Microbiology,
College of Dairy Science and Technology, Kerala Veterinary and Animal Sciences
University (KVASU), Mannuthy, Thrissur, Kerala, India
Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana,

Punjab, India
Rita Mehla, Ph.D. (Dairy Chemistry) National Dairy Research Institute, Karnal,
Haryana, India
Jagrani Minj, Ph.D. (Dairy Microbiology) Department of Food Science and
Technology, Nebraska Innovation Campus (NIC), University of Nebraska,
Lincoln, NE, USA
Food Research and Development, SBPIL, Raipur, Chhattisgarh, India
Editors and Contributors xi
M. Misha Madhavan, Ph.D. (Agricultural Extension) ICAR-National Research

Centre on Pig, Indian Council of Agricultural Research, Guwahati, Assam, India
Bulbul G. Nagrale, Ph.D. (Dairy Economics) College of Dairy Technology,
Maharashtra Animal and Fishery Sciences University, Nagpur, India
Pranali Nikam, M.Tech. (Dairy Chemistry) ICAR-National Dairy Research
Aiswarya S. Panicker, Ph.D. (Dairy Microbiology) College of Dairy Science and
Technology, Kerala Veterinary and Animal Sciences University, Idukki, Kerala, India
Dev Priya, M.Sc. (Animal Biotechnology) DST Women Scientist-C, PFC-TIFAC,
New Delhi, India
Suvartan Ranvir, Ph.D. (Dairy Chemistry) Sam Higginbottom University of
Agriculture, Technology and Sciences, Allahabad, Uttar Pradesh, India
K. G. Rashmi, M.Tech. (Dairy Technology) Department of Dairy Technology,
College of Dairy Science and Technology, Thrissur, Kerala, India
Kamna Saini, Ph.D. (Dairy Microbiology) ICAR-National Dairy Research
Arpna Sharma, M.Sc. (Animal Biotechnology) Leibniz Institute of Farm Animal
Biology (FBN), Dummerstorf, Germany
Neelima Sharma, Ph.D. (Dairy Chemistry) ICAR-National Dairy Research
Niraj Kumar Singh, Ph.D. (Animal Biotechnology) College of Animal
Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University,
Himanshi Solanki, M.Tech. (Dairy Microbiology) DFIL-CreamBell, Gurgaon,
Haryana, India
Aparna Sudhakaran V, Ph.D. (Dairy Microbiology) Food Research and
Development, SBPIL, Raipur, Chhattisgarh, India
Dairy Microbiology Department, College of Dairy Science and Technology, Kerala


Anuj Tyagi, Ph.D. (Fish Microbiology) Guru Angad Dev Veterinary and Animal
Sciences University, Ludhiana, India
Kesha D. Vankar, M.Tech. (Dairy Engineering) Brand Sales, PI Industries,
Gurgaon, Haryana, India
Aswin S. Warrier, M.Tech. (Dairy Engineering) University Dairy Plant (Kerala
Veterinary and Animal Sciences University), Thrissur, Kerala, India
Abbreviations
3D Three dimensional
AA Amino Acids
ACE Angiotensin Converting Enzyme
AFLP Amplified Fragment Length Polymorphism
AFS Automatic Feeding Systems
AI Artificial insemination
AI Artificial Intelligence
AMS Automatic Milking Systems
AMUL Anand Milk Union Limited
AoA Agreement on Agriculture
APEJ Asia-Pacific excluding Japan
ARDRA Amplified rDNA Restriction Analysis
ATMA Agricultural Technology Management Agency
ATP Adenosine Triphosphate
BAPs Bioactive Peptides
BCT Block Chain Technology
B-GOS Bimuno-galactooligosaccharide
BHA Butylated Hydroxyanisole
BHT Butylated Hydroxy Toluene
BioMEMS or bioNEMS Biomedical or Biological Micro (and nano)-Electro-
Mechanical Systems
BLG β-lactoglobulin
BMCC Bulk Milk Chilling Centre
BOD Biological Oxygen Demand
BREW Binary Runtime Environment for Wireless
BSA Bovine Serum Albumin
CAC Codex Alimentarius Commission
CARD Computer-aided Administration of Registration
Department
CDC Cold Data Center
CDC Center for Disease Control and Prevention
cDNA complementary DNA
CFU Colony Forming Unit/Millilitre
CIEF Capillary Iso Electric Focusing
xiii
xiv Abbreviations
CIP Cleaning-in-place
CLA Conjugated linoleic acid
CMC Carboxy methyl cellulose
COB Clot-on-Boiling
COD Chemical Oxygen Demand
COP Coefficient of Performance
CPPs Caseinophosphopeptides
CPR Common Property Resources
CVD Cardiovascular Diseases
CZE Capillary Zone Electrophoresis
DA Development Alternative
DAHD & F Department of Animal Husbandry, Dairying and
Fisheries
DBD Dielectric barrier discharge
ddPCR Droplet Digital PCR
DEDS Dairy Entrepreneurship Development Scheme
DGGE Denaturing Gradient Gel Electrophoresis
DHA Docosahexaenoic acid
DMC Direct Microscopic Count
DNA Deoxyribonucleic acid
DNA Chips Deoxyribonucleic Acid Chips
dPCR Digital PCR
DPP-IV Dipeptidyl peptidase-IV
DSC Differential Scanning Calorimetry
dsDNA double-stranded DNA
DST Department of Science and Technology
DTA Differential Thermal Analysis
DVD Digital Versatile Disc
EA Emulsifying Activity
EC Emulsifying Capacity
EGF Epidermal Growth Factor
ELFA Enzyme-Linked ImmunoFluorescence Assay
ELISA Enzyme-Linked Immunosorbent Assay
EM Electromigration Techniques
EMA Ethidium Monoazide
EPA Eicosapentaenoic acids
EPEC Enteropathogenic Escherichia coli
ERIC Enterobacterial Repetitive Intergenic Consensus
ESI Electrospray Ionization
ESL Extended Shelf Life
ETEC Enterotoxigenic Escherichia coli
ETP Effluent Treatment Plant
EU European Union
FAO Food and Agriculture Organization of the
United Nations
Abbreviations xv
FDV Flow Diversion Valve

FFS Form Fill Seal
FIFO First In First Out
FISH Fluorescence In Situ Hybridization
FOS Fructo Oligo Saccharides
FRISBEE tool
Food Refrigeration Innovations for Safety, consum-
ers’ Benefit, Environmental impact and Energy
optimization
FSANZ Food Standards Australia New Zealand
FSSAI Food safety and standards authority of India
FT Fourier Transform
GC Gas Chromatography
GDA Guideline Daily Amount
GDP Gross Domestic Product
GE Gel Electrophoresis
GHG Greenhouse Gas
GI Gastrointestinal Tract
GIS Geographic Information System
GMOs Genetically Modified Organisms
GMP Glycomacropeptides
GOS Galacto Oligo Saccharides
GPS Global Positioning System
GRAS Generally Recognized as Safe
GSM Global System for Mobile communication
GSO GCC Standardisation Organisation
H2S Hydrogen Sulphide
HACCP Hazard Analysis and Critical Control Points
HAE Hereditary Angioedema
HBK Hariyali Kisaan Bazaar
HC-MCC Highly Concentrated Micellar Casein Concentrate
HDA Helicase-Dependent Amplification
HEPA High-Efficiency Particulate Air
HHP High Hydrostatic Pressure
HIUS High Intensity Ultrasound
HMF Hydroxyl Methyl Furfural
HPLC High Performance Liquid Chromatography
HPLT High Pressure Low Temperature
HPP High Pressure Processing
HRM High-Resolution Melt
HRP Horse Radish Peroxidase
HTST High Temperature Short Time
HUL Hindustan Unilever Limited
HWCF Healthy Weight Commitment Foundation
IBD Inflammatory Bowel Disease
IBS Irritable Bowel Syndrome
xvi Abbreviations
IC50 Inhibitory Concentration

ICAR Indian Council of Agricultural Research
ICMR Indian Council of Medical Research
ICT Computer-based Information and Communications
Technologies
ICTD Information and Communications Technologies for
Development
IDF International Dairy Federation
Ig Immunoglobulins
IgA Immunoglobulin A
IGF Insulin-like Growth Factor
IgG Immunoglobulin G
IgM Immunoglobulin M
IMS Immuno Magnetic Separation
IoT Internet of Things
IPP Isoleucine-Proline-Proline
ISO International Organization for Standardization
IT Information Technology
ITC India Tobacco Company Ltd.
KKC Kisan Call Centres
KKS Kallikrein–Kinin System
KVK Krishi Vigyan Kendras
LAB Lactic Acid Bacteria
LAMP Loop-Mediated Isothermal Amplification
LC Liquid Chromatography
LDL Low Density Lipoproteins
LoC Lab on chips
LPH Liter Per Hour
LTLT Low Temperature Long Time
LTST Low Temperature Short Time
MALDI-TOF Matrix-Assisted Laser Desorption Ionization Time-
Of-Flight Mode
MEE Multilocus Enzyme Electrophoresis
MIC Minimum Inhibitory Concentration
MIS Management Information System
MLEE Multi Locus Enzyme Electrophoresis
MLST Multi Locus Sequence Typing
MOCIT Ministry of Communications and Information
Technology
MS Mass Spectroscopy/Spectrometry
MSNF Milk Solids Not Fat
MSP Minimum Support Price
MSSRF M.S. Swaminathan Research Foundation
MSSRF FFMA MS Swaminathan Research Foundation Fisher Friend
Mobile Application
Abbreviations xvii
MST Milli Second Technology

MVLST Multi-Virulence-Locus Sequencing Typing
NABARD National Bank For Agriculture and Rural Development
NASBA Nucleic Acid Sequence-Based Amplification
NCDC National Collection of Dairy Cultures
NDDB National Dairy Development Board
NDP-1 National Dairy Plan Phase-1
NDRI National Dairy Research Institute
NeGP-A National e-Governance Plan in Agriculture
NGOs Non-Governmental Organizations
NGS Next Generation Sequencing
NIC National Informatics Centre
NIH National Institutes of Health
NISG National Institute for Smart Government
NLM National Livestock Mission
NPBB National Programme for Bovine Breeding
NPBB & DD National Programme for Bovine Breeding and Dairy
Development
NPDD National Programme for Dairy Development
NPN Non-Protein Nitrogen
OPC Oat Protein Concentrate
OPI Oat Protein Isolate
PAFC Photo Acoustic Flow Cytometry
PCM Phase Change Materials
PCR Polymerase Chain Reaction
PEF Pulsed Electric Field
PET Polyethylene Terephthalate
PFGE Pulsed-Field Gel Electrophoresis
PHE Plate Heat Exchanger
PI Proportional and Integral
PI Isoelectric Point
PID Proportional Integral Derivative
PLF Precision Livestock Farming
PMA Propidium Monoazide
PPP Public-Private Partnership
PTHrP Parathyroid Hormone-Related Protein
QCM Quartz Crystal Microbalance
qPCR quantitative PCR
RABIT Rapid Automated Bacterial Impedance Technique
RAPD Random Amplification of Polymorphic DNA
RAS Renin–Angiotensin System
RASI Rural Access Services through the Internet
RBP Ration Balancing Programme
RCA Rolling Circle Amplification
rep-PCR Repetitive Extragenic Elements Palindromic PCR
xviii Abbreviations
RFID Radio Frequency Identification

RFLP Restriction Fragment Length Polymorphism
RIA Radioimmunoassay
RNA Ribonucleic acid
RNS Reactive Nitrogen Species
ROS Reactive Oxygen Species
RPA Recombinase Polymerase Amplification
rRNA Ribosomal RNA
RRT Resazurin Dye Reduction test
RS Remote Sensing
RT PCR Reverse Transcriptase PCR
RT qPCR/qPCR/qRT PCR quantitative Real-Time Polymerase Chain Reaction
SARI Sustainable Access in Rural India
SC/ST Farmers Scheduled Castes (SCs)/Scheduled Tribes
(STs) Farmers
SCCM Smart Cold Chain Management
SCFE Supercritical Fluid Extraction
SDA Strand Displacement Amplification
SDS-PAGE Sodium Dodecyl Sulfate Polyacrylamide Gel
Electrophoresis
SHGs Self-Help Groups
SINED Society for Innovation & Entrepreneurship in
Dairying
SKEPL Shree Kamdhenu Electronics Pvt. Ltd.
SKU Stock Keeping Unit
SMEs Small and Medium Scale Enterprises
SMP Skim Milk Powder
SMS Short Message Service
SNF Solids Not Fat
SNPs Single Nucleotide Polymorphisms
SOD Superoxide Dismutase
SPC Standard Plate Count
SPR Surface Plasma Resonance
SPS Sanitary and Phytosanitary
SSG Special Safeguard
SSHE Scraped Surface Heat Exchanger
TARAhaat Technology and Action for Rural Advancement
TBI Technology Business Incubator
TGF Transforming Growth Factor
TLC Thin Layer Chromatography
TTGE Temporal Temperature Gradient Gel Electrophoresis
TTI Time Temperature Integrator
UHPH Ultra High-Pressure Homogenization
UHT Ultra High Temperature
UNDP United Nations Development Programme
Abbreviations xix
USA United States of America

USDA United States Department of Agriculture
USFA Unsaturated Fatty Acids
USFDA United States Food and Drug Administration
UV Ultraviolet
VBNC Viable But Non-Culturable
VHF Very High Frequency
VICTERS Virtual Class Technology on Edusat for Rural Schools
VLC Viable But Non-Culturable
VOICE Vijayawada Online Information Centre
vPCR viability PCR
VPP Valine-Proline-Proline
VTEC Verocytotoxigenic Escherichia coli
WGS Whole Genome Sequencing
WHO World Health Organization
WMP Whole Milk Powder
WPC Whey Protein Concentrate
WPI Whey Protein Isolate
WSN Wireless Sensor Networks
WTO World Trade Organization
XOS Xylooligosaccharides
α-la α-lactalbumin
β-lg β-lactoglobulin
Basic Facts About Dairy Processing
and Technologies 1
Aparna Sudhakaran V and Jagrani Minj
Contents
1.1 Introduction 2
1.2 Overview of the Global Dairy Industry 3
1.3 The Impact of Dairy Processing 4
1.4 Classification of Milk Products 5
1.5 Major Dairy Processing Area and Applied Technologies 6
1.5.1 Liquid Milk Processing 6
1.5.2 Fat-Rich Dairy Product Processing Technologies 9
1.5.3 Frozen Dairy Product Processing Technologies 11
1.5.4 Fermented Dairy Product Processing Technologies 12
1.5.5 Enzyme Coagulated Dairy Products 14
1.5.6 Dehydrated Dairy Products 15
1.5.7 By-Products in Dairy Industry 16
1.5.8 Specialized Dairy Products 19
1.6 Importance of Advanced and Innovative Technologies in the Dairy
Processing Sector 21
1.7 Conclusions 22
References 22
Aparna Sudhakaran V (*)

Food Research and Development SBPIL, Raipur, Chhattisgarh, India
Department of Dairy Microbiology, College of Dairy Science and Technology, Kerala
Veterinary and Animal Sciences University (KVASU), Mannuthy, Thrissur, Kerala, India
e-mail: aparna@kvasu.ac.in
J. Minj
Department of Food Science and Technology, Nebraska Innovation Campus (NIC),
University of Nebraska, Lincoln, NE, USA
© Springer Nature Singapore Pte Ltd. 2020 1

J. Minj et al. (eds.), Dairy Processing: Advanced Research to Applications,
https://doi.org/10.1007/978-981-15-2608-4_1
2 Aparna Sudhakaran V and J. Minj
Abstract
Milk and milk products are best known for their nutritional and functional value
and are considered as an inevitable part of the healthy diet. They are widely
cherished by all, irrespective of age. The perishable nature of milk highlights the
need of various processing technologies. The innovative technologies not only
increase productivity and profitability of the business, but also saves time and
energy. The dairy processing sector involves automation for the improvement of
overall dairy products’ quality and safety as well as reduces the environmental
impact. There is a substantial improvement in unit operations like separation,
standardization, pasteurization, homogenization, condensing, drying, freezing
and packaging. These developments enhanced the production capacity, automa-
tion and hygiene. It is one of the best way to increase food safety by reducing
outbreaks and health-related issues. The incorporation of novel ingredients and
use of novel processing and packaging technologies lead to the development of
innumerable varieties of value-added products with improved sustainability.
Keywords
Dairy processing · Dairy technology · Milk products · Milk · Dairy industry
1.1 Introduction
Milk and milk products are best known for their functional value and are considered
as an inevitable part of the healthy diet. Milk is considered as our primal nourishment
till weaning and is widely cherished by all age groups. Milk and its products are rich
in almost all the nutrients such as proteins, fat, milk sugar and vitamins and thus
makes it a ‘complete food’. It is one of the most nutritious foods which provides the
energy and nutrient requirements needed to ensure proper growth and development.
Dairying is the means of livelihood since the era of Sumerian civilization
(3000 BC) (Nemet-Nejat 1998). The preservation of milk by heat, dates back to the
domestication of cow and the use of fire (Holsinger et al. 1997). The modernization
and expansion of dairy farming began in the early 1900s after the discovery of pas-
teurization by Louis Pasteur. Pasteurization is a heating process which assures the
safety of the product and extends the shelf life by killing the pathogenic and
spoilage-causing microorganisms. The milk pasteurization was first adopted in
Germany and Denmark in the 1880s and later on expanded to Europe, America and
other countries (Maslehat and Mostafavi 2018). Subsequently, in 1898 the pasteuri-
zation of bottled milk was made compulsory in Denmark (Henderson 1971; Rankin
et al. 2017). In the 1900s, the easy availability of milk led to the wider consumption,
and the major thrust was given for the hygienic production of milk. In 1909, Chicago
in the United States became the first city to make pasteurization mandatory for milk
(Rankin et al. 2017). Later on, the 1920s, 1930s and 1940s witnessed the shift from
environmental hygiene to technological developments. By 1920s itself, sterilized or
1 Basic Facts About Dairy Processing and Technologies 3
pasteurized milk became popular (Atkins 2010). The mechanization of cream sepa-
ration, bottle washing, filling and capping marked the commencement of the mod-
ern milk processing industry. This period witnessed tremendous advancements in
dairy processing with respect to the capacities handled and the profits made, but the
diversification in case of milk products was minimal. In continuation with the devel-
opments in the previous centuries, the twenty-first century has seen progress in
value addition, quality improvement, improved cold chain and logistics manage-
ment, use of novel processing methods and better analytical techniques. Now, dairy
industry is one among the most organized and established sectors among all other
food sectors. It provides the major livelihood opportunity to farmers, processors,
technologists, engineers, retailers, shopkeepers and other stakeholders in the dairy
value chain (FAO Report 2018). The upcoming trend is for the development of
novel, customizable and personalized dairy products. This chapter briefly describes
the outlines of various dairy products and its processing.
1.2 Overview of the Global Dairy Industry
The Food and Agricultural Organization (FAO) estimated the global milk output as
843 million tonnes in 2018. There is a growth of 2.2% from 2017, contributed by
production expansions in India, Turkey, the European Union, Pakistan, the United
States and Argentina (FAO Report 2018). The cow and buffalo milk are the fore-
most contributor to the total world milk production. According to Eurostat, Asia is
the leading milk producer (30%), the European Union is the second-largest (28%)
and the North and Central America is the third-largest milk producer (18%). The
remaining milk producers such as South America contribute 9%, the other European
countries have 9%, Africa has 5% and Oceania contributes 5% to the overall world
milk production. The increased demand for dairy products in developing countries
can be correlated to the population growth. When it comes to the major contributors
of the dairy industry, India is the largest milk-producing country in the world since
1998, with a production of 165.54 million tonnes in 2016–2017 as reported by the
Department of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture,
Government of India. India contributes 19% of the world milk production. The milk
production was improved by 5.6% in 2018. The backbone of dairy processing in
India is the established dairy co-operative systems for proper processing and distri-
bution of milk and milk products, expanded dairy herds and higher productivity.
The per capita availability of milk has sharply increased to 375 g/day in 2017–2018,
as compared to the average world per capita availability of 302 g/day (National
Dairy Development Board 2018).
The world milk production and processing play a key role in the manufacturing,
storage and distribution of different types of dairy products as well as management
of dairy-based data. Different processing technologies have been applied to the vari-
ous types of milk and milk products. The major processed dairy products include
market milk, flavoured milk, cream, butter, butter oil/ghee, condensed and evapo-
rated milk, milk powder, fermented milk, yoghurt, cheese, ice cream, indigenous
dairy products, etc. The major processing technologies require a strong setup for
continuous production as well as the maintenance of the final product quality.
1.3 The Impact of Dairy Processing
The immediate processing of raw milk is the preliminary and crucial step in dairying.
As milk is considered as a perfect food for humans, it also provides nourishment and
ideal growth medium for microbes. Moreover, milk production is absolutely and
inseparably linked to the environment; milk and milk products are extremely vulner-
able to the environmental contaminants. The delay in the storage of raw milk at
refrigerated conditions leads to the development of microbial load in it, which further
affects the quality of the final product. From the safety point of view also, this is very
critical as it slow downs the fast growth of microbes in milk. The cold chain manage-
ment from ‘farm to table’ ensures the safety and quality of milk and milk products.
Quality of raw milk is the most significant sole aspect that decides the quality of
products made from it. Both compositional and hygienic qualities have an impact on
final product quality. The refrigeration and processing extends the shelf life of highly
perishable milk, thereby making it available every day in the supermarkets. The lon-
ger shelf life expanded the distribution pattern of milk and milk products globally.
The useable life of milk can be extended to several days, months and years, by vari-
ous processes such as pasteurization, dehydration and fermentation. Different pro-
cessing techniques can be used to convert milk to numerous value-added products,
thereby making the handling, storage and distribution easier.
Raw milk upon reaching dairy plant is first subjected to a sequence of rapid plat-
form tests to decide on the acceptance or rejection of milk which include organolep-
tic (taste, flavour, colour) test, pH, titratable acidity, clot on boiling (COB) test,
alcohol test, alcohol alizarin test, 10 min resazurin test (RRT), direct microscopic
count (DMC), etc. Several chemical tests can be done to detect adulterants, pesti-
cide and antibiotic residues, heavy metals, etc. The other tests like methylene blue
reduction test (MBRT), standard plate count (SPC), coliform count, etc. give a bet-
ter idea of the microbiological quality and the hygienic production of milk.
Moreover, a number of physicochemical or microbiological standards/limits are
given to serve as a guideline for monitoring the quality of raw milk and processing
factors as well as the finished products. The cleaning and sanitation in milk process-
ing is also a key step in the maintenance of product quality, the effective processing,
their quality and safety to elude major outbreaks. Cleaning in place (CIP) is one of
the most adopted automated cleaning and sanitation techniques used in the various
sections of the dairy industry.
1.4 Classification of Milk Products
Codex Alimentarius defines a milk product as a ‘product obtained by any processing

of milk, which may contain food additives, and other ingredients functionally nec-
essary for the processing’ (CODEX STAN 206-1999). There are numerous catego-
ries of milk-based products which vary considerably from region to region within a
country and among the countries. There are specific traditional milk products which
use indigenous technologies to produce some region-specific milk products. This
area is so vast in its own, making it beyond the scope of this chapter. Generally,
dairy products have been categorized on the basis of treatments undergone or com-
position. The classification of dairy products can be done basically into liquid milk,
fat rich, frozen, fermented, enzyme coagulated, concentrated and dried dairy prod-
ucts, by-products and specialized products on the basis of their processing technolo-
gies and is depicted in Fig. 1.1.
Liquid
milk
Fat rich
Specialized
milk
products
products
Frozen
By
products Milk milk
products
Dried milk Fermented

milk
products products
Enzyme
coagulated
milk
products
Fig. 1.1 Basic classification of milk products

1.5 Major Dairy Processing Area and Applied Technologies
1.5.1 Liquid Milk Processing
Among all the dairy products, liquid milk takes the prime position in processing,
marketing and consumption. Milk is enriched with proteins, fat, lactose, vitamins
and minerals. According to the FAO, consumption of milk gives on average 134 kcal
of energy/capita per day and is considered as the fifth largest energy provider, and it
comes on the third position on providing the good amount of fat and proteins.
According to the FAO, liquid milk can be categorized as depicted in Table 1.1.
Consumption of raw milk is generally not considered as safe, as it may contain
physical/chemical/microbiological hazards that gain entry from the environment or
during handling. The probability of occurrence of microbial hazards in milk and
milk products is more when compared with the chemical and physical hazards. The
consequences of microbiological hazards (endogenous or exogenous) are severe
due to its ‘dynamic’ nature as they can increase or decrease with the handling and
processing conditions. The occurrence of pathogens and spoilage microbes is
directly influenced by various reasons such as hygienic practices, farm management
practices, milking methods, animal health, number of animals in the farm, size of
the farm and milking season (Griffiths 2010). Contamination of feed, accidental
consumption or veterinary drug residues causes the entry of chemical hazards into
the dairy value chain at the primary production level. The contamination from raw
milk cannot be excluded completely, and hence these may cause certain risk to the
raw milk consumers. It can be minimized by hygienic milk practices.
Raw milk reception and immediate processing is a crucial step in the entire pro-
cess, as it is practically impossible to get superior quality products from inferior
quality raw milk. Under-processed milk significantly affects the quality of products
by increasing the microbial loads in the final products. However, it leads to spoilage
and makes it unfit for the human consumption. To augment the keeping quality of
raw milk, various technologies have been applied. The basic and popular processing
technique/unit operation applied for raw milk processing is pasteurization.
Table 1.1 Liquid milk Liquid milk

categories Raw milk
Pasteurized milk
Skimmed milk
Standardized milk
Sterilized milk
Ultrahigh temperature
milk
Flavoured milk
Reconstituted milk
Recombined milk
Fortified milk
Pasteurization is named after its inventor Louis Pasteur. Milk pasteurization refers
to ‘the process of heating every particle of milk to a specific temperature, holding it
at the same temperature for a specific duration (time) and followed by rapid chilling
to less than 7 °C’ (Dhotre 2014). The International Dairy Federation (IDF) defines
pasteurization as ‘a process applied to a product with the object of minimizing pos-
sible health hazards arising from pathogenic microorganisms associated with milk,
by heat treatment which is consistent with minimal chemical, physical, and organo-
leptic changes in the product’ (IDF 1986). There can be numerous possibilities of
temperature-time combinations for heat treatment to obtain the required microbial
or chemical effects like improvement in keeping quality, destruction of pathogens or
destruction of all microorganisms. Milk pasteurization ensures the killing of all
pathogens and spoilage microorganisms with a minimal nutritional loss, but it can-
not inactivate thermoresistant spores. Alkaline phosphatase enzyme is used to check
the pasteurization efficiency from 1935 (Davis 1955). The commonly employed
methods of pasteurization of milk are batch/vat pasteurization or low-temperature
long-time (LTLT) pasteurization (63 °C/30 min) and continuous pasteurization or
high-temperature short-time (HTST) pasteurization (72 °C/15 s). In most of the
countries, pasteurization is a mandatory operation for the commercial handling of
milk. The basic processes involved in pasteurized milk processing are raw milk
reception, clarification, preheating, separation, standardization, homogenization,
pasteurization and cooling.
Nowadays, people are more focusing on the consumption of low-calorie and
low-fat foods, thereby increasing the popularity of skim milk. The dairy industry
offers different varieties of milk by altering the fat level to get non-fat milk, low or
reduced-fat milk (<0.5% fat). In 1879, Gustaf de Laval designed the centrifugal
separators which separate cream and skim milk based on their density difference by
using centrifugal force. The skim milk is usually obtained as a by-product of cen-
trifugal cream separation process by the removal of fat from whole milk. Centrifugal
separation helps in the standardization of milk.
The standardized milk is obtained by the mixing of skim milk and cream so as to
obtain the desired fat and solids-not-fat (SNF) percent in the final product. Partial
skimming of whole milk to remove part of the fat or by mixing the whole milk with
skimmed milk being done so as to adjust fat and milk solids-not-fat (MSNF) to a
predetermined value.
The extended shelf life variety of liquid milk is sterilized milk. Sterilized milk is
a commercially sterile product obtained by high-temperature long-time heat treat-
ment. The milk has been filled in airtight containers and subjected to a temperature-
time combination of 115–120 °C for 10–20 min. The process intends to kill all
microorganisms including the spoilage and pathogenic microbes. The severe heat
treatment of the milk results in a caramelized, burnt flavour and a brownish colour
(Manners and Craven 2003). The consumers including adults and children dislikes
the increased cooked flavour in the extended-shelf-life milk (Lee et al. 2017). The
trend of sterilized milk has been declined by the introduction of ultrahigh tempera-
ture processing (UHT) or ultra pasteurization of milk.
UHT milk is also a commercially sterile product with extended shelf life. The major
attraction is that it can be stored at room temperature for a longer period. The typical
temperature-time combination for UHT treatment of milk is 130–150 °C for 1–3 s
(Manners and Craven 2003). The UHT treatment can be done either by direct steam
injection or indirect heating using heat exchangers. The UHT milk overcame the draw-
backs of sterilized milk to some extent. The shorter duration minimizes the nutritional
loss, but there is slightly cooked flavour when compared to pasteurized milk.
Some consumers do not like the natural flavour of milk. So the dairy industry
introduced the flavoured milk with a wide variety of flavours like vanilla, chocolate,
coffee, strawberry, banana, etc. having matching colours to satisfy the preferences
of consumers. Flavoured milk is defined as ‘ready-to-drink product prepared from
unfermented milk of varying fat contents, mixed with several ingredients such as
sugar or other sweeteners, cocoa powder, fruit juice, coffee, aroma agents, and/or
other ingredients and additives’. According to Codex, the milk constituent in such
products has to be an essential part in terms of quantity in the final product, and
other constituents not derived from milk should not take the place, in part or in
whole, of any milk constituent (Bisig and Kelly 2016). The flavoured milk can be
processed by pasteurization, sterilization or UHT treatment.
According to Codex, reconstituted milk product is defined as ‘a product resulting
from the addition of water to the dried or concentrated form of the product in the
amount necessary to re-establish the appropriate water to solids ratio’. The reconsti-
tuted milk is obtained from whole milk or skimmed milk powder (CODEX STAN
206-1999).
According to the Codex, recombined milk product is defined as ‘a product result-
ing from the combining of milk fat and milk solids-not-fat in their preserved forms
with or without the addition of water to achieve the appropriate milk product compo-
sition’. The basic difference between reconstituted milk and recombined milk is that
the recombined milk is obtained by adding water and milk fat separately in such pro-
portions that the desired fat content is being achieved (CODEX STAN 206-1999).
Food fortification has been defined as ‘the practice of deliberately increasing the
content of an essential micronutrient, i.e. vitamins and minerals (including trace
elements) in a food, so as to improve the nutritional quality of the food supply and
provide a public health benefit with minimal risk to health’ (WHO and FAO 2006).
Fortified milk is the milk enriched with one or more nutrients like vitamins A, D, B
or C, calcium and iron to milk. Fluid milk can be fortified with both vitamins A and
D, which are defined as optional ingredients for fluid milk in the Code of Federal
Regulations (FDA 2017). The vitamins are blended with inert carriers like unsatu-
rated vegetable oil-based concentrates (Manners and Craven 2003). Fortification of
skim/reduced milk helps to replace vitamins lost during the cream separation or
heat processing. The details about milk and milk products fortification is elaborated
in Chap. 5 of this book.
Table 1.2 Fat-rich dairy Cream

products Butter
Anhydrous butter fat (butter
oil)
Clarified butter fat (ghee)
Table 1.3 Classification of cream based on fat content and processing methods

Sl. no. Fat content Processing methods
1. Cream (18–26%) Pasteurized cream
2. Single or half cream (15–25%) Ultrahigh temperature (UHT) cream
3. Light/coffee cream (>10%) Frozen cream
4. Whipped cream >28% Dried cream
5. Heavy cream >35% Cultured cream/sour cream
6. Double cream >45% Sterilized cream
7. Plastic cream >80% Clotted cream
1.5.2 Fat-Rich Dairy Product Processing Technologies
As the name indicates, fat-rich dairy products are obtained by concentrating the fat in
milk. The basic unit operation involved is the centrifugal separation of cream. Different
processing technologies have been applied for the individual fat-rich milk products.
The basic ingredient for the production of fat-rich dairy products is cream which is
obtained by physical separation processes like skimming or centrifugal separation. The
major fat-rich dairy products are illustrated in Table 1.2. The quality of butter, butter oil
and ghee directly depends on the quality of the cream from which it is prepared.
According to Codex, ‘Cream is the fluid milk product comparatively rich in fat,
in the form of an emulsion of fat-in-skimmed milk, obtained by physical separation
from milk’ (CODEX STAN 288-1976). The cream can be in different forms like
reconstituted cream, recombined cream and prepared creams (prepackaged liquid
cream, whipping cream, cream packed under pressure, whipped cream, fermented
cream and acidified cream) (FAO and WHO 2010). The cream can be classified on
the basis of fat percentage, processing techniques and end uses which is illustrated
in Table 1.3 (Deosarkar et al. 2016).
The main principle of centrifugation is to separate the two phases, cream and skim
milk, on the basis of density difference (the specific gravity of milk fat is 0.93 and
skim milk 1.036 at 16 °C). The centrifugation process allows the rapid separation of
cream from milk, and it also removes the solid impurities such as dust and debris
from the milk. The standardization of cream is generally done to get the desired fat
level in cream. The cream is generally pasteurized before it is used for the prepara-
tion of butter and other fat-rich dairy products. However, cream pasteurization
slightly differs from milk pasteurization as it requires higher pasteurization tempera-
ture than the normal milk. According to the FAO, the time-temperature combination
suggested for cream is 75 °C for 15 s (10–20% fat), 80 °C for 15 s (above 20% fat)
and 65 °C for 30 min (batch pasteurization). The greater the viscosity or sugar con-
tent in the cream, the higher will be the temperature–time combination. In case of
UHT processing of cream, 135–150 °C for few seconds is practised so as to inacti-
vate spoilage and pathogenic microorganism with due consideration given to mini-
mal chemical, physical and organoleptic changes in cream (Deosarkar et al. 2016).
Some of the unit operations employed in cream manufacturing are neutralization
(partial reduction of acidity), homogenization (reduction of fat globule size to pre-
vent creaming), ripening (fermenting cream) and whipping (incorporation of air).
Butter, a popular dairy product, is usually prepared by the churning of cream
using batch churner or continuous butter making machine. The churning process
converts the cream into fat concentrate or compact mass of small fat globules (>80%
milk fat). The phase of cream changes from oil-in-water-type emulsion to water-in-
oil phase in butter. According to the Codex definition, ‘Butter is a fatty product
derived exclusively from milk and/or products obtained from milk, principally in
the form of an emulsion of the type water-in-oil’ (CODEX STAN 279-1971). The
buttermilk gets separated during churning of fresh or fermented cream. Butter is
available in different varieties as illustrated in Table 1.4. Butter is usually used as a
spread, for cooking, baking and frying purposes.
Anhydrous milk fat and butter oil are ‘fatty products derived exclusively from
milk and/or products obtained from milk by means of processes which result in
almost total removal of water and non-fat solids’ (CODEX STAN 280-1973).
According to FAO/WHO, anhydrous milk fat should have a minimum fat content of
99.8%, and maximum water content of 0.1%. The anhydrous milk fat being pro-
cessed from deep-frozen and normal butter or processing of cream or by fraction-
ation. Anhydrous milk fat can be used as an ingredient in the manufacture of several
products like recombined milk, sterilized milk and evaporated milk, chocolate and
other fat-containing sweets, ice cream and biscuits (Spreer 1998).
Ghee is a very popular and value-added milk product in India since Vedic times
(1500 BC). Ghee may be called as a form of high clarified butter. Ghee is a product
exclusively obtained from milk, cream or butter, by means of processes which result
in almost total removal of water and non-fat solids, with an especially developed
flavour and physical structure (CODEX STAN 280-1973). Ghee can be made either
by traditional or indigenous methods or by industrial methods. Industrial methods
of ghee manufacturing are creamery butter method, direct cream method, pre-
stratification method and continuous ghee making method (Pandya and Sharma
2002). In comparison with its western counterpart, butter oil, ghee has a character-
istic flavour, golden colour and a particular physical structure. Ghee has been used
for cooking and frying purposes, in Ayurvedic formulations and for some religious
Table 1.4 Classification of butter based on salt content, end use, processing methods and acidity
of cream
Sl. no. Salt content End use Processing methods Acidity of cream
1. Salted Table butter Pasteurized butter Sour cream (pH < 5.1)
2. Unsalted White butter Cultured butter Sweet cream (pH > 6.4)
Mildly acidified cream (pH 5.2–6.3)
Table 1.5 Frozen dairy Ice cream

products Frozen
yoghurt
purposes. The characteristic aroma and flavour of ghee and extended shelf life of up
to 2 years make it even more popular among the customers.
1.5.3 Frozen Dairy Product Processing Technologies
Frozen milk products are very popular among all age groups. The most common
frozen dairy products are illustrated in Table 1.5.
Ice cream is a popular frozen dairy food made by freezing the pasteurized ice
cream mix with an agitation and incorporation of air. The basic technology involved
in the preparation of ice cream mix is blending, homogenization, cooling and age-
ing, followed by freezing and incorporation of air/whipping (in scraped surface
freezers), packaging, hardening (in air-blast tunnels or rooms, conduction freezers)
and storage at −20 °C to keep it frozen. The high solid content in ice-cream mixes
makes it essential to have higher heat treatments than the normal pasteurization
requirements for liquid milk. Incorporation of sugar, different flavours, dried fruits
and nuts, fresh fruits, purees or probiotics and variations in composition, shape and
so on can create thousands of variations in ice cream. Freezing and whipping are the
most critical unit operations in ice cream making for the development of quality,
palatability and yield of the finished products. Ice cream can be classified on the
basis of manufacture/consumption into take-home category (purchased from gro-
cery stores and consumed at home), impulse (handheld, single-serving and eat-on-
the-spot products) or artisanal products (manufactured at the site of purchase and
consumption) (Deosarkar et al. 2016). Soft-serve ice cream is stored soft-frozen and
distributed to consumers without hardening. Many new and innovative processing
technologies in ice-cream manufacture have been implemented these days. Some of
them are low-temperature extrusion technique using scraped surface heat exchanger
(SSHE) (provides stability of the shape of ice cream at storage temperatures), ultra-
high pressure homogenization (low-fat ice cream) and application of cryogenics
(three-dimensional ice-cream products) (Hansen 2004; Jones 1997).
Frozen yoghurt combines the popularity of ice cream with the health benefits of
yoghurt. The process of frozen yoghurt is analogous to ice-cream making. It involves
preparation of yoghurt mix (usually 10%) by blending yoghurt with low-fat/non-fat
ice cream followed by freezing. The extruded soft-serve frozen yoghurt is very sim-
ilar to low-fat/non-fat ice cream with yoghurt flavour (Kilara and Chandan 2013).
1.5.4 Fermented Dairy Product Processing Technologies
Fermentation of milk by means of specific microorganisms which coagulate the

milk proteins leads to a wide diversity of fermented milk products. They are also
known as cultured milk products. Fermentation of milk is known to mankind since
10,000 BC (Canadian Dairy Commission 2012) and is one of the earliest preserva-
tion techniques. The microorganisms used for fermentation are known as starter
cultures which mainly include lactic acid bacteria, yeast and mould. The main lactic
acid bacteria involved in the fermentation of dairy processes are Lactococcus,
Lactobacillus, Pediococcus and Leuconostoc species. These lactic acid bacteria can
be either used as a single strain, mixed strain or in multiple mixed strains. The appli-
cation of specific strains depends upon the desired characteristics like taste, flavour,
appearance, body and texture of the final product. Some of the lactic acid bacteria
have specific biological activities which further enhance the biofunctional proper-
ties of the fermented milk products. These days well-known identified microbial
cultures along with their techno-functional properties are utilized for the making of
fermented milk and milk products. Some of the well-known fermented dairy prod-
ucts are illustrated in Table 1.6.
Yoghurt is very popular over other fermented milk products and has gained exten-
sive consumer acceptance as healthy food and has significantly evolved over time. At
present several varieties of yoghurt like plain yoghurt, fruit-flavoured yoghurt (includ-
ing fruit-on-the-bottom and blended forms), whipped yoghurt, granola- topped
yoghurt, drinkable yoghurt, frozen yoghurt, Greek yoghurt, varying fat contents (reg-
ular, low fat, and non-fat), low-sugar yoghurt and savoury flavoured yoghurt, probiotic
yoghurt, etc. are available in the market (Aryana and Olson 2017).
The technology involved in yoghurt manufacturing is the symbiotic relationship
of yoghurt cultures, Streptococcus thermophilus and Lactobacillus bulgaricus. The
ratio for both strains must be in 1:1 for the proper fermentation and development of
acetaldehyde flavour which makes it unique from other fermented milk products.
These days development of functional yoghurt is in trend. Functional yoghurt could
be made by the addition of different components such as probiotic bacteria, prebiot-
ics, dried fruits, fruit pulps, fruits, polyphenols, phytochemicals, stevia extract, etc.
so as to promote the health benefits. Many research data have been presented for the
various functional properties of yoghurt. Functional properties of yoghurt such as
Table 1.6 Fermented dairy Fermented milk

products products
Yoghurt
Sour cream
Fermented milk
Acidophilus milk
Kefir
Kumys/Koumiss/
Kumiss
antibacterial activity (Namaei et al. 2015; Taha et al. 2017; Mohammadi-Gouraji

et al. 2019), antioxidant property (Unal and Akalın 2012; Illupapalayam et al. 2014;
Taha et al. 2017; Dabija et al. 2018; Yoon et al. 2019; Najafi et al. 2019), immuno-
modulatory property, ace inhibitory activity (Unal and Akalın 2012; Najafi et al.
2019), anti-inflammatory activity (Gobbato et al. 2008; Yoon et al. 2019), hypocho-
lesterolemic activity (Ataie-Jafari et al. 2009) and anticancerous and antimutagenic
activity (Zhang et al. 2019) have been well studied and documented. Dahi/curd is a
fermented dairy product popular in India which is almost similar to yoghurt.
Sour cream and acidified sour cream are soured by using suitable acidifiers, with
or without the use of lactic acid bacteria. The titratable acidity of both of these prod-
ucts is at least 0.5%, expressed as lactic acid. The processing of sour cream involves
the standardization of cream (min 18% fat), followed by pasteurization (82 °C for at
least 10 min), homogenization, cooling (21 °C) and starter addition. The product
will be ripened to about 0.6% acidity, packed with minimal agitation, cooled to 4 °C
and aged for 12–24 h before marketing. A stabilizer has been considered to be effec-
tive to promote a firm body in this type of sour cream (Aryana and Olson 2017).
As per Codex ‘Fermented Milk is a milk product obtained by fermentation of
milk, which milk may have been manufactured from products obtained from milk
with or without compositional modification, by the action of suitable microorgan-
isms and resulting in the reduction of pH with or without coagulation (iso-electric
precipitation)’ (CODEX STAN 243-2003). Fermented milk is available as drinks,
flavoured and concentrated forms (Stragisto, Labneh, Ymer, Ylette). Some of the
traditional fermented milks are given in Table 1.7.
Kefir is an acid alcoholic fermented milk product which has been recently gain-
ing popularity due to its health benefits. It is the traditional fermented dairy product
in the Caucasus, Tibetan or Mongolian mountains prepared mainly from cow, buf-
falo, goat or ewe’s milk (Rosa et al. 2017). It is a highly viscous acidic beverage
which contains variable levels of alcohol and carbon dioxide (Ribeiro and Ribeiro
2010). Instead of a particular microorganism, kefir grains resembling cauliflower
florets initiate fermentation which contains ‘kefiran’, a polysaccharide. It contains a
wide variety of yeasts, acetic acid bacteria, Lactobacillus, Lactococcus and
Leuconostoc in a symbiotic relation. Kefir grains can be reused after product prepa-
ration and are passed through generations to generation. The product can be made
from raw, pasteurized or UHT-treated milk (Lopitz-Otsoa et al. 2006). Kefir grains
can be industrially produced mainly by two methods—Russian method (fermenta-
tion of the grains and using the percolate) and direct to vat inoculation/set (DVI/
DVS) (cultures isolated from kefir grains) (Prado et al. 2015). Kefir has been
reported with improved digestion and tolerance to lactose, antibacterial effect,
hypocholesterolemic effect, control of plasma glucose, antihypertensive effect, anti-
inflammatory effect, antioxidant activity, anticarcinogenic activity, antiallergenic
activity and healing effects (Rosa et al. 2017). Kefir grains can act as an excellent
vehicle for the probiotic delivery (Prado et al. 2015).
Koumiss/kumiss/kumys is an acid alcoholic fermented product prepared from
mare or goat milk by yeast lactic fermentation. It is traditionally consumed in Russia
and western Asia. In Mongolia, kumys, called airag, is the national drink, and
Table 1.7 Fermented milk available in different countries

Wider region Fermented milk Country/region Type of milk
Asia
Arkhi/Airag Mongolia Mare
Dahi India, Pakistan, Nepal, Bhutan Cow, buffalo, mixed milk
Chakka/Chakah India, Pakistan, Afghanistan Cow, buffalo
Churpi Nepal, India Cow, buffalo
Koumiss Mongolia, Kazakhstan, Russia Mare
Labneh Middle East All milk
Tarag China Goat
Kurut China Yak
America
Suero costeno Columbia Cow
Boruga Dominican Republic Cow
Sour milk Nicaragua Cow
Europe
Lactofil Sweden Cow
Skyr Iceland Ewe, Cow
Ymer Denmark Cow
Kefir Russia Ewe, cow
Koumiss Russia Mare, goat
Taette/Lapp’s Scandinavia Cow
Ayran Turkey Cow, ewe, goat
Bulgarian milk Bulgaria Ewe, cow
Oceania
Acidophilus Australia Various milk
Africa
Laban zeer Egypt All types
Amasi Zimbabwe Bovine
Roab Sudan Cow, goat, sheep
Garoor Somalia Cow
Madila Southeast Africa Cow
distilled kumys, arkhi, has also been produced (Kanbe 1992; Orskov 1995; Uniacke-
Lowe et al. 2010).
1.5.5 Enzyme Coagulated Dairy Products
The major enzyme coagulated dairy product is cheese which is a fermented dairy
product. It is produced worldwide in a great diversity of flavours, textures and
forms; there are more than 1000 varieties of cheese available in the market. As per
Codex, ‘cheese is the ripened or unripened soft, semi-hard, hard, or extra-hard prod-
uct, which may be coated, and in which the whey protein/casein ratio does not
exceed that of milk’ (Codex 283-1978). Cheese can be classified based on the mois-
ture content as illustrated in Table 1.8.
Cheese is basically the protein coagulation obtained by coagulating wholly or

partly to the protein of milk, skimmed milk, partly skimmed milk, cream, whey
cream or buttermilk or any combination of these materials. The coagulation is usu-
ally done by the action of rennet or other suitable coagulating agents followed by
partially draining of the whey. The fresh cheese can be ripened either by bacteria or
mould so as to get desirable flavour, body and texture.
1.5.6 Dehydrated Dairy Products
Dehydrated milk products are obtained by partial removal of water from milk and
are one of the best methods of preservation of milk. Some of the common concen-
trated dehydrated milk products available are given in Table 1.9.
Evaporated milk is obtained by the evaporation or removal of water from the
whole milk or skimmed milk. It is also known as sterilized unsweetened condensed
milk. According to Codex, evaporated whole milk is defined as ‘Milk and cream
from which the water has been partly removed and which has been heat-treated to
render it bacteriologically safe and stable product’ (CODEX STAN 281-1971). The
classification of evaporated milk is illustrated in Table 1.10.
The sterilization process is usually done at 118–122 °C for several minutes to
ensure the complete sterilization with minimal nutritional loss. The process kills all
bacterial species, mainly the Bacillus stearothermophilus, and deactivates enzymes
and spores. Generally, evaporated milk has been sterilized in their containers (bot-
tles or cans). Commercial sterilization of evaporated milk makes it more stable at
ambient temperature. It is used in cooking and for coffee and tea making purposes.
In 1856, Gail Borden (USA) started to make the first sweetened condensed milk in
the hermetically sealed cans to explore his business in the dairy industry. It is obtained
by partial removal water from whole or skimmed milk and can be sweetened or
Table 1.8 Classification of Enzyme coagulated milk products

cheese Cheese
Fresh cheese
Very hard
Hard
Semi hard
Soft
Processed
Table 1.9 Dehydrated milk Evaporated milk

products Sweetened condensed
milk
Milk powder
Table 1.10 Classification of evaporated milk

Milk protein
Milk fat (%m/m) in milk solids-not-fat
Types (%m/m) Minimum
Evaporated milk Minimum 7.5 34
Evaporated skimmed milk Maximum 1 34
Evaporated partly skimmed milk Minimum 1– 34
Maximum 7.5
Evaporated high-fat milk Minimum 15 34
Table 1.11 Classification of condensed milk

Milk
solids Milk protein (%m/m) in
Milk fat (%m/m) milk solids-not-fat
Types (% m/m) Minimum Minimum
Sweetened condensed milk Minimum 8.0 28 34
Sweetened condensed Maximum 1 24 34
skimmed milk
Sweetened condensed partly Minimum 24 34
skimmed milk 1–maximum 8
Sweetened condensed Minimum 16 14 34
high-fat milk
unsweetened. As per Codex, ‘Sweetened condensed milk is milk product which can
be obtained by the partial removal of water from milk with the addition of sugar, or by
any other process which leads to a product of the same composition and characteris-
tics’ (CODEX STAN 282-1971). The types of condensed milk are given in Table 1.11.
Condensed milk is not a sterilized product, but the commercial sterility is attained by
the addition of sugar and heat treatment. It is usually used in cooking and baking
purposes.
Dried milk or milk powder is the product obtained by the removal of water by
heat or the suitable means to less than 5% or less moisture. As per Codex definition,
‘Dry milk/milk powder is milk and cream from which water has been completely
removed by various methods and can be in the form of powder, granules or other
solid forms’ (CODEX STAN 207-1999). It may contain added sugar or other sweet-
eners. Nowadays, spray-drying technology is used for dried milk production. Some
of the most common dried milk products like whole milk powder, skimmed milk
powder and dairy whitener are given in Table 1.12. The purpose of drying is to
increase shelf life, reduce the bulk and storage and make transportation easier.
1.5.7 By-Products in Dairy Industry
By-products of yesterday have the potential to become the major products of today.
Every component in milk should be sensibly utilized owing to its nutritive and eco-
nomic value. A dairy by-product may be defined as a product of commercial value
Table 1.12 Types of milk powder

Moisture Milk protein (%m/m) in milk
Milk fat (%m/m) solids-not-fat
Types (% m/m) Maximum Minimum
Whole milk powder Minimum 26– 5 34
maximum 42
Partly skimmed milk Minimum 1.5– 5 34
powder maximum 26
Skimmed milk Maximum 1.5 5 34
powder
Cream powder Minimum 42 5 34
Cream
Butter
Skim milk
•Casein
Cheese
Butter milk
•Edible
•Beverages Whey
•Non Edible (Industrial)
•Caseinates •Whey hydrolysates
•Coprecipitates •Whey protein concentrates
•Hydolysates •Whey protein Isolates
•Lactose
Fig. 1.2 By-products of the Dairy Industry
produced during the manufacture of the main product. The major by-products in the
dairy industry are skim milk, buttermilk and whey produced during the manufacture
of cream, butter and cheese, respectively, and are illustrated in Fig. 1.2. There are
remarkable advancements in the processing and utilization of by-products. Some
other dairy by products are casein, caseinates, coprecipitates, protein hydrolysates,
protein concentrates, lactose, etc.
Skim milk is the by-product obtained during cream separation of whole milk. It
is mainly used for the standardization process (high SNF content), consumption in
liquid form or preserved in powder form. Buttermilk is the major by-product
obtained during butter making. There are mainly two kinds of buttermilk—sweet
and sour. The sweet cream buttermilk resembles skim milk in composition and
often admixed with the bulk of skim milk before drying. Sour buttermilk is obtained
from churning of sour cream and is usually converted into beverages. Whey is the
largest by-product produced during the production of paneer, cheeses and copre-
cipitates. It is the liquid part of milk obtained after curd separation. It can be acid
whey (acid coagulated cheese) or sweet whey (rennet coagulated cheese). It is a
precious source of lactose, proteins, minerals and vitamins altogether. Whey
proteins are next to egg proteins in terms of nutritive value. Whey is usually con-
verted to whey cheese, whey drinks and fermented whey beverages, whey protein-
enriched beverages, whey-based sports beverages, alcoholic and nonalcoholic
beverages, carbonated beverages, etc. for utilizing the nutritive value of whey pro-
teins. Whey is used as the raw material for the manufacture of whey powders, lac-
tose, whey proteins (concentrates and isolates), etc. It can be preserved as condensed
whey and whey powder which can be used as an ingredient in various foods and
feed industry. Whey can also be used as fermentation media and for the production
of alcohol, methane, organic acids, biomass, etc.
Casein is the next major derived by-product that can be edible or nonedible
depending on the coagulant used. The nonedible casein known as industrial casein
has been widely used in the textile, paint, rubber and leather industry. Edible casein
and caseinates are used in many food products as a source of protein and calcium.
Caseinates are the soluble form of casein obtained by neutralizing acid or rennet
casein with alkali and then drying of the product. The commonly prepared casein-
ates are sodium and calcium caseinates along with potassium, ammonium, citrate
and magnesium caseinate. It is used as an ingredient in processed food products and
in pharmaceuticals. The milk protein concentrates (>50% protein) are prepared by
ultrafiltration/diafiltration prior to drying. Coprecipitates are the whey proteins pre-
cipitated along with casein by heating the milk to denature the whey proteins and
isoelectric precipitation of casein by acidification. It is the protein which separates
as a solid phase after the heat treatment and precipitation. The coprecipitates have
better yield and functional properties. It is used as an ingredient in various products
like processed dairy, meat, bakery and confectionaries and animal and pet foods.
Protein hydrolysates can be derived from casein and whey proteins and are used in
functional, dietetic, geriatric, hypoallergenic, nutritional and formulated foods and
high-energy supplements. Casein hydrolysates are prepared by acid, alkaline or
enzymatic hydrolysis of casein. The whey protein concentrates (WPC) (>35% whey
proteins) obtained by ultrafiltration can be used as a substitute for non-fat dry milk
(NFDM) and are a rich source of proteins, vitamins and minerals. The predigested
proteins facilitate faster intestinal absorption and reduced allergenicity.
Nowadays, modern processing techniques like ultrafiltration, reverse osmosis,
hydrolysis, electrodialysis, ion exchange, fermentation and protein fractionation are
used to convert the various by-products, especially whey into a major source of
ingredients and as a coproduct of the dairy industry. These technologies are eco-
nomically viable for the commercial utilization of by-products as well as to increase
the availability of valuable nutrients, and it also reduces the environmental pollution
originating from dairy waste. The major dairy by-products and processing methods
are illustrated in Table 1.13.
Table 1.13 The processing technologies employed in by-product utilization

Sl. Processing
no. Main product By-product method Products to be made
1. Cream Skim milk Pasteurization Flavoured milk
Sterilization Sterilized flavoured milk
Fermentation Cultured buttermilk
Fermentation and Concentrated sour skim milk
concentration
Concentration Plain and sweetened condensed skim
milk
Drying Dried skim milk or skim milk powder
or non-fat dry milk (NFDM)
Coagulation Cottage cheese, Quarg, edible casein,
caseinates, hydrolysates, coprecipitates
2. Butter Buttermilk Fermentation and Condensed buttermilk
concentration
Concentration and Dried buttermilk
drying
Coagulation Soft cheese
3. Cheese, Whey Fermentation Whey beverage, yeast whey
casein, Concentration Plain and sweetened condensed whey,
channa, whey protein concentrate, whey paste,
paneer lactose
Drying Dried whey
Coagulation Ricotta cheese, whey protein
hydrolysates, protein concentrates,
whey protein isolates, lactose and
lactose powder
Adapted from Aswin and Aparna (2018)
1.5.8 Specialized Dairy Products
1.5.8.1 Functional Foods

The consumer awareness about the health benefits of milk and milk products leads
to the increased demand for novel functional dairy products. The concept of food
for energy and nutrition is gradually shifting towards the concept of health improve-
ment and wellbeing. Functional foods are those categories of food which contain
biologically active ingredients which provide a health benefit beyond its nutritive
value (Homayouni et al. 2012). Among the functional components, probiotics and
prebiotics; soluble dietary fibre; omega-3-polyunsaturated fatty acids; conjugated
linoleic acid; plant antioxidants; isoflavones; phytosterols; vitamins and minerals;
some proteins, peptides and amino acids; and phospholipids are the most researched
areas (Bhat and Bhat 2011).
The functional dairy foods are an area getting a lot of attention, and the health
benefits associated with the presence of specific components or bacteria in the dairy
products are gradually gaining established scientific credibility (Bhat and Bhat
2011). The functional milk components significantly contribute to the prevention of
Table 1.14 Classifications of functional foods

Sl.
no. Type Description Example
1. Fortified A food strengthened with additional Milk powder fortified with
product nutrients vitamin D or A, iron
2. Enriched A food added with new nutrients or Prebiotics, probiotics
products components not normally found in a
particular food
3. Altered A food from which a deleterious Lactose hydrolysed milk
products component has been removed, reduced or Fat reduced milk products
replaced with another substance with
beneficial effects
4. Enhanced A food in which one of the components Conjugated linoleic acid
commodities has been naturally enhanced through (CLA) enhancement in milk
special growing condition, new feed through feeding of green
composition or genetic manipulation fodder
Adapted from Siro et al. (2008)
several diseases like hypertension, coronary vascular diseases, obesity, osteoporo-

sis, cancer, diabetes and some transmissible diseases. The functional dairy foods
can be categorized as given in Table 1.14.
Food fortification has been defined as ‘the addition of one or more essential
nutrients to a food, whether or not it is normally contained in the food, for the pur-
pose of preventing or correcting a demonstrated deficiency of one or more nutrients
in the population or specific population groups’ (FAO/WHO 1994). In a simple way,
fortification is the addition of nutrients, restoring the lost nutrients during process-
ing or enriching the foods. Liquid milk/dried milk is fortified with fat-soluble vita-
mins like A and D, vitamin C, iron, calcium and phosphorus. The successful
fortification depends on several factors like compatibility with the food, fortificant
and process.
Dairy products are considered to be the best vehicle for probiotics. The probiotic
functional foods can be fermented (yoghurt, fermented beverages, kefir, cheese) or
non-fermented (ice cream, spray-dried milk powders). Details about these are elab-
orated in the Chap. 6 of this book.
The consumption of milk is limited by the presence of lactose. Around 70% of
the world population is suffering from lactose intolerance (Shaukat et al. 2010). It is
due to the reduced production of the β-galactosidase enzyme levels in intestinal
walls. There are several low-lactose and lactose-free dairy products available in the
market like lactose-free milk, low-lactose fermented milk, etc. The enzymatic
hydrolysis of whey makes the effective bioremediation process as well as biomass
productions viable with the use of these kinds of whey as cultivation media (Dutra
Rosolen et al. 2015).
1.5.8.2 Designer Milk

Designer milk is the milk whose components are modified from the standard constitu-
ents of milk. The changing scenario demands to customize the constituents in milk
like humanized milk or milk with higher health benefits. The designer milk can be
produced by two approaches—nutritional management by feed modification and by
genetic variation among cattle or transgenic methods. By combining these two
approaches of nutritional and genetic interventions, researchers are developing
‘designer milk’ tailored to consumer preferences which have a dual advantage with
health benefits as well as in processing (Sabikhi 2007). Modification or enrichment in
milk/milk products can be done in several ways such as modifications in protein/
amino acid composition, modifications in fat/fatty acid profile, alteration in lactose,
humanization of bovine milk, eliminating β-lactoglobulin from milk, milk with human
therapeutic proteins, decreasing milk allergies and melatonin enriched milk, and
many more types of the modified or enriched milk can easily be obtained for the spe-
cific proposes (Umaraw et al. 2015). The alteration of primary structure of casein to
improve technological properties of milk, production of high-protein milk, the accel-
erated curd clotting time for cheese manufacturing, the increased yield and/or more
protein recovery, the milk-containing nutraceuticals and the replacement for infant
formula increase the applicability in processing and technological developments.
1.5.8.3 Organic Milk

Organic milk is produced and processed in facilities that follow organic guidelines.
Organic milk is collected from the organic farms in which livestock are raised with-
out giving antibiotics or hormonal treatment and fed with organic feeds which are
free from pesticides, synthetic fertilizers or antibiotics. The cattle are raised on pas-
tures that comply with organic standards, and milk is collected at excellent hygienic
conditions so as to avoid any chances of contamination (Aswin and Aparna 2018).
1.6 I mportance of Advanced and Innovative Technologies

in the Dairy Processing Sector
Application of advanced and innovative approaches in the dairy processing sector is

one of the best ways to increase more food safety and to reduce outbreaks and
health-related issues. The innovative technologies not only increase productivity
and profitability of the business, but it also saves the time and energy because milk
and milk products are perishable in nature and they must be processed in the short
time. The dairy processing sector involves automation for the improvement of over-
all quality of dairy products. Automation may be done in the cleaning, sterilization,
product manufacturing units, quality analysis units, etc. Implementation of innova-
tive technologies and automation in the dairy industry is not only useful for the
quality of the product, but it also reduces the environmental impact. Milk produc-
tion and processing require inputs such as electricity, natural gas, oil, coal, water,
chemicals, packaging, etc. The major outputs such as CO2, methane, effluents, salts,
nitrogen, phosphorus, refrigerants etc. could be harmful to the environment.
Advanced and innovative approaches might be helpful in the reduction of environ-
mental impact. The novel processing technologies and rapid techniques used in the
dairy fields are elaborated in Chaps. 2 and 11 of this book.
1.7 Conclusions
The processing of milk and milk products has changed from crude and elementary
procedures to more controlled procedures over time. There is a substantial improve-
ment in unit operations like separation, standardization, pasteurization, homogeni-
zation, condensing, drying, freezing and packaging. These developments enhanced
the production capacity, automation and hygiene. The incorporation of novel ingre-
dients and use of novel processing and packaging technologies lead to the develop-
ment of innumerable varieties of value-added products with improved sustainability.
The major focus now is on the additional health benefits of various dairy products
and convenience foods and customization of the products as per the consum-
ers demand. Ensuring 100% safety to dairy products still remains a challenge in the
scenario of emerging pathogens. The constantly shifting consumer requirements
pertaining to product quality, safety and novelty will direct the development of the
dairy industry.
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Non-thermal Processing of Dairy Foods
2
K. G. Rashmi and Aswin S. Warrier
Contents
2.1 I ntroduction 26
2.2 U ltrasound Technology 26
2.2.1 Applications 27
2.3 High Pressure Processing 30
2.4 Cold Plasma Processing 34
2.5 Supercritical Carbon Dioxide Technology 36
2.6 Irradiation 38
2.7 Pulsed Electric Field Processing 41
2.8 Conclusions 44
References 44
Abstract
Milk, due to its perishable nature, requires some processing treatment to ensure
its quality, safety, and enhanced shelf life. Conventionally, thermal treatment has
been the preferred processing method. Moreover, the technology of manufactur-
ing nearly all the major dairy products involves heating. However, the thermal
treatment takes a toll on the nutritional as well as sensory quality of milk. Hence,
the profound interest in novel non-thermal methods can be justified. In this
K. G. Rashmi (*)
Department of Dairy Technology, College of Dairy Science and Technology,
Thrissur, Kerala, India
A. S. Warrier
University Dairy Plant (Kerala Veterinary and Animal Sciences University), Thrissur, Kerala,
India

https://doi.org/10.1007/978-981-15-2608-4_2
26 K. G. Rashmi and A. S. Warrier
c hapter, the outlines of non-thermal techniques used in dairy processing like

high-intensity ultrasound, high pressure processing, cold plasma, supercritical
carbon dioxide, irradiation, and pulsed electric field are elucidated.
Keywords
Dairy foods · Non-thermal processing · Milk processing · Advanced technology
· High pressure processing
2.1 Introduction
Milk, due to its perishable nature, requires some processing treatment to ensure its
quality, safety, and enhanced shelf life. Conventionally, thermal treatment has been
the preferred processing method. Moreover, the technology of manufacturing nearly
all the major dairy products involves heating. However, the thermal treatment takes
a toll on the nutritional as well as sensory quality of milk. Hence, the profound inter-
est in novel non-thermal methods can be justified. In this chapter, the outlines of
non-thermal techniques used in dairy processing like high-intensity ultrasound,
high pressure processing, cold plasma, supercritical carbon dioxide, irradiation, and
pulsed electric field are elucidated.
2.2 Ultrasound Technology
Ultrasound is an emerging technology with immense potential in food processing

operations like emulsification, homogenization, mixing, milling, extraction, pas-
teurization, filtration, drying, and crystallization (Zisu and Chandrapala 2015). This
technology has even found application in equipment cleaning. Ultrasound technol-
ogy is highly efficient and operates causing limited reduction in nutrient content
ensuring better food quality and increased shelf life. This technique is also harm-
less, inexpensive, simple, rapid, and eco-friendly and uses less energy (Akdeniz and
Akalın 2019).
Ultrasound refers to sound waves having frequencies beyond the limits that can
be heard by human (20 Hz to about 20 kHz). These longitudinal pressure waves can
be classified based on frequency and acoustic intensity as high-intensity or low-
intensity ultrasound. Low-intensity ultrasound having frequencies above 100 kHz
and intensity below 10,000 W/m2 creates inadequate power to bring about cavita-
tion effect. Hence its application is limited to food analysis only. High-intensity
ultrasound (HIUS) operates between 20 and 100 kHz and intensities ranging from
100,000 to 10,000,000 W/m2, causing major changes to the food structure (Akdeniz
and Akalın 2019). HIUS is relayed in a sequence of compression and rarefaction
cycles creating acoustic cavitation. The bubbles formed due to cavitation gradually
get enlarged by rectified diffusion or bubble-bubble coalescence in a high-intensity
2 Non-thermal Processing of Dairy Foods 27
sound field (Zisu and Chandrapala 2015). These bubbles on collapsing produce
huge turbulence and formation of shock waves, generating high temperatures and
even leading to chemical changes.
The transducers are used to generate ultrasound waves which convert electrical
energy to mechanical energy in the form of ultrasonic vibrations and can be applied
to the food products directly or indirectly by using sonotrodes or ultrasonic baths,
respectively (Arvanitoyannis et al. 2017). The waves are discharged into the medium
by an emitter. The pilot-type HIUS equipments are direct type, in which the direct
dispersal of the sound energy takes place from the transducer to the sample. A horn
in the transducer amplifies the signal before it reaches the sample. In the indirect
type, the sound energy is indirectly dispersed using a coupling fluid like water
(Guimarães et al. 2019). The advantages and limitations of ultrasound processing
are enlisted in Table 2.1.
2.2.1 Applications
2.2.1.1 Microbial Inactivation

HIUS acts in numerous ways like disintegrating particles, making structural changes,
improving mass transfer rates, damaging the cell membrane, etc. The lysis of cellular
membrane of the microorganisms by ultrasound may be attributed to intracellular
cavitation, leading to localized heating, production of free radicals, and DNA dam-
age (Zisu and Chandrapala 2015; Guimarães et al. 2019). Cameron et al. (2009) have
reported effectiveness of ultrasound as a non-thermal unit operation against both
spoilage and pathogenic microbes in raw and pasteurized milk. According to Ojha
et al. (2017), gram-positive bacteria are more resistant to ultrasound, thanks to their
Table 2.1 Advantages and Advantages

limitations of ultrasound • Non thermal process with
processing numerous applications
• Improved mass transfer
• Limited chemical changes
• Low running cost
• Energy efficient
• Environment friendly
Limitations
• Equipment for industrial
scale processing is not yet
available
• Apprehension regarding
health effects of free radicals
formed
• Some food products
develop off flavors/
physicochemical defects
thick cell wall, in comparison with gram-negative bacteria. Bacterial spores and
fungi are found to be more resistant to ultrasound than vegetative cells, while aerobes
are more resistant than anaerobes, and cocci are more resistant than bacilli (Guimarães
et al. 2019). Moreover, yeasts are vulnerable to sonication due to their large size and
thereby increased surface area.
Inactivation of several enzymes including alkaline peroxidase, lactoperoxidase,
etc. has been reported to have taken place at a temperature range of 61–75.5 °C dur-
ing thermosonication (Villamiel and de Jong 2000). Bermúdez-Aguirre and
Barbosa-Cánovas (2008) reported that sonication of 24 kHz at 63 °C in UHT milk
prevented mesophilic growth higher than 2 log at refrigerated storage for 16 days.
Thermosonication of reconstituted infant formula (20 kHz at 50 °C) reduced the
microbial count by 7 log reduction after 2.5 min of treatment and also inactivated
Cronobacter sakazakii (Adekunte et al. 2010).
Cellular sonication is the creation of temporary pores on the cell membrane due
to the effect of ultrasound. At low levels, this technique can enhance the microbial
growth by improving the cell membrane permeability thereby improving the mass
transfer of nutrients. Simultaneously, there will be efficient exclusion of by-products
of cellular metabolism. In the case of a high level sonoporation, there is an increased
probability for physical disruption, alteration of cell membrane, and cellular death
by leakage of cellular content. Therefore, ultrasound method should be applied in a
controlled manner to achieve desired level of cell permeability (Ojha et al. 2017;
Guimarães et al. 2019).
2.2.1.2 Ultrasound Applications for Dairy Products

HIUS is capable of improving the viability of probiotic strains of starter cultures
used in the manufacture of cultured milk products, along with hastening the hydro-
lysis of lactose by releasing lactase enzyme and stimulating acid production, thereby
decreasing the time required for fermentation. Fermented milk products treated
with ultrasound scored better in their organoleptic as well as nutritive qualities,
which may be attributed to the increased content of oligosaccharides and bioactive
peptides and the lower content of lactose (Nguyen et al. 2009; Barukčić et al. 2015;
Huang et al. 2019).
Conversely, studies have also reported the reduction in growth of probiotics after
treating with HIUS, which may be attributed to the cell membrane becoming more
permeable or even getting damaged or oxidative degradation of membrane lipids
(Yeo and Liong 2011). Hence, this technique can also be used for production of
inactivated probiotics (paraprobiotics) or beneficial metabolites (postbiotics). HIUS
is found to stabilize prebiotics, without causing major changes in its structure.
However, some amount of hydrolysis occurs in its polymer chains leading to
increased consumption by probiotics.
HIUS can cut down the time taken for ripening of cheese, expedite the breaking
down of protein structure, and ensure improved organoleptic, textural, and nutritive
attributes. Moreover, it stimulates the growth of probiotic and starter cultures and
accelerates the ripening process by the release of intracellular enzyme (Guimarães
et al. 2019).
HIUS has been reported to reduce the fat globule size in yoghurt production, by
which the viscosity is improved, syneresis decreased, gel strength increased, and
fermentation accelerated. HIUS contributes to a more stable and firm gel formation
in yoghurt by denaturing whey proteins, dividing casein micelles, and further
recombining these protein fractions to form robust networks, thereby ensuring
firmer yoghurt (Akdeniz and Akalın 2019).
In the manufacture of ice cream, providing ultrasound treatment to mix during
freezing leads to improvement in the organoleptic and textural properties of ice
cream. This is due to the boosted ice nucleation, improved heat and mass transfer,
and formation of smaller and uniform ice crystals, as a result of ultrasonic cavita-
tion. During cavitation, the explosion of bubbles leads to localized increase in pres-
sure, which promotes ice nucleation and breaks down larger ice crystals.
The ice cream freezer is customarily a scraped surface heat exchanger, whose
freezing chamber is surrounded by the refrigerant coils. Hence the heat gradient is
in such a way that the zone nearer to the walls is cooler, leading to the formation of
ice crystals first near the wall surface. If these ice crystals are not removed immedi-
ately, the heat transfer and texture of the product is affected. The dasher provided in
the freezer serves numerous functions like improving heat transfer and preventing
formation of ice layer on walls by scraping action. HIUS can also be used for reduc-
ing the ice layer formation on the freezer wall. Ultrasound makes this happen by
virtue of cavitation, thereby permitting the elimination of scrapers and thereby an
economic benefit.
The extreme turbulence and cavitation caused by HIUS can even be used for
homogenization of milk, as these mechanisms are also believed to be responsible
for the size reduction of fat globules in conventional homogenization. The frequency
of ultrasound waves, power, and the composition of the product determine the effec-
tiveness of homogenization. Ultrasound further breaks down the milk fat globule
membrane. Cold sonication and thermosonication behave differently in regard to
their homogenizing capabilities. In cold sonication, small fat globules tended to join
together to form larger clusters up to 30 s of treatment, which were broken down on
treating for a longer time. In contrast, thermosonication produces similar homogeni-
zation effect without any cluster formation (Zisu and Chandrapala 2015).
Milk and whey have a reputation of producing foam during processing, which
causes increased fouling, thereby reducing heat transfer and affecting the quality
and safety of the product. Sonication is known for its capability to remove gases or
air entrapped in a solution. This ability can be utilized for solving the challenge
caused by foam in dairy processing. Villamiel et al. (2000) have experimentally
shown the efficacy of this treatment by removing 80% foam from reconstituted skim
milk using ultrasound at a frequency of 20 kHz. The functioning of sonic defoaming
may be attributed to formation of partial vacuum on bubble interfaces, bubble reso-
nance, flow generation due to sonication and the powerful cavitation (Mason et al.
2005).
Fouling and concentration polarization are the most demanding issues in mem-
brane processing, both leading to a reduction in flux. Ultrasound treatment has been
found to be an effective method to address these issues. It further aids in cleaning
the fouled surfaces. The cavitation caused by sonication may be considered as factor
responsible for this effect. Muthukumaran et al. (2005) opined that ultrasound gen-
erally achieves this by increasing the turbulence of the solution, thereby improving
the permeate flux. In another study, they (Muthukumaran et al. 2007) observed
ultrasound causing a rise in mass transfer rate inside the concentration polarization
layer aiding removability of the fouling cake. In their opinion, the cleaning ability
of ultrasound can be attributed mostly to acoustic streaming and turbulence instead
of cavitation which acts to a limited extent. Lamminen et al. (2004) concluded dis-
ruption of cavitation bubbles to be the causative factor of sonic cleaning.
The crystallization induced by ultrasonication is called sonocrystallization.
Ultrasound aids the increase in nucleation rate as well as crystal growth in sonocrys-
tallization, yielding a larger number of smaller crystals. Moreover, ultrasound
induces collision of existing larger crystals by a process known as sonofragmenta-
tion. Crystallization is an essential step in the commercial production of dried lac-
tose. Sonocrystallization may be effectively used in this regard.
Instantaneous dissolution is an ideal character of any powdered food products,
contributing to its commercial value. Ultrasound is capable of improving the solu-
bility by breaking down the aggregates having limited solubility, during reconstitu-
tion. Application of ultrasound to various dairy products like whey protein
concentrates, caseins (Onwulata et al. 2002), whey protein isolates (Jambrak et al.
2008), etc. has been reported to improve the solubility, while the effect was not so
pronounced in case of micellar casein and milk protein concentrates (Zisu and
Chandrapala 2015).
Viscosity is a major physical property that influences the design of pipelines,
heat exchangers, agitators, etc. in dairy processing. Casein micelles play an impor-
tant role in determining the viscosity of milk during processes like heat concentra-
tion. The viscosity and age thickening during concentration can be controlled using
sonication. Zisu and Chandrapala (2015) observed that ultrasound is capable of
causing reduction in viscosity, while age thickening was only delayed in milk-based
systems including milk protein concentrate and calcium caseinate.
2.3 High Pressure Processing
High hydrostatic pressure (HHP) processing, high pressure processing (HPP), or

pascalization of food products is a notable non-thermal process used for inactivation
of most of the microbes and some enzymes with minimal changes to the food’s
nutritional and sensory qualities. In this process, the product is applied with a high
pressure in the range of 100–1000 MPa using a liquid (usually water), in a specially
designed metallic chamber. The utilization of high pressure for microbial reduction
in milk was first explored by B. H. Hite in 1899. This technology operates based on
Le Chatelier’s principle, which states that any reaction, conformational change, or
phase transition, accompanied by a decrease in volume, is enhanced by pressure;
principle of microscopic ordering, which asserts that pressure and temperature exert
antagonistic forces on molecular structure and chemical reactions; and the isostatic
principle, according to which pressure will not damage a food product as long as it
is applied uniformly in all directions (Yordanov and Angelova 2010).
The equipment used for HPP basically consists of a cylindrical container in
which the processing takes place, which is in fact a high-pressure vessel; a system
for generating the required high pressure; pressure transmitting fluid; a mechanism
for controlling temperature; and the material handling systems (Voigt et al. 2015).
The pressure vessel is made of stainless steel in a monoblock design capable of
withstanding the extreme pressure. The pressure generation can be accomplished by
either direct method using pistons or indirect method using pumps. If the food prod-
uct being processed is in liquid state, it can itself act as the transmitting fluid, while
another liquid like water is used in case of prepacked foods. During compression,
an increase in temperature takes place due to adiabatic heating, which also takes
part in microbial destruction. The temperature control mechanism ensures that there
is no undesirable increase in temperature.
Batch, semicontinuous, and continuous systems are available for high-pressure
treatment. In the operation of a batch system, the pressure vessel is loaded with the
food packed in suitable containers and closed. The pressure transmitting fluid is
then supplied from its tank to the pressure vessel with the help of pressure generat-
ing system. Once the specified pressure is built up in the system, the product is held
at that pressure for the required time to ensure the time pressure conditions are met.
Subsequently, the pressure is released, the fluid is returned to its tank, and the food
is taken out. Continuous process can only be used for liquid foods, which flow
through a pressurized pipe system. The semicontinuous system uses a combination
of pumps, several pressure vessels, and tanks and is again suitable for liquid foods.
The increased number of pressure vessels ensures a nearly continuous output by
maintaining each vessel at different stages of operation.
HPP is gaining prominence as a substitute to thermal processes like pasteuriza-
tion and sterilization, as comparable microbial destruction is achieved with minimal
changes to the food’s flavor and nutrient content. A pressure below 420 MPa results
in inactivation of bacteria similar to that of pasteurization, while 700 MPa and above
are required for sterilization (Voigt et al. 2015). The advantages and limitations of
high pressure processing are listed in Table 2.2.
2.3.1 Applications
Huppertz et al. (2002), in their extensive review on the effects of high pressure on
constituents and properties of milk, have described the following aspects:
• Reduction in freezing point of water

• Altered mineral balance
• Denaturation and thereby reduction in solubility of whey proteins, particularly
β-lactoglobulin
• Breakdown of casein micelles along with increased solubility of caseins
• Crystallization and increased solid fat content of milk fat
Table 2.2 Advantages and limitations of high pressure processing

Advantages
• Ability to retain the freshness in flavor and nutrient content without using preservatives or
other additives
• Homogeneity of treatment independent of mass and time
• Non thermal microbial destruction
• Improved shelf life
• Clean technology, flexible system for number of products and operation
• Ability to manipulate the texture
• Less thermal degradation
• Ability to handle both particulate and liquid foods
Limitations
• Initial investment and maintenance costs are high
• Destruction of spores is not ensured without thermal treatment
• Unavailability of industrial equipment for massive processing
• Requirement of water for compression
• Pressure-induced denaturation of proteins
Parekh et al. (2017); Voigt et al. (2015); Datta and Deeth (1999)
• High stability of enzymes, which are resistant to changes in pressure

• Increased pH, reduced lightness, and reduced turbidity of milk
• Reduced rennet coagulation time and increased yield of cheese

Raw milk pressurized to 400–600 MPa has a microbiological quality similar to
pasteurized (72 °C/15 s) milk. However, HP processed milk was inferior to steril-
ized milk due to pressure-resistant spores. The microbial destruction in HPP can be
attributed to denaturation of key enzymes, permeabilization of microbial cell mem-
branes, and disruption of nucleic acids and ribosomes (Datta and Deeth 1999).
Gram-positive microorganisms, which are inactivated only at a higher pressure of
500–600 MPa, are more resistant to pressure than gram-negative microorganisms,
yeasts, and molds, which can be destroyed at 300–400 MPa. The bacterial spores
are capable of surviving extreme pressures like 1000 MPa (Smelt 1998). The
destruction of spores can be ensured by first initiating germination using heat or a
pressure of 100–300 MPa followed by HPP (Yordanov and Angelova 2010).
2.3.1.2 Effects of HPP on Milk Components

Aside from microbial inactivation, the impact of high pressure on the structure of
proteins and the mineral balance offer a wide range of applications for this technol-
ogy in dairy products. As already mentioned, high pressure causes disintegration of
casein micelles leading to an increase in number (due to fragmentation) and size
(due to aggregation) of micelles. Harte et al. (2007) have reported that addition of
whey protein can protect the casein micelles from disruption caused by HPP. Among
whey proteins, β-lactoglobulin gets almost completely denatured by a treatment of
500 MPa at 25 °C, while α-lactalbumin and immunoglobulins are only partially
denatured on treating with the same pressure at 50 °C. Hence, this technology offers
an opportunity to preserve the heat-sensitive colostrums, which are rich in immuno-
globulins. Bovine serum albumin is resistant to pressure up to 400 MPa, while the
effect of high pressure on other whey proteins is not known. Among enzymes, no
effect can be observed for lipase in milk up to 400 MPa and for plasmin up to
300 MPa. Alkaline phosphatase, having a Zp value (increase in pressure required for
a tenfold reduction in D value) of 368 MPa, is completely inactivated only at a pres-
sure of 800 MPa (Mussa and Ramaswamy 1997; Rademacher and Kessler 1997).
HPP of milk does not affect the fat globule size up to 600 MPa, while an increase in
fat globule size was observed on treatment of cream at 800 MPa for 10 min, along
with causing some aggregation without damaging the milk fat globule membrane
(Kanno et al. 1998). The free fatty acid content of milk does not increase up to
500 MPa, ruling out the chances of pressure-induced lipolysis. Lopez-Fandino et al.
(1996) have reported that neither Maillard reaction nor lactose isomerization
occurred on treating milk at 100–400 MPa for 10–60 min at 25 °C. Smaller mole-
cules such as vitamins, amino acids, simple sugars, and flavor compounds are not
affected much by pascalization (Chawla et al. 2011).
2.3.1.3 High Pressure Processing of Dairy Products

Eberhard et al. (1999) have reported that treatment of cream at 600 MPa for 2 min
improved its whipping properties, which may be attributed to the crystallization of
milk fat. There was no visible improvement below 400 MPa, while an increased
pressure led to an increase in whipping time probably due to whey protein denatur-
ation. Butz et al. (1999), who studied the effect of high pressure on a model system
close to milk fatty acid composition, reported that there was no significant effect on
oleic acid, while a small increase was observed in the autoxidation of linoleic acid
at pressures above 350 MPa.
HPP has been gaining interest in cheese making, as it can ensure the microbial
safety of cheese milk without causing the sensory or textural changes like pasteuri-
zation. But, HPP of cheese milk in comparison with pasteurization yields cheese
having a different composition and ripening. The HPP of cheese milk is found to
retain more moisture, salt, and total free amino acids in the final product, having a
firmer texture with reduced fragility. HPP has also been found to increase the yield
and reduce the rennet coagulation time, along with the obvious improvement in
microbial quality.
HPP can be applied in yoghurt manufacture either to treat the milk, or for treating
the final product, or to get improved shelf life and better textural and rheological prop-
erties. Several studies have reported that the use of pressure-treated milk yields yoghurt
with a firmer body and having reduced syneresis, while viscosity was found to increase
in case of stirred yoghurt (Trujillo et al. 2002). Udabage et al. (2010) have reported a
reduction in viscosity when HPP is performed at a higher temperature, above
90 °C. Capellas et al. (2002) have reported that the consumer preference of yoghurt
made from pressure-treated milk was more in comparison with conventionally made
yoghurt in terms of texture and creaminess, while yoghurt made from heat-treated milk
was preferred more in terms of flavor. HPP of yoghurt after fermentation can ensure
better shelf life by arresting the growth of starter cultures and other microbes, prevent-
ing further acidification or spoilage.
HPP can be favorably used for processing high-value products like functional
foods, as the cost involved doesn’t cause a serious hindrance. Kelly and Zeece
(2009) have extensively reviewed the applications of HPP in functional dairy foods
including the reduction in allergenic potential of milk proteins and preservation of
probiotic dairy foods, colostrum, etc. Fonterra Co-operative Group, New Zealand,
has obtained patent for a high-pressure-based technology for processing colostrum
and probiotic yoghurt. Unlike pasteurization, HPP at 400 MPa for 5 min could
retain the immunoglobulin-A in human milk completely (Permanyer et al. 2010).
HPP offers compelling applications in ice cream manufacture. Pressurization of
ice cream mix to 400–500 MPa could enhance the viscosity, even without any hold-
ing. Ice cream made from the mix was also found to have a higher melting resis-
tance and improved sensory quality. A patented technology involving simultaneous
pressurization and freezing has been reported to produce a smoother ice cream with
finer ice crystals (Keenan et al. 2001).
2.4 Cold Plasma Processing
The term “plasma” coined by Langmuir denotes the fourth state of matter formed
by partial or complete ionization of gases to the level that its electrical conductivity
is immense and its behavior is different from that of gases. Plasma can be naturally
found in lightning, interior of stars, etc., while it can also be made artificially by
exposing a neutral gas to a strong electromagnetic field. Based on the state of ther-
modynamic equilibrium, plasma can be classified as thermal and non-thermal
plasma. Non-thermal plasma or cold plasma, in which only the electrons are at a
higher temperature than the heavier particles (unlike thermal plasma, in which both
electrons and heavy particles have a high temperature), is of particular interest in
dairy and food processing. The term “cold” can be confusing, as it indicates room
temperature condition at atmospheric pressure and not any refrigerated conditions.
Still this technology is called “cold plasma” in comparison with the thermal plasma,
which is usually thousands of degrees above the ambient conditions.
The equipment used for cold plasma treatment generally consists of a treatment
chamber, a pumping device, gas supply and control systems, and the power supply
and transmission system (Milella and Palumbo 2014). The ionization of gases into
plasma can be achieved by several methods like thermal, electric, or laser treatment.
Hence the composition of cold plasma varies depending on many factors like the
carrier gas (air, oxygen, helium, nitrogen or argon), the plasma generator (radio
wave, microwave, plasma jet, or dielectric discharges), and the operating conditions
(pressure and temperature) (Zhang et al. 2018).
Dielectric barrier discharge (DBD) and jet plasma are the most commonly used
methods for generation of cold plasma for food applications. Dielectric barrier dis-
charge refers to the electric discharge formed between two metallic electrodes hav-
ing a dielectric barrier in between. The dielectric layer ensures that spark formation
due to arcing is avoided. Plasma jet device consists of two concentric electrodes
through which the gases are passed, while a radio-frequency power is applied to the
inner electrode igniting the gases, which emerges out of a nozzle in a jetlike fashion
(Schutze et al. 1998). The advantages and limitations of high pressure processing
are listed in Table 2.3.
2.4.1 Applications
Cold plasma is an emerging technology capable of inactivating microbes including

pathogenic bacteria, bacterial spores, fungi, and biofilms (Segat et al. 2016). The
mechanism involved in inactivation of microorganisms is the generation of various
active oxygen free radical and chemical compounds like nitrogen oxides, reactive
oxygen species (ROS), reactive nitrogen species (RNS), ultraviolet radiation, etc.
affecting the vital cellular components like DNA and proteins and causing disrup-
tion of the cell membrane (Coutinho et al. 2018; Zhang et al. 2018). Liao et al.
(2017) have reported that increased acidity and moisture content in food products
improve the efficiency of treatment with cold plasma. The targeted microbes, power,
time, carrier gas, and composition of the food being treated determine the efficiency
of microbial inactivation by cold plasma.

limitations of cold plasma Non thermal process causing little
processing heat induced changes
Fast process
Energy efficient technology
Sensory quality of the products are
preserved
Efficient antimicrobial action
Limited structural changes
Resource efficient technology
Environmentally safe technology
No damage to the quality of food
products
Limitations
High cost
Volume, size and shape of food
determines the efficiency of
treatment
Surface treatment with limited
penetration depth
Difficulty to effectively treat foods
having a rough or irregular surface
Accelerated lipid oxidation in high
fat products
Only limited number of studies regarding the application of this technique in

dairy industry has been reported so far. Gurol et al. (2012) reported a significant
54% reduction of Escherichia coli in UHT and raw milks treated with plasma for
3 min, with minimal changes in pH and color. Lee et al. (2012) found significant
reduction of E. coli and Staphylococcus aureus in cheese treated with cold plasma.
However, there were a reduction in L∗ value (lightness from black to white), increase
in b∗ value (color from blue to yellow), and a marked reduction in sensory score.
Kim et al. (2015), who studied the antimicrobial efficiency of an encapsulated DBD
plasma, reported a significant reduction in the population of E. coli, Listeria mono-
cytogenes, and Salmonella typhimurium. The effect of plasma against these three
organisms in sliced cheese and cheddar cheese was studied by Yong et al. (2015) to
yield positive results with respect to microbial reduction. The treated cheddar cheese
slices had a lower pH and L∗ value, higher thiobarbiturate content and b∗ value, and
a substantially low sensory score. According to Korachi et al. (2015), a considerable
increase was observed in the levels of 1-octanol, 2-heptanone, 2-hexenal, 2-octenal,
nonanal, and benzaldehyde in milk after plasma treatment. Segat et al. (2015)
observed changes in the structure of whey proteins on treating with plasma for
15 min. In another study, Segat et al. (2016) reported the efficiency of plasma treat-
ment to inactivate alkaline phosphatase enzyme. Sarangapani et al. (2017) used
chromatographic techniques to observe the effects of plasma on dairy fats and noted
the formation of various products of oxidation along with variations in fatty acid
contents including reduction in essential fatty acids. Coutinho et al. (2019) investi-
gated the effects of process parameters on physical, microstructural, and thermal
characteristics of chocolate milk drink on treating with non-thermal plasma. They
reported enhanced particle size, better consistency, and changed melting
properties.
2.5 Supercritical Carbon Dioxide Technology
When the temperature and pressure of a substance is raised beyond its critical point,
it shows a transitional behavior between liquids and gases. In this condition, where it
is not possible to distinguish between liquid and gaseous states, the fluid is called a
supercritical fluid. Carbon dioxide (CO2), the most popular supercritical fluid, is hav-
ing diverse applications in food processing owing to its inertness, non-corrosiveness,
fireproof nature, nontoxicity, easy availability, low price, and safety for consumption.
Carbon dioxide having temperature and pressure above its critical point (304.25 K,
7.39 MPa) is an effective extraction solvent, chemical reagent, heat transfer fluid, and
cleaning and sterilizing agent. As the critical temperature of CO2 is close to ambient
temperature, this technique is considered as non-thermal. The high solubility, moder-
ate diffusivity, and low viscosity values facilitate its suitability as an excellent agent
for extraction and microbial degradation. Furthermore, the residual CO2 after pro-
cessing can be effortlessly removed just by reducing the pressure. Sometimes, other
high-pressure techniques like liquid CO2, subcritical CO2, and dense phase CO2 hav-
ing comparable abilities are used in lieu of supercritical carbon dioxide (sCO2).
As the pressure of sCO2 is high, several effects are comparable with HPP like
changes in properties of water, proteins, and membrane materials, along with inac-
tivation of enzymes and microorganisms. The pressure employed in sCO2 (10–
20 MPa) is much lower than that of HPP. Hence, the expenditure involved is also
lower. Another advantage that sCO2 has over HPP is its adaptability to operate as a
batch, semicontinuous, or fully continuous process, while HPP usually operates as
a batch process. The equipment used for batch process comprises a treatment cham-
ber, carbon dioxide pump, pressure regulator, and a pressure release mechanism. In
a semicontinuous system, sCO2 flows continuously through the treatment chamber.
In case of a continuous system, the liquid food to be treated is mixed with CO2 and
then pressurized to the required level by passing through a high-pressure pump
(Amaral et al. 2017). The advantages and limitations of supercritical carbon dioxide
are given in Table 2.4.
2.5.1 Applications
The major applications of this technology in the food industry include fractionation,
extraction, microencapsulation, pasteurization, sterilization, and chromatographic
techniques (Amaral et al. 2017). This technique can also be used for the manufac-
ture of particulate products by a method similar to freeze-drying, known as super-
critical drying.
As reported by Bonnaillie and Tomasula (2015), carbon dioxide can act on micro-
organisms and enzymes in two ways. At an intermediate pressure and temperature, the
CO2 will replace oxygen causing reduction in pH, thereby discouraging microbial
growth or enzymatic reactions. Higher pressures of CO2, even at ambient temperature,
can cause reactions which are much harmful to microbes including spores and can
arrest enzymatic reactions. Perrut (2012) has reported solubilization, structural
Table 2.4 Advantages and limitations of supercritical carbon dioxide processing

Advantages
• Low viscosity, high diffusion and lack of surface tension ensure increased penetration of
the supercritical fluid
• No thermal degradation
• Short processing time
• Oxidation prevented due to lack of oxygen in system
• High selectivity for extraction and fractionation
• The fluids used are inexpensive
• Easy removal of residual CO2
Limitations
• High initial investment required
• Requires trained staff to reduce safety risk
• Real-time control is difficult
• Changes in phase diagram, making it complicated to predict processing conditions
changes of bacterial membrane materials, pH reduction within the cells, enzyme inac-
tivation, carbonation, salt imbalance, and extraction of certain vital components from
the cell as the mechanisms of action of sCO2. Numerous factors like the pressure,
temperature, treatment time, pH, composition and physical state of food, combination
with other treatments, and type of equipment used contribute to the inactivation effi-
ciency of sCO2.
Di Giacomo et al. (2009) studied the ability of sCO2 to pasteurize and sterilize
milk and reported that milk pasteurized using sCO2 had its shelf life increased by
35 days without any sensory deterioration, while increasing the pressure affected
the sensory score. Amaral et al. (2017) have extensively reviewed several works
related to the use of sCO2 in processing milk. Though assumptions can be made in
comparison with other techniques, the effect of this technique on the nutritional
attributes of milk is not yet completely known and requires further research. Yet,
this technique can be utilized for precipitating casein or for fractionating whey pro-
teins. Hence, sCO2 offers immense potentiality in cheese manufacture besides
ensuring microbial destruction without causing any heat-induced changes. The abil-
ity of sCO2 to fractionate lipids can be utilized in the development of low-fat dairy
products. Yee et al. (2008) have reported utilization of supercritical fluid extraction
(SCFE) in manufacturing low-fat varieties of cheddar and parmesan cheeses. In an
interesting work reported by Chitra et al. (2015), they have used SCFE for selective
fractionation of cholesterol from whole milk powder, optimizing the process
parameters.
2.6 Irradiation
Analogous to other non-thermal techniques, irradiation is a process capable of

ensuring food quality and safety without any heat-induced changes. Though the
applicability of this technology is known for more than a century, and even after
being commercially used in food processing for half a century, still a lot of appre-
hensions and misconceptions are existent among the consumers. Moreover, the full
potential of this technology has not yet been explored by the dairy industry, owing
to the multiplicity and diversity of dairy products.
In irradiation, the food to be treated is exposed to some kind of controlled ion-
izing radiation in a shielded chamber. The ionizing radiation used may be gamma
rays emitted by Co-60 or Cs-137 (radioactive isotopes of cobalt and cesium, respec-
tively, having high penetration depth), beta rays emitted using electron beam tech-
nology (having limited penetration depth) or X-rays emitted by striking an electron
beam on a thin metallic plate (having high penetration depth like gamma rays, but
can be switched off when not required like beta rays). Though the energy of these
radiations is capable of liberating electrons from atoms, it is not sufficient to make
the food radioactive. The impact of irradiating any substance depends on the type
and energy level of the radiation along with the composition, physical state, tem-
perature, and the surrounding conditions of the substance being treated (Morehouse
and Komolprasert 2004). The chemical effects of radiation can be due to the radi-
olysis itself or due to the liberated ions or free radicals.
The chemical changes taking place in food as a result of irradiation have always
raised the question about its safety among the common consumers. This concern
was seriously taken up by the researchers and decision-makers alike. International
agencies including the International Atomic Energy Agency, World Health
Organization, and Food and Agricultural Organization of the United Nations have
unanimously agreed, after stringent evaluation, that irradiation is not more harmful
than its conventional processing counterparts (Morehouse and Komolprasert 2004).
Several international and national regulatory authorities have authorized the irradia-
tion of food and have specified the necessary guidelines and restrictions for the
same. Though the regulations may vary from country to country, a dose of up to
10 kilogray (kGy) has been internationally agreed as safe. However, considering the
consumer sensibilities, radiated foods need to be specifically labelled with the
“radura” symbol along with declaring the same as a statement. The advantages and
limitations of irradiation are enlisted in Table 2.5.

limitations of irradiation • Cold process
• Effective in microbial
destruction and disinfestation
• Extended shelf life
• Due to high penetration depth,
irradiation can be used for even
packed foods
• Reduced need of preservatives
• Reduced risk of migration of
pests across the borders through
agricultural produce
Limitations
• Negative consumer perception
• Apprehensions regarding
radiolytic products and free radicals
formed
• High capital investment required
• Not all foods are suitable for
irradiation
• Some loss of nutrients like
vitamins and minerals
2.6.1 Applications
The major application of ionizing radiation in food processing is enhancing the

shelf life and safety of food by destroying microorganisms and insect pests. This
technology is mostly used for treating foods like fruits and vegetables, meat, poul-
try, fish, and seafood. Moderate doses of radiation, below 2 kGy, are found to delay
aging in fruits and sprouting in vegetable, along with ensuring destruction of insects.
Intermediate doses, from 1 to 10 kGy, match the performance of pasteurization,
while doses above 10 kGy ensure sterilization.

The intensity of microbial destruction depends on the dose of radiation absorbed by
the food product being treated. The electrons liberated due to the action of radiation
attack the chemical bonds of the microbial nucleic acids, thereby destroying the
organism. A secondary mechanism is ionization of water molecules. Irradiation
induces water molecules to release electrons, producing hydroxyl radicals and
hydrogen peroxide which have antimicrobial action.
2.6.1.2 Irradiation of Dairy Products

Some older studies have considered milk and dairy products as unlikely candidates
for irradiation, referring to the development of off-flavors or odors (Wertheim et al.
1957; Smith and Pillai 2004). However, contradictory findings showing effective-
ness of irradiation in low doses or in frozen condition have also been reported
(Odueke et al. 2016). de Oliveira Silva et al. (2015) reported that gamma irradiation
of whole milk at 2 kGy dose favorably reduced the microbial load without any nega-
tive effect on organoleptic quality, while 3 kGy dose caused rancidity. Nayeem et al.
(2018) suggested irradiation as a promising method to control milk allergy. Kaddouri
et al. (2008) reported that gamma irradiation of cow milk didn’t affect the antigenic-
ity of β-lactoglobulins. However, Gomaa and Boye (2015), who studied the effect of
irradiation on milk allergens, opined that at low doses, the effect of irradiation on
the antigenicity of allergens is determined by the food being processed and the pro-
cess parameters.
Żegota and Małolepszy (2008) studied the efficiency of decontamination of
casein and milk powder using irradiation. They found that a dose of 5 kGy was
enough to bring down the microbial count to permitted levels. Moreover, this count
was maintained even after 1 month of storage study. Ahmad et al. (2018) irradiated
camel whey powder and reported that a dose of 9 kGy caused a 5 log reduction in
microbial count, 7.4% reduction in antibody activity, and 13% reduction in immu-
noglobulin, while major whey proteins were unaffected. According to Osaili et al.
(2008), irradiation in low dose immediately after manufacture was effective in
reducing the count of Enterobacter sakazakii in infant milk food. Cieśla et al. (2006)
observed that irradiation improved the barrier properties and tensile strength of
films containing calcium caseinate, whey protein isolate, and glycerol by cross-
linking of β strands.
Kim et al. (2008) investigated the effect of gamma radiation on yoghurt and
found that a dose of 5 kGy could completely destroy coliforms and cause 6 log
reductions in aerobic count, without affecting the acidity or sensory properties
much. Ham et al. (2009) observed that there were no compositional changes to the
irradiated yoghurt samples up to 10 kGy and the sensory quality was stable up to
3 kGy. They also found that the microbial count showed a reducing trend during
storage at a constant temperature.
Several studies have been reported on the application of irradiation to cheeses.
Seisa et al. (2004) studied the effect of irradiation and ripening temperature on
cheddar cheese. Though the bacterial quality improved, the psychrotrophic count
was not affected. Irradiation caused an increase in thiobarbiturate content, while the
sensory quality remained same. Rabie et al. (2011) observed gamma irradiation as
an effective treatment for reduction of biogenic amine content in blue cheese during
storage. Application of irradiation in lower doses for microbial inactivation and
shelf life extension for numerous varieties of cheeses like mozzarella, sliced pizza
cheese, feta, camembert and brie cheeses, fior di latte cheese, etc. has been reported
(Huo et al. 2013; Kim et al. 2010; Konteles et al. 2009; Velasco et al. 2016; Lacivita
et al. 2019).
Kamat et al. (2000) observed that irradiation at a low dose of 1 kGy was suffi-
cient to bring the microbial quality of vanilla, strawberry, and chocolate ice creams
to prescribed limits, while the sensory quality was affected on treatments above
2 kGy. Jo et al. (2007) studied the irradiation sensitivity of foodborne pathogens,
namely, Listeria ivanovii, Escherichia coli, and Salmonella typhimurium in vanilla,
chocolate, and strawberry flavor ice creams. A dose of 3 kGy was enough for ensur-
ing microbial inactivation in vanilla ice cream, while 5 kGy was required for the
other two flavors. Hoseinpour-Ganjaroudi et al. (2016) reported that a dose of 5 kGy
was required to completely destroy Brucella species in ice cream.
2.7 Pulsed Electric Field Processing
The possibility of processing milk using electricity has been explored since the
beginning of the twentieth century. However, most of these early researches consid-
ered electricity only as a means to heat up the product. Pulsed electric field (PEF)
emerged as a technology only by the 1960s, when it was first suggested by German
engineer Heinz Doevenspeck, and within two to three decades, it became much
popular. PEF processing is a gentle non-thermal treatment applicable to liquid and
semiliquid foods. This technology utilizes very short electric pulses (μs) having
high voltage (generally 10–80 kV/cm) at moderate temperatures, thereby ensuring
the retention of sensory and nutritional properties together with ensuring the quality.
Even though the temperature of processing can sometimes be above ambient, this
treatment can be considered non-thermal as the temperature is usually below the
conventional pasteurization conditions. The food to be treated is placed between
two electrodes where the electric field is generated. The efficiency of treatment is
dependent on the strength of this electric field, and PEF treatments at varying
strengths have been found useful in enhancement of drying, improving the yield in
extracting, transfer of genes, pasteurization, etc. (Wang et al. 2018).
The equipment used for PEF consists of a treatment chamber containing the elec-
trodes, a pulse generation system connected to a high-voltage electric supply, pumps
to handle the liquid food, and the control mechanisms (Mohamed and Eissa 2012).
The treatment chamber can be designed to operate either as a batch system or as a
continuous system. The electrodes are connected to each other using a material hav-
ing low electric conductivity to prevent the flow of current between them. The pulses
generated are transmitted via the electrodes through the food to be treated. The elec-
tric field damages the cellular membrane, causing microbial inactivation.
When an electric field is applied, the charged particles transfer the current to
every part of the food, leading to a potential difference between the cellular compo-
nents and the surroundings. This transmembrane potential destabilizes the cellular
membrane causing formation of pores, a process known as electroporation. The
permeabilization may be either reversible or irreversible depending on the strength
of treatment. This disruption of membrane is considered as the prime reason for
microbial inactivation by PEF, though the mechanism of action of PEF has not yet
been completely deciphered. Another hypothesis supports the idea of denaturation
of proteins and phospholipids due to potential difference as a possible mechanism
(Buckow et al. 2014). The advantages and limitations of pulsed electric field are
enlisted in Table 2.6.

limitations of pulsed electric • Minimal processing
field
• Less treatment time
• Color, flavor, and nutritional
retention
• Applicable as batch or
continuous process
• Enhanced mass transfer
• No chemical reactions
• Reduced fouling
Limitations
• High initial investment required
• Treatment is applicable only to
specific food types
• Not very effective against spores
• Cannot inactivate enzymes
• Presence of bubbles reduces the
efficiency
2.7.1 Applications
The application of PEF has been explored in several fronts including microbial
destruction, extraction, pressing, osmotic dehydration, drying, and freezing
(Sitzmann et al. 2016). As already mentioned, the main mechanism of action of PEF
is through electroporation of microbial membranes. The efficacy of PEF treatment
depends on several factors like process parameters including field strength, wave
characteristics, and time-temperature of treatment; product parameters like food
type, composition, moisture content, conductivity, pH, etc.; and microbial parame-
ters like type, population, stage of growth, etc. (Wang et al. 2018).
A large number of studies have been done applying PEF to milk. Almost all of
these have concentrated on the microbial inactivation efficiency of PEF. As most of
these works have been done more than a decade back, they are not detailed here.
The extensive review by Bendicho et al. (2002) may be referred for getting an
insight. He has quoted several studies and showed the effect of PEF against numer-
ous organisms in milk like Escherichia coli, Staphylococcus aureus, Lactobacillus
delbrueckii, Bacillus subtilis, Salmonella dublin, Listeria innocua, Pseudomonas
fluorescens, Lactobacillus brevis, Streptococcus thermophilus, Lactobacillus bul-
garicus, and Saccharomyces cerevisiae. Of late Sharma et al. (2018) treated milk
with a pulsed electric field of 25.7 kV/cm for 34 μs and reported shelf life and alka-
line phosphatase levels commensurate with thermal pasteurization. Alirezalu et al.
(2019) proposed combining PEF with mild heating below pasteurization tempera-
tures as a promising alternative to thermal pasteurization at an industrial level.
Several studies of PEF treatment of beverages containing milk along with fruit
juices have also been reported (Barba et al. 2015).
Yu et al. (2009) investigated the effect of field intensity, pulse strength, and tem-
perature on the coagulation properties of milk like rennetability and firmness of
curd and found a significant response. PEF-treated milk was found to have better
firmness and rennetability in comparison with the conventional process. Sharma
et al. (2015) reported a reduction in size and increase in surface area of milk fat
globule membranes as a result of PEF treatment. Further, they observed the adsorp-
tion of plasma proteins to the surface of PEF-treated fat globule membranes. Xiang
et al. (2011) suggested the use of PEF for modifying the structure of whey proteins
for product development or improved functionality. They found the whey proteins
in PEF-treated whey protein isolates to be partially denatured, exposing the hydro-
phobic groups. Simonis et al. (2019) studied the possibility of PEF treatment for
microbial inactivation of heat-sensitive acid whey by incorporating a cooling mech-
anism. They observed significant reduction in microbial population. Another inno-
vative study by Lemay et al. (2019) involved using PEF in combination with
electrodialysis for the demineralization of sweet whey. They reported an increase in
demineralization rate by 81% and a saving in energy by 16% by the incorporation
of PEF.
Reineke et al. (2015) used a combination of PEF and thermal processing to study
the inactivation of spores in skim milk. They have reported a synergistic effect of the
two techniques to have enhanced the destruction of spores, suggesting the extension
of scope of PEF to sterilization also. Ohba et al. (2016) observed that PEF in a mod-
erate strength (8 kV/cm, 1 μs pulse) was capable of increasing the exopolysaccha-
ride production of Lactococcus lactis ssp. cremoris up to 94%. The electroporation
was slight and reversible, and there was no damage to the cell wall as the treatment
was mild. Pankiewicz et al. (2014) found that PEF treatment was capable of increas-
ing accumulation of magnesium and zinc ions in Saccharomyces cerevisiae. Later
studies have extended this technique using lactic acid bacteria also for production of
probiotic and fermented ice cream (Góral et al. 2018; Pankiewicz et al. 2019).
2.8 Conclusions
As the food habits and consumer requirements are evolving over time, newer meth-
ods of processing are also getting identified. Even though processing of foods has
been in practice for millennia, we have not yet settled with an ideal processing
technique. Advances in different areas of science and technology are contributing to
food processing also. Furthermore, a food product beyond the bounds of spoilage,
having unlimited shelf life, is still a very distant dream. From this perspective, non-
thermal technologies are indeed a step forward, though these techniques need fur-
ther improvements in terms of optimization, cost involved, etc.
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Dairy Engineering: A Keystone
to the Dairy Industry 3
Kesha D. Vankar
Contents
3.1 Introduction 52
3.2 The Beginning 52
3.3 Engineering the Dairy 53
3.4 Milk Processing 54
3.5 The Cold Chain 55
3.6 Farm Automation 55
3.7 Rheology 56
3.8 Engineering in the Dairy Plant 56
3.9 Pointers to the Future 59
3.10 Conclusions 60
Reference 61
Abstract
Dairy engineering, as the name suggests, is the application of engineering to milk
and milk products. But the interesting part comes when different products are to
be made from one base material and how the characteristics change. With mil-
lions of tons of milk being produced each day, it becomes necessary to process
using proper engineering techniques and methods before sending it to the end
user. Dairy engineering has played a key role in shaping the dairy industry. Right
from the collection of raw milk to processing and packaging, dairy engineering
has laid its foundation to make safe milk available for consumption. Along with
milk, the process of making value-added milk products requires good technology
and engineering processes behind it. Not only in product manufacturing but even
in understanding the utility requirements and in ensuring minimum losses
K. D. Vankar (*)
Brand Sales, PI Industries, Gurgaon, Haryana, India
e-mail: a16keshav@iima.ac.in

https://doi.org/10.1007/978-981-15-2608-4_3
52 K. D. Vankar
in effluent causing limited environmental impact, engineering plays a crucial

part. While dealing with machinery in the plant, dairy engineering is useful to
carry out preventive and breakdown maintenance to maintain and/or improve the
efficiency of the machines. Engineering helps in new product development to
know its properties and even to understand the product life cycle. Automation
has played an important role in fine-tuning dairy engineering. Lastly, while deal-
ing with milk and milk products, there is a wide range of temperature difference,
ranging from minus degree to ambient temperature. In order to deal with this
variance, we need to have a cold chain to retain the product properties and even
shelf life.
Keywords
Dairy industry · Dairy engineering · Milk processing · Dairy plant · Milk process-
ing techniques
3.1 Introduction
Around the globe, approximately, 150 million households are involved in milk pro-
duction. Since the last three decades, milk production has increased by 58%. It has
amplified from 522 million tons in 1987 to 829 million tons in 2019 (FAO 2019).
Milk production is carried out both in the developed and developing markets.
Developing countries dominate with the lion’s share in global dairy production.
India is the world’s largest milk producer, with 21% of global milk production, fol-
lowed by the United States, China, Pakistan, and Brazil (FAO 2019). In most devel-
oping countries, milk is produced by small-scale producers, which contributes to
household livelihoods, food security, and nutrition. Milk provides comparatively
quick returns for small-scale producers and is an important source of rural income
generation. With such a huge quantity of milk being produced, it becomes necessary
to have good technology and processes to handle this delicate product. Since the
commencement of the dairy sector, there has always been a need to develop a strong
engineering system to process the milk and also to convert excess of milk into
value-added dairy products. Many developing countries have hot and/or humid cli-
mates that are unfavorable for dairying. Engineering through cold supply chain
management system plays a vital role in safeguarding the milk’s quality. The crux
of the matter is that engineering plays a key role in the dairy industry.
3.2 The Beginning
History mentions that initially milking was done from milch animals and then con-
sumed locally. Milk has been a part of nutrition in the human diet since inception.
But with the increasing number of the population both of humans and the milch
3 Dairy Engineering: A Keystone to the Dairy Industry 53
cattle, there arose the need of preserving milk and even transporting to distant places
to make it available to a larger number of populations. But in this process of storing
milk or transporting it to far places, there was a problem, where the milk used to get
spoiled. It was then in the year 1862, when a French microbiologist named Louis
Pasteur carried out heat treatment of milk and was termed as pasteurization process.
Commercially the pasteurization machines came into the industry in the year 1895.
This was the first big leap in building blocks for dairy engineering. Once the milk
started getting processed, then came a need to convert milk into dairy products.
Traditionally, people used to make products like cheese, ghee, butter, etc. at their
home using their own methods. But then there came a need for separating cream
from milk and processing it further. It was in year 1879, when a Swedish engineer
named Gustaf de Laval introduced the centrifugal principle for separating cream.
Since then from year 1882, the separators have been commercially built and
upgraded with time. Later in the year 1899, a scientist named Gaulin invented the
process of homogenization. Later membrane filters were invented for further con-
centration of milk. With time, the excess of milk was used to get converted into
powder, for which evaporators and dryers were designed. As said by Karl Marx
“Revolutions are the locomotives of history,” there have been many pioneers in the
field of dairy engineering which has led to milk revolution in the world. With all
these locomotives, there was a revolution leading to 1st of June being celebrated as
world milk day every year.
3.3 Engineering the Dairy
There are different kinds of specializations done in the field of engineering, and
dairy engineering is one among such specialized fields. Dairy engineering deals
with the processing of milk and milk products, where the technologies are estab-
lished, and is one of the major branches of dairy processing. Here the engineers
apply the principles of science and mathematics to bring about a sustaining techni-
cal and economical solution. Usually, the engineer in this field creates product
design and concepts that are viable for the long term. The engineer helps in specify-
ing the functionalities of the product design to help avail the overall efficiency along
with the cost and safety. Once the concept of dairy is understood, the principles can
be used for the whole range of milk and milk products and can further be extended
to the food sector too. The work is not just limited to the product design, but it is
further expanded into the areas of plant layout, the maintenance part, testing, auto-
mation, production supervision and help in maintaining its quality, etc. As the dairy
engineer is well aware of the dairy sector and how the things work, the project
management can be easily done as they understand the timelines and the cost asso-
ciated with different components and its scheduled completion. A dairy engineer
can also be associated with the sales activities of the sector, as the technicalities are
easily understood by them. They can even help the consumers in the product selec-
tion, planning, and installation of the products and the usage of the equipment.
Automation and artificial intelligence are gaining entry into this field, where again
54 K. D. Vankar
the dairy engineering plays a crucial role in understanding the product properties
and quality.
3.4 Milk Processing
The things discussed till now were the outer aspects of the engineering, but dairy
engineering has a lot more to do into the industry, right from the raw material until
the final stage engineering is involved. There is much more to dairy engineering
when it comes to milk being a perishable commodity, and hence lots of processes
are developed to turn this low shelf life product into long shelf life products. Lots of
technologies have been developed to process milk and milk products. To success-
fully turn this technology into commercial production, engineering plays a vital role
to set up the equipment and run the whole plant. Making milk and milk products
requires a defined set of procedures. Few of the procedures that are necessary for the
dairy are cooling, heat treatment, fermentation, etc. Pasteurization is one such pro-
cess that is used to extend the milk shelf life. Here milk is heated at a particular time
and temperature combination which helps in removing the pathogenic microorgan-
isms and makes milk safe for consumption. Engineering design helps to ensure that
milk is heated at that temperature and held for a particular time. If that is not done,
FDV (flow diversion valve) comes into the system which does not allow milk to go
for packing. Pasteurization helps to get a shelf life of milk for 1 or 2 days. But with
the generation “Z,” where they get busy with work and other activities, there came
a need to have milk with longer shelf life. Engineering made some more innovations
in the current process, and thus a process named sterilization got invented. Here in
the aseptic ultrahigh treatment process (i.e., sterilization), milk is heated through
steam at very high temperature and held up for few seconds, ensuring that the
F-value is maintained and all the microbes are killed. This process makes milk to
have a shelf life of approximately 6 months. When we consider milk production,
there are few countries who are milk deficit and others are milk excess producing
countries. Milk is traded in terms of powder to those milk-deficit countries. Milk
when converted into powder has a shelf life of 1 year. Here drying process is
involved where milk droplets undergo flash heating at very high temperature that it
gets converted into powder. This process even helps in transporting bulk milk to
distant places without any fear of spoilage. Condensing is another process where
milk is heated in vacuum pan which helps remove all the odors and the burnt taste
from milk. This by-product is again used to make other milk products, and it can be
even used in the bakery segment, ice-cream segment, etc. These are few heat engi-
neering techniques used to convert a perishable product into long shelf life products.
Milk can also be converted into other high-value dairy products like cheese, fer-
mented milk, paneer, condensed milk, ice creams, baby food, milk powder, butter,
etc. The consumption of milk and milk products varies in different regions based on
their consumption pattern; hence there are different products and processes in dif-
ferent regions. Engineering plays a vital role in converting this milk into other
delicacies. Right from the reception of milk procurement till the final dispatch at
consumers home, the processes are engineered.
3.5 The Cold Chain
Cooling is another engineering process which is inevitable in dairying. Milk is

chilled till the time it reaches to the milk processing plant. When the milking is
done, soon it is stored in cold temperature which ensures that no other microorgan-
isms grow into it. Sometimes the distance between the milking place and processing
plant is more, and hence it is important to keep the milk chilled before it gets spoiled.
This whole farm to fork process involves cold supply chain where engineering is
involved. When processing is done, then the very next step after the processing is
chilling the products at the desired temperature or to store the products at the desired
cold temperature. For this cold storage, rooms are designed. Engineering aspects
help in understanding the heat load emitted by the product, and this will help in
determining the refrigeration capacity for the overall plant. There is lots of engi-
neering involved while designing cold storage. Say while designing cold storage for
milk, knowledge in engineering helps to determine what will be the stack height of
milk crates, what will be the air convection, if there is any requirement for air cur-
tain, etc. Similarly, while designing cold store for ice cream, some important param-
eters are considered like what is the chilling load, small heating coils to be kept to
remove defrosting, what should be the airflow in the cold storage unit, etc. Hence it
becomes important for dairy engineer to minutely check all the details making engi-
neering a key to dairy industry. The cold chain is one important engineering concept
required in temperate countries. If we talk about ice cream which is the favorite
product of many in all age range, it needs to be cooled as soon as it is produced and
kept into hardening room at around −18 °C or less. In another example, the aging of
cheese is done at a controlled temperature to develop flavor and aroma. In other
terms, the engineering involved in cooling is not just limited to storage but also in
enhancing flavors in many products. Chilling may sound a simple word, but there
goes lots of engineering into making the product chilled or say have a cold chain.
3.6 Farm Automation
Hand milking is preferred by marginal farmers with two to three cows or buffaloes.
Many times, due to a large number of milking animals, it becomes difficult to do
hand milking and even take care of hygiene. Engineering helps in making auto-
mated milking machines that can be installed in the dairy farms. Special care is
taken while engineering these machines since they come in contact with the animals
directly. Care is taken that these machines are gently fitted to the animals while
milking. Also, the whole milking plant is cleaned properly to maintain the hygiene
of animals. Thus even on farm level, the dairy engineering plays an important role
which helps the dairy industry indirectly.
56 K. D. Vankar
3.7 Rheology
Rheology is an important engineering term linked with the dairy products. Many
factors are considered while a dairy processing plant is designed, and hence rheol-
ogy plays an important role. It helps in assuring the quality of the final products. If
the flow behavior properties are not understood properly, then there are chances that
the product characteristics may be completely or partially spoiled. Practically rheol-
ogy stands for flow behavior, viscosity measurement, and determination of material
structure, which helps in process design and evaluation of product quality. Milk and
milk products possess liquid, semiliquid, and solid characteristics. Rheological
properties help in identifying such properties that help in understanding the struc-
ture and are one of the quality control parameters in the production plants. To under-
stand this structure, different engineering equations are created and worked upon it
for the quality parameters. Texture and viscosity are important parameters to under-
stand the product properties and to understand the overall plant requirements. If we
know the product rheology, then it helps us in understanding the pressure required
to pump the product from one place to another, and thus engineering can help in
design optimization of the plant. Indirectly it even helps in understanding the utility
requirements of the overall manufacturing unit/plant.
3.8 Engineering in the Dairy Plant
The dairy engineer understands the different parameters to be considered while

designing the processing facility. For example, if whipping cream is to be made,
there are different parameters involved, like the cream to be pasteurized at the initial
stage has different properties and once it is pasteurized and chilled the properties
change. It becomes necessary to understand the product properties and thus make
the component selection for the same. Component selection involves the selection
of pumps, valves, homogenizers, cream separators, etc. Homogenizer is a beauti-
fully engineered equipment, where milk fat is broken into small fat globules having
even shape in two stages and equally distributed into the milk. Homogenizer can be
called the heart of the dairy industry as many processes involve homogenization.
Even a cream separator, which is usually used to adjust the fat percentage required
in the cream to the desired level and in skim milk, can be used for further process-
ing. The cream separator also helps in butter making where high-fat cream is further
used to make butter or ghee (for Indian market). Lots of designing is involved in the
equipment, and piping is also made in such a way that the equipment can serve
multiple products. Engineering has joined its hands with microbiology in making
another equipment known as bactofuge, which primarily helps in reducing the bac-
terial load without heavy heat treatment. Providing milk with long shelf life and low
bacterial count has always been the priority of the industry. Such equipment, if used
for cheese making, can help in increasing yield, taste, and even quality. This can
even help indirectly in reducing the biological oxygen load in the effluent treat-
ment plant.
Once the rheology is understood, dairy engineering plays an important role in

material design and selection. Since we deal with a food product, it is necessary to
know the food-grade material which comes in direct contact with the product. The
material selection is made in such a way that the cleaning solution does not corrode
the material or else it compromises the product quality. Another aspect where the
inputs of dairy engineering are required is in the movement of the product on the
plant floor. Given the state of the product like solid, semisolid, or liquid, the dairy
engineer is able to understand the fluid mechanics and the transport phenomena.
Once this is known, engineering helps in designing the pipeline layout of the plant.
This helps to understand how the product properties change during processing or
during the transport of it, and hence necessary modifications can be made, like if
UHT treatment is to be done, it is done at 135–150 °C for few seconds, and the hold-
ing section of the pasteurizer is designed in such a fashion that the milk particles
don’t get burnt while the flow of the milk is maintained. If milk powder is to be
made, the pipelines for inlet of milk are designed in such a way that they help in
maintaining the force in the drying chamber, and once the milk is dried, the outlet is
also designed in a manner where the product flow is properly done. If the flow
behavior of the product is not understood properly, then products like cream and
cultured milk can get spoiled completely. Dairy engineering plays a pivotal role in
the design of equipment. Consider a simple cream cooling silo. It seems simple, but
while designing, one has to understand the properties of the product and the impact
of a faulty design. Generally during the design of a cooling vat for cream, if one
keeps the jacketed chamber where cold water flows, there are chances that the cream
may develop ice crystals on the surface that comes in direct contact with the jacket.
But if engineered properly, like if the jacketed silo is made with the provision of
water pipe, where the chilled water is not stagnant and keeps on flowing and on the
inside a cream agitator at optimum speed is kept, then the desired processing param-
eters are achieved along with the required quality. Engineering of equipment is a
critical part of dairy engineering. Equipment design helps in optimizing the pro-
cessing parameters, the product quality, simulating with other equipment, and even
in energy and efficiency optimization. Let’s say if a heating plate is to be designed
in a pasteurizer, there are many small parameters involved into it, like what should
be the surface area so that the milk is heated with optimum properties, it should be
even able to withstand the pressure of the system. Also, while designing the pasteur-
izer system, the pressure on the product side is kept higher than the utility pressure
so that the steam or water does not get mixed with the milk through pinholes, if any
are present, during any pressure drop in the system. Another aspect of dairy engi-
neering is the manufacturing of the lab equipment that is used for the quality control
check. It’s very important to understand this core property to design the testing
equipment because the product manufactured reaches the consumer and engineer-
ing plays an important part in ensuring food safety. Packaging of dairy products also
involves dairy engineering. The packaging materials are generally chosen according
to the product properties. Packaging material strength, its quality, etc. play an
important role to hold the product and even make advantage of its shape. The pack-
aging machines are engineered in such a way that it causes no damage to the product
58 K. D. Vankar
and minimum loss to the packaging material. Parameters like the temperature to be
kept in the sealing jaws used for heat sealing need to be monitored and engineered
properly. Thermodynamics and refrigeration are also important pillars that make
dairy engineering a keystone to the dairy industry. Milk and milk products are stored
at different temperatures starting from storing the processed milk at 4 °C to the ice
cream at −18 °C temperature. Or if we consider a product like cheese, it needs to be
aged at a relative humidity of 65%. So, if one knows the properties of the product
and how much heat to extract from the product to get a desirable outcome, it
becomes important to understand the laws of thermodynamics and refrigeration
mechanism. Also, for the overall utility requirements, one can know the capacity of
the boiler to be installed or the required tonnage of the cooling plant. While doing
the engineering of dairy, it’s not complete without doing the engineering of the utili-
ties. When one has to design a boiler selection, proper engineering is carried out as
to understand what is the normal steam load of the whole dairy. The boiler selection
is done based on steam load and the capacity. Based on the steam generation require-
ment, required utility line piping size is to be selected, and care is taken that proper
insulation is being done on the utility pipes so that the safety issues are also taken
cared of. When one works in a dairy plant, there is usually a heavy processing load
in the morning time, which in turn increases the load on the boiler side too. Thus,
when the boiler capacity is selected, even these parameters are considered. Similarly,
while designing a refrigeration plant, chilling load is calculated based on different
cold storages, and accordingly the refrigeration plant is designed. Once the process-
ing capacity is decided, there is another area where dairy waste is processed and
ensured that the environment is not harmed—the effluent treatment plant (ETP).
While designing the effluent treatment plant, care is taken that the processed waste
does not exceed the permitted biological oxygen demand and chemical oxygen
demand limits. In this, the products processed or manufactured in the dairy plant
also play an important role. If cheese or paneer is manufactured and the by-product,
i.e., whey, is let to flow down the drain, there is more load on the ETP plant, and
accordingly, more intense treatment of effluent will be required. Initial load of the
waste coming out of dairy impacts the energy utilized in the ETP. The engineering
part does not end here, but it also helps in understanding the product wastage and
how that waste can be utilized in generating bioenergy, which can partially contrib-
ute to the overall energy requirement of the plant. For example, the whey that is
coming out from the cheese plant can be used in generating biogas from the effluent
treatment plant and can be used in running plant components like pumps. Similarly,
the wastewater that is obtained from the dairy industry can be reused after applying
proper filtration techniques. With simple treatments itself, the treated wastewater
can be used for the irrigation of garden or lawns, thereby improving the aesthetic
appearance of the unit. By incorporating treatments like reverse osmosis, the water
can even be used for drinking. Dairy engineering also helps in conserving the energy
in dairy industry. There are areas in the plant where proper insulation to the pipe-
lines or the cold storages can help in reducing energy loss and thereby improve
savings in the industry. While cleaning the plant, one can always work on control-
ling the temperature and increasing the flow to some extent, which can help in
saving water as well as energy. If any pump is running in full capacity and we just
want it to run on part load, technologies like variable frequency drives help in ensur-
ing energy saving along with reducing the wear and tear of the pump. Engineering
also plays a key role in the maintenance of the plant. Once an equipment starts run-
ning, there are chances that it will have a breakdown some time in its functional life.
To delay the breakdown, a preventive maintenance schedule is made and thus helps
in improving the efficiency of the equipment. Last but not least, engineering is also
applied in designing the plant layout. Plant layout is very important to understand
where the necessary equipment and other associated things are to be positioned and
how it will help in improving the efficiency of the plant and the people who are
working in it. Also, the plant layout is designed in such a way that, if there is an
emergency in the plant, the employees can easily escape and safeguard themselves.
If there is a processing layout followed, it helps in minimizing the processing time
and indirectly helps in seamless production and time-saving. Similarly, in the store
section if the layout is made in the philosophy of first in first out (FIFO), then it gets
aligned with Six Sigma process which helps in reducing the variability of the busi-
ness. Thus, the dairy engineer needs to consider all these details while making the
plant layout. In a nutshell the different aspects of dairy engineering are represented
in Fig. 3.1.
3.9 Pointers to the Future
Few projects where dairy engineering has focused in moving toward the next-
generation dairy sector are:
Milk Processing
Cold chain management
Farm automation
Reology and product development
Testing and instrumentation
Utilities
Designing and layout
Fig. 3.1 The different aspects of dairy engineering

60 K. D. Vankar
• High-resolution microscopy is being used for research purpose to focus on the

rheological properties of the product. This helps in new product development
along with its quality assurance.
• During the manufacture of several dairy products, a few high-value products are
drained/lost. Separation techniques are researched, to recover such valuable
dairy ingredients and thus help in reducing the disposed waste since a significant
cost is involved in it. This also helps in reducing the environmental impact.
• Processing strategies are been developed to innovate new ingredients particularly
in dairy fat and emulsions sector.
• Engineering techniques have been researched to understand the physical and
technological properties of the products, which helps in ensuring consistently
positive organoleptic scores.
• Waste processing is one field where there is a scope of saving cost and even
improving efficiency. Research is being carried out to develop processes where
the waste is processed properly to get good output in terms of expenditure, effi-
ciency, and environmental impact.
• Energy saving is becoming a global concern. Dairy engineering research work is
going on to have clean energy utilization where technologies are made that use
green energy for production. Also, research is being carried out in technologies
that reduce the wastage and help in energy saving along with reducing the envi-
ronmental impact.
• Research is carried out in innovative processes so that new products can be
evolved and even the shelf life can be extended in a novel way.
• Pilot plants are being designed to carry out lab-scale research, and they are being
fine-tuned in such a way that there is minimal difference found between the lab-
scale and the industrial manufacturing.
• Smart packaging techniques which help in knowing the freshness of the product
are getting popular with dairy products.
• In many countries, region-specific sweetmeat industry is highly localized.
Research is being carried out to streamline these processes and have a large-scale
production with similar organoleptic properties across the country.
This list can go on indefinitely, and it is impossible to enlist all the advancements
in dairy engineering. A separate chapter has been included in this book, which
describes the recent developments and advanced researches being carried out in the
field of dairy engineering.
3.10 Conclusions
Right from milking the animals until the product reaches the customers, engineer-
ing is involved in every stage. Also, with artificial intelligence (AI) and Internet of
Things (IoT) coming in trend in the manufacturing sectors, slowly it will catch on
the dairy engineering sector too. Few food sectors have started using AI and IoT in
their smart packing systems and making the food safe for consumption. Dairy
engineering plays an important role in shaping the dairy industry and upholding its
legacy for the future.
Reference
FAO (2019) Milk production. Gateway to dairy production and products. http://www.fao.org/
dairy-production-products/production/en/. Accessed 25 June 2019
Advances in Dairy Engineering Research
Across the Globe 4
Aswin S. Warrier
Contents
4.1 Introduction 64
4.2 Advances in Farm-Level Interventions 64
4.3 Engineering in Milk Testing 66
4.4 Advances in Dairy Processing 68
4.4.1 Pasteurization and Sterilization 69
4.4.2 Cream Separation, Standardization, and Homogenization 70
4.4.3 Evaporation and Drying 71
4.4.4 Butter and Cheese 73
4.4.5 Ice Cream 73
4.5 Packaging 74
4.6 Cleaning and Sanitation 75
4.7 Dairy Effluent Treatment 76
4.8 Storage and Distribution 78
4.9 Conclusions 79
References 79
Abstract
Dairy engineering is the multidisciplinary field located at the intersection of
“dairy” and “engineering.” It involves the utilization of insights in the areas like
processing techniques, physical properties, chemistry, microbiology, engineer-
ing, economics, and management for fixing issues related with production, pro-
cessing, testing, handling, and storage of milk and milk products. This chapter
describes briefly about the advanced engineering interventions in milk produc-
tion, testing, processing and associated activities, storage, and distribution.
A. S. Warrier (*)
University Dairy Plant (Kerala Veterinary and Animal Sciences University),
e-mail: aswinswarrier@kvasu.ac.in

https://doi.org/10.1007/978-981-15-2608-4_4
64 A. S. Warrier
Keywords
Dairy engineering · Dairy processing · Milk products · Milk processing tech-
niques · Dairy equipment
4.1 Introduction
The word “dairy” can be used as a noun or as an adjective. As a noun it means an

establishment that produces processes or handles milk and milk products. As an
adjective, it can mean anything concerned with milk or milk products or the milch
animals. “Engineering” involves the application of science, technology, and math-
ematics to solve technical problems, utilizing optimum resources. Accordingly,
dairy engineering is the multidisciplinary field located at the intersection of “dairy”
and “engineering.” It involves the utilization of insights in the areas like processing
techniques, physical properties, chemistry, microbiology, engineering, economics,
and management for fixing issues related with production, processing, testing, han-
dling, and storage of milk and milk products. Within engineering, understanding in
diverse areas like chemical engineering, agricultural engineering, civil engineering,
instrumentation, material science, electrical engineering, etc. is exploited to sort out
a potpourri of challenges associated with the dairy sector. In this chapter we will be
discussing briefly about the engineering interventions in milk production, testing,
cold chain, etc. and in slightly more detail about the processing advancements.
4.2 Advances in Farm-Level Interventions
The quality of raw milk is of cardinal importance, as it will determine the quality of
final product. The quantity and quality of milk produced can only be ensured by
being attentive of various factors contributing to animal health and overall hygiene
of the production. Even though rearing animals for milk production is known to
mankind for millennia, large-scale farming or mechanization of farm activities was
unheard of till the beginning of the twentieth century. A major change in this sce-
nario happened with the introduction of milking machines and refrigeration systems
working on electricity. With the advent of automation, milk “untouched by hand” is
no more surprising to the common consumers.
Recently, the term “precision livestock farming” (PLF) is being utilized for the
use of advanced technologies for making more profitable “industrial farms” (Vessier
et al. 2017). According to Berckmans (2017), the aim of PLF is to manage individ-
ual animals by continuous real-time monitoring of health, welfare, production,
reproduction, and environmental impact. Automated and uninterrupted monitoring
is the crux of PLF. The monitoring can be done by camera and real-time image
analyses, by microphone and real-time sound analyses, or by sensors around or on
the animal (Berckmans 2017). The major interventions of PLF are in the areas of
4 Advances in Dairy Engineering Research Across the Globe 65
identification, feeding, milking, animal health, etc. These interventions make it pos-
sible to manage farms of larger size, holding an increased population of cattle, with
less number of farmers.
Cattle identification refers to accurately recognizing individual cattle using a
suitable marker (or any other methods) for increased traceability. Documentation,
traceability of products, and tracking of disease outbreaks are made easier with
proper identification. Methods like ear tags and branding, which are still widely
used, are slowly getting replaced by electronic identification systems, particularly
radio-frequency identification (RFID) which operates using radio waves. Electronic
ear tags, microchips, and electronic collars are the most prevalent automated identi-
fication methods today. Microchips containing a tiny radio transponder and antenna
are implanted below the skin of the animal (Neary and Yager 2002). Tags and collars
contain electronic information which can be read by a scanner. In a reported research
from Nigeria, Francis et al. (2018) have designed a collar consisting of a Global
Positioning System (GPS) and Global System for Mobile (GSM) module which can
communicate the coordinates of the cattle via short message service (SMS) or track
on Google Earth maps, enabling farmers to check the cattle’s positions from a long
distance and also prevent theft or restrict it from wandering beyond some fixed lim-
its. A common issue with these methods is that in all these systems the identification
is done based on devices attached to the animal and not the animal per se (Awad
2016). Biometric identifiers, such as muzzle prints and retinal imaging, are prospec-
tive methods which can precisely identify the animals instantaneously (Awad 2016).
An animal’s biometric identifier is permanent, enduring, and cannot be easily coun-
terfeited (Awad 2016). Kumar et al. (2016) have reported use of computer vision
approaches for cattle recognition using their facial images.
The health and productivity of milch animals are closely associated with the
quality and quantity of its feed. Apart from its direct influence on the productivity
of the cows, feeding contributes to a large chunk of expenditure, since more than
one fourth of labor time is taken up by this operation (Pezzuolo et al. 2016). Several
means have been identified for automation of feeding the cattle, which may be dif-
ferentiated based on three automation stages: (1) mixing-distribution, (2) filling
mixer-mixing-distribution, and (3) unloading and transport-filling mixer-mixing-
distribution (Da Borso et al. 2017). In automatic feeding systems (AFS), distribu-
tion of feeds is done automatically using conveyor belts, rail-guided wagons, or
self-propelled feed robots (Grothmann et al. 2010). Conveyor belts and chain link
tables are stationary feeding systems which are already prevalent in some developed
countries for feeding the animals in groups. Rail-guided wagons occupy an interme-
diate position between the stationary systems and the self-propelled systems. The
self-propelled system comprises automatic self-propelled mixer wagons that can
work without manual supervision (Bisaglia et al. 2010). The wagon has a block
unloading device which fills a vertical auger mixing device. The navigation system
permits easy use of these robots in the existing farm structures without any major
modifications and proper feeding of animals based on a time schedule (Bisaglia
et al. 2010). According to the companies in this business, automatic feeding can
reduce the workload while ensuring hygiene and limiting feed loss (Grothmann
66 A. S. Warrier
et al. 2010). The main advantage AFS has over conventional feeding system is that
the manual interventions required for preparing and conveying the feed are minimal
and hence the frequency of feeding can be increased (Bisaglia et al. 2012). Recently,
Wijayanto et al. (2019) have reported development of an automated feeding system
based on Internet of Things (IOT) that can be integrated harmoniously with existing
software.
In an “automatic milking system” (AMS), all the activities involved in the milk-
ing process and livestock management that were performed manually in the conven-
tional system have been automated. AMS gives priority to the cows’ milking
instinct, by permitting them to be milked whenever they feel so simply by walking
into the milking area, unlike conventional milking, where the priority is on the con-
venience or routine of the farmers (De Koning and Rodenburg 2004). AMS, also
called “robotic milking,” has a robotic arm for automatically connecting and remov-
ing the teat cups to the udder, thereby reducing the manual labor requirement sig-
nificantly (Schewe and Stuart 2015). Jiang et al. (2017) have reviewed three main
types of AMS, namely integrated AMS, industrial robotic AMS, and the automatic
milking rotary system.
Electronic identification of cattle is indispensable in a robotic milking system.
When an animal enters the milking area, a sensor identifies the animal by electroni-
cally reading its tag/transponder and is then rewarded with a preferred feed in quan-
tities relative to her production (Brouček and ToNGeĽ 2015). The robotic arm,
assisted by a 3D camera and laser technology to recognize the movement of the
animal and locate the teats, connects the teat cups and then starts milking each teat
consecutively (Brouček and ToNGeĽ 2015). Facility for cleaning the teats before
and after milking is also usually provided. When milking is complete, the teat cups
are detached and the animal is allowed to move out of the milking area (Nayik et al.
2015). Data from sensors, which monitor the milking process, are systematically
saved in a database, based on which necessary controls or adjustments in parameters
can be made by the farmer using its software (De Koning and Rodenburg 2004). The
milk collected is cooled immediately to below 4 °C and is then stored in a bulk tank.
4.3 Engineering in Milk Testing
Testing of milk can be done for determining certain physical, chemical, or micro-
biological parameters related to milk. Although many of these parameters are at
times checked for academic or industrial research and design purposes, chemical
and microbiological tests are mostly done to ascertain milk quality. Quality control
of milk includes numerous tests to assure the chemical and microbiological safety,
nutrition, and the legal requirements (Tessema and Tibbo 2009). Even though a lot
of technological advancements have happened in the physical, chemical, as well as
microbiological testing of milk, the conventional methods requiring much skills and
consuming more time and energy are still in common use.
In most of the countries, payment of raw milk to farmers is done on the basis of
volume as well as composition of the milk, though the interpretation of composition
may vary. There are payment systems which consider only fat content, total solids
content, fat and non-fat solids separately, or fat and protein separately. The automa-
tion in milk composition analysis using light scattering or infrared spectroscopy is
in vogue for more than 40 years (McGann 1978). The infrared method checks the
absorbance in the near- or mid-infrared spectra to estimate total solids. The estima-
tion is done by comparing the calculated values of fat, protein, and lactose with the
total solids detected by the standard procedure and then applying a regression cor-
relation (Barbano and Lynch 2006). Most of the researches in compositional analy-
sis of milk utilize optical methods or use some kind of electromagnetic radiations.
Techniques like Fourier transform (FT)-Raman spectroscopy (Mazurek et al. 2015;
Li et al. 2015), dielectric spectroscopy (Zhu et al. 2016; Guo et al. 2017), laser-
induced breakdown spectroscopy (Bilge et al. 2016; Moncayo et al. 2017), etc.,
alongside other spectroscopic techniques (Kala et al. 2018), have been reported for
analyzing the composition of milk.
The knowledge of acidity and pH is essential in ascertaining the keeping quality
and heat stability of milk. Automatic potentiometric titrators have been known for
many decades and have found many applications more than just determining acidity
(Pyne 1934). An automatic titrator usually consists of an electric burette, a sensor,
and a microcomputer. While titrating, addition of titrant from the electric burette is
controlled by the signal from the sensor, which is in fact just a pH electrode in case
of acidity measurement. On reaching the endpoint, the microcomputer calculates
the volume of titrant added and converts this value to a result based on programmed
formulas (GPS Instrumentation Ltd 2019). Recently, Isildak and Gones (2018) have
reported determination of pH and total acidity of milk by employing a sequential
injection analysis method, which uses a potentiometric system. Several researchers
(Manuelian et al. 2017; Calamari et al. 2016; De Marchi et al. 2009; Su and Sun
2019) have reported the use of mid-infrared spectroscopy in determination of titrat-
able acidity.
Adulteration of milk is a very troubling challenge faced by dairy industry. It not
only reduces the quality of the product but can even affect the health of the con-
sumer and the reputation of the processor. Typical adulterants found in milk include
extraneous water, sugar, fats and proteins of vegetable origin, milk from other spe-
cies, whey, urea, detergents, formaldehyde, hydrogen peroxide, salicylic acid,
melamine, etc. This is not a complete list, and newer methods of adulteration are
getting identified, while better methods for determining known adulterants are
found. Development in the field of chemometrics has made increased adoption of
instrumental methods in adulteration detection possible. Techniques like liquid
chromatography, gas chromatography, spectroscopic techniques, digital image anal-
ysis, electrochemical sensors, etc. are used for detecting milk adulteration
(Nascimento et al. 2017). An extensive review of studies in this area is beyond the
scope of this chapter. Reviews by Nascimento et al. (2017), Poonia et al. (2017),
Baiano (2017), Abernethy et al. (2016), and Kamal and Karoui (2015) may be
referred for this purpose.
Microorganisms play a crucial role in shelf life, beneficial effects, and even
adverse effects of milk and dairy products. The detection and identification of
68 A. S. Warrier
microbes within a short time is thus a thriving field in dairy research. Even though,
some amount of mechanization and automation has been used in techniques like
flow cytometry, enzyme-linked immunosorbent assay (ELISA), or polymerase
chain reaction (PCR), we cannot consider these as fully automated as their out-
comes cannot be used automatically for plant operation control (Sharpe Anthony
2018). Chemical sensors and biosensors (Poghossian et al. 2019), spectrometric
methods, PCR-based methods, multivariate techniques (Buszewski et al. 2017), etc.
are proposed as alternative faster methods. A detailed description on such rapid
methods is given elsewhere (Chaps. 10 and 11) in this book.
Rheology is the study of flow and deformation of matter. The measurement of
viscosity and other rheological parameters of practical significance is termed as
rheometry. A rheometer performs either of the following two functions, namely
applying a deformation mode and measuring the subsequent force generated or
applying a force mode and measuring the subsequent deformation (Shenoy 1999).
Rheometry has evolved from simple rotational viscometers to more advanced and
accurate Fourier transformation rheometers (Ahmed 2018). Tribology, the science
of interacting surfaces in relative motion, including the study of friction, wear, and
lubrication, is nowadays considered as a contributing discipline to rheology. The
advances in the field of rheological research, including rheometry, oscillatory shear
tests, tribology, etc., are explained in detail by Ahmed et al. (2016).
Sensory evaluation refers to evaluating a food product with the help of human senses.
Though senses are the foundation of sensory evaluation, a lot of research is going on in
instrumental analysis using machines that mimic human senses including olfaction,
taste, and mouthfeel. Nowadays, electronic nose technology is getting prevalent in
assessing food quality by sensing aroma without separating the volatile components.
Normally e-noses are made of a sampling system, a sensing system, and a pattern rec-
ognition system (Gliszczyńska-Świgło and Chmielewski 2017). Several reviews
(Gomes 2016; Shi et al. 2018) on the application of e-nose technology for milk and
dairy products have been published. Electronic tongue and bioelectronic tongue are
innovative technologies for sensing taste, which uses electrochemical sensors or biosen-
sor array to mimic the taste detection procedure of human tongue (Ha et al. 2015).
Application of electronic tongues to milk and dairy products has been reviewed by sev-
eral scholars (Ha et al. 2015; Ciosek 2016). Computer vision involves capturing, pro-
cessing, and analyzing images, thereby assessing the visual quality attributes in food
products (Brosnan and Sun 2004). Within dairy products, this technology has found
potential mostly in cheese manufacture (Lukinac et al. 2018). Di Rosa et al. (2017) have
reviewed a fusion of electronic nose, electronic tongue, and computer vision technolo-
gies for the authentication and quality assurance of foods from animal sources.
4.4 Advances in Dairy Processing
Dairy processing encompasses all those operations carried out for ensuring the
quality and safety and increasing the shelf life of milk, as well as its conversion into
various dairy products. Dairy industry, having a rich history of more than a century,
has always been progressive in adapting to newer technological developments.

Presently dairy processing is a highly organized business having huge automated
factories and operating across the international frontiers. This section will discuss
the major engineering advances related to dairy processing and associated activities
in the last few years.
4.4.1 Pasteurization and Sterilization
Pasteurization is the most prominent and universally accepted milk processing oper-
ation. Since the early twentieth century, when the regenerative heat exchangers were
introduced and the necessary time–temperature combinations were established, there
have been no fundamental changes in the pasteurization process to the present. Most
of the progress made with respect to pasteurization is in the areas of improving the
capacity, energy efficiency, hygiene, and automation, along with identifying alterna-
tive methods (Warrier et al. 2017). Milk pasteurization plants are now capable of
producing volumes of up to 200,000 L/h, with regeneration rates of 94% being pos-
sible (Goff and Griffiths 2006). Though the process of sterilization was used since
the early twentieth century, its level of popularity was not very impressive until the
ultrahigh temperature (UHT) process and aseptic packaging took the center stage.
Further researches done in this field were in the areas of fouling reduction, fixing the
flavor, etc. along with works similar to those of pasteurization.
Taghizadeh-Tabari et al. (2016) reported that using a nanofluid made by adding
titanium dioxide nanoparticles to distilled water increased heat transfer rate in plate
heat exchanger for milk pasteurization. This increase can be attributed to the higher
thermal conductivity of the nanofluid. Zhang et al. (2019) have extensively reviewed
the methods for enhancing heat transfer rate in plate heat exchangers concentrating
on the effects of corrugation geometries, parameters on the heat exchanger effi-
ciency, and application of approaches like passive surface techniques and the use of
nanofluids.
Pasteurization or sterilization of milk using methods like bactofugation, micro-
filtration, high pressure processing, pulsed electric field processing, treatment using
UV and IR radiation, ohmic heating, microwave processing, pasteurization using
supercritical fluid extraction, irradiation, electron beam pasteurization, thermosoni-
cation, etc. has been proposed as alternatives to conventional processing techniques.
However, these topics are not discussed here as a whole chapter dedicated for alter-
native processing techniques has been included in this book. On a side note, Lim
et al. (2019) reported that neither cold sonication nor thermosonication was an
appropriate technique for reducing bacterial count in fluid milk beyond standard
pasteurization and, in fact, increased counts of spore-forming spoilage bacteria.
The importance of process control in any mechanized food processing operation
cannot be overlooked. In order to improve pasteurization temperature control,
Khadir and Ringwood (2017) applied generalized predictive control as well as
model predictive control (Khadir and Ringwood 2003) to the milk pasteurization
process. In their work, the process was simulated by artificial neural network-based
70 A. S. Warrier
model, and the responses were compared with an optimized proportional integral
derivative (PID) controller behavior. Riverol et al. (2008) have discussed the appli-
cation of three advanced control technologies, namely fuzzy control, model refer-
ence adaptive fuzzy control, and neural networks in a plate heat exchanger (PHE)
used for milk pasteurization, emphasizing the usefulness of artificial intelligence in
milk process control. Pour et al. (2017) combined model predictive control with a
fatigue estimation approach to decrease the damage of parts of a pasteurizer, so as
to find a compromise between life of the machine parts and energy consumption.
Another area of interest among researchers is in utilizing renewable energy
sources to meet the energy requirement of processing milk. Geothermal energy is
the thermal energy from earth, usually extracted from hot water reservoirs. It may
be used for heating, cooling, or power generation. Yildirim and Genc (2015)
designed a milk pasteurization system, in which the whole energy requirement was
supplied from geothermal source, wherein a vapor absorption refrigeration system
was used for the chilling stage. Panchal et al. (2018) have written an extensive
review on utilization of solar energy for pasteurization of milk, in which they have
compared systems with flat plate collectors, solar photovoltaic panels used for PEF
pasteurization, Fresnel-type concentrators, low-cost system with cardboard box and
aluminum foils, evacuated tube collector, etc. Researchers from Roskilde University,
Denmark (Lybæk and Sommart 2016), have made a case study on the applicability
of biogas in dairy cooperatives of Thailand, where they have suggested use of bio-
gas for process heating and electricity generation for milk pasteurization.
Researchers from University of Tennessee and University of Purdue have devel-
oped an add-on system to the conventional high-temperature short-time (HTST)
pasteurization, which they have named low-temperature short-time (LTST) method
(Myer et al. 2016). A Millisecond Technology (MST) chamber was used for this
study, in which the milk is sprayed into a chamber maintained at a below-
pasteurization temperature and heated for 0.02 s, utilizing residual energy from tra-
ditional pasteurizer, ensuring dispensability of additional energy resources. They
claimed that using this system in-line with the traditional pasteurizer showed
increased microbial reduction and shelf life, with negligible sensory changes. Not
many compelling works related to engineering advances in sterilization have been
reported in the last few years, except for the use of various nonconventional
methods.
4.4.2 Cream Separation, Standardization, and Homogenization
Centrifugation was used as a method of separating fat and skim milk by Gustaf de
Laval in 1879, and the use of conical discs in the centrifuges for their increased
efficiency was invented in 1888 by Freiherr von Bechtolsheim. By the mid-twentieth
century, the cream separator which we commonly see now was developed. Since
then, the major development has only been the increase in its capacity. However,
minor changes made include simplifying the design and design modifications to
improve the yield and hygiene and to remove the sludge easily (Gunsing et al.
2009). On searching the Internet, we can see that a lot of patents related to centrifu-
gal separation are granted every year, most of which are obtained by separator man-
ufacturing companies. Alfa Laval Corporate AB was granted a US patent (Königsson
2017) for a centrifugal separator comprising a rotor forming a separation space with
a set of separation plates defining separation passages there between. GEA
Mechanical Equipment GmBH was granted a US patent (Quiter et al. 2017) for a
separator disk package with separator disks having labyrinth-like flow channel.
Though a lot of industrial research is carried out to improve the process of cream
separation, only very few alternatives to centrifugation have been identified, which
are not yet technologically mature. Microfiltration has been proposed (Jukkola et al.
2018) as a viable alternative to centrifugation and is reported to cause less damage
to shear-sensitive components in milk like the milk fat globule membrane.
Megasonics or the use of high-frequency ultrasound has also been proposed as hav-
ing potential for cream separation on a large scale, with minimum damage to the
natural properties (Juliano et al. 2015).
Homogenization is the mechanical process of breaking down fat globules to pre-
vent fat separation or forming a visible cream layer. This process was invented by
Auguste Gaulin in 1899. The equipment used consists of a reciprocating pump,
having 3–7 pistons, which increase the pressure of milk up to 15–25 MPa, to push
it through a narrow opening of the homogenizing valve. Innings (2015) has pro-
vided a detailed description about how the design, construction, and operation of
homogenizers have developed over time. When pressures above 100 MPa are used,
the process is called ultrahigh pressure homogenization (UHPH). UHPH, having a
deleterious action on microorganisms, is proposed (Rodarte et al. 2018) as an alter-
native to pasteurization with homogenization. Ultrasonication has also been reported
as a method of homogenization (Sfakianakis et al. 2015).
Standardization is the process of adjusting the fat content in milk to a predeter-
mined level to meet the legal requirements, for value addition, or for conversion into
products. Most modern dairy plants having significant amount of automation use a
direct in-line system with separation for standardization, in which control valves,
flow and density meters, and a computerized control loop are used to regulate the fat
content of milk and cream to required limits (Bylund 2003). Tetra Laval Holdings
and Finance SA has filed an international patent for a method and system for pro-
ducing milk products having different fat contents (Önnheim et al. 2019). In this
system, the cream and skim milk outlets are divided into two pipes each, the fat
content and flow rate of the cream/skim milk in the four streams are determined, and
with the help of a flow controller, the flow is regulated before cream and skim milk
are mixed separately to form two products having separate fat contents, without the
need of a buffer tank in between.
4.4.3 Evaporation and Drying
Evaporation and drying are important methods used in dairy industry for increased
shelf life with reduced costs of storage and transportation. Both these methods
72 A. S. Warrier
employ heat exchangers to vaporize the moisture in milk. Usually these processes
are energy intensive and require large, expensive equipment for operation.
Evaporation of milk is generally done in long shell and tube heat exchangers also
known as calandria, at a low pressure. Though not suitable for complete concentra-
tion, membrane separation is the main technology which has succeeded as an alter-
native to evaporation of milk. Drying of milk is commonly done in spray dryers and
vibrating fluid bed dryers, though roller dryers, vacuum tray dryers, freeze dryers,
etc. are also used in the dairy industry to a limited extent.
Gourdon and Mura (2017) who used a simulation tool to study the process of heat
transfer in industrial evaporators concluded that pressure drop is critical in deciding
the performance of evaporators, due to its association with the saturation tempera-
ture. Pinch analysis (Walmsley et al. 2016) and Total Site Heat Integration (Walmsley
2016) methods have been reported for designing integrated evaporation systems
with vapor recompression, for reduced consumption of utilities thereby saving
energy. A study (Haasbroek et al. 2013) on different control techniques for falling
film evaporators used in dairy industry compared techniques like linear quadratic
control, fuzzy predictive control, cascade control, and the commonly used propor-
tional and integral (PI) control, concluding cascade control to be a well-balanced
approach. Alternate methods like use of ohmic heating (Parmar et al. 2018), use of
thin film spinning cone (Centritherm) evaporator (Tanguy et al. 2015), etc. have also
been reported as having potential for the industrial evaporation of milk.
Drying is one of the most energy-intensive unit operations in dairy industry.
Hence most of the research works in this area are intended to conserve energy, along
with improving product properties and reducing product losses. Atomizer is the
heart of any spray drying unit. O’Sullivan et al. (2019) have thoroughly examined
the advances in atomization technology for spray drying, with emphasis on dairy
applications, along with reviewing different technologies for atomization, such as
rotary atomizers, pressure nozzle atomizers, pneumatic atomizers, ultrasonic atom-
izers, and electrospray atomizers, explaining in detail their working principles, suit-
ability for industries, and challenges in use. Walmsley et al. (2015) have reported a
thermo-economic design optimization of an industrial milk spray dryer exhaust to
inlet air heat recovery system, which is claimed to be economically viable for
exhaust heat recovery in an industrial case study. Moejes and van Boxtel (2017)
reviewed the energy conservation capabilities of advanced techniques like mem-
brane distillation, monodisperse droplet drying, air dehumidification, radio-fre-
quency heating, use of renewable energy sources, etc. in comparison with the
prevailing technologies. They found that the combination of reverse osmosis with
membrane distillation, monodisperse droplet drying, and zeolites consumed 4–5 MJ/
kg milk powder against 10 MJ/kg milk powder used presently. Moejes et al. (2018),
in a later research, studied a closed loop spray drying system which used a monodis-
perse droplet atomizer for reduction of fines in the exhaust air and facilitated the
recovery of latent and sensible heat from the dryer exhaust by recirculating the air.
This setup was found to consume 4.9 MJ heat per kg milk powder produced.
4.4.4 Butter and Cheese
Butter is a fat rich dairy product highly valued across the globe as a spread or a food
ingredient. Nowadays most of the butter made in large-scale dairy processing units
is in continuous butter making machines working based on Fritz process. This
involves feeding the cream into a cylinder where it is churned using rotating beaters.
The butter granules and buttermilk formed are moved to a secondary churning and
separation section where the butter and buttermilk are separated. The separated but-
ter is then washed, worked, and conveyed using a screw conveyor to a squeeze dry-
ing section, where the remaining buttermilk is removed by passing the butter
through a conical channel and perforated plate. It is then conveyed to a second
working section where salt addition and moisture adjustment take place. The vac-
uum section removes air from the butter to improve its physical properties. In the
final working section, the butter passes through a series of perforated discs where
the final adjustments in moisture and texture take place (Dairy Research and
Information Centre 2019). As this technology has been existent for more than half a
century, it has become somewhat matured through minor improvements, and
scarcely any recent developments have been reported, except for a few patents.
There is no other dairy product as popular as cheese. The art of cheese making has
been known for centuries, and over thousand varieties of cheese are made today,
across the globe. Over time, this art evolved into a technology with the understanding
of science behind cheese making and the introduction of newer devices followed by
improved mechanization and automation, even though most of the basic stages
remained the same. Use of membrane separation processes like microfiltration, ultra-
filtration, etc. in cheese making has been a noteworthy development. Numerous
equipment for cheese making including vat-based systems like single-axis vertical
vats, double-axis vertical vats, single-axis horizontal vats, and double-axis horizontal
vats, continuous systems, and machines needed for the post-vat stages have been
developed by the second half of the twentieth century (Legg et al. 2017). Legg et al.
(2017) may be referred for a detailed description of the equipment. Robotic manufac-
turing, 3D printing, etc. are the other emerging research hotspots in cheese making.
4.4.5 Ice Cream
Ice cream has evolved from a premium delicacy of the royal courts to a widely avail-
able dessert accessible to the masses. This transformation happened due to the tech-
nological developments in processing equipment and economical refrigeration
techniques. The manufacture of most types of ice creams and frozen desserts
includes mix preparation and aging, simultaneous whipping and freezing in a
scraped surface heat exchanger, addition of flavoring ingredients/fruits/nuts, pack-
aging, and hardening. The continuous freezers which are currently used are capable
of processing a volume of 3000–4500 L per hour (LPH), incorporating filtered air,
controlling the overrun precisely, and dosing particulate ingredients (Goff and
Hartel 2013). The ingredient feeders which are used now have much advanced
74 A. S. Warrier
controls that they permit automatic and precise dosing of ingredients based on the
flow rate of ice cream.
Advances in ice cream making include the slow churning process (low-temperature
extrusion) in which the ice cream coming out of freezer at −5 to −6 °C is further
cooled to −13 to −15 °C in an extruder with slowly rotating screws, to obtain a prod-
uct with smaller ice crystals and smoother texture (Goff and Hartel 2013). This pro-
cess even permits elimination of the hardening stage. Direct cryogenic freezing
using liquid nitrogen can be used for rapid hardening of ice cream of rapid freezing
of novelty bars. Addition of liquid nitrogen to the ice cream mix together with con-
tinuous agitation leads to the formation of tiny ice crystals to yield an ice cream with
a smooth texture. Pacojet®, a kitchen appliance company based in Switzerland, has
developed a process called “Pacotizing” which they claim can produce nature-fresh
ice creams. In this process, the fresh foods are frozen together with an appropriate
liquid, which is then shaved off into micro thin portions (Pacojet 2019).
Ultrasound-assisted crystallization is an innovative technique which can be used
for making ice cream having small, uniform ice crystals, where the application of
ultrasound aids inducing crystal nucleation and improving heat and mass transfer
(Zhang et al. 2015). High-pressure low-temperature (HPLT) processing at
50–100 MPa, another emerging technique used for controlling ice nucleation, has
been reported to produce a thermodynamically stable product with improved sen-
sory attributes (Soukoulis and Fisk 2016).
Three-dimensional (3D) printing was identified as a feasible approach for mak-
ing ice cream, when a group of students from Massachusetts Institute of Technology
hooked up a 3D printer with a soft-serve ice cream machine with a liquid nitrogen
cooling system as part of their graduate project (Gibbs 2014). This innovation based
on fused deposition modeling was granted with a US patent in 2018 (Bunker et al.
2018), though the technology has not yet been commercialized. Hong et al. (2017)
reported development of a 3D printing ice cream machine based on rapid prototyp-
ing technology.
The Italian ice cream machine manufacturer Carpigiani has developed an Internet
of Things (IoT)-based maintenance platform named Teorema that connects 12,000
smart ice cream machines worldwide. These machines work without any local tech-
nical support and incur reduced maintenance cost during their long lifetime (Corradi
et al. 2018). A recent entrant into the worldwide ice cream pantheon is the Thai-
style rolled ice cream, which is made by spreading the mix on a frozen steel plate
and then rolling it into swirls.
4.5 Packaging
The packaging technique and material, alongside the processing technology, are
major factors that decide the shelf life of perishable food products like milk.
Nowadays, with continuous technological progress, the packaging materials are
expected to perform many more functions than just contain, protect, and inform the
consumer. Packaging of milk has emerged from glass bottles to a wide range of
materials and forms including polyethylene bottles, jugs, and pouches, polyethylene
terephthalate (PET) bottles, laminated paperboard cartons, etc. The different dairy
products utilize a wider range of materials, forms, and technologies for their pack-
aging, depending on their diverse characteristics.
A detailed explanation regarding technological advancements in packaging is
beyond the scope of this chapter. Hence, only a few works having specific engineer-
ing interventions are mentioned. Edible packaging using polysaccharides like
galactomannan and chitosan coatings on cheese has been reported to provide better
quality, preserve freshness, and reduce the amount of packaging material (Karaman
et al. 2015). Many works related to intelligent and active packaging of milk and
dairy products have been reported. Reviews by Karaman et al. (2015), Ščetar et al.
(2019), and Yildirim et al. (2018) may be referred for a detailed reading. Most com-
monly used intelligent packages for dairy products are those with time–temperature
indicators or freshness indicators, while antimicrobial coatings were the most
widely studied active packages in recent years. Advancement in nanotechnology
has also influenced the dairy packaging research. Nanolaminates, nanocomposite
bottles, and bins with silver nanoparticles have been reported to have enhanced
oxygen barrier properties, while nanoclays and nanocrystals embedded in the plas-
tic films and bottles are shown to block oxygen, carbon dioxide, and moisture trans-
fer (Ščetar et al. 2019). Kuswandi (2017) has reviewed the recent advances in food
nano-packaging, including the application of nanotechnology in bio-based packag-
ing, improved packaging, active packaging, and smart packaging.
4.6 Cleaning and Sanitation
Proper sanitation of the contact surfaces to make them physically, chemically, and
bacteriologically clean is an essential requirement for any dairy processing unit.
Nowadays, manual cleaning of equipment is limited to very small-scale units or
some specific equipment for which circulatory cleaning is not feasible. In most of
the dairy processing units, manual cleaning has been replaced by cleaning-in-place
(CIP) systems having varying levels of automation, in which the rinsing water and
cleaning chemicals are circulated without dismantling the equipment.
Li et al. (2019) have written an exhaustive review of the different elements that
influence the efficiency of CIP in closed processing systems, in which they have
elaborated the fouling problems of processing plants, significance of suitable CIP
practices, and the forces affecting cleaning highlighting the effect of the hydrody-
namic forces. They emphasized the importance of factors like increasing turbulence
and wall shear stress of flow, application of pulsating turbulent flow, etc. to improve
the efficiency of CIP. Sadeghinezhad et al. (2015), in their review on milk fouling
on heated surfaces, have focused on the previous works on parameters influencing
fouling and its remedies. Bouvier et al. (2019) installed a device to create swirl flow
in a holding tube after plate heat exchanger and observed that the swirl flow reduced
the cleaning time, along with increasing the heat exchange capability. Palabiyik
et al. (2015) designed a novel CIP protocol to reduce environmental impact of
76 A. S. Warrier
cleaning process, which included two different cleaning stages having varying
requirements of temperature and velocity. This protocol has been reportedly effec-
tive in conserving water and electricity.
Replacing the water used for rinsing of stainless steel surfaces with ozonated
water is reported to remove residues and reduce bacterial populations and biofilms
more efficiently. Use of electrolyzed water, which is a solution produced by elec-
trolysis of water containing added minerals or salts, is another recent technology
with potential in environment-friendly cleaning and disinfectation processes
(Jiménez-Pichardo et al. 2016). Ultrasonic cleaning is another recent area of
research. Acoustic cavitation at low ultrasonic frequencies can generate strong
physical forces like shear forces, microjets, microstreaming and shockwaves, which
can deactivate microorganisms along with cleaning (Yusof et al. 2016).
Considerable amount of automation is used in the CIP process today for monitor-
ing as well as for controlling the process. Simeone et al. (2018) designed an optical
monitoring system consisting of a digital camera and an ultraviolet light source to
monitor the fouling in process tanks and to predict the appropriate cleaning time.
They also built a neural network-based decision-making support system for predict-
ing the required cleaning time. The use of nephelometers to monitor changes of
turbidity in cleaning solution has also been reported (Piepiórka-Stepuk et al. 2017)
to control the duration of cleaning programs. Úbeda et al. (2016) developed an ultra-
sonic method for tracking and removing fouling deposits in an experimental plate
heat exchanger used for heating milk. This system is reported as a sensitive and
noninvasive technique with a low cost. Abidin et al. (2015) developed a cleaning
device attached to an in-pipe robot to assist pipeline cleaning operations. The sys-
tem consisting of sensor to detect route, controller, power source, motor to rotate the
cleaning part, bearing, cleaning part, and housing was designed to work based on
Arduino software. 3D TRASAR™, a technology developed by Ecolab, is a sensor-
based diagnostic tool that verifies every CIP cycle and gives alert in case of any
deviations, along with identifying opportunities to improve CIP efficiency (Marriott
et al. 2018). The manufacturers claim that this technology improves product quality,
reduces cleaning time, reduces water consumption, optimizes cleaning cycles, and
decreases chemical usage.
4.7 Dairy Effluent Treatment
Dairy processing requires a large quantity of water. Even though the quantity varies
between individual plants and operations involved, every dairy plant drains off more
water than the milk it processes. These effluents carrying milk solids, cleaning
chemicals, etc. have high levels of biological and chemical oxygen demands (BOD
and COD), thereby making effluent treatment an inevitable activity. Due to the com-
plex composition of dairy effluent, the conventional treatment involves various
stages consisting of mechanical, chemical, and biological treatment methods. As a
wide variety of techniques are used and several subject areas are involved, dairy
effluent treatment has always been a raging topic for research. A simple search in
any search engine on the Internet will lead to thousands of recent works in this area.
Only a few of these advancements have been mentioned here.
Thermal pretreatment of dairy effluent and subsequent fermentation of coagu-
lated residues were studied by Kasmi et al. (2017) who reported that above 40%
improvement occurred in reduction of COD by treating the samples over 60 °C
temperature, which increased to 72% at 100 °C. Nanda et al. (2015) reported that
dairy effluents rich in lactose can be potentially converted to biofuels and biochemi-
cals by supercritical water gasification using a continuous flow tubular reactor.
Ozonation is another promising technology for dairy wastewater treatment.
Application of atmospheric cold plasma to dairy wastewater has been reported
(Patange et al. 2018) as a potential method for reducing the bacterial population,
without causing toxicity to any aquatic species.
Membrane filtration has been identified as a promising method of dairy effluent
treatment, as it provides good quality water which can be directly reused. Even
though this technology is known for decades, it is still considered novel and
research-worthy. Brião et al. (2019) subjected the rinse water used in dairy industry
to nanofiltration and reverse osmosis under different operating conditions, to obtain
a permeate which can be reused for cooling water and a retentate rich in milk solids
which can be processed further for recovering the solids. Aydiner et al. (2016) hier-
archically prioritized the preferability of four innovative dual membrane combina-
tions, which included novel membrane processes like forward osmosis and
membrane distillation, in comparison with a traditional ultrafiltration/reverse osmo-
sis system for environment-friendly treatment and reuse of dairy industry wastewa-
ter employing a multi-criteria decision-making tool called analytical hierarchy
process. They concluded that novel membrane technologies performed better in
treating wastewater from dairy industry by facilitating whey utilization, reducing
effluent quantity, and ensuring efficient reuse of water, along with utilizing waste
heat in case of membrane distillation (Aydiner et al. 2016).
Electrocoagulation, an electrochemical process, has recently gained a lot of
interest as an alternative to the resource-intensive conventional coagulation which
was a physicochemical process. The several leverages of this technology include
ease of operation, uncomplicated equipment, low retention time, reduced chemical
requirement, rapid sedimentation of the electrogenerated flocs, and reduced quan-
tity of sludge (Benazzi et al. 2016). Using this technique for treating dairy wastewa-
ter, Benazzi et al. (2016) reported removal efficiencies of 94% for color, 93% for
turbidity, 65% for total organic carbon, and 70% for COD, while Bassala et al.
(2017) reported removal efficiencies of 80% for COD, 98% for phosphate, and
100% for suspended solids and turbidity using a new electrocoagulation reactor
operating with 20 aluminum electrodes in bipolar configuration mode. Microbial
electrochemical systems like microbial fuel cells are another attractive option for
treatment of dairy wastewater, along with recovery of resources like electricity.
78 A. S. Warrier
4.8 Storage and Distribution
Storage and distribution form the penultimate links of the dairy supply chain. The
major concern of this stage is ensuring a stable quality till its consumption or its
expiry date. This can be ensured only by providing a stable and reliable cold chain
in case of milk and high-moisture dairy products. The processed and packaged milk
and milk products having short shelf life are usually stored in the dairy plant in cold
storages. Though the advanced researches in refrigeration, insulation, etc. may be
extrapolated to cold storages, not many remarkable changes have happened to the
dairy cold stores in the last decade. Distribution of milk and dairy products is usu-
ally done through insulated or refrigerated trucks or directly to the customer using
vending machines.
Alavi et al. (2019) proposed a method of upgradation of existing vapor compres-
sion refrigeration plants following a power regeneration concept based on bleed
loops, which can give cooling power and COP improvements up to 100% and 80%,
respectively. Singh and Dasgupta (2017) conceptualized a trans-critical CO2 heat
pump system with internal heat exchanger for the recovery and reuse of waste heat
in dairy industry having ammonia-based refrigeration. In this work, the condenser
of the refrigeration system was paired with evaporator of the heat pump maintained
at a fixed temperature. The designed heat pump was capable of producing heat at
70 °C, which can be used for preheating the water for boiler. The authors claim that
this system significantly reduces CO2 emission along with reducing the energy
consumption.
Utilization of phase change materials (PCM) for banking hot or cold conditions
has always been in vogue for balancing the electric load. Wang et al. (2017) studied
performance of a cold storage using a water/ice phase change material layer coupled
with its insulation and found that this system reduces the energy requirement along-
side balancing the electric load between peak and off-peak hours. Bista et al. (2018)
have reviewed in detail the advances in research related to use of phase change
materials in refrigeration systems, including the effects of PCM in evaporators, con-
densers, compartment, and compressor. Most of the articles they have referred sug-
gest the use of PCM in refrigeration as a viable option for energy conservation.
Leducq et al. (2015) investigated the possibility of using a PCM packaging, in com-
parison with normal packaging and insulated packaging, during long-term storage
with temperature fluctuations. They observed that PCM packaging had a significant
effect on the quality of ice cream after storage. With respect to alternative methods
of storage, Duarte et al. (2015) investigated the feasibility of hyperbaric storage of
Portuguese whey cheese. They observed that hyperbaric storage for 4 h at 100 MPa
maintained the microbial count similar to refrigerated storage, while storage at
150 MPa for 8 h cut down the microbial count to undetectable levels.
As the cold chain is only as strong as its weakest link, the monitoring of time-
temperature throughout the chain is important to find the weak links and tempera-
ture abuses. Wireless technologies like radio-frequency identification (RFID) tags,
wireless sensor networks (WSN), and time–temperature integrators (TTIs) are the
most common trackers used in food cold chain (Ndraha et al. 2018). Badia-Melis
et al. (2015) have reviewed recent advances in traceability systems applicable to
milk like isotope analysis, DNA barcoding, etc. Gwanpua et al. (2015) have devel-
oped a software named the FRISBEE tool (Food Refrigeration Innovations for
Safety, consumers’ Benefit, Environmental impact and Energy optimization along
the cold chain in Europe) within the framework of the European Union FP7 project,
which is capable of simultaneously evaluating the three major sustainability indica-
tors of refrigeration systems, viz., quality and safety of food, energy consumption,
and global warming impact. Frozen milk products like ice cream were also one of
the reference products considered for the development of this tool. Blockchain tech-
nology, the cryptographic transaction register which powers bitcoin and the likes, is
emerging as a promising system for improved transparency and operating efficiency
with reduced costs in managing supply chains (Nakasumi 2017). Internet of Things
(IoT), a system which interconnects physical objects via the Internet, is the most
fashionable area of research across various disciplines. Tsang et al. (2017) designed
an IoT-based cargo management system for providing adequate guidance for main-
taining a cold chain environment, which can ensure real-time monitoring and alert
management. The system consists of Bluetooth-operated sensors attached to every
cargo unit, fuzzy logic which inspects any changes in storage temperature or elec-
tricity consumption, and case-based reasoning which receives input from the fuzzy
logic and aids in decision support process. Tian (2017) developed a food supply
chain traceability system for real-time food tracing based on HACCP (Hazard
Analysis and Critical Control Points), blockchain, and Internet of Things accessible
to all stakeholders on the food safety status, alleviating the inconveniences associ-
ated with centralized information systems, and ensuring increased security, trans-
parency, and collaboration.
4.9 Conclusions
Being a multidisciplinary field of study, advances in any of its contributing disci-

plines influence the direction of researches in dairy engineering. As long as food
continues to be a basic requirement for life, or in more conservative terms, as long
as milk and dairy products continue to be a part of our cuisine, the importance and
scope of dairy engineering is not going to subside. Though many of the current
practices like pasteurization/sterilization using heat, spray drying, cleaning with
water, refrigerated storage, etc. are expected to stay back decades ahead, it would be
unwise to comment on the future of this discipline, considering the abundance of
research works going on and the momentum of progress.
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Significance of Fortification of Beneficial
Natural Ingredients in Milk and Milk 5
Products
Jagrani Minj and Sonam Dogra
Contents
5.2 I mportance of Fortification in Milk and Milk Products 89
5.3 Components Used for the Fortification of Milk and Milk Products 89
5.3.1 Probiotics 89
5.3.2 Prebiotics 92
5.3.3 Vitamins 93
5.3.4 Minerals 94
5.3.5 Proteins 95
5.3.6 Phytochemicals or Plant-Based Ingredients 96
5.3.7 Fruit/Fruit Pulp/Fruit Purees 96
5.3.8 Short-Chain Fatty Acids Such as Omega-3 Fatty Acids 97
5.4 Major Groups of Milk and Milk Products Used for the Fortification 98
5.4.1 Milk/Dairy-Based Beverages 98
5.4.2 Yoghurt and Fermented Milk Products 99
5.4.3 Cheese and Cheese Products 104
5.5 Fortification Challenges in the Milk and Milk Products 105
5.6 Industries Working on Fortified Milk and Milk Products 107
5.7 Future Prospects on Fortified Milk and Milk Products 108
5.8 Conclusions 109
References 110
J. Minj (*)
S. Dogra
Punjab Pollution Control Board, Zonal Office Laboratory Ludhiana, Pujnab, India

https://doi.org/10.1007/978-981-15-2608-4_5
88 J. Minj and S. Dogra
Abstract
Fortification is a process where essential ingredients like vitamins, minerals,
omega-3 fatty acids, probiotics, prebiotics and phytoconstituents are incorpo-
rated to the dairy products. The main purpose of fortification is to reduce the rate
of nutrition deficiency and related diseases among all age group of people. The
various dairy products like whole milk, yoghurt, cheese, yoghurt drinks, dairy-
based beverages, milk powder, butter, and buttermilk favor the fortification
ingredients and a great choice for the essential nutrient fortification. The need for
milk fortification with essential nutrients is required because milk and milk prod-
ucts are deficient in some nutrients or the amount is too low (iron) for the proper
nutrition intake. Several processing factors such as heating, pasteurization,
homogenization, ultrahigh temperature treatment and spray drying can also con-
tribute to the loss of some essential nutrients. Therefore, fortification of milk and
milk products was introduced to overcome the lost nutrients and to maintain its
stability at the end of the shelf life of the product. Thus, fortification could be
considered as a supportive technique to deliver the required micronutrients and
other functional ingredients and can serve as a tool to minimize nutrition defi-
ciency and related diseases.
Keywords
Milk fortification · Milk products · Milk · Functional ingredients · Health benefits
5.1 Introduction
Fortification is one of the fascinating opportunities for improving the nutritional

quality of milk and milk products. Nowadays the demands of fortified milk products
are gaining popularity due to the health-conscious consumers. Food fortification,
though sometimes considered synonymous to food enrichment, is slightly different
by definition. Food fortification has been defined as the addition of one or more
essential nutrients to a food, whether or not it is normally contained in the food, for
the purpose of preventing or correcting a demonstrated deficiency of one or more
nutrients in the population or specific population groups (FAO/WHO 1994). The
process improves the nutritional quality of food, minimizes the risks to health, and
provides public health benefit. Food enrichment can be defined as the addition of
essential micronutrients to the food to make over the lost micronutrients during
processing. According to the FAO/WHO (1994), enrichment has been used inter-
changeably with fortification.
5 Significance of Fortification of Beneficial Natural Ingredients in Milk and Milk… 89
5.2 Importance of Fortification in Milk and Milk Products
Food fortification has a significant role in the public health management since more
than 100 years. Nowadays, food fortification is an approach to eradicate the nutrient
deficiency illnesses like rickets and osteoporosis linked to the vitamin D deficiency
(Cosman et al. 2014), night blindness and xerophthalmia caused by the vitamin A
deficiency (Zayed et al. 2015), anemia due to iron deficiency (Alaofè et al. 2017),
and birth defect problems linked to the folate deficiency in the pregnant women
(Harika et al. 2017). Majority of consumer likes milk and milk products to fulfill the
essential nutritional components required by the body. Milk and milk products like
dairy-based beverages, yoghurt, milk powder, cheese, and ice cream are considered
as a delivery vehicle for the supplementation of specific ingredients and other essen-
tial micronutrients.
5.3 omponents Used for the Fortification of Milk

C
and Milk Products
To increase functionality of milk and milk products, these days fortifications of vari-
ous natural ingredients are frequently used. Ingredients which are generally used for
the fortification of milk and milk products are prebiotics, probiotics, vitamins, min-
erals, sweeteners, flavors, fruit purees or pulp, herbs, etc. Food fortification not only
improves the technological properties, but it also enhances the biofunctional proper-
ties of the products. The fortification should not alter the quality, appearance, body
and texture, and acceptability of the products. The list of ingredients used for the
fortification purposes of milk and milk products are listed in Fig. 5.1.
5.3.1 Probiotics
Value-added milk and milk products can be made by using live and active probiotic
bacteria. Probiotic dairy products not only improve the gut health but also have
many more beneficial health effects. Different probiotic bacteria have different and
unique health effects. Many promising research suggested that some of the probi-
otic bacteria have some of these potential health benefits such as preventive role in
ulcerative colitis (Gionchetti et al. 2000), urinary tract infections (Akgül and
Karakan 2018), vaginal infections (Ortiz et al. 2014), antibiotic-associated diarrhea
(Wright et al. 2015), bladder cancer (Feyisetan et al. 2011), infection caused by
Clostridium difficile (Ramai et al. 2019) and Helicobacter pylori (Chen et al. 2019),
Crohn’s disease (Bjarnason et al. 2019), and irritable bowel syndrome (IBS)
(Barajas-Nava et al. 2018).
Fermented dairy products like cheese, yoghurt, milk beverages and frozen dairy
products offer a very good delivery vehicle for the probiotics. These products not
only provide a suitable medium for the growth and development of probiotic bacte-
ria but also protect the viability of probiotic bacteria. However, it acts as a carrier for
Fig. 5.1 Components used for fortification in milk and milk products
the delivery of certain number of live bacteria in the gut. Many probiotic bacteria
have been well documented for their health benefits as given in Table 5.1. These
well-known probiotic bacteria can be used for the formulation of beneficial dairy
products.
One of the most crucial points of consideration for probiotic bacteria is that
it must be at an efficacious level in any products and that level should be maintained
throughout the shelf life of the product. Many factors can affect the survivability of
live probiotic bacteria which include storage temperature, processing conditions,
presence of inhibitory components, pH, etc. (Teixeira et al. 1995; Fowler and Toner
2005; Gill 2006; Jay et al. 2005; Nualkaekul et al. 2011; Schutyser et al. 2012;
Grattepanche and Lacroix 2013; Putta et al. 2018). Dairy products like ice cream,
fermented milk, cheese, fermented beverages, etc. can act as vehicle to deliver pro-
biotics in adequate amount. Many companies are manufacturing the probiotic forti-
fied dairy products. For example, probiotic fortified light ice cream Culture
Republick being manufactured by Unilever claims that three billion live probiotic
bacteria are present in the product (https://www.culturerepublick.com/). Similarly,
Dannon YoCream frozen yoghurt by Danone North America claims the presence of
100 million cultures per gram of the product. Cultures present in the frozen yoghurt
are Bifidobacterium, L. acidophilus, L. bulgaricus, and S. thermophilus (http://
www.yocream.com/). Viability of any probiotic bacteria during storage is an impor-
tant factor for the maximum health benefits. Many research studies have been
Table 5.1 List of some of the probiotic bacteria and their claimed health benefits
Probiotic bacteria Claimed health benefits References
L. acidophilus Antifungal activity, preventive McFarland (2007); Cortés-Zavaleta
action on irritable bowel syndrome et al. (2014); Ortiz et al. (2014);
(IBS) Phavichitr et al. (2013)
L. casei Preventive action on diarrhea Wu et al. (2013); Wright et al.
(IBS), antimicrobial and immune- (2015); Shida and Nomoto (2013);
modulatory actions Castillo et al. (2013)
L. delbrueckii Immune-modulatory action, Moro-García et al. (2013); Abedi
subsp. bulgaricus antibacterial action et al. (2013)
L. fermentum Antimicrobial action, preventive Homayouni et al. (2014); Chen
role in hypercholesterolemia, et al. (2013); Tomaro-Duchesneau
treatment in bacterial vaginosis et al. (2014)
L. plantarum Antifungal, preventive action on Cortés-Zavaleta et al. (2014); Ortiz
IBS et al. (2014)
L. rhamnosus Anti-allergic, preventive action on Licciardi et al. (2013); Wickens
bacterial vaginosis, reduction of et al. (2013); Homayouni et al.
atopic dermatitis, antidiabetic, (2014); Foolad and Armstrong
antiviral effects (2014); Panwar et al. (2014); Luoto
et al. (2013)
L. reuteri Anti-diarrheal action, cholesterol Dirienzo (2014); Szajewska et al.
reduction, treatment in (2014); Indrio et al. (2014)
gastrointestinal disorder
Bifidobacterium Cholesterol reduction, preventive Ortiz et al. (2014); Jacobs et al.
animalis subsp. action on necrotizing enterocolitis, (2013); Bordoni et al. (2013)
lactis preventive action on urinary tract
infection
B. bifidum High cholesterol reduction, Bordoni et al. (2013); Phavichitr
reduction of diarrhea et al. (2013)
B. breve Cholesterol reduction, treatment in Di Gioia et al. (2014); Janvier et al.
necrotizing enterocolitis (2014); Bordoni et al. (2013)
B. infantis Preventive action on IBS Wu et al. (2013)
B. longum Preventive action against allergic Wu et al. (2013); Yu et al. (2013);
reactions, preventive action on Schwarzer et al. (2013)
IBS, preventive action on
gastrointestinal diseases
Streptococcus Reduction of necrotizing Wu et al. (2013); Jacobs et al.
thermophilus enterocolitis and IBS (2013); Li et al. (2013)
Saccharomyces Anti-diarrheal action, treatment in McFarland (2007); Choi et al.
boulardii acute gastroenteritis problem and (2011); Ciccarelli et al. (2013);
IBS Vieira et al. (2012); Johnston et al.
(2011); Dinleyici et al. (2012); Shan
et al. (2013); Szajewska and
Mrukowicz (2005)
documented on the viability of probiotic bacteria. Probiotic strains Bifidobacterium

animalis ssp. lactis and L. acidophilus used for the fruit yoghurt preparation exhib-
ited good viability during the storage (Kailasapathy et al. 2008; Godward et al.
2000). Another popular dairy product cheddar cheese can also be considered
suitable for the probiotic delivery (Stanton et al. 1998). The viability and therapeu-
tic aspects of probiotic bacteria L. acidophilus LA-5 and B. bifidum BB-12 in white
brined cheese have also been reported (Ozer et al. 2009). Similarly, fresh cheese
made with probiotic L. salivarius CECT5713 and PS2 survived at 4 °C during
28 days of storage (Cárdenas et al. 2014). Fermented milk drinks are also consid-
ered as an excellent carrier for the probiotic delivery. The combination of three
probiotic cultures L. acidophilus, B. animalis subsp. lactis, and L. casei have been
reported for their production of probiotic milk drink (Yerlikaya et al. 2013). Many
lactobacilli and bifidobacteria have been isolated from human origin and have
exhibited probiotic properties. These potential probiotic strains can be used for the
formulation of functional fermented milk products. Awasti et al. (2016) claimed that
Indian human origin probiotic bifidobacteria strains NBIF-2, NBIF-5, and NBIF-7
can also be used for the development of functional dairy products.
5.3.2 Prebiotics
Prebiotics are dietary fiber which is helpful in the growth and promotion of probi-
otic bacteria in the gut mainly lactobacilli and bifidobacteria. The International
Scientific Association for Probiotics and Prebiotics (2016) defines prebiotics as “A
substrate that is selectively utilized by host microorganisms conferring a health ben-
efit” (https://isappscience.org/). The most common prebiotics inulin, galactooligo-
saccharides (GOS), fructooligosaccharides (FOS), lactulose, and
xylooligosaccharides (XOS) have been studied for their functional aspects (Boehm
and Moro 2008). Recently, human milk oligosaccharides (HMOs) have been recog-
nized as prebiotics and many researches are going on in this area. Meyer et al.
(2019) reported that fucosylated HMOs found to be abundant in human milk pro-
mote the growth of two commensal microbes Streptococcus oralis and S. mitis.
These HMOs could be helpful in maintaining the bacterial imbalance in the gut.
Prebiotics not only promotes the commensal bacteria, but it also protects the neona-
tal intestine via upregulation of the intestinal mucin level (Wu et al 2019a, b).
Prebiotics FOS, XOS, resistant dextrin, and polydextrose have been proven for the
elevated level of immunoglobulin M, transferrin, and immunoglobulin G. The pop-
ulation of probiotic bacteria Bifidobacterium has also been increased with the use of
these prebiotics (Xie et al. 2019). Prebiotic inulin also promotes the growth and
viability of National Collection of Dairy Cultures (NCDC) yoghurt cultures which
can be further used to make functional fermented milk products (Minj and Vij 2017).
Intake of prebiotic inulin by human subjects increased the production of short-
chain fatty acids in the feces. Prebiotic inulin not only promotes the production of
short chain fatty acids, but it also reduces the plasma insulin concentration, free
fatty acids, and glucose level (Van der Beek et al. 2018). Dietary fiber has also been
reported for the reduction of hyperinsulin levels (Provost et al. 2019). Application
of dietary fiber in obese human subjects has been reported to increase the intestinal
microbiota and acetate levels in feces, while it decreased the total bile acids and
body weight of the participants (Mayengbam et al. 2019). Another prebiotic XOS
can be helpful to reduce the fat accumulation in the mice. The effects of prebiotic
XOS have been seen in the reduction of perirenal, mesenteric, and epididymal fats,
serum leptin, and MCP-1 and also downregulated the gene expression of fatty acid
synthase. Production of short-chain fatty acids and cecum microbiota has been
modulated by the prebiotic XOS (Long et al. 2019). Similarly, the health benefits of
prebiotic lactulose have been seen in the rats by reduction of serum transaminases,
TNF-α and IL-6, and heme oxygenase-1, whereas antioxidant enzyme superoxide
dismutase (SOD) activity has been elevated in the liver (Yu et al. 2015). Many more
health benefits of prebiotics have been documented. However, prebiotic or dietary
fiber fortification could be beneficial for both the probiotic bacteria and therapeutic
purposes as well. Prebiotic components also improve the technological and nutri-
tional properties of the food (de Paulo Farias et al. 2019). The IASPP suggested
dietary recommendation for prebiotic or dietary fiber is 28 g/day (source: ISAPP).
5.3.3 Vitamins
Vitamins are very essential components for the proper development of body and
their physiological functions. Lack of vitamin supplementation may lead to condi-
tions like malnutrition. Different countries have different reports on vitamin malnu-
trition. About 195 million people are undernourished only in India which accounts
for about 24% of the undernourished population in the world (https://www.nfn.org.
in). In India, 43% of children (under age 5 years) are underweight and 48% are
stunted due to severe malnutrition (IFPRI 2016). According to WHO, the term mal-
nutrition describes three major groups of conditions:
1. Undernutrition
2. Micronutrient-related malnutrition
3. Overweight, obesity, and diet-related non-communicable diseases
Micronutrient-related malnutrition occurs due to the inadequate intake of vitamins

and minerals. Vitamin A, vitamin D, iron, zinc, folate, iodine, and calcium are the
most important nutrient components for global public health. Deficiency of these
nutrients represents a major threat to the health mainly in children, old age people,
and pregnant women. Vitamin D deficiency leads to many disease progression such
as rickets, osteomalacia, and osteoporosis. Vitamin D deficiency has also been
known for the risk factor for the development of autoimmune diseases, osteoporotic
fractures, and certain types of cancers (Holick 2002). According to UNICEF report,
two billion people are suffering from micronutrient deficiency mainly from iron or
vitamin A. The malnutrition condition is even worst in the South Asia as compared
to the other parts of the world as illustrated in Fig. 5.2.
Micronutrient deficiency can be eradicated with proper supplementation of essen-
tial micronutrients. Food products like rice, oil, wheat, and milk could be used for the
micronutrient fortification. All the vitamins are not soluble in water; some of the
vitamins such as vitamin A, vitamin D, vitamin E, and vitamin K are fat-soluble in
Fig. 5.2 Global micronutrient deficiency report for iron or vitamin A. (Source: UNICEF
report; Bresnyan and Gadha (2018); https://blogs.worldbank.org/endpovertyinsouthasia/milk-
fortification-india-journey-so-far)
nature. So, fortification with these vitamins must be a challenge in all the food prod-
ucts. However, selection of food products is one of the topmost criteria for the
food fortification. Milk and milk products are considered as one of the best compat-
ible medium for vitamin A and D fortification. Maurya and Aggarwal (2019) sug-
gested that milk product “Lassi” (a yoghurt-based beverage) can be used for vitamin
D fortification. Detailed information on vitamin D fortifications in dairy products are
given in a review by Zahedirad et al. (2019).
5.3.4 Minerals
Minerals are essential for the growth of children and teenagers. Lack of mineral
intake and low quantity of minerals in food product needs emphasize the need for
supplementation. Minerals like iron and calcium are very essential for the growth
and development of children. However, to overcome mineral deficiency, fortifica-
tion is considered as the best way to fulfill the requirements. Increasing the different
varieties of fortified milk and milk products with minerals is aimed to satisfy the
requirements of children and old age customers (Robison 2015).
Initially, mineral fortification was a challenge in dairy products. Because miner-
als like calcium could react with the milk proteins, induce coagulation (Gastaldi
et al. 1994) and sedimentation problems (Singh et al. 2007). The coagulation reac-
tion directly affects the sensory properties of the products. To overcome these prob-
lems, changes in the mode of delivery via microencapsulation or some other
technique might be more fruitful (Allen et al. 2006). Some of the minerals easily get
degraded, so microencapsulation technique could be useful for these minerals to
prevent from degradation. Iron fortification in milk with microencapsulation tech-

nique has been reported for the stability of iron and protection from degradation
(Abbasi and Azari 2011). Microencapsulation also masks the unwanted oxidized
flavor of iron in yoghurt (Jayalalitha et al. 2012). Mineral size used for the fortifica-
tion also affects the physicochemical and rheological properties of the product
(Santillan-Urquiza et al. 2017). Similarly, microencapsulated ferrous sulfate parti-
cle size has been reported to improve the retention, nutritional value, and sensory
attributes of the cheddar cheese (Ustunol 2018).
European Union (EU) with Regulation No. 1925/2006 has approved different
salts for the food fortification. Germany-based company Dr. Paul Lohmann® GmbH
KG has been known for the production of mineral salts for the food industries, phar-
maceutical industries, food supplement industries, and many other technical indus-
tries all over the world (https://www.lohmann4minerals.com/index.php/company.
html). They have recommended the following minerals for the fortification of milk
and milk products (Table 5.2 ).
5.3.5 Proteins
Although milk and milk products are rich in protein, fortification of milk and milk
products with protein can further add to its technological and functional properties
as well as nutritional value. Proteins can be either fortified in the concentrated form
or isolated form. It depends on the solubility of protein and type of food products to
be fortified. Different forms of protein such as acid casein hydrolysate, whey pro-
tein concentrate, whey protein isolate, and whey powder can be used for the food
fortification. The protein content in whey protein isolate (WPI) is approximately
90%, whey protein concentrate (WPC) contains 34–80% protein, and dried whey
powder contains 11–14.5% protein (Hoppe et al. 2008). The WPC has many func-
tional properties such as excellent solubility, foaming, and gelling properties
(Bacher and Køningsfeldt 2000; Heino et al. 2007; Svanborg et al. 2015). The whey
protein isolate has also been reported to increase viscosity and gel formation, to act
as water-holding agent, and to stabilize foams or emulsions (Kinsella and
Whitehead 1989).
Table 5.2 Dr. Paul Lohmann® GmbH KG recommended minerals for the fortification of milk and
milk products
Major mineral types
Calcium Magnesium Zinc Iron
Calcium carbonate Magnesium carbonate Zinc acetate Ferrous gluconate
Calcium chloride Magnesium citrate Zinc citrate Ferrous fumarate
Calcium citrate Magnesium gluconate Zinc lactate Ferrous lactate
Calcium lactate Magnesium oxide Zinc oxide Ferrous sulfate
Calcium phosphate Magnesium sulfate Zinc sulfate Ferric pyrophosphate
Magnesium phosphate Ferrous bisglycinate
Source: https://www.lohmann4minerals.com/index.php/company.html
Protein fortification in yoghurt could be an innovative technology for additional

protein supplement, improvement in textural properties of yoghurt, to increase the
water holding capacity and as a growth promoter for the probiotic bacteria present
in the yoghurt. Fortification of milk base with WPC affects the rheological proper-
ties, acidification profile and viable bacterial counts in the yoghurt (Marafon et al.
2011). Similarly, protein derivatives such as acid casein hydrolysate, whey protein
concentrate, whey protein isolate or tryptone have been known as growth promoter
for the probiotic bacteria (Dave and Shah 1998).
5.3.6 Phytochemicals or Plant-Based Ingredients
Natural plant-based components have always been known for enhancing overall
health. Components like polyphenol, flavonoids, phytosterols, catechin, phytoestro-
gens, phytostanols, and carotenoids have many potential health benefits (Saxena
et al. 2013; Rodriguez et al. 2006) and can be used for the fortification of milk and
milk products. These plant-based ingredients are known as potential antioxidants
that can enhance the antioxidant properties of milk and milk products. Although
synthetic antioxidants are also available for food fortification but the use of these
antioxidants can cause some toxic and carcinogenic effects (Abdel-Hameed et al.
2014). Regular consumption of natural antioxidant-rich food products can reduce
the incidences of many diseases such as cancers, cardiovascular diseases, and
hypertension. Herbs and medicinal plants are rich in antioxidants and polyphenolic
compounds which are known to improve the nutritional and therapeutic properties
of dairy products (Shori and Baba 2011a, b). Fermented dairy product yoghurt forti-
fied with crushed mint leaves have been reported to improve its sensory property
and shelf life (Kumar et al. 2013). Yoghurt fortifications with phytochemicals such
as peppermint essential oil, basil, and zataria have improved the functional proper-
ties of yoghurt (Azizkhani and Tooryan 2016). Fortification of probiotic ice cream
with phenolic compounds have been reported to increased its functional properties
(Sagdic et al. 2012). However, dairy products enriched or fortified with plant com-
ponents can act as a therapeutic agent to fight oxidative stress and related diseases.
5.3.7 Fruit/Fruit Pulp/Fruit Purees
Fortification with fruit and fruit pulp to the dairy products not only improves its
biofunctional properties, but it also contributes to the technological properties of the
product. Fruits are rich in vitamins, minerals, fibers, and polyphenols, and they are
an excellent source of antioxidants. Fortification with fruit, fruit pulp, or purees also
provides various options to the consumer to buy the products. Yoghurt fortified with
fruits improves its flavor (Ndabikunze et al. 2017), taste, and nutritional properties
(Hossain et al. 2012). Many antioxidant-rich fruits like blueberry, strawberry, rasp-
berry, cherries, apricots, pineapples, orange, and peaches are used for the dairy
product formulation (Arslan and Ozel 2012).
Fruit consumption alone or with fortified food products could be preventive in

the development of chronic diseases (Boeing et al. 2012). Yoghurt and fruit intake
have also been seen for their protective role in cardiovascular diseases (Wood et al.
2014; Mozaffarian 2016), type 2 diabetes (Ruhee and Mahomoodally 2015), and
obesity and weight gain (Mozaffarian et al. 2011). Yoghurt or fermented milk made
with fruits and vegetables has also been reported for lowest rate of hip fractures in
Swedish women (Michaëlsson et al. 2018). Similarly, fruit-supplemented probiotic
yoghurt have been documented for their antioxidant potential (Kumar and
Kumar 2016).
5.3.8 Short-Chain Fatty Acids Such as Omega-3 Fatty Acids
Some essential fatty acids such as omega-3 fatty acids have been known for their
involvement in various physiological functions like for cognitive development and
maintenance, preventive role in cardiovascular diseases and their management, anti-
inflammatory effects, anti-aggregatory action, telomere shortening, molecule
expression in plaque, hypertriglyceridemia management, antithrombotic action, and
control of high blood pressure (Weber 1989; Weber and Leaf 1991; DeFilippis and
Sperling 2006; US Department of Health and Human Services Agency https://ods.
od.nih.gov/factsheets/Omega3FattyAcids-HealthProfessional/). The polyunsatu-
rated fatty acids are classified according to their position within the fatty acid of the
first double bonds into the omega-3 and omega-6 fatty acids. The major dietary
sources for healthy omega-3 fatty acids are known as fish containing docosahexae-
noic acid (DHA) and eicosapentaenoic acids (EPA). Other dietary sources such as
seeds, nuts, and vegetable oil contain α-linoleic acids, while milk contains negligi-
ble amount of polyunsaturated fatty acids. Dietary recommendations for omega-3
fatty acids (both DHA and EPA) have been suggested as approximately 500 mg/d or
two 4-oz servings of fish per week (Kris-Etherton et al. 2009). Almost similar
dietary recommendations for omega-3 fatty acids have been reported in the Journal
of the American College of Cardiology which is 500 mg/d of omega-3 fatty acids for
adults without any CVD whereas 800–1000 mg/d for individuals with known
chronic coronary heart diseases or heart failure (Lavie et al. 2009).
However, omega-3 fatty acid supplementation could be possible with fortifica-
tion to meet the required dietary recommendations. Milk is an important food for
the children, elderly people, and pregnant women (Upadhyay et al. 2014); milk
powder can be fortified with the essential fatty acids DHA and EPA. Lane and
Derbyshire (2018) suggested that omega-3 delivery can be done by food fortifica-
tion, encapsulation, nano-emulsion technique, or topical delivery. Omega-3 fortified
yoghurt with algal oil can provide 600 mg DHA per 272 g of yoghurt which signifi-
cantly increased the DHA in plasma (McCowen et al. 2010). Fortification of micro-
encapsulated flaxseed oil powder for omega-3 supplementation in dahi (Indian
yoghurt) has been suggested for the potential delivery medium (Goyal et al. 2016).
The fortification with omega-3 must not be alter the product characteristics and rele-
vent study on omega-3 fortified milk has been done and found that it did not alter the
final product characteristic mainly in cheese and butter flavor (Jones et al. 2005).
However, consumption of omega-3 fortified dairy products could be beneficial in
cholesterol reduction which could be an additional benefit for the management of
cardiometabolic health (Soto-Méndez et al. 2019).
5.4 ajor Groups of Milk and Milk Products Used

M
for the Fortification
5.4.1 Milk/Dairy-Based Beverages
The consumption of dairy-based beverages is more popular worldwide. The dairy-

based beverage market has been forecasted to reach 13.9 billion USD by 2021
(Anonymous 2018). This forecast does not include the traditional dairy beverages
such as buttermilk, koumiss, kefir, etc. Milk-based beverages can be classified based
on their traditional values, value-added ingredients, or functional values.
Milk-based beverages especially fermented dairy beverages are an important
part of regular diet. Fermentation process is considered as one of the cheapest tech-
niques for preserving the food and improving its nutritional and sensory properties
(Gagada et al. 1999). Milk beverages favor the multiple fortifications to deliver
more than one micronutrient (Barclay 1998). Multiple micronutrients and other
Table 5.3 Basic classification of dairy-based beverages and their examples

Traditional Value-added Functional beverages
beverages beverages Supplemented or enriched products either with:
Aryan/dough Carbonated/ Bioactive compounds (i.e., ACE-I inhibitory peptides,
effervescent antioxidant peptides, antimicrobial peptides),
beverages melatonin
Plain Consumer-specific Caseinophosphopeptides (CPP)
buttermilk nutrition solution
Drinking Energy drinks/sports Conjugated linoleic acid (CLA)
yoghurt beverages
Kefir High-protein Dietary fibers or prebiotics
beverages
Koumiss Milk fat globule Fish oil
membrane beverages
Thirst-quenching Hydrolyzed whey protein
beverages
Isoflavones
Omega-3 fatty acids
Polyphenols
Probiotics
Sterols/stanols
Synbiotics (prebiotics + probiotics)
Vitamins
Minerals
functional component fortification can be more effective in improving nutritional

status than fortification with a single key micronutrients. Many trials have been
done to see the efficacy of specially formulated foods and beverages as vehicles for
the multiple fortifications. The dairy-based beverages can be classified according to
the methods used for their preparation or based on the used fortified components.
The basic classification is not established yet, but the general classification of some
of the dairy-based beverages is given in Table 5.3.
5.4.2 Yoghurt and Fermented Milk Products
Fermented milk and milk products are among highly consumed food all over the
world. Fermented milk like yoghurt can be considered as one of the best approaches
to improve the essential nutrient intake in daily consumption of foods (Preedy et al.
2013). Fermented milk yoghurt is made by the starter culture Streptococcus ther-
mophilus and Lactobacillus delbrueckii ssp. bulgaricus in 1:1 ratio. Both the strains
are responsible for the fermentation process and enzymatic breakdown of organic
substances into low molecular weight compounds. These compounds have even
more nutritional and therapeutic values than the normal milk components.
5.4.2.1 Yoghurt Fortification with Vitamins

Vitamins are very essential components required by the body and involved as a
cofactor in the metabolic pathway and also act as the oxygen transporter. Although
milk itself contains various vitamins, fermented milk and milk products like yoghurt
contain even higher amount of vitamins. Because some of the starter cultures used
in the preparation of fermented milk and yoghurt have been known for the synthesis
of vitamins like vitamin B groups.
The vitamin content in milk might be different in different species of ani-
mals (Table 5.4). The major vitamins found in milk are vitamin B1 (thiamine), vita-
min B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6
(pyridoxine), vitamin B12 (cobalamin), and a trace amount of folate and vitamin
C. However, milk is considered as a good source of water-soluble vitamins specially
vitamins B1, B2, and B12 rather than other food products. Other vitamins like vita-
min A, vitamin D, vitamin E, and vitamin K or fat-soluble vitamins are also found
in milk and milk products.
The Fortification with vitamins might be affected by the various processing fac-
tors such as high-temperature short-time (HTST) pasteurization and ultrahigh tem-
perature treatment (UHT). And stability of any fortified vitamin is an important
factor to meet the recommended dietary requirements. Many studies have been done
to check the stability of vitamins during processing and storage. A study on stability
of vitamin D have been done by Hanson and Metzger (2010) to see the effect of
vitamin D fortified up to 250 IU/serving of HTST-processed milk and UHT-
processed chocolate milk during the processing and storage. Fortification with vita-
min D from 100 to 250 IU/serving did not affect the processing treatments and the
Table 5.4 Vitamin content of milk varieties by serving; 1 serving = 1 cup (8 oz, 244 g)
Cow milk
Whole Water
(3.25% Reduced fat Low fat Goat Sheep buffalo
Unit fat) (2% fat) (1% fat) Skim milk milk milk
Amount per 8 oz serving (1 cup; 244 g)
Vitamins
Vitamin A μg 68 134 142 149 139 108 129
Thiamine mg 0.107 0.095 0.049 0.110 0.117 0.159 0.127
(vitamin B2)
Riboflavin mg 0.447 0.451 0.451 0.446 0.337 0.870 0.329
(vitamin B2)
Niacin (vitamin mg 0.261 0.224 0.227 0.230 0.676 1.022 0.222
B3)
Pantothenic mg 0.883 0.869 0.881 0.875 0.756 0.997 0.468
acid (vitamin
B5)
Pyridoxine mg 0.088 0.093 0.090 0.094 0.112 0.147 0.056
(vitamin B6)
Cobalamin μg 1.07 1.12 1.07 1.30 0.17 1.74 0.88
(vitamin B12)
Vitamin C mg 0.0 0.5 0.0 0.0 3.2 10.3 5.6
Vitamin D IU 98 105 127 100 29 ND ND
Vitamin E mg 0.15 0.07 0.02 0.02 0.17 ND ND
Sources: Adapted from USDA Nutrient Database and http://www.milkfacts.info
stability of vitamin D was not altered during the storage. There were no significant
changes have also been reported on the sensory properties of these products.
Among all fat-soluble vitamins, vitamin A and vitamin D have been recognized
for their important biological function. Deficiency of these vitamins may lead to
bone deformation, joint-related diseases, and certain types of cancer and cardiovas-
cular disorders. However, vitamin D plays a major role in various diseases and their
pathophysiological pathways. So, proper intake of vitamin D is must to avoid many
health-related problems. Milk and milk products fortified with vitamin D could be
a great source for the daily requirements. Vitamin D fortification is the primary
source to obtain the nutrition. Many countries are working on the vitamin fortifica-
tion implementation in the milk and milk products. Vitamin D fortification in milk
and milk products is mandatory in Finland since 2003. Similarly, vitamin D fortifi-
cation of 10 μg/day is mandatory in the United Kingdom for pregnant and breast-
feeding women and old age people (Spiro and Buttriss 2014). Some of the vitamin
fortified varieties of milk are given in Table 5.4.
5.4.2.2 Yoghurt Fortification with Minerals

There is no other best way to achieve mineral supplementation rather than through
food fortification. Fermented dairy product yoghurt has been known to be rich in
nutrients such as calcium and proteins. Mineral contents are less in yoghurt than
vitamins and proteins (Blanc 1981). Among all minerals, calcium and magnesium
have been found in higher amount in dairy products, whereas zinc and iron contents
are very low in milk and milk products (Mehar-Afroz et al. 2012). Fortification with
minerals could affect the rheological and physicochemical properties of the product.
Micro- and nano-size minerals of calcium phosphate, iron oxide, and zinc oxide
have been studied for their effect on rheological and physicochemical properties
during 28 days of storage (Santillan-Urquiza et al. 2017). Iron oxide fortified set
type of yoghurt showed color variations and increased during the storage. Whey
expels or syneresis has been noted higher in the micro size, whereas it was lower in
the nano-size mineral fortified set-type yoghurt. But, the whey syneresis has been
seen more in both types of fortified set type of yoghurt during the storage. Calcium
and zinc nanoparticle fortification had helped to improve the firmness and consis-
tency in set type of yoghurt. Mineral of nano-size particles have even better solubil-
ity than the micro-size minerals (Santillan-Urquiza et al. 2017). Yoghurt fortification
with seven minerals chromium, iron, magnesium, manganese, molybdenum, sele-
nium, and zinc did not affect the overall product characteristics (Achanta et al.
2007). Zinc sulfate fortification of 16 mg/kg in cheese milk did not alter the quality
of cheddar cheese (Kahraman and Ustunol 2012). Similarly, multiple micronutrient
fortification in Chhash (traditional Indian yoghurt-based drink) in the premix form
(vitamin A, vitamin D, iodine, folic acid, iron, and zinc oxide) did not affect the
physicochemical properties of the product (Gaur et al. 2019).
5.4.2.3 Yoghurt Fortification with Probiotics

Yoghurt fortification with live and active probiotic bacteria is in trend. To make
more innovative yoghurt-like products, fortification can be done with some of the
essential and biofunctional ingredients like probiotics. Yoghurt drinks are consid-
ered as one of the best medium for the formulation. A senior analyst of US food and
drink for Mintel Reports Beth Bloom reported that the popularity of yoghurt drinks
are increasing among US consumers (https://www.mintel.com). Most of the people
take yoghurt in their diet to improve the digestive health. Many companies are man-
ufacturing probiotic yoghurt. General Mills Inc. is manufacturing the strawberry
and pecan fortified Yoplait yoghurt beverages, whereas Chobani LLC is manufac-
turing fruit flavor fortified yoghurt drink by incorporating strawberry, banana, and
mixed berry flavor and mango, spinach, cucumber, and apple flavor. Fruzinga pro-
biotic drinkable yoghurt from Dairy Innovations LLC is known to boost up the
digestive health. Probiotic bacteria not only improve the digestive health but also
strengthen the immune system (Maldonado Galdeano et al. 2019). Probiotic bacte-
ria can survive in the acidic and bile salt conditions; however it helps to maintain the
population of good and healthy bacteria (Azad et al. 2018). To promote the probiotic
bacteria in the gut, prebiotic fiber can be used as fuel to further enhance and main-
tain their survivability. Greek yoghurt made with active probiotic bacteria, prebi-
otic, and stevia as a sugar replacer are manufactured by the Pillars Yogurt LLC. This
Greek yoghurt and functional dairy beverages are very delicious and are rich in
nutrients. These kinds of yoghurts are highly acceptable during snack times.
5.4.2.4 Yoghurt Fortification with Prebiotics

Application of prebiotic is not only to promote the probiotic bacteria in the gut, but
it has many other techno-functional properties. Fortification of prebiotics in fer-
mented milk product improves its sensory properties like mouthfeel and taste
(Elleuch et al. 2011), masks the sour taste of yoghurt (Irvine and Hekmat 2011),
reduces the syneresis in yoghurts (Aryana et al. 2007), and improves the rheological
and micro-textural properties (Crispín-Isidro et al. 2015) of the yoghurt. The sen-
sory properties of yoghurt can also be affected by the fat composition present in the
yoghurt. The sensory and textural properties of yoghurt can be affected by the milk
and other components and the concentration used to prepare the yoghurt. Prebiotic
inulin concentration of 40 g per liter and fructan concentration of 60 g per liter used
in the manufacture of reduced-fat stirred yoghurt significantly affect its creaminess,
flavor, viscosity, and overall acceptability (Crispín-Isidro et al. 2015).
Other biofunctional properties of yoghurt containing prebiotic have also been
reported. Yoghurt containing prebiotic is known to increase the short-chain fatty
acids in stools (Aryana et al. 2015; Hussein et al. 2014) and to improve the viability
of bacteria into the yoghurt (Capela et al. 2006; Akalin et al. 2004).
Fig. 5.3 Water-holding capacity of yoghurt made of milk enriched with oat protein concentrate
(OPC), skim milk powder (SMP), and oat protein isolate/lactose (Brückner-Gühmann et al. 2019)
5.4.2.5 Yoghurt Fortification with Proteins

Protein fortification is very common in the fermented milk and milk products like
yoghurt. Slingshot Foods, San Francisco, have been involved in the production of
milk protein concentrate fortified yoghurt drinks. Protein fortification has been
known for the improvement in the textural and sensory properties as well.
Fortification of protein might be in the form of powder or concentrate which can
improve its overall properties. Recently a paper has been published by Brückner-
Gühmann et al. (2019) on the yoghurt formulated with oat protein fractions such as
oat protein isolate (OPI) and oat protein concentrate (OPC). Their study suggested
that OPC was even better for the overall properties of yoghurt as compared to the
OPI. Oat protein concentrate not only improved the nutritional properties, but it also
improved the product quality and water-holding capacity to prevent excess whey
expulsion and sensory properties as demonstrated in Fig. 5.3. Akalin et al. (2012)
reported that whey protein concentrate and sodium calcium caseinate fortification
with probiotic yoghurt affect their sensory, textural, and microstructural properties.
Fortification with sodium calcium caseinate improves the adhesiveness and firm-
ness of the probiotic yoghurt (Akalin et al. 2012). Sodium calcium caseinate also
affects the viscosity of yoghurt during the storage (Akalin et al. 2012). Whey pro-
tein concentrate (WPC) differently affects the functional properties of yoghurt.
WPC has been reported to enhance the water-holding capacity, whereas sodium
calcium caseinate affects the textural properties resulting in a smooth, coarse, and
compact protein network (Akalin et al. 2012).
5.4.2.6 Yoghurt Fortification with Omega-3 Fatty Acids

Many health benefits of omega-3 fatty acids have been suggested by the US
Department of Health and Human Services, 2016 which included its preventive role
in cardiovascular diseases, cancer, rheumatoid arthritis, Alzheimer’s disease,
dementia and cognitive function, dry eye disease, and age-related diseases.
Milch animals like cows cannot produce the omega-3 fatty acid; they must obtain
them from their regular diets (https://biobeef.faculty.ucdavis.edu/2016/02/15/
omega-3-fatty-acids-and-milk/). So, it is obvious that milk and milk products have
low concentration of omega-3 fatty acids. The requirement of omega-3 fatty acids
could be fulfilled by consuming other food sources which are rich in omega-3 fatty
acids or through fortification of regular foods. When it comes to regular diet, milk
and milk products are very common. So, fortification of yoghurt with omega-3 fatty
acid is an attractive method to deliver the omega-3 fatty acids which ultimately
increases its concentration in the plasma lipids (McCowen et al. 2010). The dietary
recommendations for omega-3 fatty acids could be fulfilled by the fortification of
dairy products like yoghurt both set and drinkable types. Omega-3 fatty acids from
flaxseed oil microcapsules can be potentially delivered through dahi or Indian
yoghurt (Goyal et al. 2016).
Many dairy/food industries are doing research in these aspects. San Francisco-
based company Slingshot Foods manufacturing the omega-3 fortified drinkable
yoghurt with 600 mg of omega-3 fatty acid. The source of omega-3 fatty acid has
been used as a blend of almond bits, chia seeds, and rolled oats (https://
slingshotfoods.com/). The whole milk probiotic yoghurt fortified with omega-3

fatty acids are also manufacturing by Springfield Creamery (https://nancysyogurt.
com). This kind of yoghurt could be used for more beneficial and therapeutic aspects
in terms of improving digestive health as well as immune system (https://nancy-
syogurt.com).
5.4.2.7 Yoghurt Fortification with Plant-Based Components

as Natural Antioxidants
Yoghurt fortification with natural plant antioxidant is increasing these days. One
of the best advantages of using plant-based components like phenolic extracts,
polyphenols, flavonoids, catechin, etc. is its easy availability and compatibility
with fermented dairy products. These components not only affect the product
qualities but also boost the products’ functional and therapeutic values such as
ACE-I inhibitory activity (Amirdivani and Baba 2011), antioxidant activity
(Karaaslan et al. 2011; Najgebauer-Lejko et al. 2011; Aliakbarian et al. 2015;
Coman et al. 2017), anticancer activity (Wang et al. 2014; Fujiki et al. 2015; Zhou
et al. 2016), and antimicrobial activity (Cueva et al. 2010; Coman et al. 2017;
Pacheco-Ordaz et al. 2018). Fortification of plant component enhances the viabil-
ity of bacteria (Lee et al. 2006; Vasile et al. 2011; Coman et al. 2017; Pacheco-
Ordaz et al. 2018). The plant phenolic compounds’ interactions with probiotics to
boost their health-promoting properties and functionalities are described in details
in the review by De Souza et al. (2018).
Yoghurt fortification with plant components such as sterols and stanols have
been reported for their cholesterol-lowering properties (Parraga-Martinez et al.
2015). Sterols and stanols significantly reduced the high cholesterol and other lipid
profiles, and it significantly changed the low-density lipoprotein (LDL) cholesterol
profile. Similarly, plant component stanols with or without camelina oil fortified
yoghurt showed the very good cholesterol-lowering effect (Salo and Kuusisto 2016).
5.4.3 Cheese and Cheese Products
Cheese can also be used for the fortification and supplementation of essential nutri-
ents and other beneficial components. Cheese fortification improves its flavor and
appeal. Research studies suggested that low-fat cheese is a suitable product for vita-
min D3 fortification (Wagner et al. 2008) because its manufacturing process does
not affect the stability of vitamin D3 (Wagner et al. 2008). Similarly, cheddar cheese
fortified with vitamin D3 at amount 200 and 400 IU per serving was stable for
around 9 months, and cheese flavor was also accepted by the consumers (Ganesan
et al. 2011). Many other works related to cheese fortifications are going on around
the world. Fortification of cottage cheese with vitamin D without any loss in the
manufacturing process has been investigated in Saint-Hyacinthe Research and
Development Centre, Agriculture and Agri-Food Canada in Quebec (Crevier et al.
2017). The processing condition for cottage cheese making like homogenization
and pasteurization makes it more suitable for the vitamin D fortification. During
cottage cheese manufacturing, the cream has been fortified with vitamin D and
mixed with the fresh cheese curds. The loss of vitamin D has not been even detected
in the cheese whey (Crevier et al. 2017). The stability of fortified vitamin D was not
even affected at refrigeration temperature for up to 3 weeks (Crevier et al. 2017).
Cheese like cottage cheese is also an excellent vehicle for the delivery of live
microorganisms and probiotics. Cottage cheese is also known as the fresh curd
cheese because processing steps do not involve the pressing of cheese block to drain
out the whey. The dressing of cottage cheese with cream makes it distinguished
from other varieties of cheeses. Dressing of cottage cheese with cream increases its
flavor and palatability. Castro et al. (2015) suggested that cheese has better buffer-
ing capacity than other fermented dairy products and less water activity which is
suitable for the growth and activity of probiotic bacteria. However, the probiotics
delivery via different varieties of cheeses could be a great idea for the maximum
health benefits. Many cheese manufacturing industries are selling probiotic incor-
porated cheeses. Lifeway Foods sells probiotic cheese named as Farmer Cheese.
Successful delivery of active and live probiotic bacteria to the gastrointestinal tract
through the cheddar cheese could be a protective medium as compared to the
yoghurt (Gardiner et al. 1998).
Cheese fortification with protein or protein concentrate has been known for its
positive effect on curd formation, curd strength, and rigid matrix. A study suggested
that cheese made by blending with highly concentrated micellar casein concentrate
(HC-MCC) and cream improved the curd profile and also contributed to the form of
extensively cross-linked protein strands with high rigid protein matrix (Lu
et al. 2016).
Cheese fortification with polyphenols enhances the antioxidant activity of low-
fat ultrafiltrated soft cheese (Hala et al. 2010). Functional cheese containing poly-
phenolic compounds improves the nutritional value of cheese and radical scavenging
activity (Han et al. 2011). Quark cheese supplemented with ginseng extract does not
affect the overall quality attributes, but it can promote the public health as functional
foods (Kim et al. 2019). However, polyphenolic compound fortification in cheese
products might be a good approach for the preparation of better quality and func-
tionality of the products.
5.5 Fortification Challenges in the Milk and Milk Products
Fortification of any ingredient in the milk and milk products has always been tricky.
Because milk contains many components like protein, vitamin, carbohydrate, miner-
als, etc., it goes through many processing steps to convert into final products like
yoghurt, cheese, butter, milk powder, etc. Different kinds of fortification challenges
have been seen for different types of milk and milk product fortification. For example,
yoghurt manufacturing industries face the challenge of how to achieve consumer
demand for typical yoghurt flavor, taste, body, and texture. The valuable solutions to
overcome these challenges are to use protein or protein concentrate which could be
helpful in the abovementioned challenges (Peterson 2011). Proteins are known for
their different unique properties like thickening, gelling and overall texture improve-
ments. Excluding effects on physical properties, proteins are a good source of
essential amino acids (Peterson 2011).
Before fortification with any ingredients in milk and milk products, one must be
aware about the properties of that particular component like its solubility. The for-
tificant should not interfere with the original properties of the milk and milk prod-
ucts such as sensory property, textural property, overall appearance, and its shelf life.
The ultimate solution to avoid such problems is the key for the best product
development. Manufacturers must have knowledge about the components which are
going to be used for fortification, interactions with other nutrients, degradation abil-
ity, and interactions with its overall qualities. Some processing conditions directly
affect the fortified components and nutritional value. There are a lot of benefits with
mineral fortification, but some minerals like iron can pose other challenges like its
reactivity. Minerals like iron bioavailability can be enhanced by the vitamin C forti-
fication, and vitamin C can reverse the inhibiting effects of phosphate and calcium
(Lynch and Cook 1980).
The selection of ideal fortificant like iron must be done based on the following:
high bioavailability, no alteration of its sensory properties, compatibility with liquid
and solid foods, no diminishing effect on the product’s original nutritional value,
resistance to food processing, low in cost, and easy accessibility (Boccio et al. 1996).
An individual mineral fortification like iron can pose some problems on oxida-
tion, metallic taste, or discoloration. To avoid such problems, ingredient mix for-
mula can be adopted as an innovative solution (https://www.corbion.com/food).
However, formulated ingredient mix helps to minimize the cross interaction of min-
erals and vitamins in the final product. Many mineral products are available in the
market which supplies calcium-based minerals for the milk and milk product forti-
fication. Products of Purac like PURACAL XPro and PURACAL are natural and
calcium-based minerals known for their minimal impact during processing, out-
standing bioavailability, and neutral taste. These minerals are suitable for milk bev-
erage fortification. One of the best advantages of these minerals is their very fast and
complete dissolving property. Different brands of PURACAL have been suggested
for milk and milk product fortification. To fortify pasteurized milk, PURACAL®
calcium lactate has been recommended, whereas fast-dissolving PURACAL® PP
calcium lactate is recommended for yoghurt drink fortification. This calcium lactate
does not change the yoghurt flavor, and it boosts the calcium content up to 50%. For
some of the acidified milk beverages, PURACAL® XPro calcium lactate gluconate
has been recommended which contributes to smooth silky mouthfeel and no influ-
ence on its stability. Corbion supplies some of the other mineral lactates and gluco-
nates such as GLUCONAL® Zn. Corbion also supplies minerals and vitamins mix
under the brand name Nutrivan® to the North American dairy producers. Use of
Nutrivan® products has been suggested for the best performance during production
and for reducing the cost and time as well. Similarly, fresh cheese production
requires lactic acid, and brand PURAC lactic acid can be a good choice for the pro-
duction of fresh cheese (https://www.corbion.com/food). PURAC lactic acid could
be added directly to the product immediately after processing which creates a
microbiologically stable end product. Fermentation process has been known for the
development of typical flavor in the product. PURAC products have been known to
boost the typical flavor in the end products.
5.6 Industries Working on Fortified Milk and Milk Products
Many industries are working on fortified milk and milk products. The global market
of fortified milk and milk products are expanding day by day due to the demand for
food products with essential ingredients in order to fulfill the body’s physiological
requirements. Many companies such as Nestle, Amul, Arla, General Mills, Danone,
Fonterra, Bright Dairy & Food, BASF, China Modern Dairy, and Dean Foods, who
are manufacturing global fortified milk and milk products, have a competitive sce-
nario. Some of the companies manufacturing fortified milk products are mentioned
in Table 5.5.
Fortification with essential fatty acids such as omega-3 fatty acids can be easily
fortified with milk and many industries are working on it. The fortified milk manu-
facturing company fairlife LLC, Chicago, is known for manufacturing omega-3
Table 5.5 List of the fortified milk and milk products and their manufacturer
Name of the
Product name Fortified ingredients manufacturer
Reduced fat ultra-filtered milk, whole Omega-3 fatty acid Fairlife LLC,
ultra-filtered milk, chocolate whole Protein Chicago, USA
ultra-filtered milk Vitamins A, D, E, and C
Ready-to-drink energy milk beverage Caffeine, protein JoeFroyo,
Drinkable yoghurt Milk protein isolate, whey, California, USA
Probiotic cold brew active probiotic bacteria
Arabica cold brew coffee
Ice cream Cauliflower (source of folate,
fiber, and vitamins C and K)
BioKefir Probiotics Lifeway, USA
Yoplait yoghurt beverages Strawberry and pecan General Mills,
Inc.
Yoghurt drinks Mixed berry, strawberry, banana, Chobani LLC
mango, apple, cucumber, and
spinach
Probiotic drinkable yoghurt Probiotics, protein, fiber Dairy Innovations
LLC
Drinkable Greek yoghurt Stevia, prebiotic fiber, active Pillars Yogurt
probiotic culture LLC
Whole milk kefir Cream and strawberries Lifeway Foods,
Cream and coconut Inc.
Cream, wildberries, and lemon
meringue
Cream and peaches
Curb yoghurt smoothies Protein, whole grain Grupo Lala
fatty acid fortified milk. They are manufacturing three different types of omega-3
fortified milk which are reduced fat ultra-filtered milk, whole ultra-filtered milk,
and chocolate whole ultra-filtered milk. All these types of milk are rich in protein
and fortified with vitamins A, D, E, and C. These fortified milks are also well proven
for their various biological activities such as antioxidant activity and for their pre-
ventive actions against tissue and cell damages and anticancerous activity (https://
fairlife.com/). Similarly, the US-based Nestle Arlington, Va., is also manufacturing
the fortified milk for children which includes shelf-stable drink fortified with cal-
cium, vitamins A and D, and milk protein concentrate. They are also offering
Nesquik Super Breakfast protein breakfast milk drink flavored with vanilla and
chocolate. Nestle is manufacturing Boost High-Protein complete nutritional drink
for old age consumers which has been reformulated for 33% more protein per bottle
on its original formula. This nutritional drink has been mostly fortified with milk
protein concentrate and some portion from soy protein isolate, minerals, and 26
vitamins.
5.7 Future Prospects on Fortified Milk and Milk Products
Fortification of milk and milk products in the market has already been expanded.
Milk and milk products are known for their nutritional components, and it is consid-
ered as a suitable medium for fortification with health beneficial ingredients. The
main aim of fortification with vitamins, minerals, prebiotics, probiotics, plant
extracts, omega-3 fatty acids, etc. is to reduce the nutrient deficiency and related
diseases in the consumers. A wide range of fortification compatibility of the dairy
products will lead the future market of fortified milk and milk products. Moreover,
these days consumers are more aware about the product specifications and their
impacts on the body and therapeutic approaches as well. This will again boost up
the future market forecast for the development of fortified milk and milk products.
According to the Organisation for Economic Co-operation and Development, the
global consumption of milk products which was noted to be 411,173 thousand
tonnes in 2015 has immensely increased to 435,805 thousand tonnes in the year
2019. News published on 27th March 2019 in American NewsHour or via Comtex
suggested that the global fortified milk and milk product market can be categorized
into types of micronutrients like minerals and vitamins; product types such as
cheese, fermented milk, flavored milk, and yoghurt; types of product on the basis of
their consistency like liquid, semisolid, and solid dairy products; and according to
the demand of age groups which may include kids, young age groups, and old age
groups. The future market of consumer demand for functional dairy products could
be a great forecast for the demand of fortified milk and milk products i.e. 2018–2027
(https://www.globenewswire.com).
Milk and milk products are highly consumed food products across the world.
According to the American NewsHour or via Comtex, Asian countries like China
and India will have the highest market share for fortified milk and milk products.
Other countries like Europe and North America are following the Asian countries
for production and consumption of fortified milk and milk products. Increasing
health awareness and demand for specific healthy dietary supplements are the major
reasons for raising the market value for fortified milk and milk products. And it is
well known that milk and milk products are a very good medium for the supplemen-
tation of essential micronutrients and other dietary supplements.
As per Future Market Insights report on fortified dairy product market, the
worldwide sales and revenue generation by fortified milk and milk products will be
more than US$150,000 million by the end of year 2026. The importance of fortifica-
tion with essential ingredients and accepting the practice of fortification in dairy
products around the world are helping to boost up the sales of these fortified milk
and milk products. The Asia-Pacific excluding Japan (APEJ) is dominating the
global market for fortified milk and milk products, and estimated sales revenue will
be reach approximately US$ 50,000 million by 2026. According to the report, milk
powder formula is leading for the revenue generation followed by milk, flavored
milk, and yoghurt. Among the main fortified ingredients used for fortification pur-
pose, vitamin is leading followed by minerals and other components. It has also
been predicted that the market of fortified dairy products in North America will rise
at higher compound annual growth rate (CAGR) than Europe by 2026.
According to the WHO (2018) reports, vitamin A deficiency is very common
among many children. However, the demand for vitamin A fortified milk and milk
products is tremendous. Market values of dairy products are high, and yoghurt,
milkshakes, yoghurt drinks, beverages, and ice cream are the most demanding prod-
ucts which fulfill most of the nutrition. Fortifications with beneficial ingredients are
very much acceptable among these products. When it comes to the price of fortified
dairy products, consumers are willing to pay even higher price for their health.
Consumers have a lot of options to choose the brand of any particular type of forti-
fied dairy products.
5.8 Conclusions
Milk and milk product fortification with essential components is one of the best
methods to deliver the nutritional components. Dairy products are a highly con-
sumed food all over the world in all age group of people. Fortification with vita-
mins, minerals, probiotics, prebiotics, omega-3 fatty acids, and phytoconstituents
makes dairy products more valuable and appealing. Consumer awareness about the
nutrition and their importance in health leads to a demand for nutrition-rich dairy
products. Ultimately, this leads to the high market share value of fortified milk and
milk products, and an increasing revenue generation. The application of an advanced
and improved technologies in the food fortification makes it more homogenous dis-
tribution of all the fortified ingredients among each batch so that each and individual
consumer could get proper nutrition. Hence, fortified dairy product could be an
excellent approach to eradicate the hidden hunger and malnutrition problems and
other health-related problems all around the world.
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Multifunctional Aspects of Probiotics
and Prebiotics in Health Management: 6
An Overview
Kamna Saini and Jagrani Minj
Contents
6.2 W hat Are Probiotics, Prebiotics, Synbiotics, and Postbiotics? 121
6.2.1 Probiotics 121
6.2.2 Prebiotics 121
6.2.3 Synbiotics 123
6.2.4 Postbiotics 123
6.3 Probiotics: Mechanistic Actions 123
6.4 Role of Probiotics in Health Management 124
6.5 Effector Molecules from Probiotics 124
6.6 Bioactive Metabolites from Probiotics 126
6.6.1 Vitamins 126
6.6.2 Bacteriocins 126
6.7 Prebiotics: Mechanisms of Action 126
6.8 Role of Prebiotics in Health Management 126
6.9 Conclusions 128
References 128
Abstract
Probiotic bacteria have been known for their many potential health benefits
including improving healthy gut microbial populations; antimicrobial activity;
strengthening the immune system; hypocholesterolemic effects; anti-diabetic,
anti-cancerous, anti-oxidative, and anti-allergic characteristics; and also
K. Saini (*)
Faculty of Life Sciences, Institute of Applied Medicines and Research,
Ghaziabad, Uttar Pradesh, India
J. Minj

https://doi.org/10.1007/978-981-15-2608-4_6
120 K. Saini and J. Minj
p reventing different types of diarrhea and inflammatory bowel diseases. Different

probiotic strains have different or more than one unique health beneficial prop-
erty. Any disturbance in gut microbiota may lead to the development of diseases
or disease-like conditions. Although several factors such as age, diet, antibiotics,
disease status, environment, immune status, and physiology influence the healthy
gut microbial balance, diets enriched with probiotic and prebiotic can provide a
homeostasis in the gut microbial ecosystem. The combination of prebiotic and
probiotic is called a synbiotic, which further improves the healthy bacterial pop-
ulation in the gut. Metabolites produced by the probiotic bacteria have almost
similar properties like live probiotic bacteria, also known as postbiotics. However,
the probiotics, prebiotics, and their metabolites could be a great choice for health
management. This chapter deals with the understanding of the important role of
probiotics and prebiotics in the prevention and management of various diseases
and their complications.
Keywords
Microbiota · Probiotics · Prebiotics · Effector molecules · Health management
6.1 Introduction
The gut microflora plays a substantial role in metabolism, immunity, and health.
Any disturbance in the balance of gut microbiota has been emphasized as an
important factors for various lifestyle, diet-related, and metabolic disorders like
metabolic syndrome, obesity, colorectal cancer, diabetes mellitus type 2, inflam-
matory bowel disease (IBD), and so on (Bhushan et al. 2019). Several factors such
as diet, environment, antibiotics, physiology, age, disease status, immune status,
prenatal contact etc. influence the healthy gut microbial balance (Cresci and
Bawden 2015). Diet plays the major role as it provides the nutrition to gut microbes
and hence influences the gut ecosystem. Supplementing the diet with prebiotics,
probiotics, or synbiotics (probiotics plus prebiotics) has been the subject of pre-
ferred interest to improve the health or restore the gut microbial balance, supported
by several research-based studies and trials on safety aspects and many potential
health benefits. Probiotics taken in appropriate amounts benefit their hosts by dif-
ferent direct and indirect methods: competitive exclusion of pathogens, immuno-
modulation, production of growth-promoting substances such as vitamins,
antimicrobial activity by bacteriocins and bioactive peptides, and so on. Further,
prebiotics are complex nondigestible oligosaccharides that are not digested by
enzymes present in the intestine and are metabolized by probiotics and thus stimu-
late their growth (Gibson and Roberfroid 1995). This chapter’s focus is on under-
standing the role of probiotics and prebiotics in management of numerous health
conditions and diseases.
6 Multifunctional Aspects of Probiotics and Prebiotics in Health Management… 121
6.2 hat Are Probiotics, Prebiotics, Synbiotics,

W
and Postbiotics?
The terms probiotics, prebiotics, synbiotics, and postbiotics are generally defined in
many different ways, but all of these are connected with gut health and confer health
benefits to the host. These include live microbes that reside within the gut or their
bioactive products or the nondigestible substances that are metabolized by microbes
in the gut and ultimately stimulate their growth.
6.2.1 Probiotics
The joint FAO and WHO Expert Consultation (2002) on evaluation of health and
nutritional properties of probiotics in food defined probiotics as “live microorgan-
isms that, when administered in adequate amounts, confer a health benefit on the
host” (FAO/WHO 2002). Probiotics have been recognized to have a role in competi-
tive exclusion of pathogens, immunomodulation, production of bioactive metabo-
lites including bioactive peptides, vitamins, and bacteriocins in health management
(Bhushan et al. 2017; Thursby and Juge 2017). A list of common species of different
microbes that are currently used as probiotics is given in Table 6.1.
6.2.2 Prebiotics
Prebiotics are nutrients that are not easily digested but stimulate the growth of ben-
eficial microbial species in the gut and modify the gut microbiota (Rastall and
Gibson 2015). Prebiotic ingestion causes specific changes in population of gut
microbiota and mainly increases the gut microbial population, specifically
Lactobacillus and Bifidobacterium, with simultaneous reduction in harmful bacte-
rial population. Some of the commonly known prebiotics and their natural sources
are given in Table 6.2.
Prebiotics have been defined as “a non-viable food component that confers a
health benefit on the host associated with modulation of the microbiota” (FAO
2007). Gibson et al. 2004, defined prebiotic as a “selectively fermented ingredient
that allows specific changes; both in the composition and/or activity in the gastroin-
testinal microbiota that confers benefits upon host well-being and health”
(Gibson et al. 2004; Roberfroid 2007). The most recent definition says the
following criteria should be fulfilled: (1) should be resistant to digestion by gastric
acid and digestive enzymes, (2) should not be absorbed in the upper gastrointestinal
tract (GIT), and (3) should be fermented by the beneficial gut microbiota and can
stimulate their activity and growth as well (De Vrese and Scherezenmeir 2008).
Moreover, prebiotic index (absolute increase in concentration of the fecal
bifidobacteria/g of daily consumption of prebiotics) has been included in the current
definition. The oligosaccharides are low-molecular-weight carbohydrates and show
properties typical of dietary fibers (Delzenne 2003). The presence of glycosidic
Table 6.1 Common and Lactobacillus Lactobacillus acidophilus

actively used species of dif- L. rhamnosus
ferent probiotic microbes
L. sporogenes
(Kerry et al. 2018)
L. bulgaricus
L. reuteri
L. brevis
L. plantarum
L. fermentum
L. gasseri
L. crispatus
L. casei
L. paracasei
Streptococcus Streptococcus thermophilus
S. cremoris
S. lactis
S. salivarius
S. intermedius
S. diacetylactis
Bifidobacterium Bifidobacterium bifidum
B. infantis
B. longum
B. thermophilum
B. animalis
B. breve
B. lactis
Saccharomyces Saccharomyces boulardii
Saccharomyces cerevisiae
Leuconostoc Leuconostoc mesenteroides
Pediococcus Pediococcus pentosaceus
Propionibacterium Propionibacterium freudenreichii
Bacillus Bacillus cereus
Enterococcus Enterococcus faecium
Table 6.2 Commonly known prebiotics and their natural sources (Kerry et al. 2018)
Prebiotics Sources
Fructo-oligosaccharides or FOS Banana, chicory, garlic, and onion
Galacto-oligosaccharides or Human milk, kidney bean, and lentil
GOS
Inulin Banana, chicory, dandelion, garlic, and onion
Lactulose Skim milk
Resistant starch Starch-rich vegetables and fruits (banana) and whole grains
Xylo-oligosaccharides or XOS Bamboo shoot, fruits, honey, and vegetables
bonds in oligosaccharides makes them more resistant to hydrolysis by the gastric

acid and digestive enzymes, thus reaching the colon in a non-digested form where
gut microbes used it as a fermentable substrate (Nyman 2002). The gut microbes
produce many short-chain fatty acids such as acetic acid, butyric acid, propionic
acid, and lactic acid; and gases such as hydrogen, carbon dioxide, and methane gas
after fermenting nondigestible oligosaccharides which makes prebiotic a suitable
candidate for diet therapy to maintain the bowel in patients with irritable bowel
disease (Seidner et al. 2005).
6.2.3 Synbiotics
When probiotics and prebiotics are used as a combined product, it is known as syn-
biotics, and symbiotic relationship between them contributes significantly to the
host’s health. The dietary supplements increase the colonization of live and active
microbial populations in the gut (Tufarelli and Laudadio 2016). It has been sug-
gested that beneficiary effects of synbiotics are much more than individual use of
probiotics and prebiotics (Kerry et al. 2018). Therefore, current research is aimed at
producing new synbiotics to exploit further possibilities of health benefits.
6.2.4 Postbiotics
Recently, it has been suggested that nonviable bacterial products or metabolic end
products such as acetaldehydes, bacteriocins, diacetyl, ethanol, hydrogen peroxide,
and organic acids from probiotic microbes might have almost similar effects on the
signaling pathways and barrier function and are mostly considered as postbiotics
(Kerry et al. 2018). Postbiotics can be defined as nontoxic, nonpathogenic, nonvia-
ble bacterial metabolites from probiotic bacteria that can enhance barrier function
against pathogens and exert other beneficial biological activities for maintaining the
host health (Patel and Denning 2013). Postbiotics from L. rhamnosus GG have been
reported to have similar health benefits to that of live probiotic bacteria to increase
the intestinal barrier maturation in the murine model (Patel et al. 2012).
6.3 Probiotics: Mechanistic Actions
Some of the significant mechanisms of action of probiotics are competitive exclu-

sion of pathogens, modulation and increasing of gut microbial populations, immu-
nomodulation, and maintenance of the gut permeability and barrier integrity, as
specified in Fig. 6.1.
Fig. 6.1 Mechanisms of action of probiotics
6.4 Role of Probiotics in Health Management
Numerous clinical studies showed the health benefits of specific strain of probiotics
in the prevention and management of various diseases. Probiotics have been pro-
posed to be supportive in several diseases and health conditions such as colorectal
and liver cancer, obesity, rheumatoid arthritis, IBD, and hepatic disease, as outlined
in Table 6.3.
6.5 Effector Molecules from Probiotics
Probiotics and their effector molecules with their exact mode of action for a particu-
lar health claim must be authenticated with complete information. Some of the well-
recognized health benefits from probiotics Bifidobacterium and Lactobacillus
generated cellular effector molecules have been recently reviewed by Bhushan et al.
(2019). Effector molecules from L. rhamnosus GG include SpaCBA pili, p40 and
p75 (secreted proteins), lipoteichoic acid, CpG-rich DNA, and exopolysaccharides
(Bhushan et al. 2019). Probiotic effector molecule SpaCBA pili have the ability to
adhere to the mucous membrane and gut epithelial cells where they involve in the
crosstalk with innate immune cells like dendritic cells and monocytes and also
exhibits the competitive exclusions. Other probiotic effector molecules or secreted
proteins such as p75 and p40 helps in the prevention of gut epithelial cell damage
caused by the cytokine induction. They also help in the prevention of colitis and
related complications. Similarly, probiotic effector molecule lipoteichoic acid has
been reported to suppress the colitis induction, whereas CpG-rich DNA molecule
causes suppression of IgE. Probiotic effector molecule exopolysaccharides have
Table 6.3 Different types of probiotic bacterial strains along with their preventive roles in
diseases
Health condition/
disease Probiotic strain(s) Reference
Colorectal B. lactis Bl-04 and L. acidophilus NCFM Hibberd et al.
cancer (2017)
Oral mucositis Combined probiotic strains B. longum, E. faecium, and Jiang et al. (2019)
L. lactis (Bifico, Shanghai Sine Pharmaceutical Co.,
Ltd)
Liver cancer L. casei Shirota and L. rhamnosus GG Kumar et al.
(2011)
Probiotic product VSL#3 composed of L. acidophilus, Appleyard et al.
L. casei, L. delbrueckii subsp. bulgaricus, L. (2011)
plantarum, B. breve, B. infantis, B. longum, and S.
salivarius subsp. thermophilus
VSL#3, a probiotic mixture, Escherichia coli Nissle Li et al. (2016)
1917, and L. rhamnosus GG
Obesity B. animalis subsp. lactis CECT 8145 Pedret et al. (2019)
Akkermansia muciniphila Everard et al.
(2013)
L. plantarum KY1032 and L. curvatus HY7601 Park et al. (2013)
Acute diarrhea Probiotic yeast Saccharomyces boulardii Villarruel et al.
(2007); Feizizadeh
et al. (2014)
L. rhamnosus GG Aggarwal et al.
(2014)
Rheumatoid L. casei 01 Alipour et al.
arthritis (2014)
Hepatic disease L. acidophilus ATCC B3208, L. rhamnosus DSMZ Famouri et al.
21690, B. lactis DSMZ 32269, and B. bifidum ATCC (2017)
SD6576
Irritable bowel B. bifidum MIMBb75 Jacobs et al.
syndrome (IBS) (2013)
B. bifidum KCTC 12199BP, B. lactis KCTC 11904BP, Yoon et al. (2015)
B. longum KCTC 12200BP, L. acidophilus KCTC
11906BP, L. rhamnosus KCTC 12202BP, and S.
thermophilus KCTC 11870BP
also been reported for their help in mitigation of adipogenesis. Surface layer pro-
teins (SlpA, SlpB, and SlpC) from L. acidophilus NCFM modulate the intestinal
epithelial barrier by adhesion to SIGNR3 and C-type lectin receptor. Bacteriocin
Abp118 from L. salivarius UCC118 inhibits the growth of Listeria monocytogenes.
Dgk (diacylglycerol kinase) from L. reuteri 6475 suppresses the histamine produc-
tion via anti-inflammatory mechanism. Probiotic B. longum 35,624 produced exo-
polysaccharides have also been reported for their anti-inflammatory action and
anti-TH17 response (Bhushan et al. 2019).
6.6 Bioactive Metabolites from Probiotics
6.6.1 Vitamins
Probiotics have also been considered as a source of several vitamins in the small
intestine (Said and Mohammed 2006). Potential of probiotics as producers of vari-
ous vitamins, especially B vitamins, have been reviewed elaborately by Bhushan
et al. (2016, 2019). Probiotics play a significant role in complementing human’s
vitamin needs (Burgess et al. 2004).
6.6.2 Bacteriocins
The most common definition of bacteriocins is ribosomally synthesized peptides or

proteins by various bacteria that have the ability to kill or inhibit other microorgan-
isms (Cotter et al. 2013; Leroy and De Vuyst 2004). In addition to their use as natu-
ral food preservatives, these peptides have been recognized as therapeutic agents
because of their ability to kill or inhibit a number of pathogens (Perez et al. 2014).
Efficiency of bacteriocins like lacticin Q and nisin A have been reported for the
pathogen elimination (Bhushan et al. 2019). Furthermore, these bacteriocins are
better than antibiotic vancomycin in eliminating some other strains like methicillin-
resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus
faecalis (VRE) (Bhushan et al. 2019).
In addition to the above-mentioned production of bioactive molecules, probiotics
have also been recognized for the biotransformation of micronutrients such as sele-
nium, to increase its bioavailability (Saini et al. 2014, 2015; Saini and Tomar 2017).
6.7 Prebiotics: Mechanisms of Action
The most noteworthy mechanistic action of prebiotics is fermentation in the colon by

beneficial gut microbes and changes in composition of gut microflora. Usually,
microbes with only carbohydrate metabolic activity without any proteolytic activity
are beneficial. Some strains of lactobacilli and bifidobacteria have such typical meta-
bolic profile. A summary of mechanisms of action of prebiotics are shown in Fig. 6.2.
6.8 Role of Prebiotics in Health Management
It has been recognized that prebiotic stimulates growth of certain gut microbes and,
consequently, specific microorganisms support specific functions (Gibson and
Roberfroid 1995). Evidently, lactulose supports bifidobacteria and declines bacte-
roides in the gut (Bouhnik et al. 2004). Similarly, galacto-oligosaccharides have
been reported to support the growth of Bifidobacterium lactis Bb-12 (Malinen et al.
2002). Prebiotic beta-glucans have also been reported to increase the growth of
Fig. 6.2 Mechanisms of action of prebiotics
Table 6.4 Different types of prebiotics and their roles in disease prevention/management
Health condition/disease Prebiotic Reference
Irritable bowel diseases Trans-galactooligosaccharide Silk et al.
(2009)
Crohn’s disease Fructo-oligosaccharide Lindsay et al.
(2006)
Ulcerative colitis Oral oligofructose-enriched inulin Casellas et al.
(2007)
Hepatic encephalopathy Lactulose Bongaerts
et al. (2005)
Lactulose and lactitol Gluud et al.
(2016)
Diabetes Oligofructose Chan et al.
(2016)
Hypocholesterolemia Soluble fiber (Plantago ovata husk) Solà et al.
(2007)
Asthma and hyperpnea- Bimuno-galactooligosaccharide (B-GOS) Williams
induced bronchoconstriction et al. (2016)
Obesity Prebiotic-supplemented diet (10 g inulin/ Dehghan
day) et al. (2014)
8 g per day or 13.2 kcal per day of Nicolucci
oligofructose-enriched inulin et al. 2017
Prebiotic-supplemented diet with (1) 16 g Verhoef et al.
FOS per day or (2) 10 g FOS per day (2011)
Prebiotic bar (inulin-type fructans with 2 g Reimer et al.
inulin (from chicory root) + 6 g (2017)
oligofructose)
lactobacilli even more than bifidobacteria (Jaskari et al. 1998). Prebiotics confer
several health benefits on humans like protective effects to prevent colorectal can-
cer, liver cancer, diarrhea, intestinal bowel disorders, obesity, asthma, and many
more, as outlined in Table 6.4.
Although prebiotics provide several health benefits, there is a scarce number of
studies concerning screening of new potential prebiotics. Screening of new poten-
tial prebiotics is the need of this hour. However, higher quantities of prebiotic con-
sumption (i.e., more than 10 g per day) may lead to abdominal disorders, diarrhea,
flatulence, and intestinal discomfort because of fermentation of these prebiotics by
gut microbiota in the large intestine (Anadón et al. 2016). Therefore, eradicating the
side effects from prebiotics will be a big challenge for researchers.
6.9 Conclusions
The use of probiotics and prebiotics in health management is emerging as a safe and
promising therapy in different types of health conditions and diseases. Evaluation of
new strains of probiotics and new prebiotics and their potential health benefits needs
to be done by different types of clinical trials such as placebo-controlled, random-
ized, double-blind, etc. These studies will help to achieve the exact mechanism of
probiotic and prebiotic along with their beneficial health effects on human subjects.
However, it will be more fruitful to get the maximum therapeutic aspects of indi-
vidual probiotic and prebiotic components for health management.
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Research-Based Biofunctional Aspects
of Milk Protein-Derived Bioactive 7
Peptides
Suvartan Ranvir, Nancy Awasti, Pranali Nikam,

and Neelima Sharma
Contents
7.1 Introduction 134
7.2 Bioactive Peptides 135
7.3 Bioactive Peptides in Milk 135
7.4 Source of Bioactive Peptides 137
7.5 Bioactive Peptide Production 137
7.5.1 Digestion in Gastrointestinal Tract (In Vivo) 138
7.5.2 Microbial Fermentation 139
7.5.3 Enzymatic Hydrolysis 140
7.5.4 Food Processing 141
7.6 Occurrence of Bioactive Peptides in Dairy Products 142
7.7 Functions of Milk-Derived Bioactive Peptides 142
7.7.1 Antihypertensive Effects of Bioactive Peptides 142
7.7.2 Effects on Immune System 147
7.7.3 Effects on Nervous System 148
7.7.4 Antimicrobial Effect 149
7.7.5 Antioxidative Effect 149
7.7.6 Antithrombotic Effect 150
7.7.7 Antidiabetic Effect 150
7.7.8 Anticancerous Effect 151
7.7.9 Mineral Binding Effects 151
7.7.10 Miscellaneous Functions of Milk Protein-Derived Bioactive Peptides 152
7.8 Conclusions 152
References 153
S. Ranvir (*)
Sam Higginbottom University of Agriculture, Technology and Sciences,
Allahabad, Uttar Pradesh, India
N. Awasti
Department of Dairy and Food Science, South Dakota State University, Brookings, SD, USA
P. Nikam · N. Sharma
ICAR-National Dairy Research Institute, Karnal, Haryana, India

https://doi.org/10.1007/978-981-15-2608-4_7
134 S. Ranvir et al.
Abstract
Bioactive peptides are milk-derived short chains of amino acids with potential
biological activity, which are released in the gut through digestion of milk pro-
teins. Some of their beneficial effects include immunomodulatory, anticancer-
ous, antithrombotic, antihypertensive, antidiabetic, and antioxidative. Although
these peptides are made up of the same amino acids as other dietary proteins, the
specific sequence of these amino acids is responsible for such extraordinary
effects. This chapter attempts to summarize the available data on bioactive pep-
tides such as their general characteristics, production, physiological functions,
and potential applications.
Keywords
Bioactive peptides · Milk protein · Casein · Whey protein · Biofunctional peptides
7.1 Introduction
Milk is regarded as an excellent source of nutrients like carbohydrate, fat, protein,

vitamin and mineral. From few decades, many scientific studies have progres-
sively acknowledged protein as a physiologically active component in several
product matrix. Usually, bovine milk has total protein content about 3.5%, among
which casein and whey protein contributed approximately 80% and 20%, respec-
tively (Fox and McSweeney 2015). The casein and whey protein contents vary
considerably through different phases of lactation, particularly during the initial
days post-partum, and then the peak change mainly arises in the whey protein
fraction. The principal function of proteins in milk is to provide young mammals
with essential amino acids, for the development of their muscular as well as other
protein-containing tissues. Milk proteins also supply numerous biologically active
proteins (Fox and Flynn 1992) and other beneficial components, for example,
immunoglobulins (Shah and Lee 2000), metal and vitamin binding proteins,
growth factors (Pakkanen and Aalto 1997), and different protein hormones
(Walzem et al. 2002).
From past few decades several researches have demonstrated the potential ben-
efits of food-derived bioactive peptides on biomarkers linked with bone health and
metabolic syndrome (Hernández-Ledesma et al. 2014; Panchaud et al. 2012;
Udenigwe and Aluko 2012) and were also found endowed with properties like
immunomodulation, antidiabetic, antihypertensive, antiobesity, mineral binding,
antimicrobial, and antioxidative effects (FitzGerald and Meisel 2003; Korhonen and
Pihlanto 2006).
7 Research-Based Biofunctional Aspects of Milk Protein-Derived Bioactive Peptides 135
7.2 Bioactive Peptides
Bioactive peptides are definite protein fragments that have a positive influence on
the body and may ultimately influence health other than their nutritional value
(Bhat et al. 2015). Oral intake of these bioactive peptides may affect the key body
functions of digestive, nervous, cardiovascular, and immune systems. They impact
on human health due to their beneficial properties like antioxidative, antimicro-
bial, antithrombotic, immunomodulatory, or antihypertensive effects (FitzGerald
and Meisel 2003; Kitts and Weiler 2003; Korhonen and Pihlanto 2003). The activ-
ity of these bio-functional peptides depends on the inherent sequence and compo-
sition of amino acid. Generally, bioactive peptides contain about 2–20 residues of
amino acids (AA), but in few cases, they may also have more numbers of amino
acid carrying multifunctional beneficial properties (Bhat et al. 2015). For exam-
ple, a bioactive peptide (β-casein) with about 60–70 AA sequence was reported to
show multiple activities like immune stimulation and opioid and angiotensin
I-converting enzyme (ACE) inhibition. Therefore, sequence of β-casein has been
identified as a strategic zone (Meisel 1998) due its multifunctional properties.
One of the key properties of this sequence is its resistance and protection from
proteolysis due to its high proline residues and hydrophobic behavior. Another
example of milk-derived peptides with multifunctionality includes αs1-casein
fraction with amino acid sequence ranging from 194 to 199. Previous studies
reported multifunctional attributes of αs1-casein fraction that included ACE inhib-
itory activity and immune modulation.
In addition to this, peptides like opioid peptides α- and β-lactorphin were also
reported for their ACE inhibitory activity and calcium-binding phosphopeptides,
thus possessing immune modulation properties (Korhonen and Pihlanto 2003).
Based on the bioactivity of these milk components, they are mainly categorized into
four areas (Jayathilakan et al. 2018):
• Development, function, and activity of gastrointestinal tract

• Functions and development of immune system
• Infant development
• Activity of microbes together with their probiotic and antibiotic action
7.3 Bioactive Peptides in Milk
Milk comprises bioactive components in the form of minerals, lipids, casein,

enzymes, vitamins, lactoferrin, lactose and oligosaccharides, immunoglobulins,
cytokines, various growth factors, whey proteins, and derived peptides. Among all
these components, bioactive peptides are the most studied and explored component.
Bioactive peptides can be easily delivered to consumers by supplementing them in
conventional foods or as a dietary supplement and functional foods, or they can be
orally administered as a medical food. Milk components with their bioactive com-
pounds, sequence, and functional roles are described below (Tables 7.1 and 7.2).
Table 7.1 Major biologically active milk components and their functions
Milk precursors or
components Bioactive compounds Bioactivities observed
α-, β-caseins Casomorphins Opioid agonist (decrease gut mobility, gastric
emptying rate; increase amino acids and
electrolyte uptake)
α-, β-caseins Casokinins ACE inhibitory (increase blood flow to
intestinal epithelium)
α-, β-caseins Phosphopeptides Mineral binding (Ca binding; increase mineral
absorption, i.e., Ca, P, Zn)
α-, β-caseins Immunopeptides Immunomodulatory (increase immune
Casomorphins response and phagocytic activity)
Casokinins
αs1-Casein Isracidin Antimicrobial
αs2-Casein Casocidin Antimicrobial
κ-Casein Casoxins Opioid antagonist
κ-Casein Casoplatelins Antithrombotic
κ-Casein Glycomacropetide Probiotic (growth of bifidobacteria in GI tract)
α-Lactalbumin Lactorphins Opioid agonist
(α-La)
Serum albumin Serorphin Opioid agonist
α-La, β-La, and Lactokinins ACE inhibitory
serum albumin
Immunoglobulins IgG, IgA Immunomodulatory (passive immunity)
Lactoferrin Lactoferricin Immunomodulatory (increase natural killer
cell activity, humoral immune response,
thymocyte trafficking immunological
development, and interleukin-6; decrease
tumor necrosis factor-α)
Antimicrobial (increase bacteriostatic
inhibition of Fe-dependent bacteria; decrease
viral attachment to and infections of cells)
Probiotic activity (increase growth of
bifidobacteria in GI tract)
Lactoferrin Lactoferroxins Opioid antagonist
Oligosaccharides Oligosaccharides Probiotic (increase growth of bifidobacteria in
GI tract)
Glycolipids/ Glycolipids/ Antimicrobial (decrease bacterial and viral
oligosaccharides oligosaccharides attachment to intestinal epithelial cells)
Prolactin Prolactin Immunomodulatory (increase lymphocyte and
thymocyte trafficking and immune
development)
(continued)
Table 7.1 (continued)
Milk precursors or
components Bioactive compounds Bioactivities observed
Cytokines Interleukins-1, 2, 6, Immunomodulatory (lymphocyte trafficking,
and 10 immune development)
Tumor necrosis factor-α
Interferon-γ
Transforming growth
factors-α and β;
leukotriene B4
Prostaglandin E2, Fn
Growth factors IGF-1, TGF-α, EGF, Organ development and functions
TGF-β
Parathormone-P PTHrP Increase Ca2+ metabolism and uptake
Source: Adapted from Jayathilakan et al. (2018), Schanbacher et al. (1998), Park and Nam (2015)
Table 7.2 Bioactive peptides derived from whey proteins

Precursor protein Fragment Peptide sequence
α-Lactalbumin 50–53 Tyr-Gly-Leu-Phe
102–105 Tyr-Leu-Leu-Phe
142–148 Ala-Leu-Pro-Met-His-Ile-Arg
146–149 His-Ile-Arg-Leu
Bovine serum 399–404 Tyr-Gly-Phe-Gln-Asp-Ala
albumin 208–216
Lactoferrin 17–42 Lys-Cys-Arg-Arg-Trp-Glu-Trp-Arg-Met-Lys-Lys-Leu-Gly-
Ala-Pro-Ser-Ile-Thr- Cys-Val-Arg-Arg-Ala-Phe
Source: Adapted from Jayathilakan et al. (2018)
7.4 Source of Bioactive Peptides
Milk proteins like casein and whey proteins serve as a source of bioactive peptides
as they can be fragmented into numerous peptides by means of fermentation, enzy-
matic hydrolysis, and proteolysis. Each fraction and subfraction of bioactive pep-
tides with their distinctive biological assets can be found in whey or casein part
of milk.
7.5 Bioactive Peptide Production
Bioactive peptides are generally biologically inactive when present within the pro-
tein matrix; however, to make them biologically active, these peptides should be
released from their parent fragment by using three different strategies: (1) digestive
enzymes such as trypsin, pepsin, and chymotrypsin can be used for enzymatic
hydrolysis; (2) starter cultures with proteolytic behavior can be used for milk fer-
mentation; and (3) by using proteolytic enzymes derived from microorganisms
(Korhonen and Pihlanto 2003).
After liberation of these bioactive peptides from their parent protein, they
stimulate numerous physiological responses such as in digestive, cardiovascular,
immune, and neurological and endocrine systems. These bioactive peptides are
naturally present in dairy and can be widely found in numerous foods products
(especially with dairy-based ingredients). Based on their sequence and composi-
tion, different bioactive peptides may get into the small intestine where they are
either absorbed, degraded, or hydrolyzed by serum peptidases and intestinal (GI)
enzymes.
Numerous researches have examined the bioavailability and stability of bioactive
peptides derived from milk, with more focus on in vitro simulated digestive enzyme
treatments (Nongonierma and FitzGerald 2014; Stuknytė et al. 2015) or permeation
through Caco-2 cell monolayers (Shimizu et al. 1997; Vermeirssen et al. 2002;
Quirós et al. 2008; Picariello et al. 2013). These can also be produced in the gastro-
intestinal tract via milk digestion or during processing and fermentation of food
(Atanasova and Ivanova 2010). The biological activity of different peptides depends
on the composition and sequence of inherent amino acid. Bioactive peptides carry-
ing multifunctional beneficial properties generally contain about 2–20 amino acid
residues (Meisel and Fitzgerald 2003). Apart from this, these peptides also show a
wide range of other biological actions that may influence metabolic responses to
absorbed nutrients, digestion, development, resistance, and prevention against dis-
ease and growth of specific organs.
7.5.1 Digestion in Gastrointestinal Tract (In Vivo)
Usually, bioactive peptides can be released in the gastrointestinal tract by the

action of numerous digestive enzymes such as chymotrypsin, trypsin, and pep-
sin. In the presence of gastrointestinal acids mainly hydrochloric acid, these
dietary proteins go through denaturation, and acid also stimulates pepsinogen
into its active form, pepsin. Therefore, due to the action of pepsin, these proteins
are then metabolized to different amino acid chains. The resultant action of gas-
trointestinal enzymes such as trypsin, pepsin, and chymotrypsin occurs in the
small intestine, thus resulting into further hydrolysis (Korhonen and Pihlanto
2003). Numerous studies (Gobbetti et al. 2002; Meisel and Fitzgerald 2003;
Yamamoto et al. 2003; Fitzgerald et al. 2004) have reported that most of the bio-
active peptides are usually released during gastrointestinal digestion of whey
and/or casein proteins from milk and milk products. Further, proteolytic enzymes
like thermolysin and Alcalase can also be exploited in combination with trypsin
or pepsin to simulate gastrointestinal digestion and have been used to release dif-
ferent biologically active peptides such as ACE inhibitory (Vermeirssen et al.
2004), caseinophosphopeptides (CPPs) (Mcdonagh and Fitzgerald 1998),
antioxidative (Suetsuna et al. 2000; Rival et al. 2001a, b), antibacterial (Mohanty
et al. 2016), opioid peptides (Pihlanto-Leppala et al. 1994), and immunomodula-
tory (Gauthier et al. 2006a, b).
7.5.2 Microbial Fermentation
Numerous non-starter and starter cultures such as Lactobacillus delbrueckii ssp.

bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus
plantarum, Lactobacillus rhamnosus, Streptococcus thermophilus, and Lactococcus
lactis can produce bioactive peptides by their proteolytic activities during fermented
dairy product manufacturing. Lactic acid bacteria hold well-characterized proteo-
lytic system including numerous intracellular peptidases, cell-wall-bound protein-
ase, endopeptidases, tripeptidases, aminopeptidases, and dipeptidases. Extracellular
proteinases usually produce oligopeptides by degrading casein, and these long oli-
gopeptide chains are further degraded into numerous bioactive peptides by intracel-
lular peptidases released by lysed lactic acid bacteria (Zhou et al. 2019). Studies
from the last decade have demonstrated the production of numerous bioactive pep-
tides including immunomodulatory, antimicrobial, ACE inhibitory, and antioxida-
tive via microbial proteolysis (Korhonen and Pihlanto 2003; Gobbetti et al. 2004).
Fermentation using microbial culture with proteolytic attributes is one of the most
efficient approaches to enhance the bioactive peptide concentration in fermented
dairy products.
The effective release of variable bioactive peptides depends on the type of differ-
ent proteolytic strain. Based on previous knowledge, the microbial strain should not
have high proteolytic activity because to produce increased concentration of active
peptides. Right concentration and specificity of enzyme should be used, as the
increased concentration of proteolytic enzyme may result into product destruction.
For example, the concentration and production of ACE inhibitory peptides appear to
depend on a balance among their formation and breakdown into amino acids and
inactive peptides which in turn rely on various storage conditions and time
(Korhonen 2009). Numerous bioactive peptides including antihypertensive peptides
or ACE inhibitory, immunomodulatory, antimutagenic, and antioxidative peptides
have been released from milk proteins through microbial proteolysis (Korhonen and
Pihlanto 2001; Matar et al. 2003; Gobbetti et al. 2004).
The best-known bioactive peptide is ACE inhibitory peptides such as Ile-Pro-
Pro (IPP) and Val-Pro-Pro (VPP) and has been produced due to the fermentation
by Saccharomyces cerevisiae and Lactobacillus helveticus in milk. In addition to
live lactic acid bacteria (LAB) cultures, addition of LAB-produced proteolytic
enzymes has been effectively utilized to release bioactive peptides from milk pro-
teins. The production of bioactive peptides by milk fermentation employing dif-
ferent microbial cultures and their proteolytic enzymes has been detailed in
Table 7.3.
Table 7.3 Bioactive peptides released from milk proteins by various microorganisms
Precursor
Microorganism protein Peptide sequence Bioactivity
L. rhamnosus + digestion with β-CN Asp-Lys-Ile-His-Pro- ACE inhibitory
pepsin Phe,
Tyr-Gln-Glu-Pro- Val-
Leu
Lactobacillus helveticus β-CN, Val-Pro-Pro, ACE inhibitory,
k-CN Ile-Pro-Pro antihypertensive
Lactobacillus GG β-CN, Tyr-Pro-Phe-Pro, Opioid, ACE
enzymes + pepsin and trypsin αs1-CN Ala-Val-Pro-Tyr-Pro- inhibitory
Gln Arg
Thr-Thr-Met-Pro-Leu- Immune-stimulatory
Trp
Lactobacillus delbrueckii k-CN Ala-Arg-His-Pro-His- Antioxidative
subsp. Pro-His-Leu-Ser-Phe-
met
Kluyveromyces marxianus var. β-lg Tyr-Leu-Leu-Phe ACE inhibitory
Lactobacillus helveticus CP90 β-CN Lys-Val-Leu-Pro-Val- ACE inhibitory
proteinase Pro-(Glu)
Source: Adapted from Mohanty et al. (2016)
Table 7.4 Antimicrobial peptides derived from milk and their target microorganisms
Milk peptides Protease Pathogens
Isracidin αs1-CN (f1–23) Chymosin, Staphylococcus aureus
chymotrypsin
Casecidin αs1-CN Chymosin, Staphylococcus, Bacillus subtilis, Diplococcus
chymotrypsin pneumoniae, Streptococcus pyogenes
Lactoferricin B and Pepsin Bacillus, E. coli, Candida albicans, Listeria,
lactoferrin (f 17–41) Streptococci, Klebsiella, Staphylococci
Lactoferrin (f 17–41) – Proteus, Pseudomonas, Salmonella
β-Casein-derived Trypsin and Enterococcus faecium, Bacillus megaterium
peptides chymotrypsin
Source: Adapted from Mohanty et al. (2016)
7.5.3 Enzymatic Hydrolysis
The common approach to release bioactive peptides is via enzymatic hydrolysis of

protein present in milk. A combination of various proteinases (such as chymotryp-
sin, pepsin, alcalase, and thermolysin), digestive enzymes, and/or fungal or bacte-
rial enzymes has also been used to produce bioactive peptides (Mohanty et al. 2016)
(Table 7.4). Further to isolate these peptides, the hydrolysates are generally frac-
tioned and enriched by using numerous techniques like ultrafiltration, precipitation
(including salts or solvent), and chromatography. From few decades, to enrich
cationic peptides possessing antibacterial properties and to desalt whey protein

hydrolysates, negatively charged membrane materials have been used. This tech-
nique has been providing a new projection towards enriching peptides and their easy
scale-up especially for low molecular mass. Among different commercially avail-
able dairy products, cheese is considered as a major bioactive peptide source. These
bioactive peptides are known to produce during cheese ripening resulting into pro-
teolytic activity by naturally present milk microflora, peptidases, rennet, and prote-
ases. The incidence of bioactive peptide presence in various cheeses has been stated
by numerous researches, and their activity was found related with cheese matura-
tion. During cheese ripening, caseinophosphopeptides (CPPs) are produced because
of action of proteolytic enzymes, and plasmin is also produced by the
LAB. Commercially available cheese has been found endowed with numerous ACE
inhibitory peptides, plus the tripeptides IPP and VPP in physiological appropriate
concentrations. In different types of cheese, especially in Brie cheese, variable con-
centrations of β-casomorphin-7 and opioid peptide have been found ranging from
0.15 mg kg−1. Apart from cheeses, based on different types of milk, other commer-
cially available fermented dairy products like yogurts, kefir, and fermented milks
are also known to contain bioactive peptides such as antioxidant, opioid, immuno-
modulatory, and ACE inhibitory peptides (Bargeman et al. 2000; Haque and
Rattan 2006).
Digestion by pepsin and other pancreatic enzymes generally results into release
or cleavage of latent bioactive peptides from milk proteins, thus producing active
peptide fragments in the gastrointestinal tract of the individual consuming milk
products (Schlimme and Meisel 1995). The physiological effects of bioactive pep-
tides vary on their capability to reach in intact form to their target sites, resulting
from absorption of these peptides through epithelium cells of intestinal walls to the
peripheral organs (Vermeirssen et al. 2004). Several bioactive peptides like CPPs
and ACE inhibitory peptides have been commercially produced in vitro by using
gastrointestinal enzymes, such as pepsin and trypsin. ACE inhibitory peptides and
CPPs, for example, are most commonly produced by trypsin.
7.5.4 Food Processing
During product processing, proteins usually go through chemical and structural

changes which may result into production or release of active fragments in the prod-
uct. In comparison to other food processing conditions, alkali and/or heat treatment
can produce intra- and intermolecular covalent bonds that makes protein resistant
towards hydrolysis.
Such conditions may also encourage the racemic modification of L-amino acids
to D-isomers and, therefore, result into generation of peptide bonds which cannot be
digested easily.
It is worth mentioning that some food processing techniques can result into the
production of potentially active peptide sequences, which have more bioavailability
and do not occur naturally in the precursor protein. These peptides can thus be
ingested as food components in infant formula for hypoallergenic infants, which
contain partially hydrolyzed milk proteins and possess clinical applications in
nutrition.
Cheese is also known to contain phosphopeptides as a natural constituent pro-
duced during cheese ripening, whereas secondary proteolysis was found to be asso-
ciated with the formation of several ACE inhibitory peptides (Meisel 1993; Haque
and Rattan 2006).
7.6 Occurrence of Bioactive Peptides in Dairy Products
It is now well known that during milk fermentation, bioactive peptides are usually
produced due to their proteolytic activity of starter cultures. Therefore, the end
product of milk fermentation generally results into release of numerous types of
peptides with variable bioactivities. After intake of different traditional dairy prod-
ucts, numerous studies have reported specific health benefits under certain condi-
tions. For details on various bioactive peptides present in dairy products, refer
Table 7.5.
7.7 Functions of Milk-Derived Bioactive Peptides
Milk-derived bioactive peptides have many functional roles as an exogenous regula-

tory compounds. A number of studies have been done on the potential biological
activities of these milk-derived bioactive peptides. Although many research claims
that these peptides can play a role of hormones or food hormones, still more clarifi-
cation about it is needed. Biologically active peptides have been reported for their
physiological effects in in vivo conditions, where they get released by the proteo-
lytic enzymatic action. These released small bioactive peptides get absorbed by the
intestinal epithelial cells and then move to the peripheral organs at targeted sites.
Different structural peptides have different and unique biological activities. Some of
them are listed in Table 7.6.
7.7.1 Antihypertensive Effects of Bioactive Peptides
Hypertension is a known risk factor which leads to development of cardiovascular

diseases including stroke. According to World Health Organization (WHO) reports
(2016), cardiovascular diseases kill 17.5 million people every year. Many drugs are
available for the effective treatment of hypertension but can cause several adverse
side effects. Therefore, an alternative treatment can be a good choice to avoid side
effects. Natural remedies like reducing alcohol consumption and salt intake, regular
Table 7.5 Bioactive peptides identified from milk products

Product Origin Biofunctional role Reference
Cheddar cheese αs1- and ß-casein Several phosphopeptides with a Singh et al.
fragments range of properties including the (1997)
ability to bind and solubilize
minerals
Italian cheeses: ß-CN f (58–72) ACE inhibitory Smacchi and
mozzarella, Gobbetti
crescenza, Italico, (1998)
Gorgonzola
Yogurt-type αs1, ß-, and ACE inhibitory Yamamoto
products κ-CN fragments et al. (2003)
Gouda cheese αs1-αs1-CN f ACE inhibitory Saito et al.
(1–9), ß-CN f (2000)
(60–68)
Festivo cheese αs1-CN f (1–9), f ACE inhibitory Ryhänen et al.
(1–7), f (1–6) (2001)
Emmental cheese αs1- and ß-casein Immunostimulatory, mineral Gagnaire
fragments binding and solubilizing , et al. (2001)
antimicrobial
Manchego cheese Ovine αs1-, ACE inhibitory Gomez-Ruiz
αs2,- and et al. (2002)
ß-casein
fragments
Sour milk ß-CN f (74–76), ACE inhibitory/antihypertensive Nakamura
f (84–86), κ-CN et al. (1995)
f (108–111)
Dahi Ser-Lys-Val- ACE inhibitory Ashar and
Tyr-Pro Chand (2004)
Source: Haque and Rattan (2006)
exercise, and intake of nutrient-rich foods could be alternative methods to get over
from the hypertension problem. In hypertension, angiotensin I-converting enzyme
(ACE-I) plays an important role to regulate the blood pressure. This enzyme belongs
to the zinc metallopeptidases and involves in the renin-angiotensin system (RAS)
and kallikrein-kinin system (KKS). These two systems are known for increasing
high blood pressure. In RAS system, the ACE-induced cleavage of C-terminal pep-
tide of angiotensin-I releases the potent vasopressor angiotensin-II (Skeggs et al.
1956). Similarly in KKS system, the ACE inactivates the vasodilator bradykinin
through the removal of the two C-terminal dipeptides (Yang et al. 1970). The renin
cleaves angiotensinogen and releases angiotensin-I, which is further hydrolyzed by
ACE to produce angiotensin-II (a potent vasoconstrictor). Also vasodilator bradyki-
nin inactivated by ACE, which further contributing to an increased blood pressure
development of cardiovascular disease (CVD) (FitzGerald et al. 2004). The angio-
tensin also stimulates the production of aldosterone, which is another hormone,
from the adrenal cortex. In distal nephron, aldosterone helps in sodium retention,
Table 7.6 List of structural element of peptides, their biological activity, and their precursor proteins
144
Protein precursor Bioactive peptide Specific structural element of peptide Targeted health aspect Biological activity
α-Casein Casokinins C-terminal peptides, mainly Phe or High blood pressure/hypertension ACE inhibitory activity
Lys/Arg
β-Casein Lactokinins Tyr, Pro, or hydroxyl Pro
κ-Casein
α-Lactalbumin
β-Lactalbumin Lactotensin High extent of hydrophobic amino High cholesterol Hypocholesterolemic
acids effect
Lactostatin Low ratio of Lys-Arg and Met-Gly High blood lipid profile Lipid-lowering effect
contents
α-Casein – Cys, His, Lys, Met, Trp, Tyr amino Oxidative stress Antioxidant activity
β-Casein acid sequence with Pro-His-His
κ-Casein
β-Lactalbumin
κ-Casein – Asp115, Ile108, and Lys112 and some Thrombosis Antithrombotic action
sugar content Blood clotting
Plaque formation
Casein – Casoplatelins Diabetes Antidiabetic activity
Whey proteins DPP-IV inhibitory
action m
Whey proteins – – Obesity/high body weight Satiety effect
β-Casein α-Casein 3rd and 4th position aromatic amino Role in nervous system Opioid-like activity
αs-Casein β-Casomorphin acids, N-terminus Tyr Neuromodulator
α-Lactalbumin Exorphins
β-Lactoglobulin Lactorphins
Lactoferrin
S. Ranvir et al.
β-Casein Caseinophosphopeptides Phosphorylated regions Bone health including teeth Boost up of mineral
αs-Casein or CPP absorption
β-Casein Casocidin Certain amphiphilic peptide sequence, Body defense from harmful Antimicrobial action
αs-Casein Isracidin highly positive charged peptide pathogenic and disease-causing
α-Lactalbumin Lactoferricin sequences bacteria, food quality
β-Lactoglobulin
Lactoferrin
Source: Adapted from Pihlanto (2011)
7 Research-Based Biofunctional Aspects of Milk Protein-Derived Bioactive Peptides
145
which also drives blood pressure up in the kidney (Crowley and Coffman 2014).
However, ACE plays an important role in both systems which have been known for
increasing high blood pressure. Blocking or inhibition of ACE pathways by the
ACE inhibitors could control the high blood pressure and ultimately hypotensive
effect. Therefore, the existence of natural ACE inhibitors from milk and milk prod-
ucts came into streamline. Milk and milk products, especially fermented milk prod-
ucts, contain many ACE inhibitory substances, which is an opportunistic preventive
measure to control high blood pressure. Many ACE inhibitory peptides have been
found in milk proteins, especially lactokinins and casokinins derived from whey
protein and casein, respectively. These peptides have been reported for their potent
ACE inhibitory activity (FitzGerald et al. 2004; Vercruysse et al. 2005). Some other
potent ACE inhibitory peptides with tripeptide sequences like Ile-Pro-Pro and Val-
Pro-Pro have been found in the sour milk fermented with L. helveticus and
Saccharomyces cerevisiae. Soedamah-Muthu et al. (2012) also reported that low-fat
milk and dairy products reduce hypertension. The presence of ACE inhibitory pep-
tides has been reported in different sources of milk such as cow milk (Sieber et al.
2010; Rodriguez-Figueroa et al. 2012), goat milk (Chen et al. 2018), and camel milk
(Yahya et al. 2017).
The tripeptide sequences Ile-Pro-Pro, Leu-Pro-Pro, and Val-Pro-Pro have been
mostly studied for their ACE inhibitory and hypotensive activities (Nakamura et al.
1995; Masuda et al. 1996). The release of these tripeptide sequences could be pos-
sible either by enzymatic actions, during food processing, or by microbial fermenta-
tion. Milk proteins, mainly casein and whey proteins, are the main source for these
ACE inhibitory peptides. The ACE inhibitory activity of any peptides usually is
measured by the percent inhibition with IC50 values. The Japanese sour milk drink
Calpis® containing L. helveticus and S. cerevisiae has been recognized for its high-
est concentration of tripeptide sequences IPP and VPP with IC50 values 5 and
9 μmol, respectively (Nakamura et al. 1995). The hypotensive effect of these two
peptide sequences has been studied in a placebo-controlled study with hypertensive
patient. Daily intake of 95 ml of sour milk containing these two peptides for 4
weeks significantly decreased hypertension. The daily dose of ACE inhibitory pep-
tide was almost corresponding to about 2.6 mg per day (Hata et al. 1996). Similarly,
another commercial fermented product Evolus® of Valio Ltd, Finland, or Kaiku Vita
brand®, Spain, made with starter culture L. helveticus LBK-16H exhibited hypoten-
sive effect on hypertensive patients with 150 ml daily consumption. Some commer-
cial ACE inhibitory peptide products such as whey protein hydrolysate of BioZate,
Davisco, USA, and casein hydrolysate with peptide sequence FFVAPFEVFGK and
precursor protein αs1-casein f23-34 of Casein DP, Kanebo Ltd, Japan have been well
claimed for the blood pressure lowering effects of peptide (FitzGerald et al. 2004).
However, it seems that consumption of food products containing these tripeptides
directly affects the blood pressure-lowering mechanisms (Hirota et al. 2011). The
feeding of tripeptide might be toxic, but research on rabbits for 90 days of repeated
dose oral gavage test did not show any serious adverse effects (Dent et al. 2007;
Beltran-Barrientos et al. 2017).
7.7.2 Effects on Immune System
A good health requires proper functioning of the immune system because the
immune system plays an important role in the body’s defense system where it fights
against harmful cells like cancerous cells; recognizes and neutralizes the harmful
substances from the environment; neutralizes and kills harmful pathogenic bacteria,
viruses, fungi, and parasites; and removes them from the body. That is why strength-
ening the immune system is very important to avoid many diseases. Healthy foods
and lifestyle could be considered to boost up the immune system. When it comes to
healthy foods, milk and milk products are known for their nutritional and therapeu-
tic aspects. Milk and milk products contain many biologically active components
which help to strengthen the immune system. These biologically active components
might be proteins, peptides, or amino acids. Milk peptides having immune strength-
ening property are known as immune modulatory peptides. Many immune modula-
tory peptides have been found in the bovine milk and milk products as well.
Hydrolysates of major milk protein have been reported for their immunomodulatory
activity (Brandelli et al. 2015; Park and Nam 2015). The exact mechanisms of this
immunomodulatory activity are not yet clearly defined. Individual peptide has a
unique and individual structure that could affect the cellular functions which directly
affects the immune responses.
Milk protein-derived peptides have been known for their potential immune
modulatory actions, and they do not have any toxic effects, making them an alter-
native treatment for disease conditions (Agyei et al. 2019; Gokhale and
Satyanarayanajois 2014). Major milk proteins such as αs1-casein, αs2-casein,
β-casein, κ-casein, lactoferrin, and α-lactalbumin contain many peptide sequences
which exhibit immunomodulatory effect. Some of the immunoglobulin fragments
and hormones can act as immunomodulators by regulating the humoral and cell-
mediated immune functions. Immunomodulatory activity may have included
increase in immunoglobulin (IgA, IgG, IgM) production, lymphocyte prolifera-
tion, phagocytosis, proliferation of spleen cells and lymph nodes, and increase in
B-cells, T-cells, and monocytes. Casein- and whey protein-derived peptides have
been seen for these properties including antibody synthesis and cytokine regulation
(Clare et al. 2003; Gill et al. 2000). Similarly, immunomodulatory activities of
bioactive derived peptides have been seen for their specific and nonspecific immune
responses (Meisel 2004; Gauthier et al. 2006a, b). Immunomodulatory peptides
not only improve the immune system, but they also protect the body from several
diseases. Gu et al. (2018) reported that immunomodulating peptide sequence
PGPIPN could show the protective effect on acute alcoholic liver injury in the mice
animal model. Similarly, allergic reactions could be alleviated by the immuno-
modulatory peptides, and on the other side, mucosal immunity of gastrointestinal
tract gets strengthened by these peptides (Korhonen and Pihlanto 2003).
Immunomodulatory peptide releases from fermented milk have also been seen for
their antitumor effects (Matar et al. 2003).
The production of immunomodulatory peptides can also be possible with the

fermentation with lactic acid bacteria. The lactic acid bacteria present in any fer-
mented milk and milk products produce metabolites such as small peptides, amino
acids, bacteriocins, etc. These small peptides or biologically active peptides may
regulate the cytokine production and can exhibit the suppressive effects in inflam-
mation. Milk fermented with Lactobacillus has been seen for the reduction of pro-
inflammatory cytokines, mainly TNF-α and IL-6, whereas it increased the
anti-inflammatory cytokine IL-10 in the lipopolysaccharide-stimulated murine
model (Reyes-Díaz et al. 2018). Similarly, L. paracasei NCC2461 fermented milk
exhibited the immune stimulatory activity in terms of upregulation of IL-10 and
downregulation of pro-inflammatory cytokines IFN-γ and IL-4 (Prioult et al. 2004).
Immunomodulatory activity of β-casein fermented with LAB has also stimulated
the production of mononuclear cells and T-helper cells (Laffineur et al. 1996).
Cheese glycomacropeptide and its derivatives have also been known for the sup-
pressive effect on the production of IgG antibodies (Monnai et al. 1998; Manso and
Lopez-Fandino 2004). Bovine milk proteins αs1-casein and β-casein fermented with
L. acidophilus ATCC4356 and Lactococcus lactis subsp. lactis GR5 released the
immunomodulatory peptides which decreased the NF-κβ activity in the recombi-
nant Caco-2 cell layers (Stuknyte et al. 2011). In another report, bovine milk pro-
teins α- casein and β-casein fermented with L. delbrueckii ssp. bulgaricus 92059
and L. helveticus 92201 exhibited the immune modulatory effect (Kliche et al.
2017). The immunomodulatory peptides like caseinomacropeptide (CMP) have
been reported for their growth and support of lactobacilli and bifidobacteria.
Many other peptide sequences have been reported for their immune modulatory
activity. Peptide sequences Tyr-Gly and Tyr-Gly-Gly have been reported for prolif-
eration of lymphocytes, whereas β-casomorphin-7 and β-casomorphin-10 have
inhibited at low concentration to the lymphocyte proliferation (Kayser and Meisel
1996). Another important milk protein is lactoferrin which is also called iron-
binding protein. Lactoferrin-derived lactoferricin B has the ability to directly bind
to the neutrophil that exhibits opsonin-like activity.
7.7.3 Effects on Nervous System
Many studies have been done to see and confirm the important role of milk-
derived peptides in the nervous system. These peptides are also called the opioid
peptides. Brantl (1984) explained that opioid peptides are opioid receptor ligands
which are generally encrypted from human and bovine β-casein enzymatically
in vitro. Opioid receptors have been found in the central nervous system and
peripheral tissues and involve in many physiological processes (Liu and
Udenigwe 2019). Opioid peptides are of two types: endogenous and exogenous
peptides. Endogenous opioid peptides involve in the physiological processes,
while exogenous peptides are derived from food or milk proteins. These food
proteins hydrolyze by the gastrointestinal proteases and exhibit opioid-like
activity. The first identified opioid peptide from bovine milk β-casein was βb-
casomorphin-7 with peptide sequence Tyr-Pro-Phe-Pro-Gly-Pro-Ile (Brantl et al.
1981). In continuous study, βb-casomorphin-4, βb-casomorphin-5, and βb-
casomorphin-6 have also been recognized as opioid peptides (Brantl et al. 1981).
Among these peptides, βb-casomorphin-5 showed the highest opioid activity in
the electrically stimulated guinea pig ileum (GPI) and mouse vas deferens
(MVD) tests (Brantl et al. 1981). Not only β-casein-derived peptides have opi-
oid-like activity but some of the α-casein-derived peptides have also opioid
activity. The identified opioid peptide from bovine α-casein is αb-casein exor-
phins (Teschemacher et al. 1997). Some other opioid-like peptides are seen in the
whey proteins such as αb-lactorphin and βb-lactorphin from bovine αb-lactalbumin
and βb-lactoglobulin, respectively (Teschemacher et al. 1997; Garg et al. 2016).
Many more opioid peptide sequences form milk protein and food proteins are
reviewed by Liu and Udenigwe (2019). The main functions of these opioid pep-
tides are to cross the blood-brain barrier and to target the opioid receptor for
opioid-like activity.
7.7.4 Antimicrobial Effect
The first antimicrobial action of milk by lactenin against streptococci was reported
by Jones and Simms (1930). This leads to do more work on the antimicrobial activ-
ity of milk-derived bioactive peptides. Milk-derived peptides with molecular
weight below 10 kDa are mostly reported for the antimicrobial action (Mohanty
et al. 2016; Tomioka et al. 2014). Many antimicrobial peptides from milk have
been reported against many gram-negative and gram-positive pathogenic bacteria
(Mohanty et al. 2014). Antimicrobial peptides such as β-casein-derived peptides,
casecidin αs1 and κ-casein, isracidin αs1-casein (f1-23), and lactoferricin B and lac-
toferrin (f17-41) have been derived from milk (Mohanty et al. 2016). Several other
peptides known for their antimicrobial action include casocidin (Lahov and
Regelson 1996), casocidin-1 (Zucht et al. 1995), and lactoferrampin (Van der
Kraan et al. 2004). The suggested mechanisms of action of antimicrobial peptides
are bactericidal actions, hydrolysis of peptidoglycan layer, immune defense sys-
tem, metal chelation, C18 fatty acid binding, and alteration in cell membrane per-
meability (Clare et al. 2003).
7.7.5 Antioxidative Effect
Some of the milk-derived peptides have been known for their antioxidative activ-
ity. The antioxidant peptides play an important role in the oxidative metabolism
which is known for the survival of cells and causes oxidative changes by produc-
tion of free radicals. The excessive free radicals lead to oxidation of membrane
lipid, cellular proteins, enzymes, and DNA which ultimately results in shutting
down of cellular respiration and generates many diseases like cancer, diabetes,
atherosclerosis, and rheumatoid arthritis (Halliwell 2000; Abuja and Albertini
2001; Halliwell and Whiteman 2004). The main functions of milk-derived anti-
oxidant peptides are scavenging or prevention of the formation of free radicals
(Rival et al. 2001a, b; Wong and Kitts 2003) and inhibition of enzymatic and non-
enzymatic lipid peroxidation.
Milk proteins such as casein and β-lactoglobulin release the antioxidant pep-
tides in the presence of enzyme trypsin, and pepsin (Suetsuna et al. 2000; Rival
et al. 2001a, b). Antioxidant peptides from milk improve the antioxidant enzymes
such as catalase, glutathione peroxidase, and superoxide dismutase in the diabetic
rats (Awad et al. 2016). Many research works are ongoing to find out the novel
antioxidant peptides from different sources of milk. Recently, novel antioxidant
peptides RLDGQGRPRVWLGR, TPDNIDIWLGGIAEPQVKR, and
VAYSDDGENWTEYRDQGAVEGK have been purified from camel milk. The
β-casein-derived peptides NEDNHPGALGEPV and KVLPVPQQMVPYPRQ
have been identified by Homayouni-Tabrizi et al. (2016); later on, they were identi-
fied as novel antioxidant peptide sequences by Homayouni-Tabrizi (2017).
7.7.6 Antithrombotic Effect
Antithrombotic can be defined as inhibition of blood clot formation or its enlarge-

ment in the body. Thrombotic condition can be caused by the hyperreactive plate-
lets, increased level of fibrinogen, irregular fibrinolysis, etc. Milk protein-derived
peptides can act as antithrombotic agents which could be an alternate to replace
drugs because drugs always cause some side effects. The κ-casein fragment caso-
platelins is known for their antithrombotic action (Bhandari et al. 2019). Similarly,
bovine κ-casein f106–f116 inhibits the platelet aggregation and prevents fibrinogen
binding with blood platelets (Erdmann et al. 2008). Chabance et al. (1995) reported
that bovine milk has the same peptide sequence with antithrombotic actions.
Another antithrombotic peptide sequence YQEPVLGPVRGPFPIIV derived from
casein was reported by Rojas-Ronquillo et al. (2012) and Shanmugam et al. (2015).
Similarly, β-casein-derived novel antithrombotic peptide YQEPVLGPVR (PICA)
has been identified from milk protein (Liu et al. 2019). Yogurt also contains κ-casein
peptides such as antithrombotic peptide Met-Ala-Ile (Manso and López-
Fandiño 2003).
7.7.7 Antidiabetic Effect
Milk peptides with antidiabetic effect have also been reported in many studies. Milk
proteins like casein and whey protein have the ability to stimulate the insulin secre-
tion which can be beneficial for the diabetic patients (Nilsson et al. 2007). The
hydrolyzed milk protein including whey proteins shows higher insulinotropic effect
than unhydrolyzed one (Power et al. 2009; Gaudel et al. 2013; Jakubowicz and Froy
2013; Nongonierma et al. 2013). The peptide sequence Ile-Pro-Ala released from
β-lactoglobulin of whey protein is reported for their inhibition of dipeptidyl pepti-
dase-4 (DPP-IV), insulin stimulation, and reduction of blood glucose levels
(Jakubowicz and Froy 2013).
DPP-IV inhibitory peptides and α-glucosidase inhibitory effects are related to
the antidiabetic action. Milk proteins from different sources contain unique
sequences for antidiabetic activity or DPP-IV inhibitory activity. The DPP-IV
inhibitory peptides KDLWDDFKGL and MPSKPPLL were reported in camel milk
protein hydrolysates (Mudgil et al. 2018). The molecular and cellular mechanisms
of antidiabetic action by camel milk may have either direct or indirect effects on
insulin secretion by the pancreatic β-cells; or direct effects on glucose transport and
insulin receptor function in the insulin-sensitive tissues; or effects on the growth,
survival, and overall activity of pancreatic β-cells (Akli Ayoub et al. 2018). The
review given by El-Sayed and Awad (2019) elaborates some more antidiabetic
activity of milk bioactive peptides.
7.7.8 Anticancerous Effect
Certain milk proteins and their peptides may influence the carcinogenesis. In this
sequence, whey protein exerts suppressive action against tumor development
(Parodi 2007). Milk protein lactoferrin-derived peptide lactoferricin plays an impor-
tant role in the modulation of carcinogen-metabolizing enzymes, anti-angiogenesis
effects, induction of apoptosis, cell cycle arrest, anti-metastasis effects, and necrosis
and acts as an iron scavenger (Yin et al. 2013; Mader et al. 2005; Gilford et al.
2005). Bovine skim milk digested with cell-free extract from the yeast Saccharomyces
cerevisiae inhibits the human leukemia cell line HL-60 (Roy et al. 1999). CPPs
derived from casein fraction also reported for the anti-cancer activity (Meisel and
FitzGerald 2003). Many other potential anticancer peptides from bovine milk have
been reported in a review by Sah et al. (2015). However, it can be considered that
milk proteins and their peptides can play an important role in cancer prevention and
management.
7.7.9 Mineral Binding Effects
The main functions of mineral binding peptides are to form complexes by binding
with minerals like calcium and phosphorus. These mineral binding peptides have
greater anionic properties; thus, they form soluble complex and become resistant to
further proteolytic attack and prevent the formation of insoluble complexes with
minerals. Many milk protein-derived peptides like whey protein, α-lactalbumin,
β-lactoglobulin, and lactoferrin with mineral binding abilities have been reported.
Lactoferrin is one of the most important whey proteins with iron binding ability.
Casein phosphopeptides (CPP) have been reported for their affinity for iron and
calcium binding (Vegarud et al. 2000). Casein phosphopeptides αS1-casein,
αS2-casein, and β-casein with many fragments have been identified to stabilize cal-
cium and phosphate ions (Cross et al. 2007). Several studies have been done to see
the potential of CPP to enhance mineral bioavailability. The enhancement in cal-
cium absorption by CPPs was reported in postmenopausal women (Heaney et al.
1994). During tryptic digestion, CPPs are released from the N-terminus polar
domain containing cluster phosphorylated seryl residues that are responsible for the
interaction between casein, calcium, and phosphate resulting in the casein micelle
formation. Finally, these phosphorylated peptides bind to the phosphate and cal-
cium ions and increase their bioavailability (Meisel and Fitzgerald 2003). Bennet
et al. (2000) reported that CPPs to calcium ratio also affects the mineral
bioavailability.
7.7.10 Miscellaneous Functions of Milk Protein-Derived

Bioactive Peptides
Milk protein-derived peptides have many other miscellaneous functions. Anti-

inflammatory effects on macrophages of the milk protein casein are reported by
Nielsen et al. (2012). Not only casein protein exhibits anti-inflammatory effects but
whey protein like lactoferrin is also reported to have anti-inflammatory effects.
Lactoferrin takes part in the downregulation of lipopolysaccharide-induced cyto-
kine production and activates the NF-κβ activation (Håversen et al. 2002).
Also, the peptide sequence Ile-Ile-Ala-Glu-Lys identified from tryptic hydroly-
sate of β-lactoglobulin shows hypocholesterolemic activity (Nagaoka et al. 2001).
This peptide was able to suppress the cholesterol absorption by Caco-2 cells in
in vitro condition, which further demonstrated the hypocholesterolemic activity in
the rats as animal model.
Cytomodulatory effects of milk peptides have also been reported. Cytomodulatory
peptides are also known as apoptosis-inducing peptides or cell growth-stimulating
peptides or immunomodulatory peptides. The milk peptide casokinin from β-casein
has been reported for the DNA synthesis stimulating activity (Nagaune et al. 1989).
The growth-promoting peptides can act as a growth factor for the development of
the digestive tract of neonatal infants.
Anti-appetizing peptides are known to prevent obesity and weight gain by sup-
pressing the appetite. Whey protein is reported for the release of appetite suppress-
ing hormone cholecystokinin (Jayathilakan and Sultana 2018).
7.8 Conclusions
Milk protein-derived bioactive peptides have many physiological functions like

antihypertensive activity, antimicrobial, antioxidant, immunomodulatory, and min-
eral binding abilities, and anti-cancer activities. These functions of milk peptide can
be useful for the treatment of many diseases and their complications. The delivery
of these peptides could be possible either with the direct consumption of food
products containing these peptides or in a drug form. However, natural and rich
sources of these peptides are fermented milk, yogurt, beverages, etc. The well-iden-
tified peptides can also be used in the functional food product development and
health-promoting nutraceuticals along with their well-documented health effects.
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Whey: Importance and Techno-
functional Applications 8
Rita Mehla, Anuradha Kumari, Jyotika Dhankhar,
Mitul Bumbadiya, and Anuj Tyagi
Contents
8.2 P hysicochemical Importance of Key Whey Proteins 164
8.2.1 α-Lactalbumin 164
8.2.2 β-Lactoglobulin 164
8.2.3 Bovine Serum Albumin 164
8.3 Physiological Significance of Whey Proteins 165
8.4 Techno-functional Properties of Whey Proteins 165
8.4.1 Solubility 165
8.4.2 Gelation 166
8.4.3 Viscosity 167
8.4.4 Emulsifying Property 168
8.4.5 Foaming Property 168
8.4.6 Fat and Oil Binding Property 169
8.4.7 Water Holding Capacity 169
8.5 Techno-functional Applications of Whey-Based Foods 170
8.6 Source of Whey-Derived Bioactive Components 171
R. Mehla (*) · M. Bumbadiya

Dairy Chemistry, ICAR-National Dairy Research Institute, Karnal, Haryana, India
A. Kumari
Department of Dairy Microbiology, College of Dairy Science and Technology,
Kerala Veterinary and Animal Sciences University (KVASU), Mannuthy,
Dairy Chemistry, Guru Angad Dev Veterinary and Animal Sciences University,
J. Dhankhar
Food Science and Technology, MDU, Rohtak, Haryana, India
e-mail: jyotika.ft@mdurohtak.ac.in
A. Tyagi
Fish Microbiology, Guru Angad Dev Veterinary and Animal Sciences University,
Ludhiana, India

https://doi.org/10.1007/978-981-15-2608-4_8
162 R. Mehla et al.
8.7 F
unctional Properties of Whey Protein-Derived Bioactive Peptides 172
8.7.1 Antioxidant Activity 172
8.7.2 ACE Inhibition Property 172
8.7.3 Anticarcinogenic Property 173
8.7.4 Antimicrobial and Antiviral Activities 173
8.7.5 Whey Protein usage as biodegradable Packaging Material 174
8.7.6 Pharmaceutical Applications 174
8.7.7 Encapsulation Capability of Whey Proteins 174
8.8 Conclusions 175
References 175
Abstract
Whey, a protein-rich source, is generally considered as the leftover of the cheese
production. Whey proteins are usually marketed as nutritional enhancement, and
several health claims have been reported. Whey proteins are commercially avail-
able in isolate and concentrate forms and have a wide range of food applications
due to their nutritional importance, functional attributes, and cost-effectiveness.
α-Lactalbumin, β-lactoglobulin, and bovine serum albumin are the major whey
proteins. Some of the techno-functional properties of whey proteins are solubil-
ity, viscosity, gelation, emulsification, foaming, fat and oil binding, and water
holding capacity. Whey protein serves as a potent bioactive component which
imparts in various bioactivities as antioxidant, angiotensin-converting enzyme
(ACE) inhibitory, anticarcinogenic, antimicrobial, and antiviral activities. It is
also extensively used as a biodegradable packaging material, encapsulating and
pharmaceutical material. Hence, this chapter dealt with the importance and
techno-functional applications of whey.
Keywords
Whey protein · Techno-functional property · Food applications · Bioactivity
8.1 Introduction
Whey is a by-product of the cheese manufacturing process or the product obtained

after elimination of casein or fat from the milk. Whey has the significance of being
utilized both as animal feed and human dietary protein constituents. On the basis of
the type of manufacturing process used, whey is classified as acid whey and rennet
whey/sweet whey. Sweet whey or rennet whey is the by-product of rennet type of
cheese, viz., hard cheeses like Swiss or Cheddar cheese. On the other hand, acid
whey is produced during the preparation of acidic milk products such as ricotta,
cottage, and similar kind of several other cheeses (Farrell et al. 2004). Whey protein
does not undergo aggregation on renneting or acidification and remains insoluble,
without any dependence on pH (El-Desoki 2009). Sweet whey and acid whey are
almost similar in composition, as presented in Table 8.1. They differ only in acidity,
whey protein fraction composition, and mineral contents (Bansal and Bhandari
8 Whey: Importance and Techno-functional Applications 163
Table 8.1 Composition of Sweet

sweet and acid forms of whey Compositional components (g/L) whey Acid whey
Total solids 63.0–70.0 63.0–70.0
Protein 6.50–6.60 6.10–6.20
Non-protein nitrogen (NPN) 0.27–0.37 0.30–0.40
Lactose 46.0–52.0 44.0–47.0
Fat 0.20–0.50 0.30
Ash 5.00–5.20 7.50–7.90
Calcium 0.40–0.60 1.20–1.60
Phosphate 0.50–1.00 2.00
Sodium 0.50–0.53 0.50–0.51
Chloride 1.10 1.10
Lactate 2.00 6.40
pH 5.90–6.40 4.60–4.70
Bansal and Bhandari (2016)
0.40%
0.35%
0.30%
0.25%
0.20%
0.35%
0.15%
0.10%
0.05%
0.05% 0.08% 0.01%
0.05% 0.01%
0.00%
0.15%
Fig. 8.1 Protein composition of whey (% w/v) (Bansal and Bhandari 2016)
2016). The whey protein comprises alpha-lactalbumin (α-La), beta-lactoglobulin

(β-lg), immunoglobulin (Ig), bovine serum albumin (BSA), and few additional
minor proteins as presented in Fig. 8.1 and exists as compact, globular proteins
(Bansal and Bhandari 2016).
Among the various commercially available forms of whey proteins, whey protein
isolates (WPI) and whey protein concentrates (WPC) are the important ones. Spray
dried WPC contains about 25–89% of protein content, whereas WPI contains protein
content of about 90–95% with a low level of cholesterol and fat (Marshall 2004).
These whey protein source also contain many bioactive components (Huppertz et al.
2006). Whey protein contributes to multiple functionalities like digestibility, acces-
sibility, sensory attributes, and biological value (McIntosh et al. 1998). Due to the
release of potential bioactive peptides from whey proteins in the small intestine,
these are considered as an important physiological and nutritional functional supple-
ment than other supplements. The bioactive peptides isolated from the whey protein
164 R. Mehla et al.
exhibit various bioactivities such as opiate, antithrombotic, antioxidative, and immuno-

modulation (Vasiljevic and Jelen 2003).
8.2 Physicochemical Importance of Key Whey Proteins
8.2.1 α-Lactalbumin
α-Lactalbumin (α-la) constitutes ~20% of whey protein and ~3.5% of total milk
proteins. With 123 amino acid residue length, the molecular weight and isoelectric
point (pH) of α-la are ~14 kDa and 4.8, respectively. The native form of α-la is
monomeric in nature with 4-disulfide bridges between the amino acid residue of 6,
120, 28 and 111, 61 and 77, and 73 and 91. Due to the inherent stability, the disul-
fide bond between 6 and 120 is more sensitive (Considine et al. 2007). Absence of
thiol group contributes towards heat stability (Paulsson and Dejmek 1990; Monahan
et al. 1993). Between pH 5.4 and 9, it exhibits a stable configuration. One mole of
α-lactalbumin contains one Ca2+ with four Asp residues in a pocket. The Ca in the
protein maintains heat stability and promotes unfolding. It is considered as the most
heat-stable form of whey protein. At pH less than 5, Ca2+ affinity decreases as Asp
residue becomes protonated resulting in reduced heat stability and renaturation
capacity during cooling (Fox and McSweeney 1998).
8.2.2 β-Lactoglobulin
With a length of 162 amino acids and a molecular weight of ~18.3 kDa,
β-lactoglobulin is the major milk protein that constitutes about ~50% of total whey
protein and ~12% of total milk proteins. The isoelectric point of β-lg is at pH 5.2.
The milk protein β-lg has well-defined primary, secondary, tertiary, and quaternary
structures. Its tertiary structure is stabilized by two disulfide bonds between Cys
66–160 and Cys 106–119 (Thompson et al. 2009). The β-lg exists as monomer at
pH below 3.5, as octamer between pH 3.5 and 5.5, as dimer in the pH range of
5.5–7.5, and as monomer at pH greater than 7.5 (Fox and Mcsweeney 1998). At
milk pH, these exist as dimer form at ambient temperature and detach to monomer
with the increase in temperature (Croguennec et al. 2004).
8.2.3 Bovine Serum Albumin
With a 582-amino-acid length, a 66-kDa molecular mass, and an isoelectric pH of

~5.3, bovine serum albumin (BSA) has a multidomain structure (Morr and Ha 1993).
Disulfide bonds are responsible for the stabilization of its domain structure (Considine
et al. 2007). BSA contains 17 disulfides and one free sulfhydryl group at position
Cys34 (Thompson et al. 2009). The molecule has mainly helical turn, elliptical shape
with three domains (Considine et al. 2007). In the circulatory system, BSA also acts
as a carrier for the insoluble fatty acids (Juliette et al. 1995).
8.3 Physiological Significance of Whey Proteins
The presence of hydrophilic residue in whey protein provides high solubility (Lee
et al. 1992). The soluble nature of whey proteins at a wide pH range facilitates their
passage through the small intestine without being precipitated in the stomach acidic
conditions. In the small intestine, it gets hydrolyzed quickly; peptides and amino
acids get delivered fast in contrast to the casein. Furthermore, the peptides derived
from whey protein have various health benefits. The glutathione acts as an antioxi-
dant, prevents the body against the free radical injury, and also detoxifies the poten-
tial carcinogen (Parodi 2007). These whey proteins are also a good source of ACE
inhibitory peptides (Mann et al. 2019). Whey protein also endows with immuno-
logical defense to neonates mostly through immunoglobulins available in the colos-
trums (Thompson et al. 2009). It provides various other health benefits as stimulation
of fast satiation which reduces the food intake (Manski et al. 2007), improvement of
mineral amalgamation, and opioid activity (Pihlanto-Leppälä 2000).
8.4 Techno-functional Properties of Whey Proteins
Apart from the physiological significance, tremendous techno-functional properties

and pleasant sensory uniqueness of whey proteins make them ideal candidates for
various food applications. Whey protein is extracted from whey by means of differ-
ent fractionation method, primarily by membrane processing. Whey protein con-
centrate is manufactured through diafiltration/ultrafiltration by removing lactose,
water as well as minerals under mild pH and temperature during processing. Ion-
exchange chromatography or microfiltration techniques are used for the manufac-
ture of WPI. WPC or WPI powders are produced by further spray drying of retentate
(Fox and Mcsweeney 1998). The functional properties of whey protein are affected
by the different physicochemical characteristics, three-dimensional structure, shape,
size, amino acid composition, total charge, and hydrophobicity/hydrophilicity ratio.
The techno-functional properties of whey proteins are described below.
8.4.1 Solubility
Solubility is one of the crucial techno-functional properties of whey proteins. It also

significantly affects other functional properties like foaming, emulsification, as well
as gelation. On the basis of this property, it is being used in various foods and bever-
ages. Water binding property and the physical state are the important factors which
affect the solubility. The swelling and water absorption property of protein is influ-
enced by the hydration site number, confirmation, surface properties, composition
166 R. Mehla et al.
of amino acid, molecular weight, sequence, and number of nonpolar/polar groups

present in the amino acid (Cayot and Lorient 1997; Zayas 1997a). Environmental
conditions like temperature, pH, processing conditions, ionic strength, and kind of
solvent used also influence the solubility of the protein (Zayas 1997a).
The extent of protein solubility is also related to the hydrophobic and electro-
static interactions between the molecules of protein. If electrostatic repulsions
among the protein molecules dominate over the hydrophobic interactions, then sol-
ubility improved. For a protein to be soluble, the solvent-protein interaction should
be utmost. The total charge of zero at the isoelectric point (PI) leads to the domi-
nance of attractive forces and consequent molecular association of proteins result-
ing in the insolubility. The net charge of the protein goes to negative above the PI
and positive below the PI. The water-protein interactions increase at lower or higher
pH than the PI. At the pH close to the PI, the least water-protein interaction has been
observed (Zayas 1997a).
The water-binding capacity increases with the increase in the heat denaturation
of protein due to the unfolding of protein structure which exposes the hydrophilic
groups to the water (Kinsella 1984). However, protein structural state affects the
water-binding capacity. Depending upon the pH conditions, β-lactoglobulin may
exist in three structural forms, i.e., monomer, dimer, and octamer. The water-binding
ability of dimer form (at 40 g H2O/100 g protein) is lesser than that of octamer form
(at 60 g H2O/100 g protein). In comparison to β-lactoglobulin, α-lactalbumin is
more hydrated, i.e., 57 g/100 g protein. The water holding capacity of whey pow-
ders ranges from 70 to 147 g/100 g protein, and it is influenced by the drying capac-
ity and the type of whey used (Cayot and Lorient 1997). On heating protein-protein
interaction increases, leading to the aggregation and loss of solubility at
PI. Denaturation of protein by heating decreases the solubility, and it further impairs
the emulsifying property. On the other hand, these properties are improved by the
partial denaturation (Leman and Kinsella 1989; Lorient et al. 1991).
8.4.2 Gelation
Gelling property significantly affects the water holding and textural properties of
several food products. Due to both solid and fluid like characteristics, protein gels
are unique in nature. These also contribute to the stabilization of emulsion and foam
(Zayas 1997b). The gelling property is also accountable for the viscoelastic prop-
erty of the foods, i.e., increased adhesiveness, viscosity, and improved water-
retention properties. Gelation property is mainly due to the three-dimensional
structure of protein (Zayas 1997b). Cayot and Lorient (1997) observed that the gela-
tion of protein can be accomplished by mixing the enzymes or salts to a concen-
trated protein solution and also by applying the excess heat or pH. Calcium is not
only essential for the gelation of whey protein, but it also has a significant impact on
hardness and water-retention properties of the gel. With the initial unfolding of mol-
ecule followed by aggregation, the heat-induced gelation is a two-step process. With
the unfolding, buried hydrogen bonding groups are exposed resulting in increased
water binding. Heat-induced gelation is mainly caused by the hydrophobic interac-

tions and disulfide bonding (de la Fuente et al. 2002; Clark et al. 2001; Singh and
Havea 2003). pH, salt, and protein concentrations have an influence on thermal
gelation.
During gel formation, partially unfolded proteins turned into the uncoiled poly-
peptide chain which further interacts at a particular position, and it forms three-
dimensional cross-linked networks. Different factors like heating, urea, acid, and
alkali treatments can cause the native protein structure unfolding (Zayas 1997b).
Gelation is the consequences of protein-water and protein-protein interactions that
imparts to the structural rigidity. When the protein-protein interactions dominate
over the protein-water interaction, the three-dimensional structure gets distorted
and water oozes out. Gel formation involves various forces like van der Waals
forces, hydrogen and disulfide bonding, hydrophobic and ionic interactions. As the
number of –SS and –SH groups increases by denaturation, the stronger network has
appeared with the formation of new disulfide bonds. The heat-induced gel can be
reversible or thermoset and irreversible or thermoplastic. Thermoset gel is obtained
by heating which results in the increase of viscosity. On heating, the gel can be
melted and get reversible on cooling. On the other hand, in thermoplastic gel, rever-
sion or melting of the gel is not acquired under normal condition. The heat-induced
gelation involves two step processes: protein-protein and protein-water interactions
resulting in the formation of three-dimensional protein network (Zayas 1997b).
Whey protein concentrates also have cold gelling property; hence, preheating to at
least 70 °C is required for cold gelling. Whey protein concentrate is commercially
available and used in the salad dressing and mayonnaise-like products.
8.4.3 Viscosity
Viscosity is the resistance offered by the liquid to flow under shear stress or applied
force. Flowability is affected by the molecular size, shape, charge, swelling, and
solubility property of the protein molecule. Environmental conditions like concen-
tration, pH, and temperature also influence these properties. Whey protein concen-
trate gets unfolded on heating and forms aggregates on their association (Johnson
2000). It has been observed that no aggregates have been formed when α-lactalbumin
is heated alone. However, β-lactoglobulin forms larger aggregates. Smaller aggre-
gates are more soluble and able to bind with the increased amount of solvent or
water which further improves the viscosity, body, and texture of the product.
Viscosity is used in determining the thickening property of the protein and also
applied in fluid foods, beverages, and soups. Whey protein preparations are not very
viscous at high temperature also. When the pH approaches at PI, viscosity decreases
(Castle et al. 1986). In addition to its water-binding tendency, whey protein also has
the property to stabilize food preparations even after cooking (e.g., cakes, sausages,
and emulsions) when the pH approaches to its neutrality (Cayot and Lorient 1997).
168 R. Mehla et al.
8.4.4 Emulsifying Property
The emulsifying capacity may be explained by the emulsifying activity (EA) and
emulsifying capacity (EC). The emulsifying capacity is the amount of oil emulsified
by 1 gram of protein under specific condition. Emulsifying property of emulsifier is
dependent on two different factors: (1) adsorptions as a film in the region of the
globule and (2) tendency to reduce the oil-water interfacial tension. Emulsion stabi-
lization is the tendency of the emulsion to stay dispersed with no separation by
coalescence, creaming, and flocculation. The maximum interfacial area (cm2) per
1 g of protein in a stable emulsion is known as the emulsion activity (Zayas 1997c).
Emulsion capacity and stability are also influenced by the conditions of emulsi-
fication as well as protein property, and differ with the protein source, pH, amount,
viscosity, and ionic strength. The emulsifying property is affected by the globular
protein solubility, by its surface hydrophobicity, and also by the molecular flexibil-
ity (Nakai et al. 1980). Apart from this, it also depends on the protein solution,
temperature, and design of the equipment. The emulsifying property and hydropho-
bicity are strongly correlated to each other (Kato and Nakai 1980). Whey proteins
act as surfactants and decrease the oil/water interfacial tension and also form a thin
film on the droplets (Cayot and Lorient 1997). They also prevent the flocculation
and coalescence of the emulsion by emulsion stabilization. WPC showed the emul-
sion ability from 38 to 52 ml of oil per gram of protein. The relative emulsifying
capacity of the WPI and WPC from sweet whey is ~52 ml oil per gram of protein
(Kim et al. 1987). During WPC powder manufacturing process, thermal denatur-
ation of whey proteins leads to reduction of their solubility and further affects their
emulsifying property. However, partial denaturation tends to improve these proper-
ties (Leman and Kinsella 1989; Lorient et al. 1991).
8.4.5 Foaming Property
Whey proteins have the capability to form a stable foam, and this property makes
the protein more useful for the preparation of several food items. Two phases exist
in the foam: air and liquid phases. The lamellar phase is a continuous thin liquid
layer that separates the air cell. Food foams exhibit their complexity due to the pres-
ence of gases, solid, liquid, and surfactants. Products’ textural properties are largely
related to the air bubble size distribution in the foam. Even air bubble distribution
contributes to the smooth and light product (Zayas 1997d).
Foamability and whippability are the two confusing terminologies. In whippabil-
ity, foam is obtained by high-speed whipping or blending. On the other hand, when
the foam is formed by the inclusion of gas or air in the protein solution, then it is
termed foamability. Foaming property is influenced by the protein source and the
conformation of protein like shape, size, structure and composition of amino acid,
processing conditions, and also physicochemical properties (Patino and Pilosof 2011).
Foaming is a three-step mechanism. During the first step, the whey protein gets
adsorbed at the air-water interface and decreases surface tension. In the second
step, protein gets unfolded at the interface, and hydrophilic groups attached to the
water and nonpolar groups attached to the nonaqueous phase. During the last step,
the interaction of polypeptides leads to film formation with slight denaturation and
coagulation. Quick adsorption of proteins at the interface leads to the formation of
a stabilized film around the bubbles resulting in the foam creation (Zayas 1997d).
Foamability of the globular proteins is analogous to the egg white protein. Hence,
egg white protein can be replaced by globular proteins (Yang and Foegeding 2011).
8.4.6 Fat and Oil Binding Property
The fat binding properties of whey protein contribute to food’s textural and other
characteristics. In addition to flavor, it also improves the mouthfeel of the food prod-
ucts. This property is also dependent on the processing parameters like particle size,
protein source, additive composition, and temperature. Protein powder fat binding
capacity is related to the size of powder particles. Fat adsorption is the entrapment
of oil (Kinsella and Melachouris 1976). Nonpolar and insoluble protein provides
better oil binding property. So, it also contributes to the hydrophobicity/hydrophi-
licity ratio (Voutsinas and Nakai 1983). Products with low protein contribute to
better binding ability (Zayas 1997e). Heating at above 70 °C leads to the adsorption
of whey protein to the fat globule membrane. Heating causes interaction of whey
protein with the casein adsorbed on the fat globule membrane via cysteine moiety
of casein.
8.4.7 Water Holding Capacity
Water and protein interaction is facilitated by the water adsorption, binding, hydra-
tion, imbibing, and water holding properties. Water holding capacity is the reten-
tion of water on their own or under various physical treatments such as force,
centrifugal force, pressure, and heating (Zayas 1997f). It reflects the ability of the
food matrix to avoid water removal from the three-dimensional protein network
(Hermansson 1986). Thus, the water is physically entrapped in the food matrix.
Water-protein interaction is involved in the textural as well as rheological proper-
ties of the food. Many other techno-functional properties like water holding, solu-
bility, viscosity, thickening, swelling, foaming, and gelation are influenced by the
protein-water interactions. For example, during the dough and custard preparation,
swelling is an important functional characteristic. In these products, water absorp-
tion takes place but not solubilization, due to the insufficient water. Hence, the
swelling occurs which imparts to the body, viscosity, and thickening to the product
(Hermansson 1972).
Swelling property is largely dependent on other conditions. Whey protein exhib-
its less solubility because of its globular nature. On denaturation, whey proteins
absorb water as alike as casein protein, i.e., 2 g/g of protein (Walstra et al. 1999).
Water retention is significant for the functionality of protein as it influences the
170 R. Mehla et al.
sensory, color, and textural properties of the finished product. Techno-functional

and physicochemical properties of the protein-rich foods are greatly affected by the
water binding capacity and water content (Zayas 1997f). The interaction of the pro-
tein with solvent in the food system can be well understood by using absorption
isotherm, which indicates the content of water sorbed by the protein (g H2O/g pro-
tein) as a function of the relative water activity. Moisture sorption isotherm not
merely indicates the water binding capacity but also shows the stability during stor-
age and the optimum drying condition of the protein-enriched foods. Water binding
is also influenced by the protein molecule’s composition and conformation (Kumari
and Choudhary 2019).
8.5 Techno-functional Applications of Whey-Based Foods
Whey-based products have abundant applications in the foods and dairy industries.
The interest of the consumer is continuously increasing not merely due to nutri-
tional significance but also due to its functional attributes which they add to the food
products. Whey protein ingredients have been used in various foods such as fish and
meat products, bakery products, confectionery, dairy products, fruits and beverages,
medicinal and nutritional products, and also in the foods manufactured for the spe-
cial needs including dietetic foods and infant formulas (Bansal and Bhandari 2016).
The techno-functional properties of whey-based products are influenced by vari-
ous properties such as solubility over broad pH range, protein-water interaction,
foaming, emulsification, and gelation (Kilara and Vaghela 2004). Generally, WPC
35 is used as skim milk powder replacer, as its composition is similar to the skim
milk powder. WPC 35 is also incorporated as an ingredient in yogurt, bakery prod-
ucts, and fat mimetics as a stabilizer and emulsifier. Different grades of WPC such
as WPC 50, WPC 65, and WPC 80 are generally utilized in health drinks, bakery
Table 8.2 Application of whey protein ingredients and their functionality

Food commodity Application Functionality
Baked products Bread, muffins, cakes Emulsifier, nutritional, and egg
replacer
Dairy-based whey Cheese, yogurt Cohesiveness, consistency and
products nutritional
Beverages Fruit juices, soft drinks, and frozen Nutritional
beverages
Frozen products Frozen juice, ice cream, frozen Whipping, emulsifier, skim milk
dessert coatings solid replacement
Confectionery Sponge cakes and meringues Foam stability, emulsifier
Pasta preparations Imitation pasta, macaroni Nutritional and freeze-thaw
stabilizer
Infant dairy foods Term formula, pre-term formula, Nutritional
and follow-up formula
Dietetic foods Clinical foods, diets for elderly Nutritional
Kilara and Vaghela (2004)
products, protein-fortified beverages, meat soups, etc. Defatted WPC (80–85% pro-
tein) has also been used as in the toppings, whipped products, and ice creams. The
functional properties of the whey-based ingredients are summarized in Table 8.2.
Whey proteins have a tendency to form gel, and because of this gelling property
of the whey protein concentrates, they are being used in the low-fat meats (Johnson
2000). With the use of high gelling WPCs with gum and starches in the low-fat pork,
sausage develops similar texture as that of the full-fat control product. The cold gell-
ing WPCs are also available at commercial level and are being used in the mayon-
naise and salad dressings.
Protein-solvent interaction is an important property of the viscous foods like
soups, processed cheese, and dough. This property is helpful in the preparations of
low-fat and fat-free salad dressings. It improves the food texture and also decreases
the cost of the food by replacement of oil with water. In fat chopped meat and other
seafood, it is found to be significant.
Owing to better emulsifying property, WPC provides a creamy and smooth tex-
ture. Efficient diffusion with good emulsifying property is helpful in the low-fat
food applications. Even in an acidic medium such as salad dressing, emulsion sta-
bility has maintained. Substitution of egg yolk by WPC partially or fully gives alike
product quality at a lower cost. In low-fat sauces and soups, whey protein provides
a fat-like lubricity with a better mouthfeel. It also develops opacity which further
gives an excellent visual appeal to the foods (Kumari and Choudhary 2019). During
the processing of meat, proteins and other ingredients are added into the meat pieces
by vacuum or injection tumbling. By binding with fat, water, and meat pieces, whey
protein minimizes the cook shrinkage and enhances the yield.
8.6 Source of Whey-Derived Bioactive Components
Whey protein acts as a good source of bioactive peptides with various physiological
activities which imparts to the health benefits. WPI is a rich source of biologically
existing cysteine and methionine which hinder the growth of colon tumors. Due to
the bioactive functionality of whey ingredients, the whey protein-enriched food
products like flavored milk, ice cream, and pasta are preferred to be a healthy
approach for the diet. Whey protein-derived bioactive peptides have various health
benefits which impact the major organs of the human body such as cardiovascular,
immune, nervous, and digestive organs (Pihlanto and Korhonen 2003; Krissansen
2007). A study was conducted earlier which indicated that lactoferrin has marvelous
health effects for human and animals. It protects the mammary gland and gastroin-
testinal tract against infection and offers better nutrition via augmented bioavail-
ability of the amino acids and iron which has been known for the promotion of the
cell differentiation, division, and cell growth of the intestinal mucosal membrane
(Pihlanto and Korhonen 2003). Lactoferrin has been reported to be involved in the
iron metabolism and to have antimicrobial, antibacterial, antiviral, and anticancer-
ous properties (Vogel 2012). Infant formula containing lactoferrin is beneficial in
retarding the growth of coliform bacteria by the antimicrobial action (Johnston et al.
172 R. Mehla et al.
2015). Partial hydrolysis of lactoferrin leads to the formation of lactoferricin which

has antitumor activity (Gifford et al. 2005).
8.7 unctional Properties of Whey Protein-Derived

F
Bioactive Peptides
The detailed descriptions of various biofunctionalities of whey protein released bio-

active peptides are described below.
8.7.1 Antioxidant Activity
The antioxidant potential is the ability to slow down the oxidative stress and to pre-
vent spoilage in food products. Whey protein has the ability to offer hydrogen ions
to the active free radical and works as an antioxidant. Free sulfhydryl group from
cysteine amino acid residue of whey protein also combines with the free radicals
and reduces the lipid autoxidation (Qian et al. 2008). Bioactive peptides exhibit
their antioxidant activity via inhibiting the lipid peroxidation by scavenging free
radicals and metal ion chelation (Qian et al. 2008; Moure et al. 2006; Rajapakse
et al. 2005).
Composition, structure, and hydrophobicity influence the antioxidant potential
of bioactive peptides (Chen et al. 1998). Wang and De Mejia (2005) reported that
several amino acids like Tyr, Met, Trp, Lys, Cys, and His also exhibit antioxidant
activity. Amino acids with the aromatic group have the nature to donate the proton
to the free radicals (Rajapakse et al. 2005). Some of the bioactive peptides obtained
from β-lg A showed better antioxidant activity than the butylated hydroxyanisole
(BHA) like synthetic antioxidants (Hernández-Ledesma et al. 2005).
8.7.2 ACE Inhibition Property
Whey protein-based bioactive peptides exhibit ACE inhibition activity via control-
ling the hypertension or elevated blood pressure (Masuda et al. 1996; Mullally et al.
1997a; Belem et al. 1999; Pihlanto-Leppälä et al. 2000; Walsh et al. 2004).
Angiotensin-I-converting enzyme maintains the blood pressure by converting
angiotensin I to angiotensin II.
Removal of two amino acids from angiotensin I results in the synthesis of an
octapeptide called angiotensin II. Angiotensin II elevates the blood pressure, so
reducing the angiotensin II production can reduce the blood pressure. β-lg f(19–25,
142–148) are considered to be ACE inhibitory peptides (Hernández-Ledesma et al.
2007; Mullally et al. 1997a, b). α-Lactalbumin and its derived bioactive peptides
(fraction f50–52 and α-lactorphin) have also been reported to exhibit ACE inhibi-
tory activity (Nurminen et al. 2000; Pihlanto-Leppälä et al. 2000).
8.7.3 Anticarcinogenic Property
Whey protein-derived bioactive peptides protect against the mammary and colon
tumors (Hakkak et al. 2000; Rowlands et al. 2001). Antitumor effects of whey pro-
teins and their derivatives have also been reported by Parodi 1998 and Bounous
2000. WPI hydrolysates prevent the human origin prostate epithelial cell line from
oxidative stress which might induce cell death (Kent et al. 2003). β-lg and its derived
peptides can attach to the heterocyclic amines and thus act as an anticarcinogen
(Yoshida et al. 1991).
These peptides have also been reported to reduce the genesis of putative cancer
precursors in the wall of the hindgut (Mcintosh et al. 1998). α-Lactalbumin and its
derived peptides in the mammalian intestinal cell lines inhibit the cell division
(Ganjam et al. 1997). Lactoferrin shows the strong anticarcinogenic property during
tumor treatment (Masuda et al. 2000; Gill and Cross 2000).
8.7.4 Antimicrobial and Antiviral Activities
Peptides hydrolyzed from β-lactoglobulin have efficient antimicrobial property

against the different food-derived pathogens like Listeria monocytogenes,
Salmonella spp., Staphylococcus aureus, and Escherichia coli O157:H7 (Pellegrini
et al. 2003). β-Lactoglobulin fragments f(92–100, 78–83, 25–40, and 15–20) have
antibacterial property against the Gram-positive bacteria. Enzymes like pepsin,
Alcalase, and trypsin hydrolyzed β-lactoglobulin also possess antimicrobial effects
(Pihlanto-Leppälä et al. 1999; El-Zahar et al. 2004). Though native α-lactalbumin
did not exhibit antimicrobial activity against Gram-positive and Gram-negative bac-
teria (Pellegrini et al. 1999), the chymotrypsin and trypsin hydrolyzed α-lactalbumin
peptides have shown antibacterial properties. α-Lactalbumin and β-lactoglobulin
also have antiviral property (Chatterton et al. 2006; Madureira et al. 2007).
3-Hydroxyphthaloyl-β-lg, a chemically distorted β-lg, had been seen to act against
human immunodeficiency virus type 1 (Berkhout et al. 1997; Neurath et al. 1997;
Oevermann et al. 2003).
The mechanism behind the antimicrobial action of lactoferrin is to chelate the
available iron in the medium and make it unavailable for the microbial growth and
proliferation. The antimicrobial activity of lactoferrin is greatly influenced by the
concentration, extent of saturation of iron, and availability of minerals in the
medium (Payne et al. 1990). Lactoferrin has the capability to bind with lipid A lipo-
polysaccharides which can rupture the outer membrane of Gram-negative bacteria
(Appelmelk et al. 1994), leading to the deformation of the structure and destruction
of the membrane integrity.
Lactoferrin has inhibitory properties against both Gram-negative and Gram-
positive bacteria like E. coli, Shigella dysenteriae, L. monocytogenes, Bacillus sub-
tilis, Salmonella typhimurium, and Bacillus stearothermophilus (Batish et al. 1988;
Payne et al. 1990; Saito et al. 1991). Antiviral activity of the lactoferrin has been
reported by Ko and Kwak (2009). It has been seen that it attaches to the eukaryotic
174 R. Mehla et al.
cell membrane and prevents the viral substance permeation into the cell membrane
and thus inhibits the initial stage of viral infection.
8.7.5 Whey Protein usage as biodegradable Packaging Material
Whey protein-based coatings and films can be useful to reduce the amount of syn-
thetic packaging material that is essential for food preservation and can also improve
the recycling of the package. Edible films should be insipid, flexible, and tasteless
and should differ in the opacity from transparent to translucent depending on the
type of protein source, formulation, and composition (Ramos et al. 2012). WPC and
WPI are being utilized as renewable, eco-friendly packaging and plasticized edible
film (McHugh et al. 1994; Ramos et al. 2012). The solubility of edible films made
from whey proteins can be decreased by either acidifying it to the isoelectric pH or
by cross-linking of protein through the enzymatic action (Mulvihill and Ennis
2003). Whey protein can be used as a coating material in confectioneries, vegeta-
bles, fruits, and nuts. Whey protein-based coatings serve as a better improver of
wettability and water absorption and oxygen barrier properties (Dangaran and
Krochta 2009).
8.7.6 Pharmaceutical Applications
The pharmaceutical formulation requires a high specification of biodegradability,

renewability, edibility, and absorbability. WPI meets all the qualities required for
pharmaceutical formulation (Caillard et al. 2011). Currently, succinylated
β-lactoglobulin has been considered to be very potent for the controlled protection
and release of the active compounds like probiotic bacteria and riboflavin (Poulin
et al. 2011; Caillard et al. 2011). WPI has good flowability and compatibility that
are desirable characteristics of the tabletability. Tabletability is dependent on the
particle size of the whey proteins. Smaller particle size has higher compactibility.
Regular shaped whey protein particles produce the highest mechanical strength tab-
lets (Caillard et al. 2012). Lactose present in whey is also used in pharmaceutical
products (Siso 1996).
8.7.7 Encapsulation Capability of Whey Proteins
Encapsulation strategy is being currently used in the food and pharmaceutical sec-
tors. It helps in the restricted release of useful ingredient, masking of typical flavor,
and consistent distribution in the finished product. Major whey protein fractions like
α-lactalbumin, β-lactoglobulin, and bovine serum albumin have better surface-
active characteristics. Therefore, the whey protein fractions can be utilized as a
good quality emulsifier and coating materials for the encapsulation. WPC and WPI
encapsulation with the milk fat has been evaluated thoroughly (Young et al. 1993a).
Milk fat can be shielded from the surrounding environment via encapsulation strat-
egy. The incorporation of other carbohydrate material in the coating material of
WPC can enhance their encapsulation competence (Young et al. 1993b). The encap-
sulation of β-carotene in the whey protein isolate stabilized oil-water emulsion was
prepared and studied for their stability by Jiang et al. (2017). Zhou et al. (2009)
reported that the whey protein can also be used as nanoparticles.
8.8 Conclusions
Whey protein, the by-product of cheese industry, has been known for its various
functionalities as solubility, gelation, viscosity, emulsifying property, foaming
property, fat and oil binding property, and water holding capacity. Due to its func-
tional properties, it is widely used in various food products. It serves as a good
source of bioactive components. Whey proteins exhibit various biofunctional prop-
erties like antioxidant activity, ACE inhibitory activity, anticarcinogenic property,
and antimicrobial and antiviral activity. Apart from their techno-functional and bio-
functional properties, whey proteins are also used as biodegradable packaging
material and encapsulating material and are applicable in the pharmaceutical
industry.
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Overcoming the Quality Challenges
Across the Supply Chain 9
C. H. Aysha and S. Athira
Contents
9.2 H azards 183
9.2.1 Physical Hazards 183
9.2.2 Chemical Hazards 183
9.2.3 Biological Hazards 184
9.3 Sanitary and Hygiene Factors Influencing Milk Hazards 185
9.3.1 Environmental Hygiene 186
9.3.2 Personal Hygiene 186
9.3.3 Equipment Hygiene 187
9.4 Dairy Supply Chain 187
9.4.1 Quality Challenges in the Supply Chain 188
9.5 Strategies to Overcome the Quality Challenges 188
9.5.1 Traceability 188
9.5.2 Blockchain Technology 193
9.5.3 Temperature Management 193
9.6 Conclusions 194
References 195
Abstract
Milk, owing to its rich nutrient contents, is a highly perishable product. Being a
high-value product, attempts to increase its shelf life have always caught the
interest of researchers. This chapter briefly discusses various hazards, including
physical, chemical, and biological ones; factors affecting hazards in milk includ-
ing sanitary and hygiene factors like personnel hygiene and hygiene of tools; and
dairy supply chain, its quality challenges, and the strategies to overcome these
challenges. Traceability, which is the ability to track the movement of food from
C. H. Aysha (*) · S. Athira

College of Food Technology, Kerala Veterinary and Animal Sciences University,

https://doi.org/10.1007/978-981-15-2608-4_9
182 C. H. Aysha and S. Athira
its production, processing, and distribution, is explained in detail, including real-

time methods and recent innovations like blockchain technology.
Keywords
Milk products · Supply chain · Quality challenge · Dairy industry · Hygiene ·
Quality
9.1 Introduction
A substance, at any point before, during, or after processing, is referred to as “per-

ishable” if any of the following takes place during the considered period: (1) clear
worsening of its physical status by spoilage, decay, or depletion; (2) reduction in its
perceived customer value; and (3) a possibility of deterioration in its functionality.
Milk is a highly nutritious food and offers an excellent medium for the multiplica-
tion of all types of microorganisms making it highly perishable. In raw milk, there
is a high risk of spoilage as well as pathogenic microbes that can cause severe dis-
eases in consumers. To avoid such problems and to increase the shelf life of milk
from a few days to many months, various processing methods can be applied. These
diverse methods can destroy the presence of spoilage and pathogenic microbes
entering into the milk, thereby causing a severe reduction in the occurrence of food-
borne illness through milk and milk products. One of them is to adopt the adequate
cooling method to increase the keeping quality of milk for several days by retaining
the quality of milk. Similarly, heat treatment can also improve the quality and shelf
life of milk and milk products. When it comes to heat treatment, pasteurization is
considered as one of the oldest and best methods to treat milk and milk products.
Pasteurization is a heat treatment process intended for improving the shelf life of
milk that reduces the number of pathogenic microorganisms to a safe level, i.e., to a
level that does not cause any health hazard to the consumer. Further, milk can be
converted to fat-rich dairy products like butter, ghee, and butter oil and can have a
greater shelf life and higher market price. Raw materials, intermediate goods, and
final products are usually highly perishable in case of all dairy-related industries,
whether it is a small scale or even large processing industry. This factor compels the
setting up of a specific supply chain process from production to distribution. The
definition of perishability is rather vague. According to Wee (1993), “perishability
includes decay, damage, spoilage, evaporation, obsolescence, pilferage, loss of util-
ity or loss of marginal value of a commodity that results in decreasing usefulness
from the original one.”
9 Overcoming the Quality Challenges Across the Supply Chain 183
9.2 Hazards
Hazard might be anything which could adversely affect the health of consumers.
When food is exposed to any of the hazardous agents, it results in the contamination
of food and thereby causes a food safety hazard. The major food hazards can be
classified as physical, chemical, and biological. The entry of hazards into the milk
supply chain can take place anywhere whether it is from production, through trans-
portation, handling, processing, packaging, storage, or till end use.
9.2.1 Physical Hazards
Incidence of several foreign matters in milk contributes to the majority of physical

hazards to the dairy consumers. Foreign particles may include soil particles, insects,
glass or tiny metals, hair, dung particles, stones, and so on. Their existence might be
a risk of injuring one’s mouth or even deeper areas of the digestive system if physi-
cal hazards like a piece of glass are consumed by chance. So preventive measures
should be taken by the manufacturers for minimizing the threat of contamination of
milk with these physical hazards. As glass is an extensively used packaging material
for milk-based beverages, care must be taken so as to diminish the risk of its break-
age, thereby protecting the consumer health. Like glass, metal, wood, or stone can
also cause similar damages (Soman and Raman 2016).
Damages to teeth and human internal body organs may occur sometimes because
of the incidence of hard objects in milk and may lead to choking particularly among
children when they swallow it. Incidence of such hard sharp-edged objects and seri-
ousness of these hazards is more in raw milk as compared to processed milk. The
physical hazards can be easily detected and the source can be easily traced. In case
of organized or processed dairies, milk is passed through metal detectors and clari-
fiers, so the incidence of such metallic objects is lesser or absent. Detection of phys-
ical foreign objects in milk pack is determined by the installation of X-ray machines
at the end of the processing line. Such types of measures are not practical in case of
village level, small-scale unpacked milk marketing channels, where farmers handle
the sales themselves (Molins et al. 2001).
9.2.2 Chemical Hazards
The occurrence of chemical hazards, especially environment-borne chemical haz-

ards, is very difficult to avoid and control. The practice of using chemicals in dairy
farms as well as processing plants leads to the increased incidence of chemical
hazards in milk. Unhygienic and falsified practices done in the farm result in the
increased risk of chemical hazards in the milk. Potential chemical hazards in
milk and milk products include residues of chemicals used in animal production
practices (veterinary drugs, hormones, pesticides, and mycotoxins), faulty practices
like adulteration (urea, starch, neutralizer, etc.), environmental contaminants (heavy
metals, dioxins, polychlorinated biphenyls, radionuclides, etc.), chemicals used in

processing units (detergents and disinfectants), compounds formed during process-
ing (neoformed contaminants), and migrants from food contact materials.
The adulterants (detergents, neutralizers, urea, etc.) are extensively added to
milk for masking the inferior quality milk that ends up with the high-test values for
chemical substances that adversely affect all groups of the society, especially chil-
dren. Liver, kidney, and intestinal tract may get affected due to the prolonged intake
of milk with minute quantities of urea or other detergents. Various acute and life-
long diseases, ranging from diarrheal diseases to different forms of cancer may arise
due to the use of foods that are unfit for human consumption. The Indian Council of
Medical Research (ICMR) noted that the chemical hazards can induce severe health
problems, including food poisoning and other gastrointestinal problems due to
long-term consumption of detergents in milk. Proteins and body tissues can be dam-
aged as a result of intake of higher amount of alkali and several chemicals, which
leads to heart disorders, cancer, or even death.
Poor agricultural, veterinary, and industrial practices lead to an unacceptable
level of chemical hazards (residues) which will remain as hidden threats in the food
chain. It is very difficult or impossible to make milk chemical-hazard-free by vari-
ous processing methods. Chemical hazards already present in milk are usually unaf-
fected by further processing. Complete inactivation of these chemicals is not
attained by processing like filtration or thermal processing as in case of physical or
biological hazards. Chemical content in food products can be determined by tech-
niques like temperature program micro sensors or nano-sensors. These techniques
are used to ascertain chemical hazards in food and in devising the necessary preven-
tive action.
9.2.3 Biological Hazards
The biological hazards mainly include the pathogenic and spoilage-causing micro-
organisms which gain entry into milk mainly from milking animals and personnel
and through the environment. The poor food hygienic practices can be correlated
with the increased chances of biological hazards. Prevalence of food poisoning
issues in milk and milk products is the result of an increased incidence of patho-
genic microorganisms in milk. As the exposure to spoilage and pathogenic micro-
bial cells increases, the human body gets sensitized and becomes more prone to
diseases. The nutrients in the medium, amount of water present, level of oxygen,
acidity of the medium, temperature, etc., are the different factors affecting the
growth of microorganisms (Boor 1997). Bacterial types commonly associated with
milk are Aeromonas hydrophila, Bacillus anthracis, Bacillus cereus, Brucella spp.,
Campylobacter coli, Campylobacter jejuni, Clostridium botulinum, Clostridium
perfringens, Coxiella burnetii, E. coli, Cronobacter sakazakii, Enterococcus faeca-
lis, Enterococcus faecium, Legionella spp., Listeria monocytogenes, Mycobacterium
bovis, Mycobacterium tuberculosis, Salmonella spp., Shigella spp., Staphylococcus
aureus, Streptococcus pyogenes, Streptococcus agalactiae, Vibrio cholerae, Vibrio
parahaemolyticus, Vibrio vulnificus, Yersinia enterocolitica, Yersinia pseudotuber-

culosis, etc. (Hayes and Boor 2001). Not only the microorganisms but also the tox-
ins produced by some of the microbes can cause serious threats to humans.
Enterotoxins are certain toxins produced by pathogens which can cause food poi-
soning. These toxins are known to have high heat stability and may not get destroyed
during normal heat treatments, and these cause gastroenteritis in humans. Raw milk
or milk that has undergone partial pasteurization will be having higher levels of
somatic cells (Oliver et al. 2005).
The microbiological hazards are highly product specific. Various intrinsic and
extrinsic factors affect the quality and type of raw materials used in milk processing.
Raw milk and pasteurized milk are better media for the transfer of microorganisms
to humans as compared to powdered milk. Mesophilic and thermophilic organisms
are usually reported in milk and milk products (Walstra et al. 2006). The processing
steps and the storage conditions also play a crucial role in the prevalence of biologi-
cal hazards. The chances of contamination and multiplication of microorganism in
milk are rather more in tropical milk farming environment, and the microbial num-
ber can be reduced only by any suitable heat treatment techniques and good manu-
facturing practices (GMP). Poor hygienic condition is the major cause of higher
microbial counts, and there are increased probabilities of spoilage-causing microor-
ganisms and pathogens which can cause serious risk to consumers who consume
raw milk. Organized dairies give better microbial quality as compared to milk from
unorganized dairies. Poor designing of the dairy and systems leads to high bacterial
count (Iyer et al. 2010). The initial load of microorganisms can be minimized or
reduced by adopting and maintaining good hygienic practices (GHP) from the farm
to distribution stage, which could further prevent the increment in high microbial
populations (Anand et al. 2006).
9.3 Sanitary and Hygiene Factors Influencing Milk Hazards
The primary requirement for the consumers of milk and dairy products is their qual-
ity and safety. Safety of milk should be ensured at every step from the production
till final consumption. In case of presence of any physical, chemical, or microbio-
logical contamination in milk, it should be considered as unfit or unsafe for con-
sumption and should be withdrawn from the supply chain. Owing to the best
nutritional quality, majority of the microorganisms survive in milk and multiply
further; some pathogenic organisms are capable of producing hazardous toxins. The
major concern in the dairy industry is the microbial hazard as it can cause spoilage
and food poisoning and even produces heat-stable toxins. Consumption of unpro-
cessed milk will be a serious issue for public concern as it may cause severe health
problems. Hygiene, with respect to the milch animals, personnel, equipment, and
environment of the handling areas are some major factors that determine the milk
quality and safety.
9.3.1 Environmental Hygiene
As chances of contamination of raw milk are very high, it is more capable to carry
pathogens and transmit various diseases to the consumers as compared to processed
milk. Milk shall be stored and handled carefully as soon as it is secreted out from
the udder to ensure its hygiene and safety until consumption. The housing of ani-
mals and feeding procedures of cattle should be in good hygienic condition as they
affect the chemical and microbiological quality of market milk (Torkar and Teger
2008). Usage of unsafe water for pre-rinsing and washing of milking machine adds
microbial load as well as pathogenic microorganisms in unprocessed milk (Bramley
and McKinnon 1990). In rural areas, lack of knowledge regarding cleaning and
sanitizing agents for containers used during milk collection and transportation
makes the milk unsafe and low quality. Proper cleaning and sanitizing procedure
should be followed in case of cans or utensils used for transporting the milk.
Direct and indirect contamination should be avoided to maintain the quality and
hygiene. The final quality of milk depends on how consumer maintains the quality
of purchased milk by storing at the required temperature and heat treatment applied
before consumption. (Valeeva et al. 2005). The high storage temperature of fresh
milk without proper refrigeration will surely increase the microbial loads and results
in poor quality of milk. The most common processing technique employed to reduce
the biological hazard is thermal processing, like pasteurization or sterilization.
Thermal processing reduces the pathogenic organisms at the earlier stage before the
production of toxins, but the chemical hazards have little effect by thermal process-
ing. As raw milk is sold without any thermal processing, the chance of occurrence
of biological hazard is even more as compared to processed milk.
9.3.2 Personal Hygiene
Good hygienic practices are essential in clean milk production. Cleanliness or

hygienic condition of milking personnel is a major factor that affects the microbial
quality of raw milk. In order to reduce such quality deterioration of milk, the health
of the milking personnel should be ensured, i.e., they should be free from diseases
so as to reduce the risk of transmitting disease-causing microbes to milk. Information
regarding hygiene rules and regulations and standard operating procedures should
be given to those milking personnel who are working on dairy farm. Unhealthy
workers in the milking parlour are one of the most important sources of biological
hazards in milk (Lelieveld et al. 2003). In case of rural and marginal farmers, the
best method to improve the quality of milk is by creating awareness regarding the
adverse effects of various hazards. In case of the processing plant, the basic hygienic
practices shall be displayed to reinforce the practices in the daily routine of dairy
workers.
9.3.3 Equipment Hygiene
Proper sanitization of all the tools used for milk handling should be done in order to
avoid the occurrence of physical, chemical as well as biological hazards. Equipment
used for milk handling must be thoroughly sanitized before the next use. In case of
small-scale farmers, kettles and cans used for milk transportation should be ade-
quately cleaned and sanitized after each use. Disinfection of milking machine
should be done properly in case of automated milking process in large farms. In
case of a big processing plant, sanitary design plays a key role in maintaining the
equipment hygiene.
A proactive approach basically emphasizes hygiene requirements (environmen-
tal, personnel, equipment, etc.) for enabling control at “source” to be set at the
production, processing, and distribution levels to make sure that product will neither
harm the consumer nor will it get spoiled during the intended shelf life period. The
FAO/WHO has recognized Hazard Analysis Critical Control Point (HACCP) con-
cept as an effective way to control all types of hazards.
9.4 Dairy Supply Chain
The dairy supply chain starts from the producers, i.e., farmers. In developing South
Asian countries like India, milk is collected at the collection center or Village Level
Cooperative Societies (VLC). The payment is done based on the fat and solids-not-
fat (SNF) percentage of milk poured. Milk is collected and stored at Milk Chilling
Centers (MCC) in order to avoid spoilage. Milk is transported from several VLCs to
the MCC using refrigerated trucks. At MCC, chilling of milk is done below 4 °C so
that microorganisms will not be able to grow further, and so the safety of milk is
ensured until the milk reaches the processing plant, which may be distant from the
collection areas, for further treatments.
The chance of contamination of milk is higher in each and every stage of its pro-
duction. The practice of adding low-cost adulterants like glucose, sugar, starch, and
skim milk powder to increase the SNF content is extensively seen in several areas to
get more profit from the milk. Chemicals like urea are added to milk to enhance its
shelf life, which is very harmful to the human body. Implementation of an effective
traceability system should be adopted so as to determine all ways of malpractice in
various stages of milk production. In every production point either in fluid milk or
products, there are some control measure points in the production process to avoid
such contamination. It is recommended to follow a system (HACCP) for preventing
the entry of hazards in the production.
After production and packing, the products get dispatched from the production
unit. Products are directly supplied to various agents/distributors scattered across
the city. So cold chain management throughout the entire chain until the products
reach the consumers is an essential strategy for effective delivery of milk and milk
products from farm to table (van Asselt et al. 2017).
9.4.1 Quality Challenges in the Supply Chain
Inadvertent introduction of hazardous compounds into milk and milk products dur-
ing various phases of operation including production, processing, packaging, stor-
age, and supply chain poses a serious health risk to the consumer. Risk factors can
threaten product quality and safety and thus affect customers’ health (Marucheck
et al. 2011). Frequency of foodborne diseases is mounting globally, and it can lead
to disruption of the international food trade (FAO 2003). The WHO (2002) stated
that food safety and quality is not only a foremost factor that contributes to the
health and well-being of the general population but also has tremendous economic
consequences for the community. Management of these kinds of perishable foods is
very difficult as they are having very low keeping quality (Bowman et al. 2009).
Globalization has opened markets beyond international borders forcing strict imple-
mentation of food safety and quality standards by different countries to safeguard
the health and safety of the consumers (Aung and Chang 2014a).
9.5 Strategies to Overcome the Quality Challenges
Globalization of food trade has resulted in long transit times for the food to reach the
end of the supply chain, posing a major challenge for ensuring the safety and quality
of the foodstuff (Aung and Chang 2014a). As a result of increased incidents of fraud-
ulent practices and the occurrence of foodborne illness, consumers are now more
concern over the safety and quality of food. So there are some recent approaches to
overcome the quality challenges and for quality management of food across the sup-
ply chain.
9.5.1 Traceability
Globalization of the food industry has raised a number of safety and quality concerns
including intentional adulteration of food for increasing profits. However, this has
also increased public awareness of food safety and quality, leading to the emergence
of traceability (Bertolini et al. 2006). Based on ISO 9000 (2005) standards, trace-
ability is defined as “the ability to trace the history, application or location of that
which is under consideration” (Hoyle 2001). Details regarding the origin of materials
and parts, its processing history and the distribution, and location of the product after
delivery may be collected from this system. The Codex Alimentarius Commission
(Codex Alimentarius Commission 2005) defined traceability as “the ability to follow
the movement of a food through specified stage(s) of production, processing and
distribution.”
According to Bosona and Gebresenbet (2013), food traceability is “a part of
logistics management that capture, store and transmit adequate information about a
food, feed, food-producing animal or substance at all stages in the food supply chain
so that the product can be checked for safety and quality control, traced upward, and
tracked downward at any time.” An efficient traceability system should be charac-

terized by the amount of collected information (breadth), the extensiveness (for-
ward or backward) of the system to track the relevant information (depth), and the
degree of assurance to pinpoint a particular movement of a food product (precision)
to be able to balance cost and benefits (Golan et al. 2004).
The traceability systems can be divided into two types based on the functionality
logistics into traceability, which considers food as a commodity and traces only the
flow of the product across the supply chain, and qualitative traceability, which gives
information regarding the quality and safety of the product. In general, traceability
can be classified based on the position in the supply chain from which the informa-
tion is collected (Aung and Chang 2014a). The information from the supply chain
can be retrieved in either forward or backward direction. Forward traceability is
useful to locate the products, whereas backward traceability helps to find the origin
and characteristics of a product. Traceability has given various dimensions by sev-
eral researchers as given in Table 9.1.
Traceability aids in managing the supply chain, ensuring even minute deviations in
quality, early detection and faster response, operational status, reducing cost distribu-
tion system, and simplifying traceback for food safety and quality (Golan et al. 2004;
Aung and Chang 2014a). The motivating factors enforce traceability as a means to get
response for the questions like “who (i.e., actor/product), what (i.e., actor/product’s
information), when (i.e., time), where (i.e., location) and why (i.e. cause/reasons)”
with regard to food safety, quality, and visibility (Aung and Chang 2014a).
Table 9.1 Classification of traceability

S. no. Classification of traceability Reference
1. Two types Moe (1998)
• Internal traceability
• Chain traceability
2. Six types—Focusing on agricultural and food Opara (2003)
supply chain
• Product traceability
• Process traceability
• Genetic traceability
• Input traceability
• Disease and pest traceability
• Measurement traceability
3. Three types Perez-Aloe et al. (2007)
• Back traceability or suppliers’ traceability
• Internal traceability or process traceability
• Forward traceability or client traceability
4. Two types Aung and Chang (2014a)
• Forward traceability
• Backward traceability
9.5.1.1 The Association Between Traceability, Quality, and Safety

Implementation of product-tracing systems is an integral part of food safety and
quality control. A well-maintained traceability system is extremely helpful in pre-
cisely and efficiently identifying the source, extent of safety, or quality control
issues and their timely resolution (Golan et al. 2004). Between quality and safety,
safety is more often implicated by traceability. Traceability may certainly boost the
self-belief and confidence of consumers through quality and safety assessments
(van Rijswijk and Frewer 2006). Traceability is an essential component for estab-
lishing a total quality management (TQM) system by improving data collection,
production flow control, and quality assurance.
9.5.1.2 Food Contamination and Traceability

Food may be accidentally or intentionally contaminated by physical, chemical, or
microbiological hazards. Genetically modified organisms (GMOs) and radioactive
substances are some of the emerging concern. Traceability is a strategy used to
identify the source(s), production, and distribution chain of a product for determin-
ing the critical point where contamination could have occurred which leads to food
outbreaks and incidents. This facilitates the food recalling from the market to pre-
vent further illness and trace out the hazardous practices which ultimately leads to
the illness. There are some challenges in the tracebacks of fresh produce which
include difficulty in case of multiple sources of product and improper labeling and
distribution records (Aung and Chang 2014a).
9.5.1.3 Real-Time Traceability

Food supply chain management is a difficult job as there is variation in the quality of
raw materials and products occur over time; continuous monitoring is required from
the producer till the end users (Apaiah et al. 2005). The efficiency of a traceability
system mainly depends on the ability to track and trace each individual product
through continuous monitoring right from the primary production to its final disposal
by the consumer (Folinas et al. 2006). Perishable foods such as meat, fish, milk,
fruits, and vegetables pass through different hands several times before they reach
the consumer, and the quality of these foods is dependent on how they are handled.
Both logistics and qualitative traceability are mandatory for highly perishable foods
which need cold chain. The ability to collect information regarding sensitive environ-
mental factors like temperature, humidity, light, etc., for providing the real-time sta-
tus of the product quality provides substantial benefits to the food industry by
reducing the food waste and spoilage. Transparency is the highlight of real-time
traceability as the information regarding the product and process is made available to
all the stakeholders (Aung and Chang 2014a).
9.5.1.4 Traceability in the Dairy Industry

The importance of traceability in the dairy supply chain and its conceptual framework
is elaborated by Behnke and Janssen (2019). They mentioned precisely about the
Food Safety Information System which involves the application of various technolo-
gies like Internet and Web, Wireless Identification and Sensor, Location-based, and
Food safety Inernal ,

Food Safety
and quality External
Information
assurance and Chain
System
system Traceability
Fig. 9.1 Contributors to food traceability framework
Fig. 9.2 Drivers for the

analysis of transparency,
traceability, and Global trade
information flow (Adapted
from Pant et al. 2015)
Emerging
Technological
International
advancements
Standards
Complex
Prouct
consumer
characteristics
behaviours
Information and Communication technologies to facilitate internal and external trace-

ability. Food safety and quality assurance system like the good practices (GHP, GMP),
ISO standards, HACCP, and TQM facilitates chain traceability as illustrated in
Fig. 9.1.
In this framework, the entire supply chain should have both internal and external
traceability in order to achieve the whole traceability. The quality management sys-
tems should follow the current regulations enforced in the entire steps of supply
chain. Modern technology advancements should be used in the supply chain opera-
tions to enhance performance in addition to serving as a medium for easy access to
the food traceability information systems. The major drivers for the analysis of
transparency, traceability, and information flow adapted to the dairy supply chain
have been illustrated in Fig. 9.2 (Pant et al. 2015).
The dairy supply chain is a multistage process encompassing production, process-
ing, and delivery. These stages or business processes are interconnected sequentially
or parallelly with priority relationships for the production of a variety of dairy prod-
ucts. The product quality at each step depends on the quality of the prior stages; thus,
the final product quality depends on the entire supply chain. Peculiarities of dairy
supply chain can be the intrinsic product attributes like perishability, variability of
milk constituents due to species difference, breed difference, feed composition,

genetic factors, etc.; seasonal variation in milk production (lean and flush season)
etc.; extrinsic product attributes like sources, history of milk, raw materials used in
production, storage, handling and transportation conditions, processing technolo-
gies, environmental impact, etc.; and process attributes like complex production
scheduling, mixing of raw milk collected from different source, etc. (Pant et al.
2015).
Each product and process of the dairy industry have their own peculiar character-
istics which ultimately affects the final quality of the products and their demand,
and these will be the basis for the formulation of transparency framework. The
consumers depend on the combined effects of quality, price, and safety and are the
ultimate judges on the product acceptance. Food traceability has been studied on the
basis of legal and regulatory aspects. A traceability system with alphanumeric code
and Radio Frequency Identification (RFID) for cheese has been formulated which
helps the consumers to get information regarding product history by inputting a
code via a website (Regattieri et al. 2007).
9.5.1.5 Technology for Traceability

Opara (2003) elaborated the requirements of technologies for traceability as hard-
ware and software. This mainly consists of measuring instruments, identification
mechanisms, etc., under hardware, while software is comprised of information sys-
tems which are used for product identification, data acquisition, analysis, storage
and transmission, as well as overall system integration. Many recent advances like
digitalization for product identification, storage, and display; biosensors for quality
and safety assessment; and satellite technologies such as geographic information
system (GIS), global positioning system (GPS), and remote sensing (RS) can be
effectively implemented in the traceability system.
The detailed information about each step in the supply chain like the identifica-
tion of location, origin (farm, cattle), and dairy plant where it is processed, pack-
aged, and stored is required in a product traceability system for the dairy industry
(Regattieri et al. 2007). Modern molecular biology techniques, which in general can
be broadly classified as (a) physicochemical (e.g., variation in radioactive isotope
content, spectroscopy, pyrolysis, or electronic nose) and (b) biological (e.g., DNA
chips, bacterial identification), can be used in traceability systems. The combination
of these techniques will be even more valuable for data generation in the traceability
system (Aung and Chang 2014a).
9.5.1.6 Challenges Ahead of Food Traceability

Although adopting and implementing traceability systems in all aspects of the sup-
ply chain, i.e., from production, storage, distribution, till the end users, are widely
recommended, there is an existence of significant roadblocks for its effective execu-
tion. The growing concerns on food safety and quality by the consumers are also a
big challenge for the effective implementation of traceability systems. The eco-
nomic viability of implementing traceability systems in the supply chains, espe-
cially by the small-scale producers from developing countries, has been a major
barrier. However, it has to be understood that the long-term benefits derived from
implementing such systems in high-risk, high-value food products outweigh the
initial costs. Another significant challenge is the lack of a standardized format for
the exchange of traceability information between different links of the supply chain.
Data from each link must be collected, stored, and distributed in a precise and effec-
tive manner. Current practices make use of conventional and manual recordings on
papers. This is cheaper, easy for distribution and long-term storage, but limited in
accuracy and searchability. Robust, fail-proof electronic alternatives should be used
to effectively collect and authenticate information for further storing or sharing it
through the chain when required (Aung and Chang 2014a). The implementation of
a digital platform for traceability can be initially expensive (Karippacheril et al.
2017), but it has its unique advantages including internal traceability (Senneset
et al. 2007), maintenance of balance between confidentiality and traceability of each
entity in the chain (Thakur and Donnelly 2010), and easy data exchange.
9.5.2 Blockchain Technology
Blockchain is a database in which all transactions are registered in each of the dif-
ferent nodes participating in that distributed ledger. The application of blockchain
technology (BCT) for supply chain management increases its efficacy by increasing
the transparency and traceability of food products (Tapscott and Tapscott 2016). It
permits the exchange of transactional data between two or more supply chain part-
ners. Implementation of BCT in supply chain depends on the extent of protection of
sensitive information as well as that of the ambiguity of the users (Tian 2017).
Specific information in the blockchain can be kept confidential to enable effective
traceability for preventing data breach (Epiphaniou et al. 2019). BCT helps to main-
tain the quality and traceability of a particular product, enables access to informa-
tion for multiple supply chain actors with a level of confidentiality, and allows a
particular level of interaction between them which is also suitable for the dairy food
traceability (Behnke and Janssen 2019).
9.5.3 Temperature Management
As milk and milk products are perishable food commodities, their temperatures
must be maintained at safe levels across the value chain to ensure safety, known as
a cold chain (Abad et al. 2009; Aung and Chang 2014b). There will be a significant
reduction in shelf life and quality when they are exposed to extreme hot or cold
temperatures. For assuring the quality of milk and milk products, vigilant and accu-
rate temperature management throughout the product flow right from the farm to
plate is essential (Jobling 2000). For properly tracing the product across the supply
chain, various advanced technologies like sensors, radio frequency identification
(RFID), and wireless networks may be utilized. According to Terreri (2009), intel-
ligent cold chain system is based on the advanced analytics and modelling of food
safety guidelines, which assist managers in handling complex situations very effi-
ciently and timely.
Aung and Chang (2014b) came up with a system called “Smart Cold Chain
Management (SCCM) system” in which every unit of stocked items would be
attached with RFID tag, and the cold chain would be connected with a wireless sen-
sor network (WSN) system. This system integrates the data retrieved from the RFID
system as well as from the WSNs. All the collected data will be saved in a central
database at Cold Data Center (CDC) to modulate and recover all relevant details
about the products throughout the cold chain using the Internet. This system enables
to track the information both via online and offline modes for ensuring the quality
and safety of the products (Fig. 9.3).
9.6 Conclusions
In any successful business, the customer is considered as a king, and so he/she

deserves the best quality. Ensuring the quality of a product like milk (or any other
milk product) is a great challenge in itself. Tracing the quality throughout the value
chain is the toughest challenge. Hence, several innovative methods and technologies
have been utilized for this purpose. Moreover, a lot of researches are going on across
the world to identify newer methods or to fine-tune the existing ones. This is open-
ing up numerous opportunities to researchers, business firms, and those that deal
with instruments, software, etc., and in turn, they are assuring the quality of prod-
ucts to the consumers they deserve.
Internet
• Production
• Tansport through
Smart Cold Chain refrigerated chain
Management • Storage
• Distribution
• Consumers
• Stock keeping Unit

Cold Data Center • RFID system
Fig. 9.3 Components of Smart Cold Chain Management (SCCM) system (Adapted from Aung
and Chang 2014b)
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Application of Research Methodologies
in Dairy Chemistry 10
Anuradha Kumari, Niraj Kumar Singh, Sonika Choudhary,
Dev Priya, and Arpna Sharma
Contents
10.2 C hromatography 199
10.2.1 Liquid-Liquid Chromatography (Partition Chromatography) 200
10.2.2 Liquid-Solid Chromatography (Adsorption Chromatography) 201
10.2.3 Gas-Liquid Chromatography 202
10.2.4 Ion Exchange Chromatography 203
10.2.5 Size Exclusion Chromatography 203
10.3 Spectroscopy Method 204
10.3.1 Source of Electromagnetic Radiation 205
10.3.2 UV-Visible Spectroscopy 205
10.3.3 Fluorescence Spectroscopy 206
10.3.4 Atomic Spectroscopy 207
10.3.5 Infrared Spectroscopy 208
A. Kumari (*)
N. K. Singh
College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences
University, Ludhiana, Punjab, India
S. Choudhary
National Dairy Development Board Dairy Services, New Delhi, India
D. Priya
DST Women Scientist-C, PFC-TIFAC, New Delhi, India
A. Sharma
Leibniz Institute of Farm Animal Biology (FBN), Dummerstorf, Germany

https://doi.org/10.1007/978-981-15-2608-4_10
198 A. Kumari et al.
10.4 Electrophoresis Method 208

10.4.1 Isoelectric Focusing 209
10.5 Supercritical Fluid Extraction 209
10.6 Immunoassay Method 210
10.7 Thermal Analysis 211
10.7.1 Differential Thermal Analysis 211
10.7.2 Differential Scanning Calorimetry 212
10.8 Conclusions 212
References 213
Abstract
Milk is a complex food consisting of numerous constituents, mainly water, fat,

protein, lactose, and minerals. All the constituents have different chemical struc-
tures and properties. There are various techniques available to identify the constitu-
ents both qualitatively and quantitatively. The common techniques used are
chromatography, spectroscopy, electrophoresis method, supercritical fluid extrac-
tion, immunoassay, and thermal analysis. Chromatography is a separation of mol-
ecules between two different phases: mobile phase and stationary phase. It is a fast,
accurate, and sensible method. Spectroscopy is the study of the interaction of the
light with the material. The movement of a charged molecule in an electric field is
electrophoresis. Supercritical fluid extraction is a separation technique used as a
preliminary step for the isolation. Antigen and antibody interaction is the basis of
immunoassay. Thermal analysis detects the physicochemical change due to expo-
sure of temperature. Chromatography, spectroscopy, and thermal analysis methods
can detect the major constituents of milk and milk products, i.e., proteins, fat,
vitamins, and lactose. Minerals are generally detected by atomic absorption spec-
troscopy. Electrophoresis differentiates the milk proteins, and immunoassay is
used to detect pesticide, antibiotics, and microbes including pathogens.
Keywords
Dairy · Chromatography · Spectroscopy · Electrophoresis · Thermal analysis
10.1 Introduction
Milk is a very complex food consisting of more than 100,000 various molecular
species. Protein, fat, water, lactose, and minerals are the major components, and
vitamins, phospholipids, cholesterol, enzymes, and pigments are part of the minor
components (Shrivastava 2010). More than 400 different fatty acids are present in
bovine milk fat (Christie 1995). All 22 essential minerals for our diet are present in
milk (Shrivastava 2010). Sixty indigenous enzymes have been found in normal
bovine milk (Fox and McSweeney 1998a).
During analysis of milk and milk products, different constituents of milk prod-
ucts respond differently due to difference in their properties. Single method is not
10 Application of Research Methodologies in Dairy Chemistry 199
Chromaography
Thermal
Spectroscopy
analysis
Electrophoresis
Immunoassay
method
Supercritical
fluid extraction
Fig. 10.1 Research techniques generally used for chemical analysis of dairy products
enough to detect all the constituents. No universal method exists for simultaneous
detection of all constituents. Several new techniques are available now for qualita-
tive and quantitative study. These techniques work under different principle.
Figure 10.1 illustrates the different research techniques generally used for chemical
analysis of dairy products.
10.2 Chromatography
Chromatography originally consists of two Greek words, “chroma” and “graphein”

which mean color and write, respectively. Earlier, chromatography method was
used to separate the colored molecule, but further, it was used to separate colorless
substances too. Chromatography consists of a group of techniques which is used to
separate a mixture. The basic principle of chromatography is that the molecule
passed through two phases: one is mobile phase (moving phase), and the other is
stationary phase (fixed phase). The mobile phase is used for sample dissolution, and
it further passes through stationary phase. The affinity of the compound towards
stationary phase and mobile phase is the basis of separation. The sample component
of polar nature will bind to polar component of the stationary phase and will be
retained for longer time as compared to other non-polar component of the sample.
Thus, different components travel at different speeds due to their different affinity.
In the chromatography system, the stationary phase can be liquid or solid sup-
ported on a solid, and the mobile phase can be liquid or gas flowing continuously
Table 10.1 Types of chromatography

Types Liquid-liquid chromatography
Liquid-solid chromatography
Gas-solid chromatography
Gas-liquid chromatography
over the stationary phase. According to the nature of mobile phase and stationary
phase properties, chromatography can be of the following types as illustrated in
Table 10.1 (Belanger et al. 1997; Rounds and Nielsen 1998).
10.2.1 Liquid-Liquid Chromatography (Partition

Chromatography)
When the solute particle is distributed between two immiscible liquid phases (sta-
tionary and mobile phases) on the basis of their solubility, the method is called
partition chromatography. The stationary phase consists of liquid bounded with an
inert support, and the mobile phase liquid medium has different polarity from the
stationary phase to avoid mixing. When the stationary phase liquid is polar and the
mobile phase has non-polar liquid, chromatography is normal phase liquid chro-
matography (Belanger et al. 1997). If the molecule is of polar nature, it will retain
by the polar stationary phase and the non-polar molecule will be eluted by non-
polar mobile phase, in normal phase. Reversed-phase chromatography has non-
polar stationary phase and polar mobile phase. So the non-polar molecule will be
retained, and the polar molecule will be eluted faster. In general, reversed-phase
liquid chromatography is mostly used. The solute molecule partitioned between
two immiscible liquids based on their partition coefficient, so called partition chro-
matography (Belanger et al. 1997). The nature of liquids is changed by changing
the buffer pH or by using different combinations of solvents. Hydrophilic polar
matters as carbohydrates, amino acids, and plant water-soluble pigments can be
separated by the normal phase liquid chromatography. However, the lipophilic
matters as fat-soluble pigments and lipids are separated by reversed-phase chroma-
tography (Rounds and Nielsen 1998).
Paper chromatography was used earlier as the example of liquid-liquid chroma-
tography. Now, it is replaced by thin layer chromatography. The paper consisting
cellulose is the support for the liquid stationary phase. The cellulose is saturated
with water. Thus, the molecule having water-soluble nature or hydrogen bonding
capacity moves slower. In paper chromatography, the degree of retention is expressed
by retardation coefficient (Rf). Rf is the ratio of distance travelled by the component
to the distance travelled by the solvent (Belanger et al. 1997).
Paper chromatography is generally used for highly polar components as amino
acid, sugar, or natural pigments (Belanger et al. 1997). Mixture of amino acids can
be differentiated by using paper chromatography.
10.2.2 Liquid-Solid Chromatography (Adsorption

Chromatography)
Adsorption chromatography is the oldest method of chromatography. Molecules

are retained on the basis of adsorption and desorption at the surface of support
(Hage 2018). The mobile phase used is either gaseous or liquid form, and the
adsorbent which is in the solid state acts as stationary phase. Separation is accord-
ing to the adsorbate interaction to adsorbent. Adsorbate is the molecule of interest.
The adsorbent is a solid porous material with large surface area which has active
sites where adsorbate gets interacted (Belanger et al. 1997). The adsorption at the
active sites is due to weak, non-ionic force as hydrogen bonds and van der Waals
(Wilson 2010). Generally, silica gel (mild acidic), alumina (mild basic), or char-
coal (non-polar) is used as an adsorbent (Belanger et al. 1997). Silica and alumina
have hydroxyl group, and the elution order of the molecule from the stationary
phase depends on their relative polarity. The separation is based on the principle
that the most polar molecule will be retained longer at the polar stationary phase
and will be eluted last. The non-polar molecule will be eluted fast (Rounds and
Nielsen 1998). There is a competition between mobile phase and the analyte for the
active site of the stationary phase. The nature of the analyte (compound to be ana-
lyzed) affects the selection of the mobile phase. For example, alcohol will
be selected if the analyte has hydroxyl group, esters or acetone will be selected for
analyte having carbonyl groups, and for non-polar analytes, hydrocarbons as hex-
ane, heptane, or toluene may be used (Wilson 2010).
Adsorption chromatography can be carried out in a thin layer on plate or glass
column. Thin layer chromatography (TLC) is an example of liquid-solid chromatog-
raphy (Cheremisinoff 1996). TLC plates usually contain adsorbent layer spreading
over an inert support of aluminum, glass, or firm plastic film. The layer is stabilized
by a binding agent as plaster of Paris (10%) in the mixture. The sample is identified
by its retardation coefficient, which is specific for a specific compound when using
the same solvent. Thin layer chromatography can be used to separate carbohydrate
mixtures, unsaturated fatty acids, methyl esters of fatty acids, and triglycerides.
Adsorption chromatography is used for the separation of hydrophobic compounds as
lipids, fat-soluble vitamins, and carotenoid pigments (Rounds and Nielsen 1998).
10.2.2.1 High-Performance Liquid Chromatography (HPLC)

Liquid-solid chromatography was used in the early 1900s. In 1970, the usage of nor-
mal phase chromatography was decreased, and the application of high-performance
liquid chromatography with reverse phase became very common (Snyder and Dolan
2013). Silica has been considered as the most preferred column packing material over
others. This technology is developed for the use of new column and different advanced
part of instrument as high pressure pump, detectors, and sample injector. HPLC is a
modern analytical technique which works on high pressure. The HPLC provides fast,
improved resolution and more sensitive separation (Rounds and Gregory 1998). The
separation mechanism depends on the distribution of the analyte between the mobile
phase (liquid) and the stationary phase (solid). The principle of HPLC is the same as
the liquid-solid and liquid-liquid chromatography (Aniszewski 2015). The detectors

used in HPLC may be UV-visible, fluorescence or refractive index detector. The
normal phase HPLC is mainly used for the analysis of fat-soluble vitamins, phos-
pholipids, and carbohydrates (Rounds and Gregory 1998). The reversed-phase
HPLC is used for the detection of proteins, amino acids, water and fat-soluble vita-
mins, antioxidants as butylated hydroxyanisole (BHA) and butylated hydroxytolu-
ene (BHT), pigments as carotenoids, and phenolic flavor compound as vanillin
(Rounds and Gregory 1998). Heavy metals such as copper, lead, mercury, cadmium,
nickel, tin, and zinc can also be detected by reversed-phase HPLC (Yang et al.
2004). HPLC coupled with mass spectroscopy (MS) is used for the pesticide resi-
dues, and LC-MS is used for the veterinary drug residues. The concentration of
lactulose and furosine, which are Maillard browning (protein-sugar reaction) prod-
ucts, can also be detected by HPLC method (Fox and McSweeney 1998b).
10.2.3 Gas-Liquid Chromatography
In gas chromatography (GC), a sample is separated by two phases: stationary phase

and mobile phase. The mobile phase is an inert gas which passes over the stationary
phase. If the stationary phase is liquid, it is called gas-liquid chromatography, and if
it is solid, it is called gas-solid chromatography (Wang and Pare 1997). In gas chro-
matography, inert gas used as mobile phase is helium or nitrogen. The gas passes
through a column, and this column is placed in an oven, where the temperature is
controlled. The concentration of the molecule is affected by the vapor pressure of
the gas. The separation is based on the difference in boiling point (Li and Liu 2019).
The separation of the volatile component is based on their partition coefficient
between the gaseous mobile phase and the stationary phase. Partition coefficient is
the ratio of solubility of the substance in mobile phase gas carrier and the stationary
liquid phase. Gas chromatography is used for the detection of volatile compounds.
The partition coefficient is inversely proportional to the volatility of the sample, so
the most volatiles will be eluted first (Wilson 2010). The component which is parti-
tioned into the gas will be eluted first, and the one partitioned into the liquid
phase will be eluted last. Usually, the temperature of the column is maintained
between 50 and 300 °C for sample volatilization. High boiling point liquids are used
for the stationary phase as wax or silicone grease which supports the column
(Wilson 2010). Gas-solid chromatography separates the permanent gases and low
boiling hydrocarbons (Seneca 2007). In gas chromatography, a sample is injected
into the inlet and introduced into the column through carrier gas of mobile phase.
The separation takes place as the sample moved into the column and get detected by
the detector.
GC is used for the analysis of thermally stable volatile compounds. Gas chroma-
tography is generally used for the analysis of triglycerides, fatty acids, cholesterols,
vitamins, amino acids, peptides, food additives, pesticides, drugs, antioxidants, fla-
vor compounds, polychlorinated biphenyls, nitrosamines, etc. (Reineccius 1998).
10.2.4 Ion Exchange Chromatography
Ion exchange chromatography separates ion and polar molecules (Belanger et al.
1997). The stationary phase (ion exchanger) contains functional group having posi-
tive or negative charge. The ions of solution have affinity for the opposite charge of
stationary phase. The functional group of the stationary phase decides whether cat-
ion or anion will be exchanged. If the functional group of stationary phase has posi-
tive charge, it is called as an anion exchanger, and if it is negatively charged, it is
called as cation exchanger (Rounds and Nielsen 1998). The separation of cations
have been done on cation exchange resin column; however, anion exchange resin
column separates anions. As the ion is formed only in aqueous solution, the mobile
phase used is generally a buffer having opposite charge ion to the overall charge of
stationary phase. The net charge of the molecule depends on the pH; the selection of
mobile phase pH depends on the total charge of the analyte. There is a competition
between the analyte ion and the similarly charged ion of eluent to bind with the oppo-
site charge functional group of the stationary phase (Acikara 2013). At pH greater than
2, strongly acidic sulfonic group (RSO3−) of strong cation exchanger is completely
ionized. Strong basic quaternary amine group of strong anion exchanger is ionized at
pH less than 10. Weak cation exchanger contains carboxylic acid group, and weakly
basic anion exchangers contain primary, secondary, and tertiary amine groups (Rounds
and Nielsen 1998). The separation of the molecule is based on the principle that there
is an ionic bond formation between the charged group of molecules and the ion of
stationary phase. The adsorbed molecules can be eluted by altering the pH of mobile
phase or increasing the ionic strength of mobile phase (Rounds and Nielsen 1998).
Ion exchange chromatography is generally used for the separation of amino
acids, proteins, sugars, alkaloids, and metal ions. Many drugs, fatty acids, and
nucleotides can also be separated (Belanger et al. 1997; Rounds and Nielsen 1998).
The concentration of milk salts such as calcium and magnesium can also be deter-
mined by the ion exchange chromatography (using cation exchange resin) (Fox and
McSweeney 1998d).
10.2.5 Size Exclusion Chromatography
The size exclusion chromatography is a simple mode of separation in which separa-

tion of molecule occurs on the basis of their size. It is also called molecular exclu-
sion, gel filtration, and gel permeation chromatography (Rounds and Neilsen 1998).
There is no interaction between the solute and the stationary phase. The stationary
phase contains a column packing materials having pore size comparable to the mol-
ecules to be separated. When the dissolved molecules are passed through the col-
umn, the small molecules flow at a slower rate and are eluted last because they get
penetrated into the pores, whereas the larger molecules flow at a faster rate and are
eluted first because they do not penetrate into the pores (Wilson 2010; Belanger
et al. 1997). Gel permeation and gel filtration chromatography are the two types of
size exclusion chromatography. In gel permeation chromatography, hydrophobic
column packing material and non-polar mobile phase (organic solvents) are used. It
is generally used for the molecular weight distribution of synthetic polymers. In gel
filtration chromatography, hydrophilic packing materials and polar mobile phase
are used to separate the hydrophilic materials as proteins and polysaccharides.
The size exclusion chromatography is mainly used to separate the molecules
from bigger or smaller molecules. Antibodies, viruses, hormones, enzymes, poly-
saccharides, and nucleic acids can be purified and separated using this tech-
nique (Wilson 2010).
10.3 Spectroscopy Method
Light is an electromagnetic radiation consisting of bundles of energy called pho-

tons. The light gets interacted with the molecule or atom and provides structural,
compositional, and interaction details. The study of interaction of light with the
material is called spectroscopy. Table 10.2 shows the spectrum wavelength for dif-
ferent electromagnetic spectrum. There is no exact division of wavelength between
two successive spectrums. It is the electrons in atoms and molecules that typically
absorb and emit photons of light (Ocean Optics 2017). The wavelength is inversely
proportional to the photon energy, so the higher the wavelength, the lower the
energy. The energy level governs the effect of light on the matter. The wavelength
of radio wave is high, so the energy level is low and the amount of energy absorbed
is negligible (Ocean Optics 2017). Infrared affects the chemical bond which causes
change in vibrational energy. The energy of infrared is not sufficient to excite elec-
tron, but it may cause vibrational excitation of covalently bonded atoms or group
(Reusch 2019). Single carbon-carbon bond absorbs lower infrared radiation as com-
pared to double carbon-carbon bond because the double bond is stronger than the
single bond (Harvey 2000). In ultraviolet and visible region, it is the valence elec-
tron that gets affected. So on absorption of these lights, the valence electron of atom
or molecule goes into higher energy state.
The light can interact with materials in different ways; it can be reflected,
absorbed, or transmitted. In absorption spectroscopy, when the electromagnetic
wave interacts with the material, the molecule or atom goes into excited state
(higher-energy state) from ground state (lower-energy state). When the molecule of
excited state comes back into ground state, it releases the excess energy, and it is
Table 10.2 Different elec- Electromagnetic spectrum Wavelength

tromagnetic spectrums and γ ray 0.01–1 A°
their wavelengths
X-ray 0.1–10 nm
UV 10–380 nm
Visible 380–750 nm
Infrared 0.075–1000 μm
Microwave 0.1–100 cm
Radio wave 1–1000 m
Source: Penner (1998a)
called emission (Harvey 2000). The different types of energy transfer and their
application in spectroscopy are illustrated in Table 10.3.
10.3.1 Source of Electromagnetic Radiation
The source of electromagnetic radiation may be continuum or line source. The con-
tinuum source provides radiation over a broad range of wavelength; however, the
line source provides radiation of fixed wavelength (Harvey 2000). The different
sources of radiation and their application are detailed in Table 10.4.
Common spectroscopy methods used for the analysis of food products are
ultraviolet-visible spectroscopy (absorption spectroscopy), fluorescence spectros-
copy, atomic absorption spectroscopy, and infrared absorption spectroscopy.
10.3.2 UV-Visible Spectroscopy
The UV-visible spectroscopy is the most widely used spectroscopy for the analysis
of food products. The wavelength in the range of 200–700 nm is used for UV-visible
spectrum. For UV region, wavelength from 200 to 350 nm is used; however, for
visible region, wavelength from 350 to 700 nm is used (Penner 1998b). The wave-
length of UV light is colorless for human eye, and the visible region has its charac-
teristic color for different wavelength. The violet color has the smallest wavelength,
and the red color has the longest wavelength in visible region.
10.3.2.1 Basic Principle

The main purpose of UV-visible absorption spectroscopy is to evaluate the concen-
tration of the sample. The sample absorbs the light when it hits the analyte in solu-
tion form. The relation between the absorbance and the concentration is expressed
by Beer–Lambert law. According to the Beer–Lambert law, absorbance is directly
proportional to the analyte concentration. The following is the Beer–Lambert equa-
tion (Hofmann 2010):
Table 10.3 Different types of energy transfer and their application

Energy transfer Spectroscopy
Absorption UV-Vis UV-Vis spectroscopy
Atomic absorption spectroscopy
Infrared Infrared spectroscopy
Raman spectroscopy
Microwave Microwave spectroscopy
Radio wave Nuclear magnetic resonance spectroscopy
Emission UV-Vis Atomic emission spectroscopy
Photoluminescence UV-Vis Fluorescence spectroscopy
Source: Harvey (2000)
Table 10.4 Different sources of radiation, wavelength, and their applications

Source Wavelength Application
H2 and D2 lamp Continuum source from 160 to 380 nm Molecular absorption
Tungsten lamp Continuum source from 320 to 2400 nm Molecular absorption
Xe arc lamp Continuum source from 200 to 1000 nm Molecular fluorescence
Hollow cathode lamp Line source in UV-Vis Atomic absorption
Hg vapor lamp Line source in UV-Vis Molecular fluorescence
Source: Harvey (2000)
1
bc
A log
T
where A is absorbance of the sample, T is transmittance, ε is absorptivity with unit
cm−1 conc−1, if concentration is expressed as molarity, ε is molar absorptivity
cm−1 M−1, b is path length (cuvette width) in cm, and c is the concentration.
The Beer–Lambert law is applicable only for low concentration of the analyte.
The fat oxidation can be measured through anisidine value (which measures
aldehyde production by fat oxidation), peroxide value, and thiobarbituric acid by
using UV-Vis spectrometer. Uv-vis spetrometer can be used for the analysis of
carotenoid content (Nawrocka and Lamorska 2013). This method is also used for
the detection of protein by Folin-Ciocalteu method, Biuret method and Bradford
method. Milk adulteration and milk protein antioxidant activity can be checked by
using this method (Finite et al. 2013). The concentration of different milk compo-
nents as fat, protein, lactose, and water can be determined by using absorption
spectroscopy.
10.3.3 Fluorescence Spectroscopy
Fluorescence spectroscopy is a specific and sensitive method to detect the native

fluorescence or the derivatized sample (Shackman 2013). Both absorption and
emission phenomena occur simultaneously. It measures the electromagnetic radia-
tion which is released from the sample when it comes back to its ground state from
the excited state. The emission of fluorescence is only possible by the molecule at
excited state, so the sample first goes to its excited state by absorption of ultraviolet
or visible light. The absorption and emission processes occur simultaneously
(Penner 1998b). The instrument used for the fluorescence spectroscopy is same as
the absorption spectroscopy, but the optical system arrangement is different, and
two wavelengths are needed: one for absorption and one for emission. The emitted
light is at 90° angle with the excited one (Penner 1998b). The sample concentration
is directly proportional to the emission intensity.
Milk products contain some natural fluorophores or may be produced after
chemical alteration as Maillard reaction products, vitamin A, amino acids, ribofla-
vin, lipid oxidation products, porphyrins, NADH, etc. Fluorescence spectroscopy
is used to assess quality, safety, milk coagulation, and syneresis. Maillard reaction
product can be used as an indicator to differentiate among different modes of heat

treatment. Lactulose and furosine, which are indicators of heat treatment, can be
estimated by fluorescence spectroscopy (Shaikh and ODonnell 2017).
β-Lactoglobulin of heat treated milk, vitamin A and melamine content can also be
analyzed by this method (Kulmyrzaev et al. 2005; Shaikh and ODonnell 2017).
Tryptophan content is important during gel formation due to coagulation; thus,
fluorescence of tryptophan helps in structural modification in milk protein during
coagulation (Boubellouta et al. 2011). Riboflavin and tryptophan act as a tracer
molecule for controlling the curd syneresis and milk coagulation (Fagan et al.
2011).
10.3.4 Atomic Spectroscopy
There are two types of atomic spectrophotometric methods: atomic absorption spec-
troscopy and atomic emission spectroscopy. Atomic absorption spectroscopy is an
analytical method to detect the element concentration. There is absorption of
UV-visible spectrum by the free atom. It is the atom of the molecule which absorbs
the radiation, so it is needed for the sample to be in atomic state, which is achieved
by atomization. The sample is sprayed by the nebulizer into high heat of flame or
oven in order to vaporize and decompose into a free ground state atoms (Miller
1998). Generally air-acetylene flame temperature 2300 °C is used for the various
elements, which is sufficient for atomization of most elements (Cheremisinoff
2001). The concentration of the element depends on the absorption of a radiation by
the atom which is produced from a radiation source, i.e., hollow cathode lamp.
Different lamp is required for different element (Cheremisinoff 2001). Hollow cath-
ode lamp contains cathode and anode in a glass tube filled with an inert gas. Cathode
is made up of the element of interest, and anode is a nickel or tungsten wire. When
a voltage is applied, the cathode metal atoms are released, which are in excited state
and release a specific radiation when they come back to their ground state. Thus, a
hollow cathode lamp offers a radiation which is used as the sample absorption spec-
trum. A monochromator is placed between the flame and the detector. It removes the
unwanted wavelength from other wavelength emitted by the lamp source so that
only the desired wavelength reaches the detector (Miller 1998).
Atomic emission spectroscopy works under the principle that when the sample is
heated to high temperature, the atom of ground state goes into excited state, and
when it returns to the ground state, it emits energy and this emitted energy corre-
sponds to the number of atoms (Harvey 2000). The number of excited atoms are
proportional to the number of emitted energy (Hofmann 2010).
All the 15 elements required as per US RDA (Ca, K, P, Fe, Zn, Mg, I, Cu, Na, Se,
Mo, Cr, Mn, Cl, and F) can be detected from some type of atomic spectrophotom-
eter. Many other elements which are important for human health such as Ni, Cd, Co,
Si, Pb, Sn, B, V, and Li can also be analyzed by atomic spectrophotometer (Sandroni
and Smith 2002; Harvey 2000).
10.3.5 Infrared Spectroscopy
Infrared spectroscopy covers the energy level below the UV-Vis region and wave-
length range between 0.075 and 1000 μm. When the electromagnetic radiation hits
the sample, some amount of energy is absorbed and some reflected or transmitted.
Infrared measures the amount of energy absorbed. The amount of energy transmit-
ted or reflected determines the concentration of the analyte which is reverse to the
amount of absorbed energy (Wehling 1998). The energy associated with the infrared
is not enough to cause excitation of electron, but it can cause vibrational excitation
of the covalent bounded atom or molecule. There are three regions of infrared spec-
troscopy, i.e., far infrared, mid infrared, and near infrared. Infrared spectroscopy is
generally used for the structural analysis of the sample. Different functional group
absorbs different amount of radiation energy. In the mid-infrared region, the atom
goes to first excited vibrational state from the ground state, and in near-infrared
region, it goes on second excited state and mixture of these two vibrations (Rudzik
and Wust 2002).
Mid-infrared spectroscopy is used in infrared milk analyzer for fat, lactose, and
protein estimation of milk sample. Near-infrared spectroscopy is used for various
food products as milk products, grains, meat product, cereal, grains, etc. It is a rapid
method to analyze the additives (e.g., stabilizer) in dairy products. Most commonly
used MilkoScan works under infrared spectroscopy. Infrared spectroscopy can be
used as on-line method which means that the product can be analyzed during pro-
cessing. Moisture content of the powder and butter can be analyzed during produc-
tion by near-infrared spectroscopy. Almost all major components of the milk and
milk products can be analyzed by infrared spectroscopy. Mid-infrared spectroscopy
is also useful for the analysis of microbes which helps in saving time of 2 or 3 days
(Rudzik and Wust 2002).
10.4 Electrophoresis Method
Electrophoresis is a movement of a charged molecule in an electric field. The electric

current flows between the electrodes, i.e., cathode and anode. Cathode contains nega-
tive charge and donates electron, whereas anode has positive charge and takes elec-
tron. The ion which will get the electron from cathode, will be negatively charged, and
will move towards positive anode called anions. The positive ions will move towards
negative charged cathode, called cations (Dennison 2002). The migration of the
charged molecule is restricted by gel to get time to resolve. The mobility of the
particle is directly proportional to its charge and inversely proportional to shape,
size, and viscosity of the particle. A conventional electrophoresis system contains
electrophoresis unit and power supply. A constant power supply is required for the
voltage. Electrophoresis unit contains horizontal or vertical gel unit (Walker 2010).
Many molecules are amphoteric in nature; the charge on it depends on the pH of the
solvent. The buffer maintains the pH and it is required for current passing. Agarose
gel or polyacrylamide gel is mostly used for electrophoresis. Protein is separated
mostly by polyacrylamide gel electrophoresis. In undenatured or native electropho-

resis proteins separated in their native form, the separation is based on the shape,
charge, and size. Denaturing electrophoresis is another type of electrophoresis. In
sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), SDS is
used to separate on the basis of size, and mercaptoethanol or dithiothreitol reduces
the disulfide bond. Binding of protein with SDS develops negative charge, and the
separation occurs based on their size only (Smith 1998).
Alkaline urea PAGE is used to fractionate casein. The high concentration of
urea is used to disrupt casein association and allows it to migrate as monomer.
SDS-PAGE is used for whey protein fractionation (Fox and McSweeney 1998a).
Milk fat globule membrane proteins can be resolved on SDS-PAGE (Fox and
McSweeney 1998c).
10.4.1 Isoelectric Focusing
Isoelectric focusing is used for the amphoteric substances as protein, and the separa-
tion is based on their isoelectric point. It provides highest resolution among all
protein separation methods. Each protein has its specific isoelectric point. It differ-
entiates protein with difference of 0.01 pH. Isoelectric focusing uses a horizontal
gel system. A gel system is placed between the electrodes, and pH is maintained
between 2 and 10. The pH gradient across the gel results in protein movement
towards a definite pH. Protein carries positive charge below the isoelectric pH and
moves towards cathode, and above the isoelectric pH, protein carries negative
charge and will migrate towards the anode. The protein migrates along with the pH
gradient under an electric field till its total charge becomes zero; the migration stops
at its isoelectric point (Büyükköroğlu et al. 2018). Agarose or polyacrylamide gel is
used for the gel system (Otter 2003).
Milk proteins from different mammal milk as bovine, murine, caprine, and por-
cine were differentiated by using isoelectric focusing (Kim and Jimenez-Flores
1994). Whey protein and caseins can also be fractionated by isoelectric focusing
(Josephson 1972). Isoelectric focusing can be used for milk fat globule membrane
protein fractions (Fox and McSweeney 1998c).
10.5 Supercritical Fluid Extraction
Supercritical fluid extraction is a technique of separation of one constituent from

another constituent by using supercritical fluid as a solvent rather than commonly
used organic solvents. This separation can be done from solid matrix or from liquid.
It could be used as a preliminary step for the isolation of the desired component
during sample preparation. Mostly, CO2 is used as a supercritical fluid, and it offers
good recovery of hydrophobic compounds; hydrophilic compounds are rarely solu-
ble in it (Donato et al. 2015). The critical point can be explained by a temperature-
pressure phase diagram at which material in liquid phase has similar temperature,
pressure, and density as in gas phase; the material behaves as both gas and liquid phase.
CO2 has low critical temperature, i.e., 31.06 °C, and pressure (7.386 MPa) suitable for
many dairy and food products analysis (Rozzi and Singh 2002). At pressure and tem-
perature more than the critical point, supercritical fluid behaves as both liquid and gas-
like phase. Gas-like phase imparts more diffusivity and less viscosity which provides
easy penetration and fast mass transfer. The density of supercritical fluids can be
changed by changing temperature and pressure (Khaw et al. 2017). This extraction
method has several advantages over other traditional extraction method. Traditional
method uses toxic chemicals for extraction and is time-consuming with low yield.
Supercritical fluid extraction involves two steps: extraction and separation. The
liquid is subjected to temperature and pressure beyond the critical point which
depends on the molecule of interest and passes through the extractor for extraction.
In next process, the solvent and extracted molecule are separated in the separator.
Supercritical fluid is changed into gas phase by lowering the pressure. Thus, the
supercritical fluid can be recycled or released in the environment (Singh et al.
2018). Some more information about the supercritical fluid extraction has been
elaborated in the Chap. 2 of this book.
Supercritical fluid extraction method can be used for the food products analysis.
Milk fat, cholesterol, and vitamins can be isolated from milk using this method.
This method is safer than the traditional Soxhlet method for fat analysis which uses
harmful chemicals (Singh et al. 2018). The flavor compounds of milk fat can also be
separated using this method (Haan and Graauw 1990). Lipid fraction, milk fat glob-
ule membrane phospholipid and proteins can be separated by using this method.
Milk enzymes as alkaline phosphatase can be deactivated, and microbial load can be
reduced by supercritical fluid extraction (Singh et al. 2018).
10.6 Immunoassay Method
Immunoassay is an analytical method in which the sample concentration is deter-

mined by antigen (sample of interest)-antibody reaction (Darwish 2006). Antibody
is an immunoglobulin that binds to a specific antigen. Antibody is of two types:
monoclonal antibody and polyclonal antibody. The monoclonal antibody identifies
only one site on antigen, and the polyclonal antibody identifies many sites on anti-
gen. Antigen is any substance to be detected and has the ability to bind the antibody.
Labels are used to identify the reaction; labels commonly used are enzymes, radio-
active substance, and fluorescent. The immunoassay is of two types: isotopic immu-
noassay and non-isotopic immunoassay. Isotopic immunoassay works under the
principle that the radioactive labeled antigen and the unlabeled antigen of the ana-
lyte compete for the antibody and form antibody-antigen complex.
Radioimmunoassay (RIA) is a type of isotopic immunoassay. The increment in the
quantity of unlabeled antigen in the analyte reduces the quantity of bound radioac-
tive antigen due to its displacement from the antibody site. The amount of antigen
is obtained by preparing the standard curve of known concentration of pure antigen
(Dixon 1998). For labeling, generally, iodine-125 is used as an isotope which pro-
duces radiation and can be measured by gamma or beta counter (Praither 1985). The
RIA method is expensive and can be hazardous due to handling of radioactive
element. Another type of immunoassay is non-isotopic immunoassay; the type of

label used is different. Fluoroimmunoassay and enzyme-linked immunosorbent
assay (ELISA) are the two types of non-isotopic immunoassay. Fluoroimmunoassay
is similar to the radioimmunoassay; the label used is fluoropores instead of radioiso-
topes, and it is more sensitive than RIA. Fluoropores have tendency to absorb and
emit the light at different wavelength. This method uses fluorometer; however, in
enzyme immunoassay, colorimeter is used. The method can qualitate or quantify the
target sample. ELISA method uses a multi-well pate which offers a solid surface to
adsorb antigen or antibody. The label used in ELISA is an enzyme as glucose oxi-
dase, alkaline phosphatase, and horseradish peroxidase (Dixon 1998). The enzyme
catalyzes the reaction and yields a colored product from the substrate. These col-
ored products can be identified by the colorimeter.
Immunoassay method, mostly ELISA, is a cost-effective, sensitive, fast, and spe-
cific method than the commonly used microbial and chemical method in food
industry. Pesticide residue, antibiotics, and microorganisms can be detected by
immunoassay (Samarajeewa et al. 1991). Foodborne pathogens such as L. monocy-
togenes and E. coli, mycotoxins as aflatoxins, and food allergens can also be
detected. Ferritin level which indicates amount of iron in the body can also be
detected by ELISA (Bleicher et al. 2018). Whey protein β-lg denaturation can be
quantified by ELISA as the denatured and native proteins act differently to antibod-
ies. Adulteration of milk with other species (bovine milk with sheep milk) and milk
microbial enzyme can be quantified and detected; proteolysis can be monitored in
cheese and protein hydrolysates by using ELISA (Fox and McSweeney 1998a).
10.7 Thermal Analysis
Thermal analysis is a method to detect physicochemical changes occurring due to

exposure of the sample under specific temperature. Differential thermal analysis
(DTA) and differential scanning calorimetry (DSC) have a wide range of applica-
tions for the estimation of food and dairy products (Schenz and Davis 1998).
10.7.1 Differential Thermal Analysis
Differential thermal analysis (DTA) analyzes the temperature difference between

the sample and the reference when subjected to a specific heat treatment. Both the
reference and the sample are placed in the furnace and heated. The difference in
temperature is recorded. The DTA graph presents temperature difference with
respect to temperature or time (Abraham et al. 2018). Temperature difference repre-
sents endothermic or exothermic reactions occurred in the sample. Temperature dif-
ference zero represents no physicochemical changes. The greater or lesser
temperature of the sample than the reference shows exothermic or endothermic pro-
cess. The area of the graph represents the quantity of heat involved during physico-
chemical changes (Schenz and Davis 1998). In the graph, the endothermic change
represents negative side, i.e., below the baseline, and the exothermic change repre-
sents positive and or above the baseline.
10.7.2 Differential Scanning Calorimetry
Differential scanning calorimetry (DSC) analyzes the change in heat flow rate of the
sample and the reference when they are maintained in a specific temperature condi-
tion (Hohne et al. 2003). In this method, the sample needs higher or smaller amount
of heat than the reference which depends on whether the process is endothermic or
exothermic. DSC contains a sample holder, sensors, furnace, and a temperature con-
troller. There are two separate holders for sample and reference; the sample holder
is made up of aluminum pan. The reference is either alumina or empty aluminum
pan (Dwivedi et al. 2017). Each pan is placed on the heater and controlled by the
computer. The temperature of the sample and the reference is heated at a fixed rate,
10 °C/min. The rate of heating would be the same throughout the analysis. There are
different parameters that affect the analysis such as the size of the sample pan, its
material, rate of heating, furnace shape, size, sample amount, and heat capacity. The
small sample size and slow heating provide a good resolution (Schenz and Davis
1998). DSC graph shows relation of heat flux and temperature or heat flux or time.
Both DTA and DSC have nearly similar applications; however, the principle
involved is dissimilar. DTA works under the principle of temperature variation, and
DSC works under heat flow change. DSC is used in the study of protein, water,
carbohydrate, and lipid interactions in the food system. Both DTA and DSC mea-
sures the extent of freezing, heat capacity, change in phase, crystallization, pyroly-
sis, and glass transition. Polymorphism of mono- or triglycerides can also be
studied. DSC measures the amorphous lactose (Gabbott et al. 2003) and α-lactose
crystallization can also be studied (Gombas et al. 2002). DTA is used to study the
property of lactose crystal and amorphous lactose. Water of crystallization of lac-
tose and milk powder and surface water was also studied by Listiohadi et al. (2009).
Heat stability of casein can be studied using DSC (Moldoveanu et al. 2013). Spray-
dried and freeze-dried milk powder can be differentiated by using DSC. Camel and
cow milk powder can also be differentiated by using this technique (Sulieman et al.
2017). DSC can also be used for the study of denaturation of milk proteins.
10.8 Conclusions
Chromatography, spectroscopy, electrophoresis, supercritical fluid extraction,

immunoassay, and thermal analysis are the various techniques generally used for
the analysis of different constituents of milk and milk products. These methods are
accurate, fast, safe, and sensitive to analyze the sample both qualitatively and quan-
titatively. All the major and minor constituents can be easily detected by these tech-
niques. Hence, these research methods have great application in dairy fields
especially in the dairy chemisty.
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Approaches for Detection of Dairy
Microorganisms: An Update 11
Aparna Sudhakaran V and Santosh Anand
Contents
11.2 C ulture-Dependent Techniques 219
11.2.1 Microbial Plating Techniques 220
11.2.2 Biochemical Screening 221
11.2.3 Biosensors 222
11.2.4 Immunological Methods 224
11.2.5 Flow Cytometry 224
11.2.6 Adenosine Triphosphate (ATP) Bioluminescence 225
11.2.7 Electrical Impedance Measurement 225
11.2.8 Electromigration Techniques 226
11.2.9 Multivariate Techniques 226
11.2.10 Omics Techniques 227
11.3 Culture-Independent Techniques 227
11.3.1 Polymerase Chain Reaction (PCR) 228
11.3.2 Multiplex PCR 228
11.3.3 Quantitative Real-Time Polymerase Chain Reaction (RT qPCR/qPCR/
qRT PCR) 229
11.3.4 High-Resolution Melt (HRM) Analysis 229
11.3.5 Digital PCR 230
11.3.6 Extreme PCR 230
11.3.7 Reverse Transcriptase PCR (RT PCR) 231
11.3.8 Viability PCR Approach: EMA-PCR and PMA-PCR 231
Aparna Sudhakaran V (*)

S. Anand
Dairy Microbiology Department, PJM College of Dairy Technology,
Dumka, Jharkhand, India

https://doi.org/10.1007/978-981-15-2608-4_11
218 Aparna Sudhakaran V and S. Anand
11.3.9 PCR-Based Denaturing Gradient Gel Electrophoresis (DGGE) and

Temporal Temperature Gradient Gel Electrophoresis (TTGE) 232
11.3.10 DNA Microarrays 232
11.3.11 Fluorescence In Situ Hybridization (FISH) 233
11.3.12 Molecular Typing 233
11.3.13 Pulsed-Field Gel Electrophoresis (PFGE) 233
11.3.14 Random Amplification of Polymorphic DNA–RAPD-PCR 234
11.3.15 Restriction Fragment Length Polymorphism (RFLP) 234
11.3.16 Amplified Fragment Length Polymorphism (AFLP) 234
11.3.17 Amplified rDNA Restriction Analysis (ARDRA) 235
11.3.18 Repetitive Element-Polymerase Chain Reaction (rep-PCR) 235
11.3.19 Multilocus Sequence Typing (MLST) 235
11.3.20 Whole-Genome Sequencing (WGS) Techniques 236
References 237
Abstract
The detection, quantification, and isolation of microbes are routine processes in

the dairy industry which are advisable for monitoring food quality from an early
stage. The microbiological tests can be classified into culture-dependent and
culture-independent techniques. The microorganism in milk and milk products can
be detected either by the conventional culture-based approach (enumeration, isola-
tion, characterization, and identification) or through a culture-independent
approach (isolation, purification of nucleic acid, and molecular identification). The
culture-dependent methods are considered as the basic gold standard technique in
microbial detection. The culture-dependent techniques (traditional approaches)
rely on the isolation procedure from complex food matrices and the growth of
microbial colonies on selective agar to detect and quantify viable microbes and
identify microbes mainly depending upon their morphology and/or biochemical
characteristics. The culture-independent techniques circumvent the steps of enu-
meration and characterization of microbes rather based on nucleic acid-based iden-
tification. In culture-independent techniques, nucleic acid (deoxyribonucleic acid
(DNA)/ribonucleic acid (RNA)) is isolated and purified from the dairy matrix, and
different molecular approaches are applied to identify and quantify the pathogens.
This chapter gives a brief overview of the latest rapid microbiological methods.
Keywords
Culture-dependent techniques · Culture-independent techniques · Polymerase
chain reaction · Biosensors
11.1 Introduction
Being a highly nutritious food, milk supports the growth of both beneficial and
non-beneficial microbes. The microbes present in milk can affect the food safety of
the milk products, cause dairy-borne outbreaks, and can interfere with the quality of
11 Approaches for Detection of Dairy Microorganisms: An Update 219
dairy products. The processing techniques and other safety practices considerably
enhance the safety of the products. The detection of microbial risks at any stage of
processing ensures safety as well as helps to reduce the economic loss and main-
tains the credibility of the food processing industry. Stringent regulations help to
ensure good quality and safer milk and milk products.
Microbiological tests are either quantitative (number of organisms per unit
weight of sample) or qualitative (to detect whether a particular organism is present
in a known weight of the sample). The detection, quantification, and isolation of
microbes are routine processes in the dairy industry which are advisable for moni-
toring food quality from an early stage. The conventional method relies on the
incorporation of a known amount of food sample in a growth medium such that
microorganisms replicate and form visible colonies. Even though these methods
demand a lot of time and energy, they are still preferred due to their simplicity,
adaptability, and affordability.
Several researchers are working on different cost-effective, sensitive, accurate
methods and fast approaches to determine the beneficial microbes or the microbial
risks in dairy products and to identify the etiological agents during the milk/dairy-
borne outbreak. Moreover, they help to explore the microbial diversity in various
dairy products. The microbiological tests can be classified into culture-dependent
and culture-independent techniques. The microorganism in milk and milk products
can be detected either by a conventional culture-based approach (enumeration, iso-
lation, characterization, and identification) or through a culture-independent
approach (isolation, purification of nucleic acid, and molecular identification).
11.2 Culture-Dependent Techniques
The culture-dependent methods are considered as the basic gold standard technique
in microbial detection. The culture-dependent techniques (traditional approaches)
rely on the isolation procedure from complex food matrices and the growth of
microbial colonies on selective agar to detect and quantify viable microbes and to
Advantages Less expensive

• Nucleic acid is present in
limited quantity or of inferior Facilitates the
quality obtainable from the detection of
milk/product matrix injured cells
Culture-Dependent Approach • Presence of high amounts of

Polymerase Chain reaction Limitations Time-consuming
(PCR) inhibitors in the Tedious
milk/product matrix Less efficient
• The target organism is too low Chances of bias
in number toward cultivable
organism
Fig. 11.1 Culture-dependent approaches—advantages and limitations

identify microbes mainly depending upon their morphology and/or biochemical

characteristics. The methods are more affordable for the assessment of the micro-
biological quality of milk and milk products. The culturing stage facilitates the
detection of bacteria even when smaller numbers are present or in case of stressed
or damaged cells by cell recovery through enrichment (nonselective and selective)
process.
The culture-dependent approaches are time-consuming, material consuming,
less efficient, and tedious and are biased to the detection of microbes that can grow
well in laboratory conditions (cultivable). The chances of misinterpretation of
microbial diversity will be more when the culture conditions are poor or the number
of organisms is less or microbiologically complex (Hugenholtz et al. 1998). Fusco
and Quero (2014) elaborated the conditions in which culture-dependent and inde-
pendent approaches were to be used. The culture-dependent approach is used in any
of the following situations as illustrated in Fig. 11.1.
The various culture-dependent methods are based on culturing of microbes
which are discussed briefly below.
11.2.1 Microbial Plating Techniques
Microbial cultivation in specific media is the most commonly used culture-

dependent method for enumeration, detection, and isolation of microbes. It almost
completely depends on the growth of microbes on specific or selective agar. The
isolation of microbes from the complex food matrix can be accomplished by pure
culture techniques, but the culturing process usually gets accompanied with limita-
tions like the inability to distinguish between the microorganism under study and
the endogenous microbes, false-positive detection, matrix-dependent complica-
tions, and non-recovery of viable but nonculturable (VBNC) cells (Sohier et al.
2014). The accepted method of sampling usually involves pre-enrichment, enrich-
ment, selective plating, and isolation of microbes, which are subsequently con-
firmed by biochemical and serological tests. It generally takes about a week to
obtain a valid result for a specific organism (Rajapaksha et al. 2019). Plating on
selective and chromogenic agar media improves the selection efficiency of microbes.
The synthesis of certain chemical compounds like enzymes or metabolites during
growth in the medium facilitates the detection of microbes. The selectivity of the
medium can be improved by compositional variations of agar medium and varia-
tions in temperature and pH or by addition of antibiotics or carbon sources in the
medium (Saccaro et al. 2011). Subsequently, in chromogenic media, an ideal chro-
mogenic substrate should be hydrolyzed to release a colored product. A wide range
of chromogenic culture media has been made commercially available which is
based on enzyme activity like β-galactosidase and β-glucosidase activities provid-
ing a useful tools for the detection of undesirable and desirable microbes in the
dairy industry. The major drawback of traditional culturing methods is that they are
effective in detecting only a particular group of microbes in the sample. Moreover,
the culturing techniques require a considerable number of consumables, glassware,
and laboratory equipment. To overcome these shortcomings, a fast, cost-efficient,

software-based miniaturized plating method was reported with droplets of bacterial
cultures. The photographs of the bacterial droplets obtained using a digital camera
and further processed with “ImageJ” software were used for the estimation of viable
counts (Rajapaksha et al. 2019).
11.2.2 Biochemical Screening
Examining the nutritional and metabolic proficiencies by a biochemical screening

of an isolate is a common approach for the determination of its genus or species-
level identification. Many metabolites, specific to the target organism, are screened
through catabolic reactions between microorganisms and components of the sub-
strate to retrieve a data. Further, the matching of generated data with the available
extensive database helps in the identification. Biochemical testing can be automatic,
semiautomated, and miniaturized systems that takes short analysis time, and these
methods can concurrently determine results for many microorganisms while retain-
ing accuracy. While dealing with organisms in dairy microbiology, Vitek 2 Compact
(bioMerieux), BD Phoenix (BD Diagnostics), Analytical Profile Index (API) 50
CHL (bioMerieux), and API 20E (bioMerieux) are some common methods as given
in Table 11.1, which are used for analyzing the identity of microbes (Franco-Duarte
et al. 2019).
A study by Chifiriuc and coworkers indicated even closely related E. coli strains,
including the enterohemorrhagic E. coli O157:H7, were identified using a biochip
based on direct differential carbohydrate recognitions developed on a gold surface
which is based on surface plasmon resonance imaging (Bulard et al. 2015; Chifiriuc
et al. 2017). Intense research in the field of culturing of microbes developed several
rapid techniques that overcome the limitations of conventional culture-based tech-
niques. Some of the promising approaches are discussed briefly below.
Table 11.1 Developed biochemical systems for the identification of microorganisms in the dairy
industry
Microorganisms Biochemical methods References
Enterococcus spp. Vitek 2 API 20 STREP Chabros et al. (2015)
Lactobacillus spp. API 50 CHL Sunthornthummas et al. (2017)
Streptococcus spp. API 50 CHL Kaktcham et al. (2019)
Pediococcus spp. API 50 CHL Singla et al. (2018)
Yeast API 32 C Phoenix Pavlovic et al. (2014)
Candida Vitek 2 Rosenvinge et al. (2013)
Psychrotrophic microorganisms API Vithanage et al. (2014)
Staphylococcus spp. API 20 STAPH Koop et al. (2012)
Gram-negative bacteria Vitek 2 GN Crowley et al. (2012)
11.2.3 Biosensors
Biosensors consist of a receptor of biological origin for sensing and a transducer

(chemical or physical) for the detection of specific compounds. The physicochemi-
cal change produced by the specific interactions between the compound and the
sensing element is identified and quantified by the transducer, permitting instanta-
neous measurements both quantitatively and qualitatively (Fusco and Quero 2014).
The major attraction of biosensors is that they are sensitive indicators, which pro-
vide results directly for complex sample analysis with limited pretreatment like
purification (Mortari and Lorenzelli 2014). The biosensors can be broadly classi-
fied: (1) on the basis of the biological material as whole cell (i.e., microbial biosen-
sors), antibody (i.e., immunosensors), or nucleic acid (i.e., DNA-aptamer-based
sensors) (Alhadrami 2018) and (2) physicochemical change by the transducer as
electrochemical, optical, acoustic, piezoelectric, gravimetric, thermometric, or calo-
rimetric biosensors, etc. (Alhadrami 2018; Poghossian et al. 2019), as illustrated in
Table 11.2.
Many biosensors are portable analytical devices with simple instrumentation and
compact design, which can be used for on-the-spot test analysis. These devices have
the advantage of their simplicity, real-time analysis, high sensitivity, miniaturization
apparatus, speedy detection time, multiple analysis, and low cost, hence widely
Table 11.2 Classification of biosensors

Recognizing element Immunosensors Antibodies
Protein receptors Metabotropic receptors
Ionotrophic receptors
Enzymes
Whole cell Microbial sensors
Mammalian cells tissue
Nucleic Acids Hybridization
DNA-aptamers based biosensors
Signal transduction by transducer Electrochemical Amperometry
Conductimetry
Impedimetry
Potentiometry
Optical Absorbance
Fluorescence
Phosphorescence
Bio/Chemiluminescence
Reflectance
Raman scattering
Refractive index
Mass sensitive Surface acoustic wave biosensor
Microcantilever biosensors
Thermal Thermomechanical,
thermoresistive, thermocouple
Source: Alhadrami (2018)
Table 11.3 Some commonly used biosensors for pathogen detection in milk
Sample
and
volume Biosensor Biorecognition Detection
Microorganism used (transducer) element limit References
Escherichia 100 μL Electrochemical Horseradish 100 cells/ Laczka et al.
coli milk (amperometric) peroxidase mL (2011)
(HRP)
antibodies,
aptamer
Escherichia 2 mL Electrochemical Sandwich 6 CFU/mL Zelada-
coli skimmed (potentiometric) assay, HRP Guillén et al.
milk antibodies (2010)
Staphylococcus 50 μL Electrochemical Competitive 1 CFU/mL de Avila
aureus raw milk (amperometric) assay, HRP et al. (2012)
antibodies
E. coli O26 2 mL Optical (SPR) Antibody based 25 cells/ Waswa et al.
skim mL (2006)
milk
Salmonella 1 mL Mass sensitive Bacteriophage 5000 CFU/ Lakshmanan
typhimurium UHT (magnetoelastic) mL et al. (2007)
skim
milk
E. coli 10 μL Mass sensitive Antibody based 53 CFU/ Shen et al.
O157:H7 milk (piezoelectric mL (2011)
quartz crystal
microbalance)
Source: Mortari and Lorenzelli (2014)
a
CFU/mL colony-forming unit/milliliter
used in the determination of foodborne pathogens. Electrochemical, optical, and

mass-sensitive biosensors are the popular biosensors in dairy research as detailed in
Table 11.3 (Mortari and Lorenzelli 2014).
The electrochemical biosensors are the most popular biosensors used in milk
microbiology. Commonly, amperometric or potentiometric transducers are used for
pathogen detection in milk samples (de Avila et al. 2012; Muniandy et al. 2019).
They are inexpensive and unaffected by sample turbidity in milk (Mortari and
Lorenzelli 2014). The electrical current developed by the enzyme (alkaline phos-
phatase or horseradish peroxidase) oxidation of a substrate is monitored, and the
strength of the signal is proportional to the microbial load (Mortari and Lorenzelli
2014).
Optical biosensors are based on various properties like light absorbance, chemi-
luminescence, fluorescence, light polarization and rotation, surface plasmon reso-
nance (SPR), and total internal reflectance (Terry et al. 2005). Fluorescence and
surface plasmon resonance (SPR)-based methods are gaining popularity due to their
excellent sensitivity (Rajapaksha et al. 2019). Some of the commercially available
optical biosensors are BioFlash® (Rider et al. 2003), BIACORE Q, and Spreeta™,
which have been adopted for the identification of foodborne pathogens (Mortari and
Lorenzelli 2014).
Mass-sensitive biosensing devices commonly use piezoelectric quartz crystal
microbalance (QCM) and wireless biosensing magnetoelastic transducers for milk
microbiology testing (Shen et al. 2011). Mass changes even at nanogram levels can
be easily detected by QCM sensors.
Micro- and nanofabrication technologies are gaining momentum, and they help
in the miniaturization and designing of lab on chips (LoC) or biochips which are
based on Biomedical or Biological Micro (and Nano)-Electro-Mechanical Systems
(BioMEMS or bioNEMS) (Fusco and Quero 2014). LoC systems made up of
numerous microscale reaction chambers and channels are used to prepare samples
and to deliver analytes (e.g., bacteria, DNA, etc.) toward miniaturized embedded
sensing sites (Charbon 2008). Culture-based (based on changes in microenviron-
ment) and culture free techniques (based on antigenic properties of microorganism)
can be applied for LoC (Abbasian et al. 2018). Nanoparticle-based biosensor strate-
gies are nowadays used in the detection of foodborne pathogens (Bozal-Palabiyik
et al. 2018). The nanobiosensors have high selectivity and sensitivity and have the
scope for economical on-site analysis (Zhao et al. 2014; Ankireddy and Kim 2019).
11.2.4 Immunological Methods
Several rapid immunological methods rely on antigen-antibody specificity. The tar-

get part of antigen binds with the available epitopes of antibody. Some of the rapid
techniques based on immunological reactions include antibody-based microarray
typing platform, immunomagnetic separation (IMS), ATP bioluminescence immu-
noassay, immunoprecipitation reactions, enzyme immunoassays, enzyme-linked
immunosorbent assay (ELISA) (which uses antibodies and senses the color change),
enzyme-linked fluorescence assay (ELFA) (which uses antibodies and senses the
fluorescence—VIDAS system), multilocus enzyme electrophoresis (MEE), etc.
(Chifiriuc et al. 2017; Hameed et al. 2018).
11.2.5 Flow Cytometry
Flow cytometry is an optical detection-based method for analyzing individual cells

or a low number of cells in complex matrixes. The sample is injected into a liquid in
which microorganisms are suspended and then forwarded to the detection area
(laser beam). When the microbes passing through the orifice get detected by the
laser beam, the laser gets scattered into different patterns which are detected by
photocells. The scattering pattern determines the intrinsic properties of the microbes
and helps to determine the size, shape, and number (Rajapaksha et al. 2019). To
increase the specificity, photoacoustic technique is adopted in flow cytometry. The
photoacoustic flow cytometry (PAFC) is stimulated by light and senses the emitted
photoacoustic signals. These acoustic longer wavelengths are unaffected toward
scattering and weakening in biological media, thereby limiting autofluorescence of

background. The photoacoustic technique can be accepted to differentiate and rec-
ognize pathogenic microorganisms (He et al. 2015).
11.2.6 Adenosine Triphosphate (ATP) Bioluminescence
Adenosine triphosphate (ATP) is found in all living organisms and is the energy
currency of the cell. It is an excellent marker for viability detection as ATP is directly
correlated to the number of viable populations. ATP bioluminescence is based on
the enzymatic reaction of luciferin/luciferase in the presence of ATP, oxygen, and
magnesium. The emitted light is in the wavelength of the visible spectrum and a
peak at 562 nm (Shama and Malik 2013) which is measured by the luminometer.
ATP bioluminescence is used for rapid detection of contamination (presence/
absence test) in ultra-high treatment (UHT) products (Shama and Malik 2013) and
is helpful in the implementation of hazard analysis and critical control points
(HACCP).
D　 Luciferin + ATP + O2 → Oxyluciferin + AMP + PPi + CO2 + photon

LuciferaseMg2+

ATP detection methods can provide viable cell counts within a short time about
1–4 h, but specificity is less. The major shortcoming of the ATP detection method is
the occurrence of nonmicrobial or extracellular ATP creating a background signal
and an overestimation of the contamination (Poghossian et al. 2019). The presence
of somatic ATP in food samples may hinder the detection (Nemati et al. 2016). For
increasing accuracy, sample treatment to remove background ATP is needed. The
variability in the ATP content of microbial cells depending on the type and physio-
logical state also affects the results, which is also a drawback. Moreover, the quench-
ing of emitted light by the food matrix is another problem faced in ATP measurement
(Poghossian et al. 2019). ATP-based biosensors are now an area of interest to
develop an inline/online monitoring in the food industries.
11.2.7 Electrical Impedance Measurement
Microbial growth can induce changes in electrical characteristics of the medium

which is the basis of electrical impedance method (Wei et al. 2019). The change in
electrical conductivity, specifically the resistance to conductivity—impedance, is
measured. Some of the commercialized conductance/impedance-based instruments
are Bactometer™ (bioMerieux), Malthus™ (Malthus Instrument), Rapid Automated
Bacterial Impedance Technique (RABIT) system™ (Don Whitley Scientific), and
BacTrac™ microorganism growth analyzer (SyLab, Purkersdorf) (Bancalari 2017).
The instruments can operate in any of the three modes: conductance, impedance,
and capacitance. The major advantage of these systems is the measurement can be
correlated to the viability of the cells. The combination of electrical impedance with
biological recognition technology has contributed to the development of impedance

biosensors, which considerably reduced the detection time to approximately 30 min
compared with the traditional methods (Yang and Bashir 2008; Wei et al. 2019).
11.2.8 Electromigration Techniques
Electromigration techniques, which utilize the movement of ionized molecules

under the effect of a homogeneous electric field. The separation of a molecule is
based on their electrophoretic mobility (Buszewski et al. 2017). The cell surface
components like polysaccharides, phospholipids, membrane proteins, and receptors
create a z-potential on the surface of the cells. The most commonly used electromi-
gration techniques include gel electrophoresis (GE), two-dimensional GE, capillary
zone electrophoresis (CZE), capillary isotachophoresis, and capillary isoelectric
focusing (CIEF). Microbial growth can be monitored by measuring the z-potential
as the composition of the surface of living organisms can change during the growth
phase of the organisms. The main shortcoming of electrophoretic separation is that
microorganisms tend (positive charge groups) to agglomerate and interact with the
internal surface of the capillary (i.e., adhesion to the silanol groups). An electropho-
retic technique can also be used for the quantitative determination of microorgan-
isms and can be used to differentiate between Gram-positive and Gram-negative
bacteria and monitors the growth changes.
11.2.9 Multivariate Techniques
With the advancements, several techniques including the ionization and separation
can be combined with mass spectrometry (MS) such as gas chromatography (GC),
liquid chromatography (LC), matrix-assisted laser desorption/ionization time-of-
flight mode (MALDI-TOF), electromigration techniques (EM), and electrospray
ionization (ESI) (Franco-Durate et al. 2019), or combination with molecular tech-
niques like PCR electrospray ionization (ESI) MS, PCR-microchip capillary elec-
trophoresis (MCE), PCR-capillary electrophoresis (PCR-CE), capillary isoelectric
focusing (CIEF), etc. (Buszewski et al. 2017) are gaining popularity. MALDI-TOF
MS is the modern next-generation rapid tool used for the precise identification and
classification of microorganisms. The basic principle is the application of short laser
pulses for the gentle ionization of intact microbial cells and then acceleration of the
particles in a vacuum using an electric field (Sandrin and Demirev 2018). Unique
molecular fingerprint (spectra profile) of the microorganism as a result of microor-
ganism ionization is then compared against a database for the complete identifica-
tion (Buszewski et al. 2017; Sandrin and Demirev 2018). The major concern in the
identification is the quality and accuracy of the available databases. The amalgama-
tion of different techniques helps to overcome the drawbacks of individual tech-
niques, thereby enhancing the specificity and efficiency of detection.
11.2.10 Omics Techniques
Omics technologies have immense potential for a deeper understanding of the

microbial communities, microbial behaviors, their interactions, and cellular path-
ways. The evolving omics technologies like genomics, proteomics, transcriptomics,
metabolomics, etc., have revolutionized our approaches to the detection of food-
borne pathogens (Chawla et al. 2018). The screening of new probiotic or other
potential cultures can be done by recognizing the biomarkers through omic tech-
nologies. It can be used for assessing the physiological states of the microbes, to
improve functionality during industrial processes (Thierry et al. 2011). Every single
omic technology presents a very vast reading in itself, making it impossible to cover
in this chapter. Before reaching the practical uses, many technical and computa-
tional issues need to be addressed to deliver all stages of information (Chawla et al.
2018).
11.3 Culture-Independent Techniques
Over the past years, there is a radical change in the emergence of culture-independent
techniques. The limitations of culture-dependent approaches are overcome by
the culture-independent techniques—mainly the molecular techniques. The culture-
independent techniques circumvent the steps of enumeration and characterization
of microbes rather based on nucleic acid-based identification. In culture-independent
techniques, nucleic acid (deoxyribonucleic acid (DNA)/ribonucleic acid (RNA)) is
isolated and purified from the dairy matrix, and different molecular approaches are
applied to identify and quantify the pathogens (O’Sullivan et al. 2013). The useful-
ness of the molecular-based method depends to a great extent on the efficiency of
the isolation and purification of the DNA (quality and quantity of nucleic acid)
obtained from the food matrix.
The nucleic acid-based detection is based on the hybridization of target DNA
with specific DNA probes. Recent advances in molecular approaches provide rapid,
accurate, specific, sensitive, reliable, high-throughput, and non-biased strategies to
detect the microorganisms in dairy foods even their presence is at very low num-
bers. The detection of both viable and nonviable bacterial cells or even the damaged
or completely lysed cells could be done by the molecular techniques. Molecular
techniques provide more accurate taxonomic information about a particular strain
which plays a key role in accurate identification and surveillance of pathogens in
the dairy food. The genus, species, and strain level identification can be done by
targeting the specific regions of the bacterial genome. The availability of suitable
DNA extraction and purification protocol is the critical step in a culture-independent
approach. The major limitations of the molecular technique are the expensive equip-
ment and reagents required and high technical skill required to conduct the experi-
ments. Some important molecular techniques applied in the dairy industry are
briefly mentioned below.
11.3.1 Polymerase Chain Reaction (PCR)
Polymerase chain reaction (PCR) is one of the cutting-edge molecular techniques

that amplifies a specific DNA sequence (nucleic acid, or cDNA generated from
RNA as template) by using primers (universal, species specific, or genus specific)
by mimicking the in vitro replication conditions in the presence of thermostable
DNA polymerase (O’Sullivan et al. 2013).
PCR is a very promising, popular, and rapid molecular method for detecting and
identifying microorganisms in a variety of foods including dairy (Auvolat and Besse
2016). For bacterial identification, the 16S rRNA gene or the 16S/23S spacer region
is specifically targeted due to its highly conserved and variable regions (Ndoye et al.
2011; Quigley et al. 2011). The microbial diversity can be studied by the amplifica-
tion and sequencing of 16S rDNA and the intergenic spacer region. The sequencing
of the 16S rDNA helps in the assessment of the diversity of the community in terms
of the number of different species.
Even though discovered in the 1980s, PCR-based techniques are being regularly
used for the detection of pathogenic and spoilage microbes as well as for the char-
acterization of microbiota in the food products (Rajapaksha et al. 2019). The
International Organization for Standardization (ISO) has also recognized PCR for
the detection of foodborne pathogens (ISO 2005a, b, 2006a, b, 2011a, b). PCR is
also used to confirm characteristic colonies from agar plates, as specified by the ISO
(ISO 2007). The major limitations of PCR are the requirement of expensive instru-
ments and reagents, the presence of PCR inhibitors (proteins and fat) in the food
matrix may affect the results, and it cannot differentiate between viable and nonvi-
able cells (Cancino-Padilla et al. 2017).
11.3.2 Multiplex PCR
An advancement in PCR is the multiplex PCR which involves the simultaneous

detection or amplification of multiple target sequences in a single reaction by using
different primers for each target. The 16S rDNA and 23S rDNA variable gene
sequences, as well as the 16–23S rDNA intergenic spacer region, are focused on the
taxonomic identification of microorganisms. In dairy products, multiplex PCR has
been effectively used for the simultaneous identification of various lactic acid bac-
teria and variants of enteropathogenic Escherichia coli, to differentiate among
Listeria species, and to characterize strains of Staphylococcus aureus concerning
enterotoxin type (Naum and Lampel 2016). Since multiple species are targeted at
the same time, there are considerable time savings, less effort, and less risk of cross-
contamination (Cancino-Padilla et al. 2017).
Deeper insights into the mechanism of nucleic acid amplification lead to gaining
popularity of isothermal based nucleic acid amplification techniques as the need for
costly equipment like thermal cycler is avoided. This isothermal amplification of
nucleic acid occurs at constant temperature and can even be conducted in a water
bath. Some of the popular methods include nucleic acid sequence-based amplifica-
tion (NASBA), loop-mediated isothermal amplification (LAMP) (based on
auto-cycling strand-displacement amplification by B. stearothermophilus DNA

polymerase), strand displacement amplification (SDA), multiple displacement
amplification, rolling circle amplification (RCA), ramification amplification,
helicase-dependent amplification (HDA) (which mimics replication fork mecha-
nisms), or recombinase polymerase amplification (RPA) (which uses recombinase–
primer complexes), beacon-assisted detection amplification, etc. (Fusco and Quero
2014; Buszewski et al. 2017).
11.3.3 Quantitative Real-Time Polymerase Chain Reaction (RT

qPCR/qPCR/qRT PCR)
The quantitative PCR (qPCR) is considered as second-generation PCR and is an

advancement over conventional PCR due to real-time quantification, automation,
and in situ analysis (Riyaz-Ul-Hassan et al. 2013). In this technique, detection,
quantification, and analysis can be done simultaneously; during DNA amplification,
the fluorescent signals produced by DNA dyes and/or fluorescent probes in the reac-
tion mixture helps in real-time monitoring of PCR products. This technique permits
rapid identification and quantification of bacteria and revolutionized the molecular
approaches to food safety (Quigley et al. 2012; Naum and Lampel 2016). The qPCR
for the identification of foodborne pathogens in foodstuff has been approved, and
guidelines have also been developed by the ISO. The qPCR technique can be used
to quantify probiotics and to study the associations of starter cultures in fermented
milk products (Herbel et al. 2013; Miller et al. 2012).
In the dairy industry, qPCR and PCR techniques are more feasible to implement
since they are quick and cost-effective and do not need much skill to perform when
compared to other molecular techniques. Both culture-dependent and independent
approaches can be used in RT PCR. The efficiency of this method is severely affected
by the effectiveness of the protocol used for the isolation and purification of micro-
bial genomic DNA (Fusco and Quero 2014). The MIQE guidelines help to overcome
this difficulty by ensuring reproducibility and comparability of the data of qPCR
(Bustin et al. 2009). The major limitations are the high equipment cost and the lim-
ited sensitivity of qPCR in the different food matrix. It is not suitable for the detec-
tion of low number of microbial populations in the food (Auvolat and Besse 2016).
11.3.4 High-Resolution Melt (HRM) Analysis
High-resolution melt (HRM) analysis is an improvement of the PCR technique

based on melting kinetics of DNA. In high-resolution melt (HRM) analysis, the
amplification of the target sequence is performed in a real-time qPCR instrument in
the presence of a saturated fluorescent double-stranded DNA (dsDNA) intercalating
dye. The specific melting temperature (Tm) and melting profile of PCR amplicons
are analyzed for the identification of microbes (Agrimonti et al. 2019). In a study
conducted by Forghani et al. (2015), multiplex qPCR with HRM analysis was used
for the specific simultaneous detection of Bacillus cereus, Listeria monocytogenes,
and Staphylococcus aureus in the milk sample. It is a precise, sensitive, fast, consistent,
and cost-effective post-PCR method that can be used to study even a single nucleo-
tide polymorphism, to distinguish closely related species, to identify pathogen, and
to detect the food allergens (Franco-Duarte et al. 2019).
11.3.5 Digital PCR
Digital PCR (dPCR) or droplet digital PCR (ddPCR) is the third-generation PCR
which is an advancement of qPCR. In digital PCR measurements, the reaction mix-
ture is divided into a numerous separate small volume reaction (millions of indi-
vidual partitions), such that there is either zero or one copy of target DNA molecule
will be present in any individual reaction. The partition of samples can be done by
different platforms like Microwell plates, capillaries, oil emulsion, and arrays of
miniaturized chambers with nucleic acid binding surfaces. In digital PCR, the sam-
ple is apportioned into nanoliter-size samples by droplet generator and encapsulated
into water droplets, which are surrounded by the oil phase. Each partition is a sim-
ple endpoint reaction. Following PCR reaction, the number of “positive” partitions
(in which the sequence is detected) versus negative partitions are determined to
quantify the number of copies of a DNA molecule in the original sample (Naum and
Lampel 2016). The main advancement of digital PCR over qPCR is that it can per-
form absolute quantification without the need of a standard curve (White et al.
2009). It is less sensitive to PCR inhibitors and can also be multiplexed for the
simultaneous detection of several organisms (Hua et al. 2010; Zhong et al. 2011).
The dPCR can be used for the detection or quantification of microbes in food. The
absolute quantification of viable lactic acid bacteria in fecal samples were detected
by dPCR (Gobert et al. 2018). It is used to detect the incidence of even a single cell
of foodborne pathogens like E. coli O157:H7, Salmonella (Jany and Barbier 2008)
and Bacillus cereus in milk (Porcellato et al. 2016) and to detect the pathogens in
soft cheese (Gobert et al. 2018). Other examples like Bifidobacterium and
Lactobacillus in the breast milk of healthy women have been detected by the droplet-
based digital PCR (Qian et al. 2016).
11.3.6 Extreme PCR
Recent advancement in PCR is the integration of extreme PCR and high-speed

melting with real-time fluorescence monitoring for detection and genotyping.
Recently, Myrick et al. (2019), integrated the extreme PCR (30-s cycles) and high-
speed melting of PCR (5-s) which can be performed in about 1 min. Specific, effi-
cient, high-yield PCR with 0.42-s cycles (35 cycles in 14.7 s) has been reported by
Farrar and Wittwer (2015). This can be achieved by increasing the rate of reaction
by increased polymerase and primer concentrations as well as by increasing the
conduction rate by physical translocation of capillaries between water baths (Farrar
and Wittwer 2015; Myrick et al. 2019). To reduce time, two-temperature cycling
(simultaneous primer annealing and polymerase extension stages) is performed.
Extreme PCR will be a suitable method for the rapid detection of microbes
(Rajapaksha et al. 2019).
11.3.7 Reverse Transcriptase PCR (RT PCR)
RT PCR is a variant of PCR which mainly targets mRNA for amplification. RNA is
converted into complementary DNA (cDNA) by reverse transcriptase and further
amplification by either traditional PCR or reverse transcription (RT) PCR. RT PCR
is one of the common amplification method used to detect RNA for cell viability
assessment purposes (Davis 2014). Ribosomal RNA (rRNA) is stable for at least
48 h after the cell death, whereas messenger RNA (mRNA) is rapidly degraded and
has a short half-life that makes it a good indicator of cell viability. RT PCR is used
to measure the stress response of microbes that they may encounter during food
production/storage/distribution/consumption, to analyze the modulation of the
expression of the genes involved, and to detect virulence factors (Fusco and Quero
2014). RT PCR is also used to detect the viability of microbes in food (Davis 2014).
The major limitations are the difficulty to extract RNA molecules, high-quality
RNA requirement, highly sensitive, and the need of a very precise sample
preparation.
11.3.8 Viability PCR Approach: EMA-PCR and PMA-PCR
The viability PCR (vPCR) approach is used to quantify the viable and dead cells. In
viability PCR, the selective penetration of DNA intercalating agents like propidium
monoazide (PMA) and ethidium monoazide (EMA) is done into dead cells in which
membranes are compromised. This results in covalent binding with the DNA form-
ing a stable monoadduct and thereby subsequently prevents the amplification of
DNA by PCR. The cross-linked insoluble complex of EMA or PMA complex
formed in dead cells gets eliminated together with cell debris, which results in
selective extraction of DNA only from viable cells. The free unbound EMA or PMA
remaining in solution is also photolyzed and converted into hydroxylamine, which
is no longer capable of covalently cross-linking to the DNA (Fusco and Quero 2014;
Davis 2014). Both EMA and PMA are useful for the detection of the physiological
status of VBNC pathogens. Lopes and Maciel (2019) reported that PMA was more
effective than EMA in eliminating the dead cells.
Several pieces of the research reported that EMA-PCR and PMA-PCR showed a
very good correlations with the plate counts and viability especially in the quantifi-
cation of viable cells in probiotic products or viable LAB in the fermented milk
(Matijasic et al. 2010; Meng et al. 2010; Fittipaldi et al. 2012). PMA RT PCR is a
promising method which can be applied in the field of probiotic and pro-technological
bacteria (Meng et al. 2010; Sohier et al. 2014) and for the detection of pathogens in
milk and dairy matrices (Soejima et al. 2012). The major drawbacks are the chances
of insufficient membrane damage leads to error in the detection and difficulty to
distinguish membrane-damaged dead and injured/stressed cells.
11.3.9 PCR-Based Denaturing Gradient Gel Electrophoresis

(DGGE) and Temporal Temperature Gradient Gel
Electrophoresis (TTGE)
Denaturing gradient gel electrophoresis (DGGE) and temporal temperature gradient

gel electrophoresis (TTGE) methods use the sequence-specific separation of PCR-
amplified 16S rDNA fragments (Naum and Lampel 2016). The migration distance
of the PCR products in gels is used for the species differentiation. The extraction of
nucleic acids (DNA or RNA) and amplification of 16S rRNA genes followed by an
analysis of PCR products by a genetic fingerprinting technique are performed in
DGGE. DGGE is based on electrophoretic separation of DNA molecules in poly-
acrylamide gel that is the same length but has different nucleotide sequences
(Chifiriuc et al. 2017). The contrasting melting behavior is due to the differences in
GC content result in different denaturation properties. PCR-amplified double-
stranded DNA is subjected to electrophoresis under denaturing conditions (achieved
by a solvent gradient urea and formamide) (Naum and Lampel 2016). DGGE band-
ing pattern helps to visualize microbial genetic diversity and also helps to determine
the abundance of predominant microbial community members. DGGE has been
applied in food microbiology to investigate mainly the ecology of fermentation,
spoilage, and safety as well as product-specific microbial diversity (Cocolin et al.
2013).
A temperature gradient rather than a solvent gradient is used to denature the
DNA in temporal temperature gradient electrophoresis (TTGE). TTGE is used to
identify dairy microflora in various commercial dairy ecosystems, including com-
mercial starters, fermented milk samples, and different types of cheeses (Ogier et al.
2002). TTGE has been recognized as a simple and powerful method for identifying
species of bacteria even with single nucleotide polymorphism (Naum and Lampel
2016).
11.3.10 DNA Microarrays
DNA microarray consists of an arrayed series of covalently bound (on glass, silicon
chips, or even to microscopic beads) defined oligonucleotides known as probes. The
DNA array is suitable to the probe in a solution containing a mixture of labelled
nucleic acids and the binding of these “targets” to the “probes” by hybridization
(Cancino-Padilla et al. 2017). The hybridization of the immobilized probe-target
DNA is generally detected by employing fluorescence, and quantification can be
done by the amount of fluorescence generated by using the software. The target is
usually a specific gene or unique region of the bacterial chromosome or plasmid
which serves as a marker for specific bacterial strains or allelic variants of a gene
((Naum and Lampel 2016). DNA microarray can quantify the expression levels of
large numbers of genes simultaneously or genotype multiple regions of a genome
(Chifiriuc et al. 2017). Biochip using DNA hybridization is a robust technique for
the detection, typing, and identification of organisms as well as for analyzing
single-nucleotide polymorphisms (SNPs). The major limitation for its practical

application is the high cost of equipment.
11.3.11 Fluorescence In Situ Hybridization (FISH)
Fluorescence in situ hybridization (FISH) is a nucleic acid-based hybridization

technique for molecular typing. FISH involves the hybridization of fluorescently
labelled oligonucleotide probes with the target DNA (Bottari et al. 2006; Franco-
Duarte et al. 2019). It combines the simplicity of microscopy and the specificity of
DNA hybridization (Wei et al. 2019). FISH uses fluorescently labelled rRNA tar-
geted probes which helps in the detection of metabolically active probes directly
from food matrices.
11.3.12 Molecular Typing
Typing is used to discriminate between different bacterial types within a species.

Conventional typing is based on phenotypes, such as serotype, biotype, phage type,
bacteriocin typing, or antibiogram (Sabat et al. 2013). Later on various methods of
typing are being used like sodium dodecyl sulphate-poly acrylamide gel electropho-
resis (SDS-PAGE) of cellular and extracellular components, multilocus enzyme
electrophoresis (MLEE), mass spectrometry (MS), matrix-assisted laser desorption/
ionization-time of-flight (MALDI-TOF) MS, Infrared or Raman spectroscopy,
Fourier transform Infrared spectroscopy, etc. (Fusco and Quero 2014). Several
molecular methods are now used for the typing which reformed the identification of
bacterial types and subtypes.
11.3.13 Pulsed-Field Gel Electrophoresis (PFGE)
In pulsed-field gel electrophoresis (PFGE), the restriction endonucleases digest the

total DNA and generate a few numbers of large DNA fragments. The fragments are
separated during electrophoresis by the alternating electrophoretic current—
“pulsed” in different directions periodically across a time scale to resolve the large
size of DNA molecules. The advantages of PFGE are that it exhibits high discrimi-
natory power that is effectively differentiating at the strain level and it has intra-
laboratory reproducibility. PFGE has been described as the gold standard for
subtyping the genus and species to provide further discrimination among bacterial
pathogens (Sabat et al. 2013; Adkins et al. 2016; Cancino-Padilla et al. 2017). It is
the most frequently used molecular subtyping method for surveillance and outbreak
detection (Deng et al. 2016). The major limitations of PFGE are that it is labor-
intensive, time-consuming, and expensive; more technical expertise is required; and
there might be chances of interference with the food matrix. PFGE was employed
by PulseNet (www.cdc.gov/pulsnet), which is a network of health and food regula-
tory laboratories created by the Centers for Disease Control and Prevention (CDC)
for typing foodborne pathogens, better tracking, and earlier detection of possible
common source of outbreaks (Fusco and Quero 2014).
11.3.14 Random Amplification of Polymorphic DNA–RAPD-PCR
Random amplification of polymorphic DNA (RAPD) is done by a short universal ran-

dom arbitrary primers (8–12 nucleotides long) that binds nonspecifically to the tem-
plate bacterial DNA with low stringency. In RAPD amplification, random, repetitive
regions of the template DNA generates a unique profile, i.e., RAPD fingerprints in
agarose gel electrophoresis which is used for bacterial identification. It can also be used
as a tool to identify the genetic variability of microorganisms by focusing on DNA
polymorphisms (Franco-Duarte et al. 2019). It is used to type the lactic acid bacteria,
to monitor population dynamics in food fermentation, and to estimate the diversity of
lactic acid bacteria (LAB) in numerous foods. RAPD-PCR has been succeeded by
more effective typing techniques due to its low reproducibility and repeatability.
11.3.15 Restriction Fragment Length Polymorphism (RFLP)
Restriction fragment length polymorphism (RFLP) includes the amplification of a

target region by PCR followed by restriction enzyme digestion of the PCR product.
The agarose gel electrophoresis is used to generate a unique pattern of bands by the
separation of DNA fragments for each specific bacterial strain (Franco-Duarte et al.
2019). It is a combination of restriction endonuclease analysis of chromosomal
DNA (REA) with Southern blot hybridization (Fusco and Quero 2014; Chifiriuc
et al. 2017). It uses the presence of variations (polymorphisms) in homologous
DNA sequences to identify bacterial strains.
If a probe targeting rRNA conserved region (16S or 23S rRNA) is used, RFLP is
known as ribotyping (Grimont and Grimont 1986). The inherent ability of single-
stranded DNA molecules to hybridize with one another is the basic principle of
ribotyping. Ribotyping rRNA-based phylogenetic analysis is used for the bacterial
identification and characterization. The highly conserved region 16S rRNA and its
gene polymorphism give an idea regarding the evolutionary lineage of the bacterial
species, its classification, and taxonomy. Ribotyping mainly involves the restriction
digestion of genomic sequence of interest, followed by Southern blot transfer and
hybridization with probes (labelled 16S, 23S, or 5S rRNA gene probes), and analy-
sis of ribotype RFLP bands (Franco-Duarte et al. 2019). RFLP and automated ribo-
typing helps in the identification or typing of bacterial strains including pathogens
and spoilage causing microorganisms in food.
11.3.16 Amplified Fragment Length Polymorphism (AFLP)
Amplified fragment length polymorphism (AFLP) is almost similar to the RFLP. In

AFLP, usually, a pair of restriction enzyme is used to fragment genomic DNA. In
AFLP, double-stranded adaptors are specifically ligated to one of the sticky ends of
restriction fragments followed by selective amplification of fragments with ligated
adaptors. The amplification is achieved by using primers that are complementary to the
adaptor sequences but also have a certain unique nucleotide sequence which results in
selective amplification of few numbers of restriction fragments (Sabat et al. 2013;
Franco-Duarte et al. 2019). AFLP fingerprints generated in gel electrophoresis from

a single bacterial genome can be analyzed using a computer-assisted comparison of
generated high-resolution banding patterns. The determination of genetic related-
ness or dissimilarity up to strain level (Mortimer and Arnold 2001) can be deter-
mined by AFLP.
11.3.17 Amplified rDNA Restriction Analysis (ARDRA)
Amplified rDNA restriction analysis (ARDRA) is based on the rDNA-based finger-

printing. A fingerprint after the amplification of bacterial rRNA gene(s) by PCR and
digestion with restriction endonuclease enzyme(s) is then resolved by the gel or
capillary electrophoresis to obtain a fingerprint. It is used for the taxonomic identi-
fication of bacterial populations (Ranjard et al. 2000).
11.3.18 Repetitive Element-Polymerase Chain Reaction (rep-PCR)
Repetitive extragenic palindromic element PCR (rep-PCR) is a PCR-based molecu-

lar typing method. In rep-PCR, the primers hybridize the noncoding intergenic
repetitive sequences distributed across the genome and DNA between adjacent
repetitive elements. Depending on the distribution of the repeat elements, multiple
amplicons are produced which are then separated by the gel electrophoresis. The
banding pattern (fingerprints) determines the genetic relatedness between the ana-
lyzed bacterial isolates. The “enterobacterial repetitive intergenic consensus”
(ERIC), the “repetitive extragenic palindromic” (REP), and the “BOX” sequences
have been used successfully for typing (Versalovic et al. 1994; Sabat et al. 2013).
These methods are more reliable and robust than RAPD-PCR to type the pathogens
(Fusco and Quero 2014).
11.3.19 Multilocus Sequence Typing (MLST)
Multilocus sequence typing (MLST) was developed using the concept of multilocus
enzyme electrophoresis (MLEE) (Selander et al. 1986). Multilocus sequence typing
(MLST) involves the PCR amplification and sequence analysis of usually seven
housekeeping genes (internal sequence of 450–500 bp). The seven species-specific
genomic loci present in all strains have been targeted in the MLST (Chifiriuc et al.
2017). The housekeeping genes in most of the bacteria have enough variation within
to provide many alleles per locus to generate billions of distinct allelic profiles.
Based on the combination of allelic profiles generated, the sequence type is deter-
mined. The allelic sequence is then compared with the online database for the deter-
mination of the genetic relatedness between bacterial strains within a species as
well as MLST maps to track the isolates (Sabat et al. 2013). MLST results are
highly reproducible, unambiguous, and the most reliable. To improve the discrimi-
natory power of MLST, multi-virulence-locus sequence typing (MVLST) is per-
formed which is based on virulence genes. To improve the effectiveness of MLST
with the combination of metagenomics, MetaMLST has been developed (Zolfo
et al. 2017).
11.3.20 Whole-Genome Sequencing (WGS) Techniques
The next-generation sequencing (NGS) technology can sequence the complete

microbial genome and gave a deeper insight into understanding microbes at molec-
ular levels. A recently popularized and affordable and promising tool for bacterial
genotyping is whole-genome sequencing (WGS). The complete nucleotide sequence
of a genome is constructed by assembling multiple short sequence reads based on
overlapping regions (de novo assembly) or comparisons with previously sequenced
“reference” genomes (resequencing) (Sabat et al. 2013). The WGS provides insights
into bacterial typing, evolutionary lineages, pathogenic identification and character-
ization, and identification of the source of contamination (Deng et al. 2016; Franco-
Duarte et al. 2019). The biggest challenge is the identification of the nucleic acid
signature for each microorganism. 16S rRNA gene sequencing is considered the
most accurate method and deemed the gold standard for the identification of micro-
organisms at the species level (Deggim-Messmer et al. 2016). With the advent of
newer sequencing platforms, the price for genome sequencing is not becoming a
limiting factor. Genome sequencing can become a routine surveillance system
shortly in near future (Chawla et al. 2018).
11.4 Conclusions
In recent years, we have witnessed considerable technical improvements in existing

approaches for the detection of microbes. A plethora of rapid methods are available
for the detection, identification, and typing of milk- and dairy-related microbes. The
conventional methods are still preferred due to affordability, reliability, and user-
friendliness. There is a major trend toward the automation of different methods,
increasing the throughput, resolution, and accuracy. With the advancement of smart-
phones in our everyday activities, the practicability of application of the smartphone
apps for real-time detection of microbes is under investigation.
There is no single method that is suitable for the detection of microbes. In recent
years, there has been an increase in the use of a combination of analytical tech-
niques for the faster and more effective identification of microorganisms. Each
method has its advantages and disadvantages. The major drawbacks of molecular
approaches are expensive equipment and reagents and the requirement of skilled
staff. The combined use of various advanced and basic techniques help in better and
precise identification of bacterial isolates than a single method. Rapid progress in
the field of molecular biology has enabled the development of new methods like
WGS. However, these high-throughput methods are still not affordable for routine
analysis. Moreover, the high cost associated with implementation, technical com-
plexity, and validation difficulties are major concerns for the popularization of these
techniques. The enormous data generated by high-throughput technologies should
be properly interpreted and saved for the future applications.
Rapid detection and identification of microorganisms is a challenging and impor-
tant aspect of the dairy and food industry. The accomplishment of conclusive results
in a very short time, online microbial detection, and real-time and online monitoring
in industries are some areas that have great potential for the further research. It can
be summarized that rapid methods and automation should be continue as an ever-
evolving field of research to develop potable, cheap, easy-to-use, on-line or on-site
devices for the future. However, the main issues to be considered are the scale-up of
developed methods and their regulatory requirements.
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The Role of Yeast and Molds in Dairy
Industry: An Update 12
Nancy Awasti and Santosh Anand
Contents
12.2 U se of Yeasts and Molds in Dairy-Based Fermented Products 245
12.2.1 Yeasts in Fermented Products 245
12.2.2 Molds in Fermented Products 246
12.2.3 Enzyme Production and Dairy Waste Management 246
12.2.4 Use of Yeasts as Probiotics in Dairy Products 247
12.3 Use of Yeast and Molds in Utilization of Dairy-Based Byproduct such as Whey 247
12.4 Dairy Product Spoilage by Yeasts and Molds 249
12.4.1 Defects Caused by Yeasts and Molds 250
12.5 Sources of Contamination and Control Strategies 252
12.5.1 Good Manufacturing Practices 254
12.5.2 Decontamination and Purification of Air 254
12.5.3 Disinfectants Against Fungal Spoilage 255
12.5.4 Low-Temperature Storage 255
12.5.5 Antifungal Agents and Chemical Preservatives 256
12.5.6 Packaging and Physical Methods 257
References 258
Abstract
Over the past few decades, understanding of yeasts and molds diversity in envi-
ronment has grown appreciably. Due to high adaptation capability of these fungal
members, they are ubiquitously found in the environment. Decades of research
N. Awasti (*)
e-mail: nancyawasthi10@gmail.com
S. Anand
Dairy Microbiology Department, PJM College of Dairy Technology,
Parkheta, Jharkhand, India

https://doi.org/10.1007/978-981-15-2608-4_12
244 N. Awasti and S. Anand
helped humans to recognize and differentiate them into good or bad ones with
their ability for use as food products. During evaluation, milk or their structural
ingredients were found as one of the efficient matrices to extract profitable things
rather than signifying them as illness or spoilage entity. Over the past few years,
commercial cultures of yeast has become increasingly popular to produce fer-
mented milk, cheese and ethanol from whey and lactose solutions. Several mold
species also find their use in the development of different dairy products. Different
mold species are used for ripening of various types of cheese (e.g., Roquefort,
Camembert), enzyme production, and dairy waste management. However, fungal
spoilage is among one of the major challenges for food and dairy industry and
continues to grow with increased demand and production of food and dairy prod-
ucts. Systematic approaches like application of good manufacturing process, new
biopreservation techniques, rapid detection methods, and upgraded packaging
methods were surely able to reduce their inhibitory effects in the future.
Keywords
Yeast · Molds · Starters · Dairy · Utilization · Spoilage
12.1 Introduction
Yeast and mold are microorganisms of great importance to the food industry. In the
modern world, both yeasts and molds are equally been used for preparing various
nutritious food products. Yeast and mold are physiologically different from bacteria
and comparatively both are more commonly considered as a sign of foodborne ill-
ness, but pioneering knowledge towards identification techniques of the species and
their metabolites provided greater opportunities to make use of it. Years of research
recognized both yeasts and molds to contain members of beneficial as well as spoil-
age nature. Those species that conform to the criterion of being generally recog-
nized as safe (GRAS) are selected for product processing with further assessment of
their profitable properties in terms of process performance and impact on final prod-
uct quality.
Milk consists of an aqueous colloidal suspension of proteins, fat, and carbohydrates
with various vitamins and minerals, which makes it extremely nutritious food. Milk
matrix provides favorable niche for their growth. In milk-based product, utilization of
either organism for beneficial purposes needs an expertise or more commonly it was an
artisan stuff. The fermentation methods for producing yeast- or mold-based dairy prod-
ucts are extremely old and refined by producers over a long time period after experience,
trial and error. Ultimately, the manufacturers successfully calibrated the condition and
fermentation process using yeasts and molds to achieve the desired product, and still
modifications were implemented timely. Over the past few years, commercial cultures of
yeast has become increasingly popular to produce fermented milk, cheese and ethanol
from whey and lactose solutions. Apart from producing dairy products and their
12 The Role of Yeast and Molds in Dairy Industry: An Update 245
byproducts, few mold species are also used in the development of various mold-ripened
cheese (e.g., Camembert, Roquefort), enzyme production, and dairy waste management.
The present chapter aims to highlight the specific constructive as well as destructive roles
of yeasts and molds in dairy sector.
12.2 U
se of Yeasts and Molds in Dairy-Based Fermented
Products
Microorganisms are vital component of dairy starter varieties and play an important
role in the preparation of dairy fermented foods. They are used as both starters and
secondary flora. In modern dairy industries, yeast and molds find their application
both as starters and secondary flora (Fröhlich-Wyder et al. 2019). All yeast-lactic or
mold-lactic fermented milk products like kefir and viili, respectively, are majorly
demanded products based on both groups of microorganisms as starters. Consequently,
secondary flora used in the development of various cheese varieties like Roquefort,
Camembert, Brie, and Tilsit portrays interactions of yeasts and molds.
12.2.1 Yeasts in Fermented Products
Yeasts are unicellular organisms in fungal group. These yeast species are usually
exploited as a monoculture bioprocessing organism and generally used to produce
fermented beverages and fermented dairy and food products like cheeses, breads,
dairy drinks, and other products. Historically, yeasts are the most common microor-
ganisms exploited by man. In dairy industry, yeast utilization spans from the pro-
duction of fermented alcoholic beverages and smear-ripened cheeses to probiotics.
Yeasts contribute a prominent role in the production of alcoholic beverages since
ancient times, due to the ability to accumulate high levels of ethanol and to produce
aroma compounds. Kefir, koumiss, and Acidophilus-yeast milk are typical examples
of such beverages. Kefir grains contain yeasts like Saccharomyces, Kluyveromyces,
Candida, Cryptococcus, Pichia, Torulaspora, and Mycotorula species, which may
also vary according to region (Bengoa et al. 2019). Koumiss, another traditional
drink from Central Asia which belongs to this category, contains lactose-fermenting
(Torula koumiss) and non-fermenting yeasts (Saccharomyces cartilaginosus).
In cheese, community of yeasts reflects cohabitation ecosystem with bacteria
and filamentous fungi. Yeast present in raw milk or in processing environment usu-
ally colonizes the cheese surface and forms a biofilm complex with other microor-
ganisms. The presence of yeast substrates like nitrogen compounds and lactic acid,
produced during proteolysis and glycolysis by starter cultures of lactic acid bacte-
ria, helps in the establishment of yeast. According to recent report (Monnet et al.
2015), yeast species can deaminate or decarboxylate amino acids and oxidatively
metabolize lactic acid to the corresponding products, which lead to surge in
pH. Yeast species significantly contribute to the appearance and flavor of cheeses
like Reblochon, Gruyere, Tilsit, and Munster. The yeast species differs with
preference and type in cheese manufacturing. Yeasts such as Debaryomyces hanse-

nii (surface ripened), Geotrichum candidum (filamentous yeast in white mold
cheese), Saccharomyces cerevisiae, and Kluyveromyces lactis (in blue-veined and
white-mold soft cheeses) are most common species (Cantor et al. 2017; Irlinger
et al. 2017). In specific case such as production of acid-curd cheeses, few yeast spe-
cies like Candida krusei and D. hansenii (German-Harzer and Czech Olomouc
cheeses) can be added to milk that serve as a rapid yeast-driven fermentation
(Fröhlich-Wyder et al. 2019).
12.2.2 Molds in Fermented Products
In dairy industry, molds are used in preparation of surface-ripened and internal-

ripened cheeses. These molds are an integral part of cheese-making process, not
actually as starters but in conjunction to starter cultures. The main role of molds
during cheese manufacturing is to produce desirable flavors, appearance, and tex-
ture of finished cheese. Internal mold-ripened cheese such as Stilton, Roquefort,
Gorgonzola, and Danish Blue has blue veins over the curd particles inserting inside
while surface- or external-ripened cheeses such as Brie and Camembert have molds
grown on surface as fuzzy mass. Common molds like Penicillium, Cladosporium,
Mucor, Geotrichum and Sporotrichum are commonly used in cheese making. Under
controlled condition, molds release their lipolytic and proteolytic enzymes which
on substantial action produces ammonia, amines, and other desirable flavor com-
pounds (Desmasures 2014). Various flavoring agents such as variety of methylke-
tones are generally produced after hydrolyzing milk fatty acids by lipases (produced
via β-oxidation pathways).
Other than cheese making, molds are involved in mold-lactic fermentation for pro-
ducing fermented products. A traditional example of this fermentation is viili, a Finnish
fermented dairy product. Mold G. candidum is commonly used with initial lactic starter
Lactococcus lactis biovar diacetylactis for the desired fermentation. Growth of G. can-
didum gives velvet-like appearance to the product (Beniwal et al. 2019).
12.2.3 Enzyme Production and Dairy Waste Management
In the dairy business, yeasts and mold species play a significant role in enzymes
production and are used to develop and boost organoleptic properties like color,
flavor, and aroma of milk products. Enzymes such as aminopeptidase, proteases,
lipases, lactoperoxidase, transglutaminase catalase, etc. are extracted from molds
like Aspergillus, Kluyveromyces, Rhizopus, and Saccharomyces spp. The major
application is recognized in dairy industry for cheese ripening, hydrolyzing whey
protein and flavor development. Furthermore, proteases were also used for the pro-
duction low allergenic milk proteins to use them as an ingredient in infant formula
and beverages (Feijoo-Siota et al. 2014).
Dairy waste water generally contains high levels of organic loads and other pol-
lutants that are harmful to the environment. Dairy industries produce wastes such as
effluents and sludge during manufacturing of different dairy products and usually
contain high levels of oils, proteins, fatty acids, grease, nitrogenous compounds and
other organic matter (Porwal et al. 2015). Studies reported that yeasts (Candida,
Saccharomyces, and Cryptococcus strains) and molds (Aspergillus niger, Mucor
hiemalis, and Galactomyces geotrichum) increased the degradation process and
reduces the cost in biological treatment (Kushwaha et al. 2011; Goli et al. 2019).
12.2.4 Use of Yeasts as Probiotics in Dairy Products
Many yeast species like Kluyveromyces lactis, Torulaspora delbrueckii, Saccharomyces

boulardii, K. lodderae, Yarrowia lipolytica, and K. marxianus have been reported to be
endowed with potential like tolerance to acidic and other severe conditions of gastroin-
testinal track and other probiotic traits (Vandenplas and Benninga 2009). Among all
yeast species, S. boulardii is one of the most explored and popular probiotic yeasts. S.
boulardii was discovered by Henri Boulard in 1923 from the lychee skin (Litchi chi-
nensis, a Southeast Asian fruit), and further research on this species evolved into
assessment of its bio-therapeutic attributes (Sazawal et al. 2006). Along with evolving
evidence of its high value, the research acquired rapid momentum into its identification
and characterization after its isolation from numerous food matrices. It plays a role in
the management of several types of diarrhea and other gastrointestinal issues. Therefore,
it has a growing range of applications in preventing and curing conditions, for example,
candidiasis, inflammatory bowel disease (IBD), and blastocystosis.
It is a nonpathogenic and distinctive yeast supplement that has the ability to sur-
vive high acidic gastric liquids and enzymes and is uninhibited and unaffected by
most antibiotics. According to the study of McFarland (2010), S. boulardii exhibit
advantageous health effects via different mechanisms, such as by inducing growth
of probiotics, production and release of antimicrobial toxins, and digestive enzymes
(sucrase, lactase, and maltase); by amplifying the simulation and production of
immunoglobulins, short chain fatty acids, and other immunological factors, and
growth of intestinal cells; and also by reducing pro-inflammatory cytokines. It is
generally administered in lyophilized powder made in skim milk powder. The
lyophilized powder of S. boulardii has been used as an additive in food and only
limited research is reported, such as additive in yoghurt (Karaolis et al. 2013) or
fermentation of raw materials of vegetable (Campbell et al. 2016).
12.3 U
se of Yeast and Molds in Utilization of Dairy-Based
Byproduct such as Whey
Utilization of whey is considered as a challenge and is an effluent with high organic

load corresponding to higher chemical and biological oxygen demand. Organic load
of whey from different sources may contain variable amounts of milk carbohydrates
such as lactose and other sugars in small amount, proteins and lipids, respectively.
These dairy effluents are usually produced during processing of yoghurt, butter, ice
cream, and cheese. Therefore, the variable nature of such effluents may pose severe
environmental risk (Rivas et al. 2011). The current scenario of dairy effluent utiliza-
tion is challenging and requires a deep understanding of different constituents based
on their source of production. To deal with these dairy effluents, different biological
and physiochemical processes are being used (Kushwaha et al. 2010). In compari-
son to other dairy effluents, cheese effluent represents a significant impact on envi-
ronment because of its higher organic load (Prazeres et al. 2012) and is regarded as
most contaminated waste produced during production of cheese (Rajeshwari et al.
2000). Approximately 4.5–6.0% of lactose and 0.6–1.1% proteins are present in
cheese whey.
Cheese whey can be used to produce bioethanol, crystallized lactose, glucose,
galactose, and energy sources such as hydrogen gas, lactic acid, etc. The bioreme-
diation of polluting cheese whey is alcoholic fermentation, and it is an interesting
alternate to produce ethanol from lactose and other whey-based fermented drinks.
There are many commercially available microorganisms that can be employed to
produce or ferment cheese whey into valuable byproducts. Several microbes such as
yeast and bacteria have been frequently reported to produce ethanol via lactose
fermentation (Guimarães et al. 2010). Since the 1940s, a significant increase in
production of ethanol has been reported by using yeast fermentation. But there
exists very less information on different species and strains of yeast that can ferment
lactose rather than assimilating it inside their cell (Fukuhara 2006). As per previous
report (FAO 2012), the annual production of whey was 160 million tons which
means approx. 8 million tons of lactose. According to Ling (2008), the United States
during 2006 has utilized 55% of whey to obtain lactose. During the past 3 decades,
several studies have addressed production of ethanol from lactose after hydrolyzing
it to glucose and galactose by using yeast fermentation. Yeast species that are asso-
ciated with ethanol production belongs to Kluyveromyces and Candida species, e.g.,
K. fragilis, K. marxianus, and Candida pseudotropicalis.
Besides various laboratory-based research, there are few cases of ethanol pro-
duction at industrial scale from permeate and whey by using Kluyveromyces spe-
cies. Apart from Kluyveromyces species, K. lactis is the most explored and exploited
species of yeast (Breunig et al. 2000). The potential of lactose utilization by
Kluyveromyces lactis has been employed for ethanol and heterologous proteins pro-
duction by using cheese whey (Fukuhara 2006; Van Ooyen et al. 2006). The genes
involved in lactose metabolization are LAC12 and LAC4 gene that produces lactose-
hydrolyzing enzymes such as β-galactosidase and lactose permease. Apart from lac-
tose hydrolysis, K. lactis has been also exploited for other biotechnological
applications. One of the significant advantages of using Kluyveromyces species is
that few species like Kluyveromyces marxianus can grow and ferment lactose and
glucose at elevated temperatures of >40 °C (Kourkoutas et al. 2002a; Nonklang
et al. 2008). Besides production of ethanol, as per few studies, Kluyveromyces spe-
cies is also involved in production of alcoholic beverages from cheese whey such as
whey wines, distilled drinks, and low alcohol content drinks (Kosikowski and
Wzorek 1977; Kourkoutas et al. 2002b; Dragone et al. 2009).
Among lactose-fermenting yeast, Candida pseudotropicalis strain is one of the
most efficient microorganisms to produce ethanol from lactose in whey. But at higher
lactose and ethanol concentration, inhibition of both strains of Kluyveromyces and
Candida were reported (Szczodrak et al. 1997). Therefore, industrial processes usu-
ally use yeast strains that are more tolerant to ethanol and possess good fermentative
capacity. Usually, Saccharomyces cerevisiae is the most preferred species for com-
mercial production of ethanol at industrial scale. Apart from tolerance against etha-
nol, Saccharomyces cerevisiae is GRAS for human consumption and can produce
more than 20% (v/v) ethanol with ability to rapidly grow at anaerobic conditions
(Antoni et al. 2007; Cot et al. 2007; Ishtar Snoek and Yde Steensma 2007).
Saccharomyces cerevisiae is one of the most studied species of Saccharomyces and
has been used as a biomass in animal feed (Bai et al. 2008). The advantage of using
Saccharomyces over Kluyveromyces species is catabolism of galactose via Leloir
pathway by using GAL2 gene encoding galactose permease (Nehlin et al. 1989). But
in order to produce ethanol, lactose is usually hydrolyzed using enzymatic activity of
β-galactosidase to produce mixture of glucose and galactose. In presence of glucose,
Saccharomyces cerevisiae and few other strains of yeast prefer to use glucose instead
of galactose until it is exhausted in the medium and this phenomenon is called as
catabolite repression. Thus, the drawback of Saccharomyces cerevisiae to prefer glu-
cose over galactose and additional step of β-galactosidase activity makes it more
challenging to utilize it for industrial processes (Gancedo 1998; Mehaia and Cheryan
1990). Previous studies (Bailey et al. 1982; Oda and Nakamura 2009) reported use of
2-deoxyglucose, an agent to produce mutant strain of Saccharomyces cerevisiae and
Kluyveromyces marxianus to utilize glucose and galactose simultaneously.
12.4 Dairy Product Spoilage by Yeasts and Molds
With growing demands of food and dairy production, spoilage, food waste, and
losses became significant issue for economy and human health. Every year about
1.3 billion tons of food is spoiled or wasted before consumption, and food loss due
to fungal deterioration is estimated between 5 and 10%. The losses and waste due to
spoilage by fungal group occurs mainly due to gaps between production and con-
sumption (FAO 2017). The spoilage by these microorganisms is associated with
their nutritional needs, various enzymatic activities, and the potential to grow at
refrigeration temperatures, acidic environment, low water activity, and high salt
concentrations (Garnier et al. 2017). Various varieties of fungal species have been
identified and isolated from milk and milk products. In Ledenbach and Marshall
(2009), fungal spoilage is mostly experienced when dairy products are used with
either nuts, sugar, or fruits, thus providing nutrients to fungal growth and
fermentation.
In comparison to other food products, milk and milk products are less suscepti-
ble to spoilage, as they are mostly held at refrigerated conditions and are also
thermally treated using either pasteurization or other heat treatment techniques.

Despite all these, substantial number of yeast and mold species can live through
these processing steps and thus can grow in milk products. This astonishing resis-
tance and adaptation potential of fungal species allows it to use various substrates
like lipids, organic acids, carbohydrates, as well as proteins that are generally pres-
ent in dairy and milk products (Huis in't Veld 1996). With reference to unwanted
fungal species, their existence in milk products may result in numerous changes in
product, e.g., detectable fungal growth on surface, off-flavor and odors caused by
fungal metabolite production, and a change in color and texture (Ledenbach and
Marshall 2009). Few heterogeneous groups of yeast species such as Debaryomyces
hansenni, Saccharomyces species, and Candida species are involved in spoilage of
chesses, yogurt, and other milk products and are resistant to heat processing.
Few species of yeast like Brettanomyces can form volatile phenolic compounds
that may lead to development of off-flavors in fermented dairy products (Steensels
et al. 2015). Most mold species grow at very low water activity (0.7–0.8) on dried
foods and can grow at wide pH range from 3.0 to 8.0. Different mold species have
variable range of optimal growth requirements like temperature and other nutri-
tional needs and grow at refrigeration temperatures. Mold species are usually sensi-
tive to heat treatments, and they cause spoilage by producing resistant spores and
toxic mycotoxins (Pitt and Hocking 2009). Molds and yeasts are among main spoil-
age microorganism found in acidic milk products like cultured milks (sour cream,
yoghurt, and buttermilk) inhibiting most of the bacterial population (Mayoral et al.
2005).
12.4.1 Defects Caused by Yeasts and Molds
Various defects caused by yeast and mold species include off-flavors, texture, and
organoleptic changes (Snyder and Worobo 2018). Yeast spoilage of milk products
can be easily identified due to visible alterations on surface of dairy product such as
browning, and defects like toad skin mainly caused by Yarrowia and Galactomyces
species. Spoilage from yeast may also result into nonvisible but detectable altera-
tions, e.g., off-flavor, change in texture, or off-odor due to formation of carbon diox-
ide, proteolytic and lipolytic enzymes, and other volatile compounds (alcohols,
aldehydes, ketones, and esters). Although in relation to spoilage of dairy products
from yeast, only few studies have been reported. A study by Ledenbach and Marshall
(2009) defined that visible changes by yeast spoilage can be detected when their
population or log numbers are equal or more than 5.0–6.0 log cfu/g. Several studies
have reported the production of biogenic amines during cheese production. As an
example, studies done by Linares et al. (2012) and Roig-Sagués et al. (2002)
reported production of tyramine and histamines from traditional Italian cheese.
These biogenic amines are nitrogenous compounds and play an essential role in
human health, but their higher production may also cause toxicity. It is worth men-
tioning that not all species of yeast produces biogenic amines, but few species like
Galactomyces geotricum do possess the potential to degrade tyramine (Leuschner

et al. 1998).
Over the years, numerous studies have reported different types of spoilage from
yeast in dairy and dairy products. A comprehensive review by Garnier et al. (2017)
has elaborated the spoilage by yeast and mold species with more emphasis on their
diversity. In relation to yeast spoilage, more than 60 species have been linked with
the spoilage of milk and dairy products. Among various genera of yeast species,
Candida is one of the most commonly reported. About 24 different species of
Candida are linked to spoilage of dairy products such as hard or semi-hard cheeses
and unripened dairy products. Besides Candida species, other yeast species associ-
ated with spoilage include Saccharomyces, Debaryomyces, Yarrowia, Crytococcus,
Rhodotorula species and others (Table 12.1). All abovementioned species are linked
with the spoilage of yoghurt, blue-veined cheese, fresh unripened cheese, butter and
margarine, raw milk, and other dairy products (Fadda et al. 2004; Fleet 1990;
Minervini et al. 2001; Pitt and Hocking 2009; Suriyarachchi and Fleet 1981; Von
Neubeck et al. 2015).
Mold spoilage in dairy products is mostly related with vacuum packed cheese,
heat-treated milk, and milk products and other cheeses. Generally, mold spoilage
results into visible changes on surface of the product or in cheese package such as
moldy growth in wrinkles and folds of the plastic film (Hocking and Faedo 1992).
Despite causing visible alterations on dairy products, molds are also linked with the
production of off-flavors due to production of ketones resulting in rancid flavor in
butter, whereas other species of mold like Penicillium and Cladosporium species
leads to off-flavor due to formation of geosmin and 2-methyllisobornol linked with
strong earthy and musty notes (Pitt and Hocking 2009). In comparison to yeast,
Table 12.1 Defects caused by different yeast and mold species in dairy products
Fungal species
Defect Description involved References
Thread mold Fungal growth in vacuum Mycelial growth of Hocking and
packaging, often associated several common Faedo (1992)
with cheese molds: Phoma,
aspergilli, penicillia
Browning Visible changes on product Yarrowia lipolytica Carreira et al.
reaction surface caused by formation of (2001)
pyomelanin
Toad skin A visible change in texture on Galactomyces Carreira et al.
product surface geotrichum (2001)
Rancidity Caused by production of Cladosporium Pitt and Hocking
ketones herbarum (2009)
Musty or Caused by formation of Penicillium and Pitt and Hocking
earthy flavor 2-methlisoborneol and geosmin Cladosporium species (2009)
Plastic-like or Metabolites like trans-1,3- Mold species Sensidoni et al.
“kerosene” pentadiene or trans-piperylene (1995), Sensidoni
off-flavor produced from sorbate salts in and Narvhus
cheese and margarine (1996)
mold spoilage in dairy product is more prominent and about more than 100 species
of molds have been identified to cause spoilage in dairy products (Table 12.1).
Among all molds, Penicillium genus with approximately 40 species and Aspergillus
genus with nearly 10 species are most frequently reported to cause spoilage of milk
and dairy products.
Apart from causing spoilage in hard and semi-hard cheeses, Penicillium species
are also associated with fresh and mold-ripened chesses, butter, yoghurt, blue
veined, as well as ewe, buffalo, or goat milk cheeses. Few species of Penicillium
species such as P. antarticum, P. fellutanum, and P. charlessi are well adapted to
extreme conditions and are called as xerophilic species and have been reported to
cause spoilage of different cheese (Garnier et al. 2017; Hocking and Faedo 1992).
Unlike yeast, few mold species like Aspergillus, Fusarium, Penicillium, and
Hamigara genera have been frequently isolated from heat-treated milk and milk
products such as cream cheese, pasteurized milk, and other heat-treated dairy prod-
ucts (Garnier et al. 2017; Hocking and Faedo 1992; Jodral et al. 1993). The occur-
rence of these mold species in heat-treated dairy products is mainly due to two
reasons, i.e., thermal resistance in mold species and post-pasteurization contamina-
tion of heat-treated products during packaging or bottling (Jodral et al. 1993).
12.5 Sources of Contamination and Control Strategies
Studies have reported various sources of fungal contamination and infection through
the complete chain from farm to final product including unhygienic conditions,
equipment, and addition of non-dairy ingredients. Regardless of milk from different
species of animals, raw milk usually comprises about 3.0–5.0 log cfu/mL fungi,
with a greater number of yeast cells as compared to fungal spores (Lavoie et al.
2012; Marín et al. 2015). A recent study reported that retarded or reduced growth of
yeast population during refrigerated holding of milk is rapidly outnumbered by the
psychrotrophic population such as Bacillus and Pseudomonas (Awasti et al. 2019;
Von Neubeck et al. 2015). Besides few fungal species, other species of molds and
yeasts are not thermally resistant and can be inactivated by pasteurization. Thus,
contamination from fungal species mainly occurs during product manufacturing or
processing. Dairy plant air is among most prominent source of variable fungal spe-
cies and results in contamination of different milk and milk products. Previous
study (Beletsiotis et al. 2011) compared fungal counts of indoor and outdoor air
samples of dairy Greek dairy plant. As per their analysis, they reported higher fun-
gal counts in outdoor air samples (362.3 cfu/m3) with respect to indoor air samples
(266.2 and 69.8 cfu/m3). A study done by Temelli et al. (2006) reported cheese rind
as a source of spoilage for Turkish white cheese production via cheese slicer and
cheese ripening room. Recent study (Bokulich and Mills 2013) identified cheese
brining tank as major source of contamination and fungal counts of approx. 9.0 log
cfu/cm3.
Nowadays, fungal spoilage is among one of the major challenges for food and
dairy industries and continues to grow with increased demand and production of
food and dairy products. Milk and milk products can be spoiled at any stage of pro-
cessing and thus require protection from spoilage during milking, transportation,
storage, processing, and distribution. With increase in dairy demand and consump-
tion of dairy products, we need more firm techniques that can completely prevent or
eradicate fungal spoilage from dairy environment as well as product processing
equipment. The requirement of extended shelf-life of products is expanding due to
their growing demand throughout the world. Thus, the need of food products with
extended shelf life has been a driving force for development of various preservation
methodologies. Preservation mainly comprises of chemical, biological, or physical
barrier that fight against spoilage causing microorganisms as given in Fig. 12.1.
Fungi as a group are one the most resistant microorganisms that can overcome
various microbial control and prevention strategies used by the dairy and food indus-
tries worldwide. Various traditional methods used in combination are referred to
hurdle technology. Traditional methods usually include thermal treatments, addition
of antimicrobials, chemical preservatives, air filtration and purification, reduction of
water activity by means of drying or addition of salt or sugar, modified atmospheric
packaging (Westall and Filtenborg 1998), high pressure processing, and refrigeration
(Brul and Coote 1999; Ross et al. 2002; Stratford et al. 2014). The use of preserva-
tion techniques such as heating, addition of chemical preservatives, and antimicro-
bial compounds has been shown to reduce the food poisoning outbreaks, but these
techniques are also known to be associated with nutrition loss and organoleptic
changes in final product. Several prevention and control strategies have been reported,
and many a times, a combination of both approaches are usually used to reduce or
prevent fungal contamination. Control strategies mainly focus on using methods
which either inhibit or slows down the microbial growth and multiplication such as
use of low temperatures, chemical preservatives, and others. Whereas, preventive
techniques usually involve, methods that either avoid contamination and recontami-
nation, or by using physical barriers along product processing and packaging via
Techniques against spoilage

Fungi
Control Methods Preventive methods
Hazard Analysis Critical

Inactivation Methods Inhibition Methods
Control Points (HACCP)
Modified
Thermal Bio- Chemical Atmospheric Good Manufacturing and
Fermentation
treatments preservation preservatives Packaging Hygienic Practices
Addition of Air Filtration

High Pressure
Bioprotective
Processing (HEPA Filters)
cultures
Pulse Electric Addition of Aseptic Techniques

Field Fermentates (Processing & Packaging)
Fig. 12.1 Prevention and control methods to eradicate, prevent, inhibit, or control fungal
spoilage
means of aseptic technique, air filtration, Hazard Analysis and Critical Control Points
(HACCP), good hygienic and manufacturing practices. Different factors such as
product composition, water activity, critical temperature limits, etc. must be consid-
ered before selecting any prevention or control technique.
Studies have reported that after combining one or more abovementioned tech-
nologies, the rate and frequency of fungal spoilage issues can be reduced in dairy
products. But with increase in fungal resistance towards chemical preservatives,
thermal treatments and increase in consumer demand for clean or natural food have
resulted researchers and food industry to find an alternative control strategy.
Therefore, this situation has resulted into development of new techniques such as
use of bioprotective culture (Delavenne et al. 2015) to control or inhibit fungal
population or by using statistical optimization techniques (Awasti et al. 2019) to
understand the behavior of spoilage organism. This section summarizes our present
knowledge on different techniques and emerging methods that can be used to con-
trol and prevent spoilage causing fungal species.
12.5.1 Good Manufacturing Practices
The success of any dairy spoilage prevention strategies depends on initial concen-
tration of microbes and methods applied during dairy and dairy product processing
and manufacturing. To reduce microbial contamination such as fungal spoilage, the
production and packaging of dairy products must be done in aseptic and hygienic
environment (Rysstad and Kolstad 2006). Good manufacturing practices should
always be followed by good distribution practices which include implementing
practices that retard or inhibit microbial multiplication and growth. Good manufac-
turing practices include selecting good quality of raw material with no or least
microbial population and assessing and preventing any cross-contamination during
each stage of product manufacturing (Loureiro and Querol 1999). The success of
these practices relies on implementation of good hygienic practices and HACCP
system during manufacturing of product until consumption. Keeping microbial
quality of dairy products in mind, each stage of product processing should be moni-
tored and controlled for any risk from microbes.
12.5.2 Decontamination and Purification of Air
Environmental air is regarded as a potential source of fungal spores to dairy envi-

ronment and products. Therefore, efficient technique should be used to reduce or
keep out the fungal spores from area close to product processing. According to
Beletsiotis et al. (2011), fungal indoor spores can be reduced by 30 times by instal-
lation of high-efficiency particulate air (HEPA) filters. A recent review by Varga and
Szigeti (2016) summarized the significance of gaseous ozone in reducing mold and
yeast population by 10 times in cheese storage and ripening room. Concerning the
cleanliness of indoor air, other methods should always be kept in mind such as
controlling pressure of air from dirtiest to clean rooms. Therefore, technologies like
clean room, ultraclean, or aseptic techniques can be used to reduce fungal contami-
nation. Most importantly, few species of fungal species are able to grow in humid
regions of any dairy and food processing environment such as corners of walls,
floor, equipment and ceiling. Therefore, these areas should be disinfected and
cleaned properly by using different disinfection and aseptic techniques.
12.5.3 Disinfectants Against Fungal Spoilage
Dairy industries have been using various disinfectants to reduce fungal contamina-
tion such as quaternary amines, alcohol, iodophors, aldehydes, chlorine-based
agents, peracetic acids, or hydrogen peroxide. But the effectiveness of these clean-
ing agents on fungal spores usually varies and mostly depends on various factors
like time, concentration, and type of disinfectant and type and species of fungal
spores. For example, a previous study (Korukluoglu et al. 2006) reported that
Aspergillus niger respond differently to various concentrations and type of disinfec-
tant. They showed that A. niger showed resistance against peracetic acid but was
sensitive to alcohol. Similarly, a previous report (Bundgaard-nielsen and Nielsen
1996) revealed the variation in tolerance of different strains of Penicillium species
towards quaternary ammonium compounds. In addition, Martin and Maris (2012)
showed reduction in fungal contamination when two or more disinfectants such as
formic acid and hydrogen peroxide were used in combination.
12.5.4 Low-Temperature Storage
In dairy and food industry, control of temperature plays a significant role in reduc-
ing contamination from microbes. A recent finding from Awasti et al. (2019)
reported reduced multiplication and germination of microbes at lower temperatures.
Although opting for low or freezing temperatures does not kill or eliminate micro-
organisms, their growth and multiplication can be reduced or retarded. Usually,
milk and milk products are held at 0–10 °C in farm or dairy plant silos. As most of
the fungal species can survive and grow at low temperatures and thus, they usually
falls under the category of psychrotrophic range. A study done by Gougouli et al.
(2011) investigated the impact of low temperatures on 12 fungal species during stor-
age of yoghurt. In addition, they also reported −6.7 and 9.6 °C as a minimum growth
temperature for P. commune and A. niger. It is worth mentioning that storage tem-
perature plays a significant role, but type of species, initial population level, and
properties of product can also be a contributing factor towards fungal spoilage.
Therefore, further study is required to investigate the impact of all these factors by
using either predictive mycology tools or predictive optimization strategies.
12.5.5 Antifungal Agents and Chemical Preservatives
Besides abovementioned methods, antifungal agents and chemical preservatives are

widely used in food and dairy sector. They serve as a food additive to control or
inhibit contaminating fungal species and other microbes. There are several antifun-
gal agents that are used in industry; these include weak acids like benzoic, sorbic,
and propionic acid or their sorbate and benzoate salts. Few preservatives like sorbic
and benzoic acid show inhibitory activity only at certain optimal pH range and are
between 4.5 and 5.5. They mainly act on microbes by entering the cell membrane
and cause the buildup of protons and anions inside the cell. Accumulation of pro-
tons and anions decreases the pH of cell and may cause inhibition of metabolic
reaction or disruption of cell membrane which finally results into death of cell (Brul
and Coote 1999). In addition to this, Garnier (2011) compared concentration inhibi-
tion of various weak acids and determined their minimum inhibitor concentration
(MIC) against different fungal species. They reported variable MIC of fugal species
against different acids. Therefore, the inhibition, resistance, and adaptation of fun-
gal species towards weak acid depend on many mechanisms. Based on species type,
several studies have reported various mechanisms which include cell wall structure
maintenance, alteration in cell wall composition or plasma membrane, metal metab-
olism, activation and formation of shock proteins, reduced molecule entry, and acti-
vation of membrane transporters such as ATP transporters to transport ions out of
the cell membrane (Brandão et al. 2014). Other resistance mechanisms include
increase in buffering capacity and breakdown of weak acids such as few fungal spe-
cies can break down sorbate salts into a volatile compound like pentadiene or trans-
1,3-pentadiene resulting into off-odor and plastic- or kerosene-like flavor in dairy
product (Casas et al. 2004). Therefore, exact resistance or adaptation of fungal spe-
cies is not clear, and more studies are required in the field of mycology and their
physiological and molecular adaptation.
So far, besides weak acids, natamycin a polyethylene antibiotic is currently used
on cheese surfaces to inhibit fungal spoilage in dairy products (De Oliveira et al.
2007; Resa et al. 2014). Natamycin is also termed as pimaricin and is an antibiotic
agent primarily formed by Streptomyces natalensis. The mode of action of natamy-
cin on fungal is by inhibiting the fusion process of vacuole by interacting with
ergosterol. Natamycin can also be used in packaging film or coatings to prevent
fungal contamination in cheese. A combination of chitosin film with natamycin as a
packaging material has shown improved quality of semi-hard cheese (FDA 2015).
In addition, current study reported tolerance and adaptation of fungal species against
natamycin when incubated or exposed to it for long duration. Previous studies have
reported MIC against natamycin as negligible, ranging from 0.04 to less than
0.2 mg/cm2 for Cladosporium species and Y. lipolytica, whereas for other yeast spe-
cies, MIC for natamycin was found variable (Stern 1978; Streekstra et al. 2016). As
per code of federal regulation, in the United States, the concentration of natamycin
in cheese should not be more than 20 ppm (Bourdichon et al. 2012).
Nowadays, consumers are more concerned about their health and usually focus
on a product that is least treated by using chemical and physical treatments. Thus,
this is the probable reasons why among few emerging methods, the use of fermen-
tates, bioprotective cultures, and lactic acid bacteria (LAB) is gaining more atten-
tion (Dalié et al. 2010). Several studies have reported antimicrobial and antifungal
properties of LAB and Propionibacteria (Aljewicz and Cichosz 2015; Awasti et al.
2016). LAB have been used as a starter culture to produce numerous dairy products
like yoghurt, several types of cheese, and other dairy products. LAB is generally
recognized as safe (GRAS) and mostly produce numerous antifungal metabolites
(Inglin et al. 2015). LAB is mostly preferred due to absence of antibiotic resistance
and biogenic amines and thus regarded as safe for human consumption. Currently,
several commercially available LAB bioprotective cultures for dairy products are
being used and mainly include various strains of Lactobacillus and Propionibacterium
strains. The mode of action of these microbes is because of the production of anti-
fungal metabolites such as reuterin, acids (lactic, propionic, and acetic acids), diace-
tyl, cyclopeptides, organic acids (4-hydroxybenzoic acid, vanillic acid, succinic
acid, azelaic acid, and many more), and fatty acids (decanoic acids and
3-hydroxybenzoic acids). These strains were found resistant against species like
Aspergillus, Candida, Yarrowia, Crytococcus, Rhodotorula, and Kluyveromyces
(Garnier et al. 2017).
12.5.6 Packaging and Physical Methods
Apart from the abovementioned treatments, high-pressure treatment can be utilized

to inactivate or kill spores and vegetative cells of yeast and mold species. A current
study by Evert-Arriagada et al. (2014) demonstrated the effectiveness of high-
pressure processing (HPP) in increasing the shelf life of cheese like fresh cheese.
They also demonstrated that HPP of dairy products can inactivate mold spores and
yeast cells and thus can also be used as a product processing step to extend shelf life
of milk products. In support to this, studies done by Mainville et al. (2001) and
Atalar (2019) showed complete inactivation of yeast cells from kefir when HPP
treatment of 400 MPa was used for 15–30 min with no significant textural changes
in lipid and protein structures. Therefore, with increase in preference for more “nat-
ural” products, it is worth mentioning that HPP can be among one of the most
accepted techniques by the consumers.
Various other treatments that are commonly used to inactivate or reduce the
microbial population included modified atmospheric packaging and heat treatments
like thermization, ultra-high temperature, and pasteurization (for detailed informa-
tion, see reviews by Garnier et al. (2017) and Hotchkiss et al. (2006)). Other tech-
niques such as pulsed electric fields (PEF) treatment, two-pass cavitation (Awasti
2019), and ultrasonication for few seconds to minutes represent another emerging
technique for inactivation of microbes. In the future such techniques can represent
as a mainstream process in dairy and food industry, with more emphasis on fluid
dairy products (see current reviews of Buckow et al. (2014), Sutariya et al. (2018),
Wan et al. (2009)). Different strategies such as optimization methodology or other
physical techniques should be used to prevent fungal spoilage in dairy products and
thus further are required to understand the behavior of these organisms in dairy
matrix. A recent finding from Awasti et al. (2019) showed that the population of
microbes can be controlled by optimizing raw milk holding conditions and under-
standing behavior of spores can improve dairy product spoilage, and thus in the
future, such study should be done for focusing on prevention of spoilage from fun-
gal strains.
12.6 Conclusions
Yeast and mold species do not cause spoilage of either raw or pasteurized milk, but
their presence may act as a source of contamination for other dairy products such as
fermented milk and cheese. Air stands out as one of the major sources of contamina-
tion and dispersion of fungal spores in dairy environment. Thus, by implementing
good manufacturing and distribution practices, reducing the number of contami-
nants in dairy environment, and identifying the source of contamination are some of
the best ways to prevent dairy spoilage issues during product manufacturing. To
prevent further fungal spoilage, current methods should be combined with emerging
technologies such as cavitation or ultrasonication or by using predictive mycology
tools or predictive optimization strategies during dairy product manufacturing.
Based on different product matrix, further knowledge and research are required to
understand contamination sources and types of fungal species associated with vari-
ous dairy products.
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Dairy Industry: Hurdles Ahead
in an Economic Perspective 13
Denny Franco and Bulbul G. Nagrale
Contents
13.2 Production 265
13.3 Consumption 265
13.4 Milk Price 267
13.4.1 Raw Material: The Essential Ingredient 267
13.4.2 Wage and Productivity 268
13.4.3 Feed and Fodder Shortage 269
13.4.4 Credit Availability 270
13.5 Value Chain 270
13.5.1 Milk Processing Industry 271
13.5.2 Export-Import Market 271
13.5.3 New Product Development 274
13.6 Marketing and Pricing 275
13.6.1 World Price and Elasticity 276
13.7 Policy Interventions 276
13.7.1 International Trade 277
13.7.2 Trade Agreements 277
13.8 Sustainable Dairy and Greenhouse Gas Emissions 279
13.9 Food Safety Measures: Boon or Bane 279
13.10 Packaging the Emerging Issue 280
13.11 Culmination Policy 280
References 280
D. Franco (*)
WTC, ICAR-IARI, New Delhi, India
B. G. Nagrale
College of Dairy Technology, Udgir, Maharashtra Animal and Fishery Sciences University,
Nagpur, India

https://doi.org/10.1007/978-981-15-2608-4_13
264 D. Franco and B. G. Nagrale
Abstract
Dairy has an important role in the livelihood of many countries. Dairy is the single
major sector which contributes the maximum to global agricultural gross domes-
tic product (GDP). High perishable nature of dairy products creates different chal-
lenges in the marketing value chain. The world dairy sector is highly competitive
and also highly distorted due to high government interventions in the sector. The
cost of cultivation of milk and processing cost of dairy products are the deciding
factor of the competitiveness in the global market. The new product development
and value chain improvements are essential in the changing food habit scenario.
The developing and developed world has a structural difference in dairying point-
ing out the inappropriateness of one common policy for the whole world.
Keywords
Dairy · Marketing · International trade · Processing · Packaging
13.1 Introduction
The world population is touched around 7.6 billion in 2018 and growing hastily
with future predictions of eight billion in 2025. The quantity and quality of food is
the concern of the country leaders to feed these populations under the worst regime
of malnutrition and poverty. Dairy is the sector which has to play an important role
in nutrition and poverty alleviation at global level.
Dairy production is the main livelihood for the small and marginal farmers in the
developing world. Dairy ensures quick and continuous income for these marginal-
ized category people. According to the Food and Agricultural Organization (FAO),
approximately 150 million populations are actively involved in dairy activities.
Around 750–900 million peoples are living in families where dairy is the part of
livelihood, which constitutes 14% of the total world population (Hemme and Otte
2010). Apart from this livestock is the labour-intensive sector that has huge poten-
tial to provide employment for the rural poor. It is estimated that the one million
litres of milk production under small-scale dairy farms have the potential to gener-
ate approximately 200 labour days per year in comparison to 5 labour days in the
intensive large-scale dairy farms of developed countries lauds the importance of
dairy on employment generation opportunities in the poor countries. Thus, the
increasing trend of milk production in devloping countries also sounds bet-
ter employment oppurtunities generated for their livelihood.
The dairy is the sector which is fetching more revenue to the many countries
through export and helps to maintain the trade balance positive. The cattle are the
main source of livelihood to the rural area in most of the developing and underde-
veloped countries. Developing and undeveloped countries are facing challenges like
malnutrition; especially under-nutrition and unemployment. Under such a scenario,
the dairy sector has a jovial role to play and is being the solution. Milk and milk
13 Dairy Industry: Hurdles Ahead in an Economic Perspective 265
products are the rich sources of easily available protein and vitamins to defeat the
malnutrition. Demographically, women population have greater participation in the
daily dairy activities that help to empower them financially secure. Elemently dairy
sector is interwoven with the socio-economy of rural people. The transportation,
marketing both in forward and backward linkages along with a processing, will
make a multiplier effect on the employment generation. These aspects clearly indi-
cate the importance of the dairy sector in the economy of every country.
13.2 Production
Globally the countries like India, USA, Pakistan, China, Brazil and Germany are the
forerunners of cattle milk production. World milk production has a drastic increase
from 392 million tonnes to 827 million tonnes from the year 1970 to 2017. The
growth rate of world milk production was estimated at around 1.8–2% per annum
for the next few decades. The growth in the production has many reasons such as
better institutional support, genetic improvement programmes, development of
infrastructure and increased livestock population. Meanwhile, the future predictions
estimated that the further increase in production is mainly from the developing
countries. The Asian countries have to play a major role in increased production
especially the countries like India and Pakistan (FAO 2019). The milk production in
the developed countries and the developing world has structural differences. The
herd size per farm of the developed world is quite high compared to the developing
countries where subsistence level dairying is predominantly practising. The average
herd size of the world is 2 herd per farm and has the capacity to provide 11 litres of
milk per day. In the developing countries, along with the low herd size, the yield of
cattle is also quite low and the increase in production mainly due to maintenance of
higher cattle population. The growth rate of developing nations showing the higher
magnitude than the developed nations rate because it starts from the base level. The
different studies opined that the herd size of the developed world will start to show
negative growth but have maintained competitiveness in the market because of the
increased per cattle yield (Hemme and Otte 2010). India is the leader of total milk
production with the production of 176 million tonnes followed by the USA and
Pakistan as detailed in Table 13.1. The production was miserably less in the earlier
stage and was picked up by the different government-supported programmes in the
main Asian and African countries.
13.3 Consumption
Coming to the consumption side, 75% of the per capita consumption in the develop-
ing countries is in the fresh raw milk format (OECD/FAO 2018). Average per capita
consumption of milk and milk products in the world is approximately 113 kg milk
equivalent per year (Jan et al. 2018). The consumption varies from a mere 30 kg to
more than 300 kg per person per annum. Finland is the leader in per capita
consumption followed by Sweden and the Netherlands as given in Table 13.2.
266
Table 13.1 Milk production across countries (million tonnes)

1970 1975 1980 1985 1990 1995 2000 2005 2010 2015 2017
World 391.95 424.73 465.82 512.98 542.53 540.07 579.31 648.22 724.45 801.13 826.75
India 20.8 25.6 31.56 44.02 53.68 65.377 79.66 95.62 121.85 155.6 176.2
The USA 53.07 52.34 58.24 64.93 67.01 70.44 76.02 80.25 87.52 94.64 97.76
Pakistan 7.45 8.19 9.01 10.86 14.72 19.01 25.57 29.44 35.49 41.59 44.29
China 1.96 2.37 2.93 4.76 7.04 9.46 12.3777 32.02 41.16 36.28 34.87
Brazil 7.42 10.05 12.06 12.57 15.08 17.13 20.53 25.53 30.96 34.86 33.74
Germany 28.18 28.75 32.1 33.63 31.34 28.63 28.35 28.48 29.65 32.71 32.69
Russian Federation 0 0 0 0 0 39.31 32.28 31.15 31.84 30.79 31.18
France 22.85 24.772 27.89 28.4 26.81 26.09 25.74 25.771 24.21 25.93 25.26
New Zealand 5.99 6.1 6.7 7.88 7.51 9.29 12.24 14.64 17.01 21.94 21.37
United Kingdom 12.97 13.93 15.97 16.02 15.25 14.84 14.49 14.47 14.07 15.32 15.26
Netherland 8.24 10.22 11.79 12.53 11.23 11.29 11.16 10.85 11.81 13.55 14.54
Poland 14.96 16.38 16.49 16.44 15.84 11.64 11.89 11.95 12.3 13.25 13.7
Argentina 4.19 5.65 5.31 5.64 6.28 8.777 10.12 9.91 10.63 12.06 10.1
Mexico 4.11 6.24 7.23 7.47 6.27 7.54 9.44 10.03 10.89 11.61 11.99
Australia 7.76 6.7 5.57 6.23 6.46 8.46 10.85 10.13 9.02 9.49 8.8
Switzerland 3.2 3.4 3.6 3.87 3.88 3.93 3.91 3.96 4.11 4.07 3.92
Source: NDDB (2019)
D. Franco and B. G. Nagrale
Table 13.2 Per capita milk Finland 361.19

consumption (kg/year) Sweden 355.86
Netherland 320.15
Switzerland 315.78
Greece 314.69
Montenegro 305.87
Lithuania 303.00
Denmark 295.62
Albania 281.17
Romania 266.19
Luxembourg 265.9
Kazakhstan 262.61
Norway 261.52
France 260.48
Italy 256.10
Source: Sawe (2018)
The studies opined that the increase in the per capita income of most countries will
further increase the demand for dairy products in the future. To satisfy this increased
demand, the dairy sector should grow around 2% annually. Some countries are self-
sufficient in the production to meet the growing consumption through increased
production like India but not all. But the demand in the Middle East, Asian and
African countries is higher than the production which clearly indicates the growth
of imports on those regions and positively signals requirement of more number of
processing industries in the near future.
13.4 Milk Price
The cost of milk production heavily skewed to the inputs used and the cost of inputs.
The processing industries will lose competitiveness if the price of milk is costlier
compared to the contender.
13.4.1 Raw Material: The Essential Ingredient
Competitiveness of the dairy industry is based on the cost of production. The inter-
national trade countries who earn from the rising demand are those that have a mini-
mum cost of production. The low cost of production leads to higher competitiveness
in international trade. Globally, countries are heavily subsidizing the input cost and
veterinary services for enjoying the lower cost of production to promote export.
Meanwhile, the yield of cattle is the main lacunae for developing countries to enter
into the world trade with holding major share. The yield of developed countries is
four to five times higher than others. The higher yield of cattle makes such countries
to invest more in the dairy sector than the others. Low milk productivity and high
cost of production pulled the developing countries to be in the vicious cycle. The
global average cost of production of the milk was calculated as $40.2/100 kg milk
in 2017. The devaluation of currencies by some countries, high feed cost and infla-
tion rates were causes for the higher cost (Katrin et al. 2018).
The studies opined that the feed price will rise to about 50% over the base price
of the year 2006 and causes a rise in the price of milk. The demand for food grains
and fuel crops to meet the food and energy requirement in the future will create
further pressure on the fodder cultivation and land price drastically. The govern-
ment’s efforts to reduce the cost of production by providing good health-care prac-
tices, better breeding programmes, infrastructure development and better
management practices of cattle animal can help to reduce the cost of per litre of
milk. The milk feed price ratio, one of the choice indicators of the cost of produc-
tion, is around 2.5 for the countries where intensive dairying is practising and 1.5
for the developing countries where the cheaply available plant residues is the main
source for cattle feed. Feeding the high productive cattle with compound feed is the
reason for higher milk feed ratio which clearly indicates the real advantage of inten-
sive farming over the subsistence level of farming.
13.4.2 Wage and Productivity
As labour intensive sector, wage is one of the major constituent in variable cost of
dairy farms. The labour wage of the industrialized countries was quite higher than
the rest of the world. However, the high wage rate is economically normalized with
higher labour productivity in those countries. Most of the developing countries with
cheaply available labour provides the competitiveness to those countries in the
international market, but the recent trends on the raising wage rate of these develop-
ing countries are the concern over the greater advantage that they were enjoying
historically. Thus, high wage rate normalized with higher productivity causes the
industrialized countries a greater advantage in the near future. The small herd size
with low-yielding cattle population jointly increases the marginal cost of milk pro-
duction. The marginal and small farmers are managing their farm in the lowest
production cost side by utilizing the locally available resources. The proper organi-
zation of the cheaply available domestic resources will make them stand straight
towards to the highly competative capital-intensive dairy farms. The average milk
production cost of developed countries was estimated 56% higher than the develop-
ing world. The low cost of milk production in the developing countries might be the
reason for why they increased their share in total global production (Hemme and
Otte 2010). The comparison of cost of production of different countries were
detailed in the Table 13.3 indicates that majority of developing countries having low
level of cost of production and have economic advantage over the developed
countries.
Table 13.3 Cost of production: a comparison among major producer

Cost of production (US
cents/kg of milk) Countries
<18 Poland, Argentina, Pakistan, Vietnam, New Zealand, Brazil (larger
farms), India (larger farms), Chile (smaller farms), Australia (smaller
farms)
18–28 Estonia, Czech Republic, Brazil, Bangladesh, China, Thailand, Brazil
(smaller farms)
28–35 Spain, Denmark, Ireland, United Kingdom, Hungary, USA, Germany
(larger farms), Netherland (larger farms), Israel (larger farms)
35–45 Austria, France, Sweden, Netherland, Israel (smaller farms)
>45 Switzerland, Norway, Finland, Canada, Germany (smaller farms)
Source: Hemme et al. (2004)
13.4.3 Feed and Fodder Shortage
The increasing demand for dairy products indirectly emphasizes the need for more
and more feed and fodder. Increased demand for dairy products can be met by
either increased number of cattle population or productivity enhancement. Feeding
this large number with balanced diet becomes a challenge. The production of milk
and crossbred cattle rearing solely depends on the availability of green fodder and
concentrates. However, the farmers maintain the herd size which was not able to feed
on the available supply of green fodder. For making the dairy an economically more
attractive sector, the production of milk has to be enhanced with lesser cost, which
is possible only through providing sufficient feed in both quantity and quality. The
land availability for the cultivation of green fodder became questioned in the sector
where marginal and small farmers were the major shareholders. The changed crop-
ping system from subsistence level to market-oriented intensive system created the
shortage of balanced fodder. The issue was further intensified by changing cropping
pattern from subsistence to cash crops where crop residue feeding was the major
source for feeding. The livestock population growth trend over the years indicated
that there is a decline in the number of indigenous breeds and an increase in the
crossbreds and buffaloes. The commercialized peri-urban dairy sector requires
more amount of green fodder-based food ration to capitalize on the genetic potential
of crossbred animals. The study about the demand and supply of forage from big-
gest cattle populated country, India shows that there is a huge gap of 25% and 12%
deficit in the availability of green and dry forage, respectively, will exist by the year
2030 (Dikshit and Birthal 2010). The FAO report on the dairy industry pointed out
that the drought in many countries affects the green fodder availability and directly
reduces the milk output and badly affects the dairy industry. The increased demand
of feed and fodder can be met by the genetic improvement of forage crops, timely
availability of forage crop seeds, development of water-efficient varieties, proper
extension activities of developed technologies, wasteland utilization for raising the
forage crop, balanced pasture land grazing and well-managed common property
resources (CPR).
13.4.4 Credit Availability
Credit availability is the backbone of any industry for further development. But in
the dairy sector, credit is a persisting challenge. The small and marginal farmers
who were poor in resources have limitations to get the loan from the banks. The
poor availability of loan makes less investment in the sector hinder further invest-
ment. The Self-Help Group (SHG) has huge hope in financing the dairy sector at the
producer level. SHG-linked microfinance system promotes the thrift and credit
facility to the small and marginal farmer. The support through the microfinance
protects the farmer from the distress sale at the adverse time. The SHG movement
in the African and Asian continent has a great role to play in the near future. Apart
from the production side, the processing industry also requires a huge investment.
Many governments supported the dairy industry through subsidies and tax exemp-
tion for the establishment of the processing plant. Cooperatives and farmer producer
organizations are a good model for the world to overcome the huge initial capital
requirement. The modernizations of dairy plants start in the twentieth century and
earnings momentum in the twenty-first century. Establishment of the dairy plant
creates more job opportunity within the country.
13.5 Value Chain
The value chain of both inputs and outputs are an important component of any
industry and needs more attention in the dairy sector. Developing world dairy sector
has achieved higher positions because of the marketing infrastructure which they
developed in recent decades. The need for well-established organized sectors came
at this point. The cooperatives and producer organizations played a greater role in
development. To produce and add more value to each activity, the education of the
farmer is very important. Proper extension education has to provide for the dairy
farmers regarding cattle shed management, cattle health management and market-
ing strategies along with value addition. The cooperative movement and different
extension programs in different countries helps to educate the farmer in the need for
clean milk production. Apart from the extension education, cooperative movement
and farmer produced organizations also helps to procure of milk from each corner
of the country in a systematic way. The production, procuring, processing and mar-
keting channels are very important for the success of the dairy sector where highly
perishable commodities are handling. Most of the developing and underdeveloped
countries have meagre development in dairy infrastructure development and are still
facing the problems in their value chain. Chilling plant in the main collection cen-
tres will help to improve the shelf life of milk to a considerable level. Even though
the marketable surplus of small and marginal farmers is quite low, the creation of
proper collection centres can account for their production to the marketing.
13.5.1 Milk Processing Industry
Industrializing the dairy through the processing industry is the connecting link
between the primary agriculture sectors and the secondary industrial sector. Globally
the milk and milk product value contributes 8.9% to total agricultural products in
2015. In some countries, the contribution of milk and milk products ranges more
than 20% to the total value of agricultural products like New Zealand, India,
Switzerland, Finland and Latvia. The increased per capita income and hastily
increased urban population further increases the demand for dairy processing. Some
countries are self-sustainable to meet the growing demand by increasing milk pro-
duction like India, but most of the Asian and African countries demand to overcome
their supply potential needs to import. This situation further intensifies the impor-
tance of processing globally.
13.5.2 Export-Import Market
The growth in production accounts mainly due to the increase in the cattle popula-
tion of developing countries like India and Pakistan along with the increased pro-
ductivity from the European Union and the USA. Apart from these the efficiency
escalation process of the milk collection process and the integrated dairy production
systems become the driving force for the increased production. However, the pro-
duction in countries like China and Ukraine decreased due to the industrial restruc-
turing and downscaling of the small-scale firms (FAO 2019). The world is trading
only 7% of the total world production. Melamine crisis of China in 2008 soared the
demand for imports of dairy products as the people who lost their belief in the
domestic market boosted the international trade especially milk powder trade. The
world scenario of dairy export and import market are detailed in Table 13.4.
Table 13.4 World export-import market: top countries and their share
World’s total milk exports World’s total milk imports
Thousand tonnes Thousand tonnes
Country milk equivalents Percentage Country milk equivalents Percentage
European 20,504 27.42 China 14,615 19.50
Union
New 18,748 25.07 Mexico 4202 5.61
Zealand
The USA 11,778 15.75 Algeria 3835 5.12
Belarus 3789 5.07 Russian 3700 4.94
Federation
Australia 3055 4.09 Indonesia 2981 3.98
Argentina 1996 2.67 Saudi 2790 3.72
Arabia
Uruguay 1556 2.08 Philippines 2501 3.34
World total 74,781 100.00 World total 74,967 100.00
Source: FAO (2019)
Consumption of dairy products has been increased steadily and results in the
growth of export and import market. The study shows that the import market marked
the growth of 187% from $15 billion US dollars in 2005 to $43.2 billion in 2014.
Similarly, exports grew 175% in value in the same period (Davis and Hahn 2016).
The USA was the largest importer of milk products in the dairy industry is shifted
to China in recent years because of the melamine crisis. The USA imported milk for
further processing and exported it to the world market after value addition. The
well-established processing industry of America flourished through value addition
and fetch considerable earning via trade. But the changed scenario made China the
largest importer followed by Mexico, Algeria and Russian Federation, respectively.
Although the importing status was changed because of demographic reasons, the
suppliers in the international market remain relatively unchanged. New Zealand is
the largest exporter country of dairy products in volume followed by the USA and
Belarus. The international market traded around 75 million tonnes of milk and milk
products in 2018 and shows a positive trend in growth. The European Union and
Oceania countries are the main shareholders of the export market. Countries like
New Zealand and the USA together hold more than 40% of the total export market.
The country China, the present largest importer of dairy products tries to increases
the production and chances to becomes self-sufficient in the near future, may affect
international trade prices of dairy commodities negatively.
World dairy product market traded many commodities in which more than half
of the trade constitutes cheese, whey, non-fat dry milk and butter. Cheese alone
contributes around 24% of the total traded value. The largest importers of cheese (in
volume) in 2018 were Japan followed by Russia and the USA as detailed in
Table 13.5. The largest suppliers of cheese were the European Union, the USA and
New Zealand, respectively. The USA alone enjoy the share of 14% of total cheese
export. Cheese consumption rises rapidly in the East Asian countries because of the
rising per capita income and changing diet preferences. The world cheese market
handles 25 million tonnes of cheese by volume per annum. The mozzarella cheese
prepared from buffalo milk gaining momentum in the market because of its various
health benefits, is a good sign for India and Pakistan where the main buffalo popula-
tions are existing.
Table 13.5 Major Cheese import-export countries with their share

Cheese import Tonnes Percentage Cheese export Tonnes Percentage
Japan 285,701 11.25 EU 832,678 32.39
Russian Fed. 263,151 10.36 The USA 350,240 13.63
The USA 175,839 6.92 New Zealand 324,171 12.61
Saudi Arabia 170,400 6.71 Belarus 210,253 8.18
China 156,396 6.16 Australia 172,520 6.71
Korea Rep 123,850 4.88 Egypt 101,000 3.93
Mexico 122,975 4.84 Saudi Arabia 72,300 2.81
World 2,539,609 100.00 World 2,570,548 100.00
Source: FAO (2019)
Skimmed milk product (SMP) is the second-largest dairy commodity traded in

the world, which accounts for 11% of the total trade (Davis and Hahn 2016). The
main importing countries were Mexico, China and Algeria which altogether import
around 31% of the total imports. The main suppliers were the European Union and
the USA which together hold half of the total export.
The world trade in the butter is on the reverse path. Both the volume and value of
butter trade were increasing but at a lower rate. China is the largest importer of but-
ter products which holds 16% of total market import, and New Zealand is the largest
supplier to the international market which holds 50% of the total butter market. The
major export and import of share of different countries in skimmed milk powder
(SMP), whole milk powder (WMP) and butter are given in Tables 13.6, 13.7, and
13.8 respectively.
The growth rate of the main dairy product in the international market indicates
the SMP has an expanding market with 32% growth as illustrated in Fig. 13.1. Even
though the share of cheese is the highest in trade, its growth rate is only 16% in the
global market.
Table 13.6 Major SMP import-export countries with their share

SMP imports SMP exports
Mexico 360,360 14.23 EU 834,092 32.23
China 309,003 12.20 USA 716,193 27.67
Algeria 166,851 6.59 New Zealand 362,989 14.03
Indonesia 161,796 6.39 Australia 155,567 6.01
Philippines 159,120 6.28 Belarus 119,948 4.63
Malaysia 128,631 5.08 Mexico 104,600 4.04
Russian Fed 87,302 3.45 Canada 65,806 2.54
World 2,532,539 100.00 World 2,587,940 100.00
Source: FAO (2019)
Table 13.7 Major WMP import-export countries with their share

WMP imports WMP exports
China 648,270 25.92 New Zealand 1,380,672 56.18
Algeria 311,780 12.47 EU 333,630 13.57
UAE 144,000 5.76 Uruguay 143,459 5.84
Saudi Arabia 138,200 5.53 Argentina 135,433 5.51
Sri Lanka 87,288 3.49 UAE 6400 0.26
Bangladesh 82,000 3.28 Oman 55,200 2.25
Oman 81,351 3.25 Australia 55,081 2.24
World 2,501,150 100.00 World 2,457,740 100.00
Source: FAO (2019)
Table 13.8 Major butter import-export countries with their share

Million milk Butter Million milk
Butter imports equivalent Percentage exports equivalent Percentage
China 146,568 16.20 New 458,631 49.96
Zealand
Russian 88,416 9.77 EU 158,357 17.25
Federation
The USA 58,186 6.43 Belarus 89,913 9.80
Saudi Arabia 50,852 5.62 The USA 51,719 5.63
Australia 39,930 4.41 Ukraine 30,383 3.31
Egypt 34,600 3.82 India 29,594 3.22
Iran 34,481 3.81 Australia 16,315 1.78
World 904,778 100.00 World 917,920 100.00
Source: FAO (2019)
35%
32%
30%
25%
21%
20%
17%
16%
15%
10%
5%
0%
Butter /- oil Cheese SMP WMP
Fig. 13.1 World: dairy product output growth 2010/2017. (Source: World Dairy Situation Bulletin
2018)
13.5.3 New Product Development
Milk products can be broadly classified as fermented and non-fermented products.

Fermented products are mainly used for confectionary food preparation and curry
making. The fermented products have a huge market in countries like India and
Pakistan. The cheese, butter, liquid milk, milk powder and baby foods are examples
of non-fermented products. The consumer preferences and perceptions towards
dairy products are intermittently changing. The new product development needs
greater attention and should be supported by proper research and development.
Different dairy products were trying to establish market space historically. But
many products failed due to the lack of consumer appealing marketing strategies.
The world is more health-conscious and searching for the alternatives of the low
calorie and low-glycemic food to fight against the non-communicable lifestyle dis-
eases like obesity, cardiovascular diseases and diabetes. The research and develop-
ment in processed food suggest that dairy products have great potential to produce
low-calorie products. The growing health concern of the citizens is likely to increase
the demand for the probiotics. The production of cheese with less fat content can
minimize obesity risk.
The dairy is acting as the connecting link between agriculture and industry by
providing the raw material. The new product development is the essential require-
ment of the industry on the changing dietary habits. The production of casein, lac-
tose, lipids and whey protein from milk is the emerging industrial requirement. The
confectionery and bakery industry heavily depend on the dairy for the raw material.
The new products like cheese spread and mozzarella cheese have huge market
potential. The new food categories like nutraceuticals, medicinal foods, therapeutic
food and superfood are the areas where the new product can be introduced.
Fortification of milk has an untapped area and needs special attention in product
development. The milk-based products can act as a substitute for the soft drinks in
health-conscious world.
13.6 Marketing and Pricing
The dairy is the sector where the product is facing a high range of volatility and
perishability without processing. The marketing of the product is facing a number
of problems like a long marketing channel, high-price spread, poor infrastructure
development, issues on fat-based pricing, lack of coordination among the different
departments and poor extension services. The lengthening of the marketing channel
through a large number of middlemen pushes the real producer share from the con-
sumer rupee at a lower level. The major chunk of the profit was swallowed by the
middleman and week bargaining power of farmers was exploited. Private, coopera-
tives and farmer producer organization were working in the procurement of milk in
many countries. But there is scope for efficiency enhancement and increased system
transparency. Research shows that only 8% of the new products are reaching into
the market because of poor marketing strategy.
Different countries have a different level of interventions in the pricing of milk.
The objective of every policy is to protect both farmers and consumers. The pro-
ducer should get maximum benefit without affecting the consumer interest to get a
better consumer surplus. Seemingly both are contradicting each other, but both tail
and head should meet appropriately for better policymaking. Contrasting to agricul-
tural commodity Minimum Support Price (MSP), no MSP is providing milk in
India. However, the solid (fat and SNF)-based price system is administered.
The cost of prodcution of different dairy products directly depends on the price
of raw milk, which is the major input for the processing industry. The direct fixed
cost has a less role in the processed products in the long run. Over the time period,
the milk procurement price increased in nominal scale, not in real terms. The hike in
the price of milk can increase the profit margin of the farmers directly, and more
products can be ensured. But in general, the cost of production is always increasing
along with the hiked support price make the profit margin of the farmer in the mea-
gre level. The feed companies increases the price of inputs along with the hike in
milk prices reduces the margin of profit and lower the milk/ feed ratio demotivates
the dairy farmers from further investment.
The pricing of milk has a different way like volumetric pricing, fat-based pricing and
two-axis pricing system. Most of the countries are following the two-axis pricing sys-
tem where both fat and SNF (soilds not fat) were taken into consideration. The goverments
are regulating the milk price through different mechanisms like administered price, input
subsidies and quota system are followed in different countries. Instead of minimum support
price like crops, the dairy sector following the administered price support system for fixing
the milk price. The cooperatives and government institutional system help to implement the
administered price. While the market signals are not quickly affecting the fixing of milk price
in the short run is the limitation of such systems. The market-based approach is one of the
best alternatives which signals quickly changes in the market prices as like the other agricul-
tural commodities where the demand and supply will decide the price. The future marketing
and hedging will help the farmer avoid risk in market-based pricing system and let the
demand and supply fix the price. But the development of such markets requires more mar-
ket intelligence and market information was a constraint.
13.6.1 World Price and Elasticity
For long time the milk and milk products were considered as inelastic good where
the price changes may not make the quantity purchased. But recent studies show
that the demand is more price elastic in nature, especially for milk products. The
expenditure on the dairy products is quite inelastic in nature which means the price
change never makes changes in the total expenditure on dairy products. The changes
in price are adjusted by changes in quantity only. The elastic nature of pricing of
dairy products pointed to the pattern has to follow in the pricing of dairy products.
The price should be fixed at minimum level to fetch more market space at which
consumer have to enjoy maximum surplus of satisfaction. The world price of milk
is showing a decreasing trend because of both demand and supply-side factors. In
recent years, China is reduced their import of dairy products especially the SMP and
WMP and Russian Federation embargo on the import from many countries reduces
the demand in world trade. At the same time the production has been increased
because of higher contribution from developing countries and the USA along with
removal of quota system in european union motivates to prodce more reduces the
world milk price level at a minimum.
13.7 Policy Interventions
The entry of private firms in the processing industry was restricted in different ways
in many countries. In India, up to the 1990s, the processing sector was the monop-
oly of the cooperative sector through a restricted licensing system. The
liberalization, privatization and globalization policies attract more foreign invest-

ment. The installation of more number of dairy modern plants invites more competi-
tion through the private sector. The modernization and huge investment boosted the
processed milk product market. But the unregulated entry of private firms and com-
petition among the firms leads to the entry of adulterated/contaminated milk in the
food chain. The frequent market check and strong legislation should be made to
prevent such anomalies. A high level government regulations were distorted the
global dairy market at high level.
13.7.1 International Trade
The rise in per capita income will be boosting the consumption of milk products.
There is a huge demand for dairy products in the middle-income and higher-income
countries, where the lifestyle changes the consumption pattern of the dairy products.
Studies suggest that the increase in income of Asian and African cities leads to the
establishment of more hotels where the milk products like cheese and butter were the
main ingredients of food. India is the largest producer as well as the consumer of milk
and contributing lesser surplus for the international market. However, the countries
like the European Union, the USA, Australia, New Zealand and Argentina were con-
tinuing as the major international traders, while China, Philippines and Bangladesh
were the largest importers of dairy products. The main exporter European Union’s
share is expected to increase from 24 to 28% by the end of 2027 (OECD/FAO 2018).
The price trends over the years have a crucial role in dairy product marketing. The
fluctuations will cause a cob-web phenomenon in supply and demand. The dairy
product price has a nominal increase in their future forecast. The real price is flattened
and not going to raise in the near future. The price of the product may increase, but the
increased cost of production on the other side will neutralize the effect of price rise.
The self-sufficiency rate of different regions shows that the regions like Asia,
Africa and Central America have been in the deficit side. The Oceania countries
have surplus production which is almost double the actual domestic demand. The
huge marketable surplus of these countries pushes the countries to dump the prod-
ucts into the world market. But the huge demand from the other parts of the world
has enough purchasing power to purchase. So the chance for lowering of the price
is very narrow and gives considerable space for the other trading countries to com-
pete. The self-sufficiency rate of different countries is given in Table 13.9.
13.7.2 Trade Agreements
Out of the many factors, the trade agreement between the country and group of
countries makes high distortions in the trade. Among agriculture sectors, dairy
industries remain the most distorted one because of high tariff rate and quota sys-
tems traditionally followed. Even after the World Trade Organization (WTO) agree-
ment and Agreement on Agriculture (AoA), many countries policies left dairy
Table 13.9 World self-sufficiency rate (per region)

Region 2010 (%) 2017 (%)
Asia 93 90
Europe 105 110
EU 107 113
Non-EU 100 102
North America 102 109
South America 102 100
Africa 88 84
Central America 82 79
Oceania 311 289
Source: World Dairy Situation Bulletin (2018)
largely untouched. Meanwhile, many countries changed their non-tariff system to

tariff-based quota system and subsidized their export to protect own interests. Under
WTO the countries agreed to reduce the level of aggregate support measures, but
most of them untouched dairy sector in reductions and reduce the rates of other
commodities to comply with the reductions. Establishing import restrictions or
export subsidies on the basis of milk equivalents would allow these measures to
have roughly the same net impact on markets. Sanitary and phytosanitary (SPS)
measures were regulated in the agreements in that direction. But some countries are
raising the standards which were not acquirable for the developing and developed
countries which will act as a trade barrier. Dumping of cheaply available products
in the developing countries by the developed countries pulled them down. The
pieces of evidence show that the European Union and the USA dumping their highly
subsidized dairy products in the developing countries (World Bank 2005) lost the
competitiveness of the domestic sector.
The free trade agreements and bilateral agreements in different regions are not
coming into consensus in the dairy sector. The chance for continuing quota and high
tariff rates are still existing in the world to protect own interest. The agreements of
Oceania countries, who are handling major trade share like Australia and New
Zealand with other countries and trade blocks expect a low rate of tariff cut in the
near future. Under the WTO agreement, all countries are supposed to reduce the
trade barriers to make a single global free market. But the rates and the amount to
deduct were decided at a base period of 1986–1988 levels. The developed countries
which are the main traders in the world market already achieved a higher level of
support at that period where the developed world support was only a meagre level.
Most of the countries use Special Safeguard (SSG) mechanism provided under the
WTO for controlling the import of products. It is expected that if all the trade restric-
tions like a tariff, domestic support and export subsidies were removed by all the
countries, the world milk price will rise (Ohlan 2006). The export from the develop-
ing countries will increase substantially and boost investment in the sector.
13.8 Sustainable Dairy and Greenhouse Gas Emissions
We are living in a world where climate change is a topic for debate. In the wake of
the present climate change scenario, the contribution of the dairy sector and its
magnitude to the total greenhouse gas (GHG) emission need to be addressed to
maintain the sustainability issue. The calculation of GHG in the dairy sector is a
complex process under many assumptions and generalizations. The GHG emission
analysis starts from ‘cradle to retail’ like emission associated from feed production,
manure management, transportation, processing, packaging and many more. The
dairy processing sector emissions particularly related to product processing and
transportation alone account 1328 million tonnes CO2 globally with an average of
2.38 kg CO2 per kg of milk and milk products. It accounts for 2.7% of global total
anthropogenic emissions of 2007 (Theun et al. 2013). The emission of GHG vary-
ing with the products depends upon the level of energy required. The International
Dairy Federation (IDF) noted the 1 kg fresh milk production caused 1.2 kg CO2 −eq
emission and the same in the cheese produces 8.8 kg CO2 −eq GHG gases. The
emission from the dairy processing industry accounts 5% to 13 % of total GHG
emssion of dairy indutry as whole. The energy consumption and packaging is the
main contributor of GHG from the processing industries. Practising of efficient low
emission energy sources and nature-friendly packaging materials will be the mitiga-
tion strategies. The low emission technologies can be converted into the earning
from the carbon credit market.
13.9 Food Safety Measures: Boon or Bane
Food safety is important for the health of citizens. The tightened food laws were
used as a trade control measured by different countries. The countries like Denmark
introduced a tax for saturated fat-containing products as the part of their cardiac
vascular disease reducing measures affects the cheese and butter industry badly
(Joshi 2014). The EU proposal to indicate the frozen material separately on the
package draws the menu cost for the trading countries. The frequency of evolving
new regulations and restrictions are a big hurdle in the dairy sector.
Importing counties are very much cautious with respect to the hygienic and sani-
tary measures practised in the exporting countries. Many countries are assuring
these through audit and frequent visit to the processing factories. Buyers are
demanding the Hazard Analysis Critical Control Point (HACCP) certification or
compliance for the trade. HACCP proposed by Codex Alimentarius Commission
(CAC) is a good manufacturing practice that assures food safety in the manufactur-
ing sector. The investment required to get the HACCP certification is a big hurdle in
most of the poor countries.
13.10 Packaging the Emerging Issue
Packaging in the dairy sector is a realm of concern for product marketing. Protecting
the product and marketing of the product largely depends on the material used for
the packaging. The non-biodegradable materials were traditionally used for the
packaging. The disposal of the used material became environmental issues in the
developed countries, and governments are cautious about banning non-degradable
materials. The high cost of substitutes became the challenge in most of the export-
ing countries. The 3R principles like reduce, reuse, and recycle have to practice in
the packaging component to overcome such issues. The researches on packaging
material have to be supported by the government to solve the issues and make the
packaging sustainable.
13.11 Culmination Policy
The regional taboos and customs also have an influence on the wellness of the sec-
tor. The cow is the part of the religious sentiments and the culling of non-productive
cattle is considered as a sin in India. The protest incidents made the transaction and
the marketing of cow very difficult and made the cattle market in doom status. The
secondary income from the cattle selling was completely stopped and made the
reinvestment in the dairy sector a vicious cycle. The institutional laws and regula-
tions should be made clear to make the cattle market more dynamic. Strayed cattle
start to create problems for the pedestrians and travellers in most of the metro cities.
The problems of strayed cattle need to be addressed and awareness campaign should
be done regarding the issues by taking all the stakeholders into confidence.
13.12 Conclusions
The dairy has a triplicate effect in the nation progress like employment generation,
ensuring food security and poverty alleviation. The sector is showing a positive
growth rate and faces the number of challenges. The international trade is highly
distorted due to high political interventions in the sector needs to be minimized. The
research and innovations are needed to be practised in the field level through the
proper extension for the wholesome development of the sector. The country-based
policy will be more appropriate for development than the one common policy for
the world.
References
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19(B)
Dikshit AK, Birthal PS (2010) India’s livestock feed demand: estimates and projections. Agric
Econ Res Rev 23(1):15–28
FAO (2019) Overview of global dairy market developments in 2018. Dairy market review
Hemme T, Otte J (2010) Status of and prospects for smallholder milk production—a global per-
spective. Food and Agriculture Organization of the United Nations, Rome
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milk production worldwide. IFCN Dairy Research Network, Kiel
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Dairy Declaration of Rotterdam showing progress. IDF dairy sustainability outlook 1
Joshi RM (2014) India’s dairy exports: opportunities, challenges and strategies. In: National
Seminar on Indian Dairy Industry, pp 20–36
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Network, Kiel
NDDB (2019) National Dairy Development Board Statistics, Gujarat, India. https://www.nddb.
coop/information/stats/across
OECD/FAO (2018) Dairy and dairy products. OECD-FAO agricultural outlook 2018-2027,
pp 163–174
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Economic Review 48(3):505–5014
Sawe BE (2018) Countries who drink the most milk. WorldAtlas. https://worldatlas.com/articles/
countries-who-drink-the-most-milk.html
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Sustainable dairy production. Wiley, Hoboken, pp 9–29
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Brussels
Novel Milk and Milk Products: Consumer
Perceptions 14
Anuradha Kumari, Himanshi Solanki,
and Aparna Sudhakaran V
Contents
14.2 Current Scenario 284
14.3 Changing Food Habits of the Population and Need of Novel Foods 286
14.4 Trends in Dairy Products 286
14.4.1 Clean Label Foods 287
14.4.2 Functional Milk Products 288
14.4.3 Probiotic Dairy Foods 288
14.4.4 Fortified Milk Products 289
14.4.5 Low-Calorie Dairy Products 290
14.4.6 Low-Lactose Dairy Products 291
14.4.7 Reduced Sugar Products 292
14.4.8 Low-Cholesterol Dairy Products 292
14.4.9 High-Protein Dairy Supplements 292
14.4.10 Organic Milk 293
14.4.11 A1 and A2 Milk 293
A. Kumari (*)
H. Solanki
DFIL-CreamBell, Gurgaon, Haryana, India
Aparna Sudhakaran V

https://doi.org/10.1007/978-981-15-2608-4_14
14.4.12 Designer Milk 294

14.4.13 Extruded Dairy Foods 295
References 296
Abstract
The opinion about and the value of milk have evolved gradually over time.
Earlier people were paying for the food, but now they are ready to pay for the
value of the food. The traditional values like price, taste and convenience are
overtaken by the evolving drivers of health and wellness, safety and social
impact. Increased prevalence of metabolic disorders and ageing population
demands with the need for innovations of inculcating health and wellness in
dairy products beyond the standard product attributes. Some of the recent trends
with dairy consumers are increased demand for healthy and clean labels, reduc-
tion in added sugar, demand for new flavours, environment-friendly nutrition,
organic milk, high-protein dairy supplements, designer milk, etc.
Keywords
Milk products · Novel milk products · Consumer perceptions · Food habit · Milk
14.1 Introduction
The opinion about and the value of milk have evolved gradually over time. The tradi-
tional view of nutritional benefits mainly intended for infant has moved to improve the
wellness as well as nourishment both for children and adults. The diet pattern is
changing from customary food to processed and value-added food products due to
growing economy, urbanization, awareness and consciousness about health-related
issues. Earlier people were paying for the food, but now they are ready to pay for the
value of the food. The change in food pattern is likely to happen more in an urban area
than in the rural area. In developing countries, urbanization is a big social transforma-
tion having a noticeable influence on the diet of the population. Rural population rely
on their own food production; however, the urban population purchase more pro-
cessed food than fresh food. In industrialized societies, the food consumption is for
health concerns, sustainability and convenience (Grunert 2013).
14.2 Current Scenario
According to the Food and Agricultural Organization (FAO), “Milk is one of the
most widely produced, and valuable, agricultural commodities worldwide”. The
world milk production in 2018 was about 838 Mt, with India being the largest milk
producer with 174 Mt (OECD-FAO 2019). Most of the milk produced
14 Novel Milk and Milk Products: Consumer Perceptions 285
% Milk production contribution
Cow milk
Buffalo milk
Goat, Sheep and Camel

combined milk
Fig. 14.1 Milk production contribution by various species
was contributed by the cows, while a meagre contribution was by the species like

buffalo, goat, sheep, yak and camel which is illustrated in Fig. 14.1. According to
the FAO, milk is the fastest growing agricultural commodity with an expected rate
of 1.7% p.a. (expected to reach 981 Mt by 2028) in the following decade.
There are more than six billion consumers of milk and milk products, with many
of them from developing countries (FAO 2016; Kapaj and Deci 2017). Most of the
dairy production is consumed in the form of fresh dairy products. In the coming
decade, the world per capita consumption of fresh dairy products is expected to
grow by 1.0% p.a. due to the higher per capita income growth, especially in India,
whereas overall per capita demand for fresh dairy products are showing a declining
trend in Europe and North America (OECD-FAO 2019). By volume, liquid milk is
the most consumed dairy product throughout the developing world. In urban areas,
the demand of liquid milk is more, whereas in the rural area, it is mainly for fer-
mented milk. According to a survey conducted by Paraffin et al. (2018), urban peo-
ple are consuming 2.8 times more fresh milk than non-urban people. The age group
of more than 30 is consuming three times more milk than under 30. Urban people
are likely to buy directly from kiosks and farms two times more than their non-
urban counterparts because it is convenient or cheap; however, non-urban popula-
tion are procuring fresh milk five times more through vendors because of cheap rate.
Youngsters are buying milk three times more from supermarkets because it seems
safe for consumption. Urban people are considering the quality, nutritive value,
appearance, taste, freshness and accessibility more than non-urban population.
Women are placing two times more important to the nutritive value. Even though its
cooked flavour, UHT milk has gained popularity due to its increased shelf life with-
out refrigeration (Chandrapala and Zisu 2016). The consumers enjoy the attributes
of mouthfeel, flavour profile, texture and appearance of the milk dominant in their
countries (Chandan 2015). Dairy products are a major vehicle for bioactive func-
tional ingredients; these bioactive ingredients target improvements in health-related
areas such as immune response, bowel health and protection against particular
types of diseases (Hoque et al. 2018).
14.3 C
hanging Food Habits of the Population and Need
of Novel Foods
Diet-related problems either due to undernutrition or overnutrition or lifestyle-

related disease are increasing day by day. There is a direct relation between the diet
and the health. According to the WHO report, 462 million of global population are
underweight and 10.8% population are undernourished. Over 1.9 billion adults are
obese or overweight. In India, 61 per cent of deaths happened due to non-
communicable diseases (WHO 2017). There are vivid nutritional status in different
countries, with some of them facing problems of undernutrition (like Myanmar,
Cambodia, etc.), while some are facing problems of overnutrition (like Singapore
and Australia).
The global prevalence of anaemia is 47.4% in preschool-aged children, 41.8% in
pregnant women and 30.2% in non-pregnant women (Badham et al. 2007). In India,
the occurrence of anaemia is 79% in children of 6–35 months, 55% in women of
15–49 years and 24% in men of 15–49 years of age (NNP 1993; NFHS-3 2005–
2006). There is a micronutrient deficiency in about two-thirds of the Indian popula-
tion (Rao et al. 2018). More than 60 countries are being affected by vitamin A
deficiency. According to Allen et al. (2006), approximately 2.5–5 years age group
have more prevalence of vitamin A deficiency and those kids are becoming blind
every year; 50% of them are dying of vision loss under the age of 12 months.
The transformation in socio-economic conditions lead to diseases of affluence
such as obesity, cancer, hypertension, diabetes, alcoholism and cardiovascular dis-
ease. These may occur simultaneously with undernutrition due to deficiency in macro
and micronutrients and protein malnutrition (Wahlqvist and Wattanapenpaiboon
2005). These problems cannot be solved simply by increasing the production rather
by introducing novel food pattern which meets the social condition to the surround-
ings. This has attracted researchers to develop newer foods which meet the consumer
requirements for every age group. Recently there is a mounting demand for custom-
ized dairy products by using genetic or nutritional modification.
14.4 Trends in Dairy Products
There is a higher level of food consumption outside the home and increasing trend in
online and mobile shopping which demand the need for convenient dairy foods. The
traditional values like price, taste and convenience are getting replaced by factors like
health and wellness, safety, social impact and experiences. Improvement in socio-
economic conditions will persist the drive for the demand for dairy products (Lagrange
et al. 2015). Increased prevalence of metabolic disorders and ageing population
demands the need for innovations of inculcating health and wellness in dairy products
beyond the standard product attributes. The consumers now have strong desires to get
information throughout the food chain including the production methods and compo-
nents of the food products. There is a visible trend in environment-friendly products
in buying patterns of consumers. The preferences, attitudes and behaviour of consum-
ers towards dairy products consumption have changed significantly across the coun-
tries (Francesconi et al. 2010). A recent study by Atabek and Atabek (2019) reported
that there is a considerable effect of mass media on perceptions regardless of their
socio-demographic differences. The more educated, wealthier, younger regular dairy
consumers have lower levels of negative perceptions about milk and milk products.
Some of the recent trends with dairy consumers are increased demand for healthy and
clean labels, reduction in added sugar, demand for new flavours, plant-based products
and environment-friendly nutrition (Park 2018). In this chapter, we will focus on some
recently developing areas in the dairy industry.
14.4.1 Clean Label Foods
Nowadays consumers are very conscious regarding the components and ingredients
used in the food products (Euromonitor International 2016). In 1980s, clean label
(clean/natural/ free from/no additives/no preservatives/no chemicals) appeared when
consumer started to avoid food with label containing the E-numbers due to its negative
health effects (Joppen 2006). Nowadays, consumers are very conscious about the com-
ponents and ingredients used in the food products. The demand for the clean label led
to the innovation in functional and organic foods which is mainly driven by the con-
sumer health consciousness. There is no legal or regulatory definition of clean label.
The clean label ensures consuming food in the most natural form with minimal pro-
cessing. Table 14.1 illustrates the multidimensional aspect of clean label. The clean
label ensures consuming food in the most natural form with minimal processing.
The food products which contains the label like natural, organic logos and free
from artificial ingredients are generally considered as clean label foods by the con-
sumers (Gelski 2016). Asioli et al. (2017) made an attempt to define the clean label
foods as the cleanliness of product by assumption and through inference by looking
both the front and back label (Asioli et al. 2017). They concisely described the
major factors that influence consumer motives such as health consciousness, intrin-
sic or extrinsic product features and sociocultural factors. Some of the companies
Table 14.1 Various dimen- Clean label Natural

sions of clean label No additives
No artificial ingredients
All natural ingredients
Minimally processed
No artificial preservatives
No artificial colour
No allergens
No genetically modified foods
are working on it such as Kraft Heinz (removal of artificial colours from macaroni
and cheese), Nestle Dreyer’s Slow Churned Ice Cream (reduced the ingredients and
removed artificial colours and flavours), High fructose corn syrup and GMO,
Cheesemaker Nuestro Queso (uses only rBST free milk) and WhiteWave Foods’
International Delight brand (Simply Pure-coffee creamers) (https://blog.market-
research.com).
14.4.2 Functional Milk Products
From decades consumer has grown with a concept considering “food as medicine”
which helps in accepting the health-promoting effects of foods in our lifestyle.
Health-promoting effect of foods has developed a new concept, called functional
food, which contributes beneficial health effect beyond their nutritive value. Despite
economic challenges, consumer demand for functional food is increasing day by
day. Increase in health awareness, the rising cost of health care, media interest for
novel food, mindset to use natural things rather than medicine, the expectation at
greater cost and increase in life expectancy are the contributors of the growing
demand of functional foods (Kapsak et al. 2011; Kraus 2015). The basic rationale
behind the functional foods is “self-care” through foods. Milk and milk products are
good sources of functional components like probiotics, vitamins, macro- and micro-
nutrients, bioactive components, conjugated linoleic acid, antioxidants, etc. The
consumers generally do not feel functional food as a special category when com-
pared to their counterparts (Oliveira et al. 2016). Health claims are significant in the
case of functional foods to communicate their health benefits. Food labels contain-
ing the health claims in legible form, nutritional composition and production pro-
cess can augment the consumer perceptions for functional foods (Bech-Larsen and
Grunert 2003; West and Larue 2004). The functional foods which are more natural
are more acceptable and healthier carriers (Grunert 2010). If a product is having a
positive health image, it can be positively evaluated with a health claim. For exam-
ple, yoghurt with the halo of the healthier image is more promising carrier than most
popular ice cream (Grunert 2010; Siegrist et al. 2015).
14.4.3 Probiotic Dairy Foods
Within the functional foods, probiotics are the most profitable categories. Probiotics
are living microorganisms which contributes to various health-promoting effects as
they have a beneficial effect on inherent gut microflora and protect them from
pathogenic infection. Probiotics retard the growth of pathogens by generating anti-
microbial substances, immunomodulatory effect, lactase production, intestinal pH
modification, binding pathogens, and production of growth factors and nutrients
(Amara and Shibl 2015). Probiotics have been reported to improve immunity, bal-
anced gut microflora, retard pathogens, prevent diarrhoea, lower serum cholesterol,
improve lactose intolerance and reduces colon cancer (Begum et al. 2017). The
probiotics contribute around 60–70% of the whole market of functional food
(Kołożyn-Krajewska and Dolatowski 2012). The probiotic functional dairy foods

can be fermented or non-fermented. From the past decades, probiotic bacteria have
been added into a variety of dairy products like flavoured milk, ice cream, frozen
dessert, milk powder, baby food, cheese, whey beverages, buttermilk and sour and
normal cream (Begum et al. 2017). Among the probiotic carriers, fermented milk is
considered as the most popular delivery vehicle.
14.4.4 Fortified Milk Products
More than two billion population of worldwide has micronutrient deficiency. In

the developing countries, about 3.5 billion people are affected by the iron deficiency
(ACC/SCN 2000). In India 185 million populations are not receiving enough nutri-
ents. More than 70% of Indian kids under 5 are deficient in vitamin D and about 57%
of children are suffering from vitamin A deficiency. (Bresnyan and Gadha 2018).
Two billion people, i.e. more than 30% of the global population, are suffering from
anaemia. Infants and preschoolers are at major risk, especially in developing coun-
tries (Menchawy et al. 2015). Food fortification is a preventive, simple and low-cost
method to combat nutritional deficiency. One glass fortified milk (320 g) offers 47%
and 34% of the recommended daily allowance of vitamin D and A, respectively.
Vitamin A and D are commonly added to liquid milk and milk powders in the
United States as fat-soluble vitamins are separated along with the cream or fat dur-
ing the production of low fat or skim milk. Some of the US dairies fortify milk with
vitamins E, C and calcium; in addition to vitamins D and A. Nearly all the homog-
enized liquid whole milk and evaporated milk of US are fortified with vitamin D,
which resulted in a noticeable decrease in rickets (child) and osteomalacia (older
people) (Guo 2011). The cow milk contains 47–105 IU (1.2–2.6 μg) of vitamin D
per litre of milk which is very low as per the recommended daily allowance
(400 IU/10 μg). Hence, the cow milk is generally fortified with 400 IU of cholecal-
ciferol per litre. This can be achieved by providing vitamin D-enriched yeast to the
cow, followed by UV irradiation. Currently, the maximum level of vitamin A and D
fortification in liquid milk is 2000 IU (606 μg) and 400 IU (10 μg) per litre, respec-
tively. In the United States minimum, 100% and a maximum of 150% of the required
level of fortification is permissible for liquid milk (Guo 2011). As per report given
by Kruger et al. (2010), vitamin D deficiency has been decreased from 70 to 22%
and 20 to 0% in Indonesian and Filipino women, respectively.
In the case of European countries, around 25–50% of additional vitamin A in the
food are obtained by fortification. In various countries, it is mandatory to fortify
milk; however, some countries voluntarily add micronutrients to low fat or free fat
liquid milk and milk powders (Raunhardt and Bowley 1996). Milk powders and
flavoured milk powders are generally fortified with vitamin D and A, iron and cal-
cium (Singh et al. 2007). Iron is fortified in different dairy products as cheese, infant
foods, yoghurt, flavoured milk and whole milk. Calcium is generally added in milk,
ice cream, yoghurt, cottage cheese, desserts and sour cream (Gerstner 2002; Pirkul
et al. 1997; Klahorst 2001). High-calcium milk drinks are fortified with vitamin D,
zinc and magnesium. Vitamin D3 fortification and quantification have been found in
cheese, yoghurt and ice cream (Kazmi et al. 2007). More details about the milk
and milk product fortification with vitamins and minerals are given in Chap. 5 of
this book.
In 2016, Food Safety and Standards Authority of India (FSSAI) released stan-
dards for fortification of milk; after that low fat, skim milk and whole milk are being
fortified with vitamin A and D. In 2017, a collaborative initiative milk fortification
project of World Bank with Tata trusts and NDDB has explored the prospect of huge
scale milk fortification in India. According to NDDB, 25 milk producer companies,
unions and federation from 20 different states are producing 55 lakh litres of forti-
fied milk per day. One million tonnes of milk have been fortified till date (www.
thehindubusinessline.com 2019). Food fortification is a potent solution to eradicate
the nutrition-related deficiency. Mother dairy of India has initiated the milk fortifi-
cation and selling 25 lac litre fortified milk per day. According to Rhythma (2017),
50% of country milk will be fortified by 2019 and 80% by 2021. The traditional
food fortification processes such as cross-breeding are more acceptable over the
genetic modification process (West et al. 2002; West and Larue 2004). The intake of
iron-fortified milk by schoolchildren resulted in about 27% reduction in iron defi-
ciency (Menchawy et al. 2015). Essential micronutrient zinc and iron were fortified
in milk and found to be effective in enhancing the growth and physical conditions
(Lestari et al. 2009). Milk may enhance the bioavailability of folic acid (Swiatlo
et al. 1990; Wigertz 1997). Milk fortification with folic acid could be done to
decrease the deficiency of folic acid or folate (Verwei et al. 2003).
14.4.5 Low-Calorie Dairy Products
For the wellbeing of consumers the Healthy Weight Commitment Foundation

(HWCF) have passed a resolution in 2010 that about 1.5 trillion calories will be
reduced from market by 2015. Robert Wood Johnson Foundation from the United
States has declared the reduction of 6.4 trillion calories by 2012 which is equal to a
decrease in 78 calories per person/day in the USA (Gable 2014). Healthy Weight
Commitment Foundation is now linked with greater than 250 companies, associa-
tions, retailers and non-profit organizations. The increased prevalence of metabolic
disorders led to the category of low fat and low added sugar products.
To combat the problem of obesity, the company started producing low-calorie
milk products. There is a shift in the preference of full-fat milk towards low-fat
products. Due to the demand, varieties of low-fat milk products like fat-free, 0.5%
fat, etc. are being produced. According to CODEX (CAC/GL 23-1997, rev. 2008),
nutritional claims for solid foods, the low fat and free fat products should have
maximum 3 g and 0.5 g fat per 100 g, respectively, and for liquid foods low fat and
free fat should have maximum 1.5 g and 0.5 g fat per 100 mL, respectively, compli-
ance with low level of saturated fat. The energy (calorie) value in solid foods claim-
ing as low and it should have maximum 40 kcal (170 kJ) and for liquid foods the
energy value as low and free means maximum 20 kcal (80 kJ) and 4 kcal per 100 mL,
respectively, compliance with a low level of saturated fat.
When compared to 25 years ago, the consumption of reduced fat or non-fat milk
has been increased from 14 to 65%. Flavoured milk is a beverage capable of captur-
ing the market held by carbonated drinks owing to its nutritive value. It is the second
most popular dairy product and is available in a plethora of flavours, colours and
varieties which attract the younger generation to experiment with new flavours and
colours (Sethi et al. 2016).
The trend in the industry is to replace the fat and sugars with an ingredient or
prevent the absorption and digestion by using nanoparticle which modifies ice
cream and chocolate that can even reduce the amount of fats and sugars which body
can absorb (Chandrapala and Zisu 2016).
14.4.6 Low-Lactose Dairy Products
Lactose intolerance among the world’s adult population is around 75 per cent
(Silanikove et al. 2015). It is the condition caused by the inability to either digest or
absorb lactose due to genetic reasons or decrease in the intestinal lactase after wean-
ing (resistance lactase pathophysiology or due to damaged lining of the intestinal
epithelial cells) (Tomar 2014). The National Medical Association reports that
lactose-free dairy products are a perfect substitute and are better suited for lactose-
intolerant people than lactose-rich dairy products. Among the top popular health
claims, the lactose-free claim is gaining more popularity among consumers (http://
www.mintel.com/global-new-productsdatabase). Lactose-free is now becoming a
preferred choice for the consumers due to the wide availability of good-quality
lactose-free dairy products. Even when a single member of the family is lactose
intolerant, the consumers are switching to lactose-free dairy which is stimulating
the sales in this segment (Dekker et al. 2019). In order to avoid dairy products elimi-
nation, the consumers are switching to the available alternatives by consuming
lactose-free products which are treated either by neutral lactase or by lactase sup-
plementation (acid lactase) (Suri et al. 2019; Dekker et al. 2019). Low-lactose or
lactose-free dairy products can create a positive health appeal among lactose-
intolerant people as they can relish the nutritional benefits of dairy products. Potable
milk is the largest category of lactose-free dairy followed by lactose-free yoghurt
and lactose-free cheese (https://www.euromonitor.com/). The preference for sweet-
ness generated in milk (pasteurized or UHT) by lactose hydrolysis is especially
appreciated in Latin and Asian countries but disliked in traditional milk-drinking
areas like the Northern Europe and North America (Dekker et al. 2019). There is an
added advantage of calorie reduction also. Flavoured lactose-free milk is in low
calorie and has lesser problems related to Maillard-related off-flavours and
browning.
The National Medical Association reports that lactose-free dairy products are the
most perfect substitute for regular dairy products among individuals with lactose
intolerance. There is a more positive response from children as they preferred lactose-
free cow’s milk over soy beverages (Bailey et al. 2013). But the higher prices for lac-
tose-free products can create an economic burden to the low income consumers, and
there are chances of avoiding such specialized foods from the diet (Suri et al. 2019).
14.4.7 Reduced Sugar Products
Sugar plays a key role not only in flavour of the product but also affects the body,
texture and colour development of the products. The amount of sugar in the products
are replaced by alternative sweeteners like natural and artificial non-nutritive sweet-
eners for making them attractive among the diabetic and health-conscious popula-
tion. The “reduced sugar” can be used to describe the foods that contain at least 25%
less sugar than the reference amount. In general, consumers select the “no sugar
added” claim to the “reduced sugar” claim, most likely because they assume that
other ingredients are added in place of sugar in the latter (Patterson et al. 2012;
McCain et al. 2018). Many consumers still prefer the taste of artificial non-nutritive
sweeteners. The sugar reduction can be done either by substitution, direct reduction
of sugar, ultrafiltration or by lactose hydrolysis methods. Among them, sugar substi-
tution using non-nutritive sweeteners has been a successful approach and is mainly
preferred by the consumers. McCain and coworkers have provided an expansive
review on this topic (McCain et al. 2018).
14.4.8 Low-Cholesterol Dairy Products
Ghee (clarified butter) contains 0.2–0.4% of cholesterol. The increase in cholesterol

level leads to the coronary heart disease. According to the World Congress of
Cardiology (2014), estimated that heart disease will be the main reason for 40% of
the total death in India. Due to an increase in risk, consumers are now very con-
cerned about the consumption of cholesterol products. As dairy products contribute
to saturated fatty acids and cholesterol, consumers are now very concerned about
the consumption pattern. To overcome this problem, many researches are going on
to develop the low-cholesterol ghee. Kumar et al. (2010) prepared the low-
cholesterol ghee using β-cyclodextrin which reduces 90% of the cholesterol. The
cholesterol reduction causes an increase in HDL (good cholesterol) by around 60%
and 75% of LDL (bad cholesterol) reduction (Meghwal et al. 2015). Not in refer-
ence, cholesterol-reduced butter was also prepared by using β-cyclodextrin (Kim
et al. 2006). According to CODEX (CAC/GL 23-1997, rev. 2008), nutritional claims
for a solid product claiming as low cholesterol or free cholesterol should have maxi-
mum 0.02 g and 0.005 g cholesterol per 100 g, respectively, and for liquid foods, it
must be maximum 0.01 g and 0.005 g cholesterol per 100 mL, respectively.
14.4.9 High-Protein Dairy Supplements
The gym culture has led to the formulation of a new category of dairy products rich
in protein targeting the appearance and health-conscious consumers. Dairy proteins
have unique characteristics like easy digestibility and satiety benefits, complete
amino acid profile, etc. making it the “gold standard” in proteins. High-protein diets
are increasingly suggested for fitness and weight management, muscle loss occur-
ring with ageing and blood-sugar and high-blood pressure control maintenance
(Gryson et al. 2014; Banovic et al. 2018). Proteins are sometimes added addition-
ally in products like protein-enriched snacks and yoghurts.
There is an increased demand for sports nutrition dairy products having casein
and whey proteins as key ingredients. The casein and whey proteins can be cheaper
alternatives to overcome the malnutrition. Whey protein concentrates are the key
ingredient in performance (sports) beverages and nutritional supplements. Whey
proteins with its nutritional benefits cost its efficiency and sustainable production
technologies will drive the growth of the dairy industry.
14.4.10 Organic Milk
Organic foods are a trending as fast-growing segment among dairy foods. According
to the Organic Food Production Act 1990 of US Department of Agriculture (USDA),
cattle that yield milk certified as organically raised naturally, not exposed to hor-
mones and antibiotics, fed with organic food for a whole year before certification
and provided with organic grazing area. Consumer strongly believes that they are
consuming a healthier product, while the composition of organic and regular milk is
similar in compositions of protein, carbohydrate, mineral and vitamins (Wisconsin
Milk Marketing Board, WMMB 2012).
There is conflict in evidence about the increased amount of conjugated linoleic
acid content in organic milk; however, the omega-3 fatty acid and polyunsaturated
fatty acids are higher than the regular milk (Ellis et al. 2005). The increased market-
ing of the organic product is mainly due to the consumer emotions for animal wel-
fare and better health (Croissant et al. 2007), and consumer considers the organic
product as a better ethical option (McEachern and McClean 2002). As per the study
conducted by McEachern and McClean (2002), more than half of the consumers
that favour organic farming is superior for animal well-being (64%), conventional
dairy farming is very intensive (53.5%) and some are not accepting animals to give
hormone for higher milk production (68.5%). Though a greater number of consum-
ers are still buying regular milk, a great number of populations are believing organic
product as extra valuable and ready to pay an extra amount for that. Since 1993,
organic milk is making an image in the market over conventional milk (Dimitri and
Greene 2000). The sale of organic milk product was 15.8 million dollars in 1996,
which increased up to 104 million dollars in 2000 as reported by the US Department
of Agriculture (2012). USDA report stated that there was a decrease of 2.8% in
regular milk in 2011 compared to 2010.
14.4.11 A1 and A2 Milk
Recently A2 milk is promoted as a healthier option than the general A1 milk. The
milk protein beta-casein differentiates the milk into A1 and A2, which is mainly due
to the different amino acid at 67th position. A1 milk protein can yield beta casomor-
phin-7, and some company claims that A2 milk is healthier and A1 milk may have
adverse health effects. However, on the basis of scientific research, in 2009,

European Food Safety Authority (EFSA) stated that there is an inadequate evidence
which claims the negative effect of regular milk (European Food Safety Authority
2009). Despite the absence of certain scientific claim, many companies are boosting
A2 milk marketing and claiming better milk with no ill health effects (Satyanarayana
2018). Indian indigenous buffalo and cow breed produces A2 milk. In India, several
dairy companies have started to supply A2 milk as brands Desigo, Amul Deshi,
GoShrushti, Haritas, etc. (Satyanarayana 2018). In 2000, New Zealand started A2
milk company to produce A2 milk. In Australia, A2 milk is the fastest-emerging
milk brand and the growth of revenue from A2 infant nutrition is 35.3% (https://
thea2milkcompany.com/our-businesses/). In China, A2 milk has launched in 2013
and set a strong position in the market. A2 milk was launched in 2015 in the United
States and successfully growing at a rate of 118% among consumers. They are sell-
ing A2 milk as A2 milk with the label “easier on digestion”, reduced-fat A2 milk,
low-fat milk and chocolate reduced-fat milk. This milk is gaining a great level of
customer reliability (https://thea2milkcompany.com/our-businesses/).
14.4.12 Designer Milk
In the era of functional foods, the demand for tailor-made milk is on the rise. The
milk is designed in such a way that its properties will be modified from the standard
constituents of milk according to the need of the customers. As per the diet and
health requirements or the processing/technological developments, the designer
milk can be developed. Some of the applications in diet and human health include
increasing the content of unsaturated fatty acids (USFA) in milk fat, reducing the
lactose content, humanization of bovine milk or eliminating β-lactoglobulin from
milk, melatonin enrichment, etc. (Umaraw et al. 2015). The alterations in the pri-
mary structure of casein, enhancing the protein content in milk, increasing the curd
clotting time, increased yield and/or more protein recovery, milk containing nutra-
ceuticals and replacement for infant formula, etc. are some of the processing and
technological developments could be done by designing the milk. The customiza-
tion or designing the milk can be achieved by either incorporating required func-
tional ingredients directly into fluid milk/dairy products or by modifying the feed
formulation of the lactating animals to get the desired milk compositions.
The designer milk is compositionally advanced that it provides health and thera-
peutic benefits. Animals capable of providing milk having components like insulin,
human plasma proteins, drugs and vaccines have already been developed. Intensive
researches have been done to reduce the saturated fats in milk, lactose and lacto-
globulin as well as to increase the protein content, conjugated linoleic acid (CLA)
and omega-3 fatty acids in the milk (Singh et al. 2019). Transgenic goats are capa-
ble of producing anti-thrombin containing milk, rabbits that can produce Ruconest,
a recombinant C1-esterase inhibitor which is used in the treatment of hereditary
angioedema, etc. have gained the approval of USFDA (Kling 2009; Nature
Biotechnology 2014).
14.4.13 Extruded Dairy Foods
In the worldwide agri-food sector, extrusion technology is gaining popularity with

the enormous opportunity in food innovation. Extruded cereals dominate a signifi-
cant distribution of the snack foods in the market. Extrusion cooking technologies
are used for cereal and protein processing in foods; new opportunities in this seg-
ment of food are by incorporating dairy ingredients in breakfast cereal foods and
snack food (Rao et al. 2018). In broader terms, the nutrient densities of snack foods
are low, which stigmatized these products in the category of “junk” foods. Inclusion
of dairy-based ingredients such as whey proteins, caseinates and other milk proteins
helps to improve the textural as well as nutritional properties of the extruded prod-
ucts (Guha et al. 2003). With the development of extrusion technology, special
attention is focused on the enrichment of extruded products with different types of
proteins and their protein fractions mainly with the addition of whey protein con-
centrate (WPC). This enhances the nutritional value and functionality of the prod-
ucts. The amalgamation of whey protein into extruded-extended products has gained
appeal due to the possibility of producing a high-protein snack food that will fit into
the functional food category. Due to the short holding time in the extrusion process,
a partial amount of protein denaturation occurs, thereby altering the overall texture
of the products, with whey proteins retaining the nutritive and functional properties
of the product (Brncic et al. 2011). WPC as an important source of proteins and
minerals is considered as one of the highest-superiority components for possible
extrude enrichment. Incorporation of different amount of WPC in corn flour as raw
material is helpful during extrusion cooking in order to improve the nutritional-
textural-mechanical properties of the manufactured product (Brncic et al. 2011).
14.5 Conclusions
In order to commence a new product in the market, it is essential to first comprehend

consumer’s perception and their necessities. People are increasingly aware about
their health and health information can be effectual in altering consumers’ demand
for milk and dairy products. Milk and dairy products are especially rich in bioactive
components, and this one state is where the scientific nutrition judgment of food
products coincides with consumers’ perception. Hence, it will gradually influence
food choices through changes both in individual activities and in the regulatory
structures. The producers will innovate new products according to the changing
perceptions. Therefore, stakeholders must take initiatives to propagate all requisite
information, specifically the nutritional facts that can help to increase the level of
belief regarding their marketed milk; the strong belief in milk’s goodness is a pow-
erful platform. But traditional products are not enough; consumers want beverages
that can deliver the greatest health benefits. Various brands worldwide are already
presenting that there are huge opportunities for innovative and healthier new prod-
ucts in the dairy sectors that will meet the shifting desires.
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39d76187c7abf78f7becca068ddadec3.pdf. Accessed 4 Sept 2019
Novel Dairy-Based Drinks: Changing
Scenario 15
Swathi P. Anand and Nancy Awasti
Contents
15.2 C hanging Regulatory Scenario 302
15.2.1 How Did the Changing Regulations Support Innovations in Dairy Drinks? 303
15.3 Dairy Drinks and Beverages 303
15.3.1 Classification of Milk-Based Drinks 304
15.3.2 Innovative and Novel Dairy-Based Drinks 304
15.4 Key Players in Dairy Beverage Market 309
15.5 Major Components of Dairy-Based Drinks 310
15.5.1 Milk Protein 310
15.6 Novel Stabilizing Additives Used in Milk-Based Drinks 313
15.6.1 Pectin 313
15.6.2 Carrageenan 314
15.6.3 Carboxy Methyl Cellulose 315
15.6.4 Modified Starch 315
15.6.5 Xanthan Gum 317
15.6.6 Agar 318
15.7 Novel Processing Techniques: Pathway Towards the Extended Shelf Life of
Dairy-Based Drinks 318
15.7.1 High Hydrostatic Pressure Technology 319
15.7.2 Ultrasonic Processing 320
15.7.3 Pulsed Electric Field Processing 321
References 322
Abstract
Dairy processing witnessed a series of developments in the fields of ingredient
and processing technology in the last few years. The development of novel addi-
S. P. Anand (*)
New Product Development (Dairy), Britannia Industries Ltd., Bengaluru, India
N. Awasti

https://doi.org/10.1007/978-981-15-2608-4_15
302 S. P. Anand and N. Awasti
tives such as gellan gum, carboxymethyl cellulose, different grades of carra-

geenan and pectin has extended the scope of novelties in dairy-based drinks. This
has enhanced the feasibility of processing techniques for processing liquid dairy
including drinks. With these developments in ingredient technology, technolo-
gists are able to use different processing techniques to develop fresh as well as
ambient dairy drinks. Apart from the heat treatments such as pasteurization and
sterilization, now we are able to utilize cold processing techniques such as high
hydrostatic pressure technology, ultrasonic processing, pulsed electric field pro-
cessing, etc. also in dairy-based drinks. These techniques will increase the scope
of innovation and also help in conserving the nutritional quality of products.
With these advancements, we are now able to witness a wide range of novel
drinks in the market such as whey drinks, carbonated dairy drinks, functional
drinks, dairy-based energy drinks and even live probiotics in processed dairy
beverages.
Keywords
Dairy drinks · Heat treatments · Extended shelf life dairy drinks · Whey-based
drinks
15.1 Introduction
The momentum of food industry is and was always influenced by the changing
consumer perceptions. There have been tremendous market swings in the last
20 years, from homemade healthy foods to healthy and tasty packaged foods and
from premium creamy rich dairy foods to healthy low fat-sugar-sodium foods.
These changes have provided wide scope for the food technologists to experiment
and provide delightful yet ‘healthy’ concepts even among dairy foods. These
changes in consumer perspectives have led to increased diversity among the pack-
aged and processed foods, especially dairy-based drinks. The last decade has been
seen for a large number of diverse dairy drinks hitting the markets, which includes
short shelf life fresh milk and fermented drinks, extended shelf life drinks, dairy
fusion drinks with cereals, malt, fruits and seeds, on-the-go breakfast drinks, sports
drinks, fortified drinks, etc. The changes happening in various fields like dairy pro-
cessing, product development, analytical methods, regulations, nutrition, etc. have
their own shares in the evolution of dairy market.
15.2 Changing Regulatory Scenario
In recent times we witnessed a variety of amendments happening in regulatory pur-

view, including US Food and Drug Administration (USFDA), Food Standards
Australia New Zealand (FSANZ), GCC Standardization Organization (GSO) and
15 Novel Dairy-Based Drinks: Changing Scenario 303
Food Safety and Standards Authority of India (FSSAI). The regulations are now pro-
viding a wider environment for innovations and emphasis on healthy eating as well.
The traffic light labelling, Guideline Daily Amount (GDA) labelling, health star rat-
ing, etc. are such initiatives.
15.2.1 How Did the Changing Regulations Support Innovations

in Dairy Drinks?
The use of food additives is very important when it comes to any shelf-stable pro-
cessed food. Although the technologies such as ultra-high temperature (UHT) or
flash heating are helpful, even those require the use of stabilizers in the form of food
additives. Carrageenan for neutral milk products and pectin for acidic milk products
are the most common stabilizers which have been in use since time immemorial. Up
to a certain period, only these stabilizers were allowed to be used when it comes to
processed milk drinks. But now the regulatory scenario has changed immensely,
and a wide range of food additives are allowed in milk products, which includes
stabilizers, preservatives, emulsifiers, acidity regulators, thickeners, etc.
15.3 Dairy Drinks and Beverages
Currently milk-based beverage market is facing challenges in terms of acceptance

and preferences by consumers and is lagging in sales in comparison to milk and
yoghurt market. However, there have been numerous innovations to produce
milk-based beverages, and nowadays it is among one of the fastest-growing dairy
segments. Buttermilk produced as a by-product of butter production is considered
as one of the oldest acid milk-based beverages. The changing preferences and
requirements with increase in purchasing power, has led to increase in growth of
dairy beverage industries worldwide. As per recent report by Steyn (2019), the
global market for dairy beverages is expected to be attaining a market value of
approximately 91 billion USD by 2024. To serve the growing demands and prefer-
ences of consumers, several dairy-based drinks have been introduced in the mar-
ket by the industries. Due to presence of numerous customers and vendors and
limited hurdles for the entry of new dealers in the dairy beverage market, there
exists a monopolistic competition between them. Therefore, to overcome these
challenges, the need and preferences of consumer and their geographic reach must
be well focused.
Overall, the dairy beverage market has been divided based on region, prefer-
ences, distribution channel, packaging type and type of dairy beverage such as whey
based, fermented milk, milk, kefir and milk and whey-based probiotic drinks, but-
termilk and others. This chapter summarizes few dairy-based drinks and beverages
and their key players or vendors that are currently available in the market.
15.3.1 Classification of Milk-Based Drinks
The classification of milk-based drinks can be done on the basis of their pH values
and shelf life. Table 15.1 represents the classification of milk-based drinks in detail.
Based on pH values, dairy-based drinks can be classified as:
1. Milk-based drinks: It includes flavoured milk, milk shakes, etc. which have a
neutral pH ranging from 6 to 7.
2. Fermented milk drinks: It includes drinkable yoghurt, lassi, buttermilk, acidified
milk, etc. which have an acidic pH ranging from 3.5 to 5.
Based on shelf life of milk-based drinks, it can be classified as:
1. Fresh/cold chain drinks: This includes low heat treated products such as pasteur-
ized flavoured milks, non-heat treated fermented milks, etc. which will have a
short shelf life of less than 1 month. These products ideally require proper cold
chain distribution and storage under refrigeration (2–8 °C).
2. Extended or long shelf life/ambient drinks: This includes sterilized milks, UHT
drinks, etc., which will have a longer shelf life of about 3–9 months. These prod-
ucts are shelf stable and thus provide flexibility in distribution and storage.
15.3.2 Innovative and Novel Dairy-Based Drinks
15.3.2.1 Functional Milk Beverages

The normal milk-based drinks can be divided into flavoured milk and plain milk.
Functional milk beverages can be made by using plain milk or flavoured milk along
with additional functional ingredients. As its name indicates ‘functional’ which
explains itself, it has specific functional components which must be beneficial for
health. The demand of functional milk beverages in market is mostly driven by the
increased popularity and interest of consumer towards the health benefits of the
dairy-based beverage. As of 2017, numerous dairy-based drinks have been launched
Table 15.1 Classification of milk-based drinks

Acidic pH (3.5–5) acidified/fermented
pH Neutral pH (6.6–7) milk based milk based
Shelf life
Fresh refrigerated Low heat treated flavoured milks Drinking yoghurts, lassi, buttermilk
storage short low heat treated milk shakes not heated after fermentation
shelf life
Long life UHT heat treated flavoured milks Drinking yoghurts, lassi, buttermilk
ambient storage and milk shakes heated after fermentation
extended shelf
life
in different countries, and out of them, only eight were highlighted in Mintel Blog,
2018. Among those eight drinks, an iconic turmeric ginger protein drink, USA was
referred as year’s big trend, golden milk. Turmeric ginger protein drink was found
endowed with approximately 130 calories with 20 g proteins and only 3 g of sugar.
The manufacturer claimed this drink is free of GMOs, lactose and artificial ingredi-
ents. It has also been claimed to be clean with premium ingredients and was made
from milk of grass-fed cows. In addition to this, a US-based company (Fairlife,
Chicago) launched lactose-free, ultra-filtered milk with 15 g of high-quality protein.
Fairlife claimed that their ultra-filtered milk has 60% less sugar and 50% more cal-
cium than regular milk. In June 2016, Cavin Care Pvt. Ltd. successfully launched
dairy-based drink in three different flavours, whereas recently in December 2018,
ITC limited (India) expanded their dairy beverages by launching milk-based bever-
age named Sunfeast Wonderz milk.
Because milk-based beverage is one of the best suitable medium for the supple-
mentation of several functional ingredients such as proteins, minerals, bioactive
components, plant extracts, polyphenols, omega-3 fatty acids, etc, these compo-
nents have been known for the boost up of human health. The consumption of milk-
based beverages is increasing day by day because of its nutritional and functional
properties by their functional ingredients (Farah et al. 2017). The milk-based bever-
ages could be low calories, zero calories, low fat/lactose/carbohydrates and low
cholesterol beverages, which are the current demand of health-conscious
consumers.
15.3.2.2 Whey-Based Flavoured Beverages and Drinks

Among various functional beverages present in the market, whey-based drinks in
combination with different varieties of fruit fermentations could be considered as
a novel functional beverage. The fruit usually contains nutraceutical components,
and after combining it with whey, it usually strengthens its functional value (Özer
and Kirmaci 2010). The functional property of this whey-based juice drink can be
changed by formulating the type of fruit juice in it. Few fruits such as strawberries
and blueberries are known to prevent oxidative stress due to the action of flavanols,
catechins, anthocyanins and phenolic acids in it. These components have the abil-
ity to scavenge free radicals and other reactive oxygen species (Zafra-Stone et al.
2007; Brambilla et al. 2008; Granato et al. 2018). However, it reduces the progres-
sion of oxidative stress-induced risk of numerous diseases like cancer-, obesity-
and cardiovascular-related issues. Apple juice being rich in polyphenols, phenolic
acids and flavonoids such as procyanidins, chlorogenic acids, phloretin glycosides
and quercetin glycosides were found associated in reducing risk of chronic disease
by several mechanisms including antiproliferative, cell signalling and anti-oxida-
tive effect (Hyson 2011). A recent study (Ferreira Wessel et al. 2016) on pear juice
demonstrated its oxygen quenching abilities and antioxidant potential due to pres-
ence of components such as quercetin, hydroxy-cinnamic acids, catechins and
other polymeric units. Several studies have focused on the development and pro-
duction of whey-based drinks with various fruit formulations and exploited the
combination of sweet rennet-based whey and acid whey (Djurić et al. 2004;
Goudarzi et al. 2015). A study done by Rizzolo and Cortellino (2018) replaced
whey with Ricotta-cheese whey and produced pectin free drinks including apple,
blueberry and pear. Apart from these whey-based drinks, especially energy drinks
also have a huge ability to provide consumer a nutrient-rich healthy choice at
affordable cost. Usually, whey-based energy drinks are fruit flavoured to effec-
tively mask the flavour as well as odour of fresh whey. Naik et al. (2009) prepared
a watermelon-flavoured whey-based energy drink and found it was acceptable in
terms of texture, flavour and colour at refrigerated conditions. A study done by
Divya and Kumari (2009) prepared drink with guava flavoured with paneer whey
and found it acceptable for consumption even after 45 days of storage. Recently,
Kumar (2017) developed nutritious beverage using whey and pineapple juices and
found it was stable for up to 2 months. It is worth mentioning that whey-based fruit
beverages and drinks are excellent source of energy, nutrition and good therapeutic
properties and therefore can be consumed by a wide range of consumers including
children and old people. For detailed review on product, application technologies
and health benefits of milk and whey-based beverages, see chapter from Guneser
et al. (2019).
Fortification of whey-based beverages with prebiotics and fruit flavour is also an
innovative idea in the new product development industries. The addition of prebi-
otics in the whey-based beverages not only improves its physico-chemical proper-
ties but also contributes to functional characteristics and promotes the growth of
probiotic bacteria. The prebiotic xylooligosaccharide or XOS concentration
1.25 g/100 mL has been reported as an interesting ingredient to improve the func-
tional properties in terms of anti-diabetic, anti-hypertensive and antioxidant activ-
ity in the strawberry- flavoured whey-based beverages (Souza et al. 2019).
Functional whey beverages have been prepared by blending pineapple juice with
paneer whey and cheese whey at different concentration 10, 20 and 30% to study
their physico-chemical, nutraceutical and microbial properties for 60 days. The
overall study reports suggested that paneer whey-based pineapple beverage had an
excellent nutraceutical quality and nutritional values (Baba et al. 2016). The nutri-
tional components of paneer whey and cheese whey have been studied by Goyal
and Gandhi (2009) and explained that paneer whey contains significantly higher
concentration of minerals such as calcium, chloride, potassium and sodium.
However, paneer whey-based beverages could improve the nutritional value of the
developed beverages.
Whey-based non-alcoholic whey drink ‘Acidowhey’ is prepared by the fermen-
tation with L. acidophilus and L. bulgaricus which is also reported for their health
benefits (Hati et al. 2013). Other whey-based beverages such as whevit, whey-based
soups, whey-based lassi, etc. have also been reported by Hati et al. (2013).
15.3.2.3 Carbonated Energy Boosting Drinks

Ravindra et al. (2014) demonstrated the use of carbon dioxide in extending shelf life
of cardamom-flavoured dairy drink. A yoghurt-flavoured carbonated drink, i.e. lotte
milk, was launched in America. This drink was made up of yogurt and carbon diox-
ide producing cultures, to give consumer a feeling of soda beverages. This innova-
tive drink is originally from South Korea and now is also available in the United
States. A combination of creamy texture of dairy and carbonation produced using
dairy cultures was found acceptable. Similarly, a carbonated lactic drink was pre-
pared in Japan and was named as Calpis soda rich. This product was prepared using
lactic acid bacteria, whey powder and skim milk powder. It was launched in
December 2012 and accounted for about 86% of total sale of carbonated dairy-
based soda drink between December 2016 and November 2017.
In Austria, a company named Milfina launched ultra-heat treated flavoured milk
by using partially skimmed milk. It is known to contain matcha, calcium-bound
dextrose and guarana and was claimed to contain health-promoting attributes with
lactose-free characteristics. According to a survey, 73% of German consumers pre-
ferred to use this drink due to its beneficial attributes.
15.3.2.4 Dairy-Based Probiotic Drinks

With the changing scenario towards nutrition and health, there is increased demand
of food that can optimize the wellness. Probiotic-based dairy beverages usually fall
under the category of functional food and are classified as fortified, whole or
enriched food that provides human body with positive health benefits (Hasler 2002).
Based on the product matrix, probiotic-based beverages can be categorized into two
groups, i.e. probiotic milk-based beverages and probiotic whey-based beverages, as
detailed below:
Probiotic Milk-Based Beverages

These are usually made up of milk from cow, sheep, mare or goat, and several other
traditional dairy-based beverages have been reported. Among traditional dairy bev-
erages, kefir is believed to be the first known functional dairy beverage and is asso-
ciated with longevity of Caucasian people. For more details on kefir, readers can
refer a study from Özer and Kirmaci (2010). In addition to kefir, Koumiss is another
dairy drink that was originated in ancient time. Koumiss has been found to associate
in curing numerous illnesses linked to stomach and liver and even tuberculosis
(Guzel-seydim et al. 2009). Often, kefir and koumiss are called as ‘probiotic’ prod-
ucts, but as per existing regulations and lack of science evidences, they cannot be
considered as probiotic products unless proven. Therefore, in this regard, both fall
in the category of traditional functional dairy beverages.
In addition to yoghurt, probiotic yoghurt drink has attracted a wide attention and
reached a high market success during the last two decades. The detailed description
on commercially available milk-based beverages containing probiotic microorgan-
ism is given in a review by Turkmen et al. (2019). Acidophilus milk produced using
Lactobacillus acidophilus comes with a slightly thick texture and tangy flavour. But
due to the production of organic acid by Lactobacillus acidophilus strain, the shelf
life of acidophilus milk is short. Therefore, a sweet acidophilus milk was prepared
using Lactobacillus acidophilus NCMF strain with extended shelf life of up to
14 days (Suarez et al. 2003). A study done by Shiby and Mishra (2013) showed an
extended shelf life of acidophilus milk for up to 28 days when freeze-dried concen-
trated cultures were used.
Bifidus milk made from heat-treated milk and Bifidobacterium bifidum is another
fermented dairy drink that was developed in the early 1950s (Gürakan et al. 2009).
Another skim milk-based probiotic dairy beverage is Yakult and is a Japanese origi-
nated fermented drink. Yakult is made up of skim milk, sugar and skim milk powder
and is fermented using Lactobacillus casei Shirota. Apart from Yakult, many well-
known probiotic drinks such as Biola, YOMO, LGG+, Gefilus and Yoplait 360 are
available in the market, and all these products contain Lactobacillus rhamnosus GG
(LGG).
Currently, a dairy drink made from milk 100% pasture raised and grass-fed cows
was launched in the United States. This yoghurt-based drink was claimed to contain
20 billion cells of probiotic cultures including BB-12 probiotics, nutrition and 12 g
protein. This full-fat yogurt drink was found endowed with high source of ribofla-
vin, vitamin C, biotin, calcium, vitamin A, iron, vitamin B6 and potassium. This
drink was called as dreaming cow lush peach, ginger, pumpkin and carrot grass-fed
yogurt drink. A similar product with lychee and mangosteen flavour (Mr. Probiotic)
was launched in China. It was claimed to contain approximately 100 billion of pro-
biotic strain of Lactobacillus strain LGG. This culture is known for its probiotic
properties and in promoting intestinal functions. With growth in health-related con-
cerns and benefits for consuming health-promoting drinks, there will be an expected
increase in demand of these drinks worldwide.
Probiotic Whey-Based Beverages

Whey is a by-product of cheese and paneer industry. The perfect utilization of this
by-product could be done by making various whey-based beverages. Whey contains
many nutritional components which can be utilized for the promotion of health
benefits. Nowadays demands for probiotic products are high because of its thera-
peutic aspects. So, utilization of plain whey beverages could be done by innovative
techniques. This may include addition of some specific functional ingredients like
probiotic bacteria, bioactive peptides, proteins, etc. The preparation of whey-based
beverages using probiotic bacteria has many health benefits. As probiotic bacteria
itself exhibit many health promotional effects, addition of probiotic bacteria in the
whey-based beverages could be even more fruitful. Examples of probiotic whey-
based beverages are butter milk, sour milk, flavoured milk, milk smoothies, etc.
Majorly two different types of whey are used for the preparation of probiotic whey-
based beverages: acid whey and sweet whey.
Probiotic fermented beverages based on acid whey have been prepared by
Skryplonek et al. (2019). Probiotic cultures Bifidobacterium animalis ssp. lactis
BB-12 or Lactobacillus acidophilus LA-5 have been used for the preparation of
probiotic acid whey beverage. The developed acid whey beverage has been seen for
the good carrier for probiotic bacteria and their viability was also maintained up to
8 log cfu/mL during the storage period. However, this product can be used for a
therapeutic purpose (Skryplonek et al. 2019). Similar study reported on fermented
probiotic whey beverage containing B. animalis BB-12 and L. acidophilus LA-5
was made by rennet whey (Drgalić et al. 2005). Whey-based beverages supple-
mented with probiotics improve the health promotional properties, textural proper-
ties as well as flavour of the products. Some of the whey-based probiotic beverages
such as Activia, Actimel, Chamyto (Nestlé), Danito (Danone), Vigor-Club (Vigor),
Yakult and Sofyl (Yakult), etc. are commercially available in the market (Katz
2001).
Sweet whey is prepared by rennet coagulation. The acidity of sweet whey is less
than the acidic whey. That is why it is sweeter and tastier than acid whey. Whey-
based beverage made by rennet whey and milk along with probiotic bacteria and
yoghurt bacteria supported the viability of bacteria and product acidifying capacity
(Almeida et al. 2009). Similarly, 30–40% of rennet whey and milk with 1–5% pre-
biotic oligosaccharide fermented with L. acidophilus, S. thermophiles and
Bifidobacterium spp. prepared and found that there was no any significant influence
on the viability of used cultures and fermentation pattern (de Castro et al. 2009). In
another study, rennet whey-based probiotic beverage has been prepared by using
yoghurt bacteria and probiotic bacteria L. helveticus. The fermentation process was
carried out at 42 °C and found to be most appropriate (Bulatović et al. 2014a). When
shelf life study was done for 22 days, this developed product exhibited the better
performance via high cell viability and stability till long storage. Further study has
been done by the addition of probiotic strain Lactobacillus johnsonii and prebiotic
inulin in the beverage, which again improved the nutritional value of the beverage
(Bulatović et al. 2014b). Similar study has been conducted by Bulatović et al.
(2014c) to see the textural and sensory attributes and stability of probiotic bacteria
in the fermented whey-based beverage. For this, they prepared the fermented whey-
based beverage using probiotic strain L. rhamnosus and enrichment with 30% of
skim milk. The developed product had desirable textural characteristics and sensory
attributes. The shelf life of this probiotic whey-based beverage was up to 20 days
with high level of live probiotic bacteria (Bulatović et al. 2014c). However,
probiotic-enriched whey-based beverage has nutritional as well as therapeutic val-
ues. Goyal and Gandhi (2008) reported that whey-based beverages have anti-
diarrhoeal property.
15.4 Key Players in Dairy Beverage Market
Some of the main players that are currently in the global dairy beverage market are
Organic Valley (USA), Fairlife (USA), Lactalis group (France), Arla Foods amba
(Denmark), ITC limited (India), Nestle SA (Switzerland), The Whitewave Foods
company (USA), Kefir company (New Zealand), Dean foods company (USA) and
many more. Based on region, the global dairy beverage market is divided into North
America, Asia Pacific, Europe and the rest of the world. In coming years, among all
these regions, North America is expected to lead and dominate the dairy beverage
market as Canada and the USA are major consumers of ready to drink and dairy
beverages. With change in eating habits and shift towards ready to drink beverages,
beverages enriched with nutritional and therapeutic values are in trend. So, the
demand for these types of beverages could be expected to grow in the future market
in the Asia-Pacific region. Further this will lead to the expansion of dairy-based
beverage industries.
15.5 Major Components of Dairy-Based Drinks
15.5.1 Milk Protein
The composition of milk depends on various factors such as genetic factors (genet-
ics, species and breed), age of the animal, seasons, lactation period, environmental
factors and feed and physiological factors. However, the average composition of
milk is as follows: 86–88% water, 3–4% fat, 3–4% protein, 4–5% lactose, <1% ash
and 9–11% milk solids-not fat (MSNF). Milk being the major component in milk-
based drinks owes to more than 50% of its composition. Thus milk as a component
greatly contributes to the body and texture of the finished product. Milk protein is
the component which greatly contributes towards the stability, texture and mouth-
feel of milk-based drinks. Moreover, the health benefits of milk proteins also play
an important role in the growth of this food category.
Long shelf life drinks are majorly subject to UHT processing, wherein the prod-
uct is subjected to heat treatments higher than normal pasteurization to make it
sterile and safe. The novel technique of UHT heat treatment and aseptic filling has
led to a revolution in the processed milk drink industry, capturing a major market
share. Keeping the micellar structure intact is the major challenge when it comes to
processed dairy drinks, may it be milk or fermented milk-based product. The UHT
heat treatment temperatures range from 90 to 140 °C depending on the pH of fin-
ished product.
Stability of milk-based beverages could be increased by some food additives
including heat treatment. Some majorly used stabilizers are pectin, carrageenan,
cellulose gum, microcrystalline cellulose, modified starch, etc. These stabilizers are
described in details, in the following sessions.
15.5.1.1 Chemistry Behind the Heat Stability of Milk Proteins

The major processing that happens in the dairy industry is heat treatment. Heat
treatment of milk and milk products are very essential to ensure food safety. The
common heat treatments encountered in dairy processing are listed in Table 15.2
(Davies and White 1966; Singh 2004).
Compared to many other fluids, milk is heat stable to some extent. This means
that the emulsified/colloidal state of milk protein remains intact during heat treat-
ment. However, numerous physiochemical changes occur during heating which
influences the nutritional and sensorial properties of the product. Let us discuss
some of these changes in detail:
Table 15.2 Heat treatments used in dairy industry

Heat treatment Conditions Primary objective
Thermization 65 °C × 15 s Destroy temperature-sensitive organisms
such as psychrotrophs, prerequisite for
low-temperature storage
Pasteurization Destroy all pathogens and also spoilage
(a) LTLT (low 63 °C × 30 min causing microorganisms, to render shelf life
temperature long time) as well as safety for human consumption
(b) HTST (high 72 °C × 15 s
temperature short
time)
Forewarming for 90 °C × 2–10 min, To increase the stability of milk proteins for
sterilization 120 °C × 2 min the upcoming sterilization (intense heating)
Sterilization Attain commercial sterility and obtain a
(a) UHT (ultra-high 130–140 °C × 3–5 s shelf-stable product with long shelf life
temperature)
(b) In-container 110–
115 °C × 10–20 min
Fig. 15.1 Inactivation of enzymes during heat treatment (Fox et al. 1998) (t’: Inactivation time for
enzymes, time required for browning, time required to convert 1% lactose to lactulose, coagulation
time)
Effect on Milk Fat

The heat treatment of milk and milk products leads to the disruption of fat globule
membrane. This results in coalescence of fat globules during direct UHT treatment.
Further at temperatures >70 °C, whey proteins on the milk fat globule membrane
denature, leading to the presence of free amino acids such as cysteine. This can
cause off flavour due to the release of H2S. These amino acids also contribute
towards Maillard browning with lactose. When the product is not subject to homog-
enization, they can show ‘oiling off’ after UHT. This is due to the release of non-
globular free fat up on heating. This phenomenon is the main cause of ‘Cream Plug’
defect in UHT products.
Effect on Enzymes
The indigenous enzymes get denatured during the heat treatment. Figure 15.1
clearly describes the effect of temperature and time on the enzymes activity.
Effect on Lactose
The effects of heat on lactose include isomerization, optical rotation and formation
of volatiles. At high temperatures especially at alkaline conditions, lactose epimer-
izes to fructose, leading to the formation of lactulose [4-O-β-d-galactopyranosyl-d-
fructofuranose]. Lactulose concentration in milk is the highest during indirect UHT
treatment. Another change noticed in milk while heating is the drop of pH or
increase in titratable acidity. This occurs due to the formation of acid (majorly for-
mic acid) from lactose. In commercial production of milk-based drinks, acidity
regulators such as phosphates are added to the product to address this pH drop.
Maillard browning is one of the major concerns on heat-processed dairy prod-
ucts. Maillard browning is the reaction between sulphur containing amino acids
with sugar to form brown compounds called melanoidins. This is undesirable not
just because of the discolouration but also because of the strong flavours from
polymerization products such as hydroxyl methyl furfural (HMF). The involvement
of amino acids such as lysine, cysteine, etc. reduces the bioavailability of proteins.
This also reduces the protein digestibility.
Effect on Proteins
Whey proteins are the most heat-labile proteins in milk because of the extended
secondary and tertiary structures. The order of heat stability is as follows:
α-lactalbumin (α-La) > β-lactoglobulin (β-Lg) > bovine serum albumin
(BSA) > immunoglobulins (Ig) (Fox et al. 1998). At temperatures higher than 75 °C,
the proteins form sulphydryl-disulphide linkages that affect the heat stability and
rennet coagulation. This effect is very critical in renneted milks/drinks. The release
of H2S from proteins results in cooked flavour in the finished product, which may
not be desirable. Cross-linkages happening during heat treatment can reduce the
solubility, digestibility and bioavailability of proteins.
Approximately 70% of milk proteins are casein. These are not susceptible to
thermal denaturation. This property of casein supports heat treatment of dairy drinks
without any concerns. However extreme conditions of processing can result in

aggregation, de-phosphorylation, disulphide bridging, etc.
Although high heat treatment imposes various negative impacts in milk-based
drinks, it is inevitable for achieving long shelf life and food safety. Technologists
have found a way out by finding suitable temperature-time combinations where the
necessary microbial inactivation can be achieved and the negative impacts can be
minimized. To reduce the negative effects of heat treatment on milk constituent dur-
ing dairy-based drink preparation, various stabilizing additives can be added in the
product formulations. Addition of stabilizers and hydrocolloids in dairy-based whey
beverages are very significant from physical quality point of view. These stabilizers
improve the textural properties such as viscosity, prevent sedimentation and separa-
tion, delay age gelation, etc. Sensory properties such as mouthfeel, flavour and taste
are also affected by the fortification with stabilizers, emulsifiers and hydrocolloids
in the milk-based drinks.
15.6 Novel Stabilizing Additives Used in Milk-Based Drinks
As discussed in the previous sessions, stability of milk-based drinks during heat

treatment is greatly influenced by the right selection of stabilizers. These stabilizers
also contribute considerable towards the flavour, texture, mouthfeel and viscosity of
the finished product. Moreover, consumers always expect a consistent, homoge-
neous, milk-like, stable product without any flocculation, coagulation and gelation,
when it comes to milk-based drinks. Let us now discuss about some of the stabiliz-
ers commonly used in milk-based drinks, their structural and functional properties
and how they contribute towards stability of milk proteins in milk-based
beverages.
15.6.1 Pectin
Pectins are most commonly used stabilizer when it comes to low pH products. It is
commercially used to prevent protein destabilization, syneresis, gelation, etc.
Pectins are hydrocolloids formed by esterification of saccharides to form homoge-
neous galacturon chain (Yoo et al. 2006). The extent of esterification greatly influ-
enced the functional properties of pectin. Based on this pectin can be low methoxy
or high methoxy pectin (Fig. 15.2).
The casein micelle in milk maintains the stable state due to stearic repulsive
forces among themselves (Tromp et al. 2004). In fermented milk drinks, milk pH is
lowered either by fermentation using suitable starter culture or by direct acidifica-
tion. The lowering of pH to around 4 affects the native structure of casein micelle.
[4)-αGalA-(1,4)- αGalA-(1,4)- αGalA-(1,4)- αGalA-(1,4)- αGalA-(1,4)- αGalA-(1,4)- αGalA-(1,4)-(1,]n
Fig. 15.2 Homogalacturonan backbone of pectin structure

This is due to the change in the conformation of k-casein (De Kruif 1998). The
k-casein chains in milk (pH 6.7) have high entropy which is attained to the protru-
sion of the chains from surface. But as the pH drops, these protrusions led to
agglomeration of the casein and this gets destabilized. The addition of pectin to the
milk forms non-adsorbing polymer at the neutral pH of about 6.7 but, as the pH
drops, helps in stabilization by adsorption (Fig. 15.3).
In milk-based drinks, proteins are usually diluted compared to that of milk. In
such systems, pectin forms loops around the casein micelle by electrostatic interac-
tions. Various blocks of pectin loops get adsorbed around the casein micelle and
results in distribution of charges, thus stabilizing the protein structure. This mecha-
nism of protein stabilization by pectin is irreversible by any kind of thermal
processing.
15.6.2 Carrageenan
Carrageenans are linear sulphated polysaccharides. These are commonly utilized in

neutral milk-based drinks. K-carrageenan has only one sulphated group per disac-
charide and is the most studied stabilizers for milk proteins. This is normally used
in milk systems to improve the viscosity and gelation (Fig. 15.4).
The protein-stabilizing function of k-carrageenan is explained by two mecha-
nisms. The first one is based on the electrostatic interactions. The k-carrageenan
molecules are negatively charged and thus adsorbs to the positively charged regions
of k-casein. This system prevents the phase separation and thus stabilizes the milk
protein. The second theory is based on gel formation. As per this theory,
k- Casein in milk k- Casein as the pH lowers Stabilisation of casein by pectin,

also showing added pectin providing protrusions
Fig. 15.3 Schematic representation of protein stabilization by pectin
Fig. 15.4 Structure of k-carrageenan (De Ruiter and Rudolph 1997)

Fig. 15.5 Structure of

CMC
k-carrageenan forms a weak gel keeping the casein micelles in suspension and thus
providing overall stability of the system (Spagnuolo et al. 2005).
15.6.3 Carboxy Methyl Cellulose
Carboxy methyl cellulose (CMC) is an anionic polysaccharide derived from cellu-

lose. CMC is used in fermented/acidified milk drinks to prevent flocculation of milk
proteins. Majority of CMC used in food industry are as salts of sodium. The use of
CMC has profoundly been increased in South East Asian countries such as China,
Malaysia and Indonesia. CMC is comparatively cheaper and hot and cold soluble
(Fig. 15.5).
CMC molecules adsorb into casein micelle at lower pH <5.2. This adsorption
leads to formation of bridges among the neighbouring casein micelles. And this
bridging results in proper stabilization of milk proteins and thus maintains the sus-
pension even at low pH. The CMC in the milk serum also contributes to the texture,
viscosity and mouthfeel of the CMC-based drinks, providing a thick body to the
drinks. However, the viscosity achieved by the addition of CMC systems is highly
dependent on the thermal processing and shear (Tasneem et al. 2014).
15.6.4 Modified Starch
Starch sourced from maize, tapioca, sago, etc. are extensively used to impart thicker
and viscous rheology to milk products such as custards, puddings, etc. A recent
trend has been observed in South East Asian countries with thicker low pH fer-
mented drinks containing modified starch. This aims to provide a delightful and
unique experience to the consumers.
Modified starch is mostly used in combination with other stabilizers, which are
commonly k-carrageenan in case of neutral products (milk based) and pectin in case
of acidic products (acidified or fermented milk based). The increase in viscosity is
achieved by the modified starch because of its water binding by extensive amylase-
amylo pectin network formation which can be termed as its self-gelling property
(Tasneem et al. 2014). This gelation is negatively impacted by heating and shear,
which can disrupt the network. Thus the combination with pectin or k-carrageenan
helps to make the modified starch network process stable, when used at a commer-
cial scale (Table 15.3).
Table 15.3 Modified starches commonly used in food (Chen et al. 2018)
E INS
Type of modified starch number number Function class
Dextrin, roasted starch white and 1400a Carrier, emulsifier, stabilizer,
yellow thickener
Acid-treated starch 1401a Emulsifier, stabilizer, thickener
Alkaline-treated starch 1402a Emulsifier, stabilizer, thickener
Bleached starch 1403a Emulsifier, stabilizer, thickener
Oxidized starch E1404 1404 Emulsifier, stabilizer, thickener
Starches, enzyme treated 1405a Emulsifier, stabilizer, thickener
Etherified starch
Hydroxypropyl starch E1440 1440 Stabilizer, thickener
Esterified starch
Distarch phosphate E1412 1412 Stabilizer, thickener
trimetaphosphate
Starch acetate esterified with E1420 1420 Emulsifier, stabilizer, thickener
acetic anhydride
Starch sodium octenyl succinate E1450 1450 Stabilizer, thickener
Cross-linked starch
Monostarch phosphate E1410 1410 Emulsifier, stabilizer, thickener
Etherified + Esterified modified starch
Phosphated distarch phosphate E1413 1413 Emulsifier, stabilizer, thickener
Acetylated distarch phosphate E1414 1414 Emulsifier, stabilizer, thickener
Acetylated distarch adipate E1422 1422 Emulsifier, stabilizer, thickener
Hydroxypropyl distarch E1442 1442 Anticaking agent, stabilizer,
phosphate emulsifier, thickener
Acetylated oxidized starch E1451 1451 Emulsifier, stabilizer, thickener
Starch aluminium octenyl E1452 1452 Anticaking agent, carrier,
succinate stabilizer
15.6.4.1 Gellan Gum

Gellan gum is called as the ‘Universal gelling agent’. It is formed as a result of fer-
mentation by the bacteria Sphingomonas sp. Chemically, they are repeating units of
glucose, rhamnose and glucuronic acid. Gellan gum can be high acyl, which is the
native form, or low acyl. These impart different textural properties to the end prod-
uct and hence find wide applications in dairy drinks. The gel formed by high acyl
gellan is soft and elastic, whereas the one formed by low acyl gellan is heat stable,
firm and brittle. The chemical structures of high acyl gellan and low acyl gellan are
given in Fig. 15.6.
Gellan gum form ‘fluid gels’, which are networks formed by the association of
gellan gum molecules. The viscosity of this fluid gel is inversely proportional to
shear and agitation. This network of gellan gum exhibits very good suspension
properties, which can be utilized in dairy drinks with solid inclusions such as cere-
als, nuts, fruits, etc.
A
CH3
C O
0.5
O
CH2 COO–M+ CH2OH
O O O
O O
O OH O OH
O CH3
OH OH
O OH OH OH OH n
C CH2OH
CH2OH COO–M+ CH2OH
O O O
O O
O OH O OH
O CH3
OH
OH OH OH OH n
OH
Fig. 15.6 Structure of gellan gum (Valli and Clark 2009), (a) High acyl gellan, (b) Low acyl
gellan
15.6.5 Xanthan Gum
Xanthan gum is a polysaccharide produced by Xanthomonas campestris. It is used

as a stabilizer as well as a thickener due to the exceptional rheological properties
imparted to the food. It is cold and hot soluble. When used in drinks and beverages,
they form pseudo-plastic fluids and the viscosity directly dependent on applied
shear. This pseudo-plastic behaviour makes it ideal for commercial use due to high
mixability and pumpability. Moreover xanthan gum contributes to good sensorial
properties such as thick texture and rich mouthfeel. Xanthan gum exhibits good
suspension in beverages, possibly due to the three-dimensional network formed by
the associated chains (Katzbauer 1998).
Xanthan gum is widely used in food applications such as beverages, salad dress-
ings, ready-to-cook foods, baked foods, cake mixes, etc. The excellent suspension
of fruit products such as crushes, pieces and pulps for a long period of time obtained
with xanthan gum makes it the primary choice in beverages and drinks especially
where there is a fusion of dairy and non-dairy ingredients such as cereals, fruits,
vegetable, seeds, fibres/prebiotics, etc. The thick and rich mouthfeel obtained with
xanthan gum makes it an ideal choice for low fat/low calorie/no or low sugar drinks,
wherein the texture contributed by fat and sugar can be compensated with the addi-
tion of xanthan gum. The high water binding capacity of xanthan gum is the major
contributor towards enhanced mouthfeel. Another interesting application of xanthan

gum is in milk replacers for calves.
15.6.6 Agar
Agar is a linear polymer with excellent gelling properties. It is predominantly used

as culturing medium for growing microorganisms, due to its gelling properties. The
gelation of agar happens due to the change of confirmation while cooling. It is
widely used in dairy applications such as yoghurts and drinks, due to viscosity
enhancement, gelling properties, synergism with other hydrocolloids and low cost.
High gel strength of agar makes it unsuitable for direct acidified drinks; however,
nowadays agar with low gelling strength is available, which can be utilized even in
direct acidified drinks.
15.7 N
ovel Processing Techniques: Pathway Towards
the Extended Shelf Life of Dairy-Based Drinks
Consumer demands high-quality foods which are fresh, safe and nutritious and have
long shelf life. Traditional technique used in dairy processing is to make the food
safe for consumption by providing heat treatments. The temperature and time of
heat treatment depend on the various factors such as nature of the product, pH,
sugar/salt content, added preservatives, other ingredients, microbial susceptibility,
etc. When it comes to dairy drinks, a higher heat treatment temperature is utilized
for neutral products and the temperature of heat treatment is comparatively less for
acidic products. This is in line with the hurdle technology. In extended shelf life
(ESL) drinks, the common technology utilized is UHT heat treatment and aseptic
filling.
One of the commonly used technologies is Tetra Brik carton packaging, which
involves the flash heat treatment of products in a sterilizer, followed by aseptic fill-
ing in a form fill seal (FFS) machine with nitrogen flushing. This technology pro-
vides an extended shelf life ranging 4–6 months depending on the processing
parameters and product formulation. This technology has been adopted by majority
of the dairy industry for the production of ambient dairy drinks, both milk-based
and acidified/fermented milk-based dairy drinks. But some of the components of
milk are heat labile such as some vitamins, minerals and amino acids. The effect of
heat treatments can have adverse effect on the nutrition profile of end product.
Moreover, the increasing emphasis on energy conservation can also pose a negative
impact on thermal processing, which utilizes considerable amount of energy.
The extensive innovations in the field of dairy processing have led to the discov-
ery of various ‘cold processing methods’ which are non-thermal processing tech-
niques rendering safety and nutrition, unlike the thermal processing. These
techniques have gained wide attention owing to the fact that they provide safe
(destroys pathogenic microorganisms), nutritious (nutrients are intact) and sensori-

ally superior (retains freshness) products. Moreover, this is a step towards sustain-
able food processing as a large amount of energy can be conserved by adapting
these non-thermal processing techniques. The detailed information on non-thermal
processing of dairy products is elaborated in Chap. 2 of this book.
15.7.1 High Hydrostatic Pressure Technology
High hydrostatic pressure technology or HHP equips higher pressures, as high as

6000 times the atmospheric pressure to process foods. This is commonly used to
process jams, jellies, fruit syrups, juices, ham, salsa, meats, salad dressings, oysters,
poultry, etc. The processing pressure normally varies in between 300 and 700 mPa.
Most of the vegetative cells get destroyed at pressures of 400 mPa or above. The
main advantages of HHP are as follows: (a) it works for both solid and liquid prod-
ucts and (b) the process is independent of mass and time and thus delivers consistent
and uniform processing.
The operation of this HHP technique is based on two principles:
15.7.1.1 Le Chatelier’s Principle

As per the Le Chatelier’s principle, when a system in chemical equilibrium is acted
upon by an external pressure, temperature or reactant, the system counteracts the
effect by shifting the equilibrium. Thus when there is an external pressure, it will be
accompanied by a phase transition and/or a change in molecular configuration of
the system which result in the enhanced volume reduction.
15.7.1.2 Isostatic Principle

As per isostatic principle, the pressure transmittance in a food system will be inde-
pendent of the geometry and size of the food. Thus the transmittance of pressure
will be uniform and instantaneous (Chawla et al. 2011) (Fig. 15.7).
The HHP processing of foods is done in a pressure chamber. The commonly used
medium for pressure transmittance is water. The pressure is applied to the product
for a desired time, after which the chamber is depressurized by releasing the
pressure-releasing valves. The time required to attain the desired pressure in the
chamber is negatively impacted by air presence in the chamber, as air is more com-
pressible than water. As the pressure in the chamber increases, the temperature
inside the chamber also increases slightly due to the increased friction between the
molecules. The application of pressure is always isostatic to ensure uniform pro-
cessing inside the chamber.
Although HHP has all the above-discussed pros, it definitely has some cons as
well:
Fig. 15.7 Typical HHP system for packaged food (Chawla et al. 2011)
1. The food which is to be processed by HHP should have considerable moisture as

the basic process involved is compression.
2. It is difficult to be applied for ‘protein-rich’ products, since the proteins gets
denatured and this can have negative impact on the appearance of the finished
product.
3. The product should be structurally stable to withstand the processing conditions.
Otherwise undesirable textural changes can happen.
15.7.2 Ultrasonic Processing
Ultrasonication is an emerging technology in dairy processing. Most of the studies

regarding ultrasonication are a batch or laboratory scale process. But the last decade
saw drastic developments which have led to an increased availability of continuous
commercial scale ultrasonicators for food and dairy products. Ultrasonication in
simple terms is the passing of food product through a vessel that contains a sonicat-
ing device. Ultrasonication is not only effective in destroying pathogens, but it also
positively helps in the emulsification and extraction processes (Ashokkumar et al.
2010).
The destruction of vegetative cell happens from the high-frequency sound waves
(>18 kHz) when passed on at high intensity. This results in mechanical as well as
chemical impacts. The mechanical impact happens especially when the medium
contains some solids. When there are solid particles in the medium, it develops
vibrations and this causes mechanical impacts but in case of liquid medium, ultra-
sound results in acoustic streaming. This results in enhanced mass transfer. In liq-
uids there are microbubbles which are inherent. When ultrasound passes through
this liquid medium, the bubbles get enlarged by bubble diffusion and coalescence.
When these bubbles cross the threshold, they collapse leading to intense shear and
turbulence within the bubble as well as in near areas. This threshold is many a times
referred to as cavitation threshold. The collapse of the cavitation bubble can also
lead to slight increase in temperature and pressure of the medium. One point of
concern in ultrasonication is the release of OH− radicals. This can reduce the natural
antioxidant properties of the food. To prevent this, ultrasonicated foods are treated
with permitted antioxidants such as ascorbic acid, which has the capability to scav-
enge the free radicals. Ultrasound also helps in degassing and emulsification, which
can be utilized in beverage and dairy drink categories.
Ultrasonic processing even now is an emerging technology when it comes to
dairy drinks. Although considerable interventions with this technique have done in
spray drying and homogenization, it possesses wide scope in other dairy applica-
tions as well.
15.7.3 Pulsed Electric Field Processing
Treatment of milk with electricity is not new. Ohmic heating was first evaluated in
milk system wherein the temperature of milk was increased by electrical resistance.
This is also called as electrical pasteurization. There are studies which elaborate on
the lethal effects of high voltage (3000–4000 kV) on vegetative cells. All these treat-
ments were still based on heat generation and thermal processing. Many studies on
treatment of food with pulsed electric field have been seen in last the decade.
Pulsed electric field (PEF) processing is done at moderate temperature and the
food is exposed to high electric intensity with short electric pulses. This destroys the
pathogenic microorganisms and also inactivates the enzymes and at the same time
safeguards the freshness of the food. The mechanism of pathogen destruction by
PEF is electroporation. Although the exact mechanism of electroporation is still
unclear, the PEF causes disruption in the lipid/protein structure of cell membranes
leading to the formation of pores. This can lead to leakage of cellular components.
The intensity of microbial destruction is directly proportional to the field intensity.
The effectiveness of PEF on microbial destruction also depends on the medium,
treatment temperature and PEF operating conditions (Table 15.4).
Table 15.4 Factors affecting the microbial inactivation by high-intensity PEF (Bendicho et al.
2002)
Microbial reduction (log
Media Bacteria Treatment conditions reductions)
Simulated milk Escherichia coli 7 °C, 36 kV/cm, 16 2–3
ultrafiltrate pulses
Simulated milk Escherichia coli 20 °C, 36 kV/cm, 8 2.5
Simulated milk Escherichia coli 33 °C, 36 kV/cm, 8 2.5
Simulated milk Escherichia coli 7 °C, 36 kV/cm, 64 4
Simulated milk Escherichia coli 20 °C, 36 kV/cm, 64 5
Skim milk Salmonella dublin 30 °C, 25 kV/cm, 1
100 pulses
Skim milk Salmonella dublin 50 °C, 25 kV/cm, 2
100 pulses
Whole milk Listeria 25 °C, 30 kV/cm, 2.5
monocytogenes 400 pulses
Whole milk Listeria 50 °C, 30 kV/cm, 4
monocytogenes 400 pulses
15.8 Conclusions
The changes in consumer perspective drive food innovations. The advancements in

ingredient and processing technology and regulatory scenario are the key factors
affecting food innovations. Recent developments in technology and modifications
in regulations have led to the developments of novel concepts in dairy-based drinks.
Hot and cold processed dairy drinks are pretty common and popular now. The
advancements in cold processing techniques such as high hydrostatic pressure tech-
nology, ultrasonic processing, pulsed electric field processing, etc. have increased
the scope of innovative drinks. However, heat treatments such as pasteurization,
sterilization, etc. remain to be the most popular techniques in the industry. The
developments in the field of ingredient technologies are leading to novel additives
for enhancing protein stability, which are the key driver for this popularity. These
innovations helped the technologists to create novel milk, fermented milk and
whey-based drinks. This will not only enhance the dairy drinks category but also led
to cross innovations such as dairy-cereals, dairy-pulse and on-the-go breakfast
concepts.
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Extending the Horizons of Dairying
to the Common Man: An Indian 16
Perspective
Aiswarya S. Panicker, M. Misha Madhavan,
and Himanshi Solanki
Contents
16.2 E lements of Extension 329
16.2.1 Extension Agent/Worker 329
16.2.2 Agencies of Extension in India 330
16.2.3 Major Continuing Extension Programmes in India 331
16.3 Information and Communications Technology (ICT) in Dairy Extension 331
16.3.1 ICT for Agriculture in Rural India 332
16.3.2 Integrated Citizen Service Centre 334
16.4 Entrepreneurship Development in Dairying in India 340
16.4.1 Dairy Entrepreneurship Development Scheme (DEDS) 341
16.4.2 Technology Business Incubator in Dairying 342
16.5 Government Initiatives in Animal Husbandry and Dairy Sector in India 343
16.5.1 National Livestock Mission (NLM) 343
16.5.2 National Dairy Plan Phase-1 (NDP-1) 344
16.5.3 National Programme for Bovine Breeding and Dairy Development
(NPBB and DD) 346
16.6 Gender Mainstreaming in Dairying 346
16.6.1 Gender Issues in Dairying 347
16.6.2 Dimensions of Women Empowerment 347
16.6.3 Role of Self-Help Groups (SHG) in Women Empowerment 348
16.6.4 Strategies for Women Empowerment in Dairying 348
A. S. Panicker (*)
College of Dairy Science and Technology, Kerala Veterinary and Animal Sciences University,
Kolahalamedu, Idukki, Kerala, India
M. Misha Madhavan
ICAR—National Research Centre on Pig, Indian Council of Agricultural Research,
Guwahati, Assam, India
e-mail: misha.m@icar.gov.in
H. Solanki
DFIL-CreamBell, Gurgaon, Haryana, India

https://doi.org/10.1007/978-981-15-2608-4_16
328 A. S. Panicker et al.

References 349
Abstract
The extension is the sine qua non for the expansion of knowledge in the dairy
sector to the doorsteps of farmers to improve their livelihood. Along with the
development of suitable technologies, proper extension activities have to be car-
ried out to bring the desired effect in dairy production and processing from the
grassroots level. In India, the first-line extension system and front-line extension
system are altogether bringing a lot of initiatives for the success of lab to land
process. The different Information and Communications Technologies (ICT) ini-
tiatives in the dairy sector were discussed to throw some light on the advance-
ment of technology transfer mode. Dairying is a sector where entrepreneurship
has an immense scope. The actions taken by the government to promote entre-
preneurship in dairying are commendable and are instrumental in maintaining
India as the largest milk producer in the world. Ambitious programs initiated by
the government like ‘National Dairy Plan’ contributed to extending the new hori-
zons of dairying to the common man. The basics of gender mainstreaming in
dairying and the gender issues in dairying were discussed to enlighten the need
for women empowerment in the dairy sector by means of self-help groups and
others.
Keywords
Dairy extension · Dairying in India · Communication technology ·
Entrepreneurship development · Women empowerment
16.1 Introduction
The extension is the practice of working among the rural people to develop their
livelihood by helping farmers to enhance their agricultural yield. Thus extension is
an informal educational method aimed at rural population to change farmers’ out-
look towards agriculture by contributing extended help and efforts to resolve their
field problems. Extension programme helps to enhance the competence of the
farmer in farming, enhancing agricultural production and livelihood of the entire
family. Extension with its way of improving physical and economic achievements
also helps to promote the growth of the community as well. Extension workers
bridge the gap between researchers, industries and farmers and play a vital role in
developing and underdeveloped countries. They may connect with the rural people
to get a better understanding of the problems and to resolve the problems by finding
the best solutions. Thus it is apparent that extension is not a single-period activity,
but a practice which occurs over a period of time. Thus, an extension is a process of
16 Extending the Horizons of Dairying to the Common Man: An Indian Perspective 329
educating, supporting and preparing the rural people, to deal with their problems
more effectively.
16.2 Elements of Extension
The four main elements of the extension education process are the improvement of
knowledge and skills of farmers, imparting technical information and advice to
farmers, the formation of an organization for farmers and motivation and building
of confidence. Since farmers have a different understanding of agricultural and
farming practices, the extension activities can help them be aware of latest knowl-
edge and skills. The transmission of valuable knowledge to farmers and their family
members is a crucial part of extension activity and the key liability of an extension
agent. One should find out the area in which farmers lack the skills or knowledge
and arrange suitable learning programs for the same for making themselves
sufficient.
The extension assists farmers by providing advice and information in taking
decisions and facilitates them to make them into action. A simple example includes
the data related to prices and available nearby markets, or about the accessibility of
types of machinery and raw materials for farming. Another one is the technical
advice about farm production activities to get better or uphold production. The tech-
nical advice includes the value addition of agricultural produce, for example, the
production of milk products from milk and their marketing.
In many instances, farmers need an organization in order to stand for their inter-
ests and to take collective actions, for example, a collective group to provide green
grasses to help dairy farmers during the drought season. Therefore, an extension
agent needs to develop and build up a farmer to farmer link by collaborating the
nearby farmers and to establish organizations; thus making any organizational setup
should be done in consultation with the local farmers and these organizations will
help in extension activities more easily in the future and treat as an essential core
channel for spreading awareness and knowledge.
16.2.1 Extension Agent/Worker
The most important activity of an extension worker is to motivate and encourage the
farmers by highlighting their capabilities, improving decision-making skills and
facilities to improve their economic status. An extension worker has to work inti-
mately with farmers, encourage them to take new initiatives and to involve in exten-
sion activities. Extension activities should motivate the farmers who are diffident
and unaware of the evolving new technologies. The real dairy extension service in
terms of marginal dairy farmers is to make them adapt good practices in farming as
well as motivate for value addition of milk to make milk products get maximum
benefits. Thus, the extension service should be intended to improve dairy farming
methods by supporting farmers through instructive action and technologies,
increasing the production capability and profits, and, thereby, facilitating farmers to
enhance their quality of livelihood. An effective dairy or livestock extension
includes:
(a) Helping farmers to discover their constraints allied with production and market-
ing followed by solving through facts, reporting and sharing of information and
skills.
(b) Assisting farmers for using the available technologies and uphold services
through the capacity edifice.
(c) Promote farmers’ organizations, by providing a platform for value addition, and
make sure to improve their productivity and profitability.
(d) Capacity building and improvement of skills of farmers to encourage them to
adopt excellent and apt production practices for improved production with low
risk.
(e) Make connection with different information sources on innovations in agricul-
tural, novel technologies and demand-supply-related market information such
as product prices.
A wide range of extension teaching methods and techniques are in common use,
and they are employed suiting to the situation. Extension methods can be broadly
classified into individual contact, group contact and mass contact. Individual con-
tact methods include farm visits, home visits, office calls, telephone calls and let-
ters. Group methods include speeches and talks, demonstrations, excursions and
group discussions. Newspapers, radio, television and other printed forms are the
major mass methods used in the extension.
16.2.2 Agencies of Extension in India
The agencies involved in dairy extension activities can be divided mainly into three:
public and private extension services, mass media and information technology cen-
tres. The public extension services imply State Government-approved Animal
Husbandry/State dairy development departments, agriculture extension depart-
ments, Agricultural Technology Management Agency (ATMA), Panchayat Raj
level institutions (village, block and district panchayats), Agricultural/Veterinary
and Animal Sciences Universities/Colleges, Krishi Vigyan Kendras (KVKs), Indian
Council of Agricultural Research (ICAR) extension centres, etc. The private exten-
sion services consists of farmers’ cooperatives/federations, producer groups, pro-
cessing/marketing agencies, self-help groups (SHGs), private vets, nongovernmental
organizations (NGOs), input suppliers groups which are feeds, vaccines, seeds,
tools, medicines and microfinance, etc. The newsletters, private cable networks,
radio, television, print media, electronic network: mobile, private portals, local
wireless loops, email/Internet, farm advisory centres and information hubs are
included in the mass media and information technology centres.
16.2.3 Major Continuing Extension Programmes in India
A lot of extension programmes are going on in India. Some of the major ongoing
extension programmes in India are listed here:
(a) State Animal Husbandry and Dairy Development Departments: veterinary clin-
ics/hospitals, dairy extension service units, dairy extension training centres,
milk quality control offices, etc.
(b) Animal Husbandry/Dairy Extension Programme: time-dependent treatment,
data compilation, awareness classes, milk quality checking campaigns, training
programmes on value addition of milk, exhibitions (Ksheeramela/dairymelas),
etc.
(c) Dairy cooperative societies/dairy processors/marketing agencies.
(d) Agricultural Technology Management Agency (ATMA): key area of focus on
crop production and introduction of new fodder crops to the farmers.
(e) Krishi Vigyan Kendras (KVKs): focus on technical skills/amenities improve-
ment in animal husbandry/dairying.
(f) Veterinary/Dairy Colleges/Agricultural/Universities: focus on authentic prob-
lems of livestock farmers.
(g) Nongovernmental organizations (NGOs) and input supply agencies.
16.3 I nformation and Communications Technology (ICT)

in Dairy Extension
In rural development, nothing is more important than the transfer of useful ideas.
The challenge of extension is to convey a clear meaning of ideas to the audience, to
get accepted and to motivate farmers to adopt them in life. To a large degree, the
success of extension lies in communication. However, communication is the
exchange process of ideas, opinions, evidence or intuitions among two or more
individuals to build a common thought, intent and message to deliver. It is a con-
scious attempt to give and gain information. Good communication is the essence of
good extension work and extension educators are efficient communicators.
ICT is the amalgamation of technologies and the methods to allocate and com-
municate the needed information to the target spectators to enhance their participa-
tion. Computer-based information and communication technology is helpful in
capturing, storing, processing, retrieving and transmitting data and further commu-
nicating the results either in the form of replica or aspect or in mutual form through
computers. Thus the base of information and communication technology is a mutual
form of combined aspects of computers and multiple information systems to deliver
the desired solutions to the users.
ICT enables the dispersal of information at the exact time and this revolution
simplifies the accessibility to information in price-efficient manner. There were dif-
ferent areas in which ICT for developmental interventions can be made. This
includes agriculture and related fields, climate change and environmental changes,
Table 16.1 Factors limiting the adaptation of information technology by marginal farmers
Limiting factors 1. Illiteracy
2. Poor financial conditions
3. The high cost of equipment
4. Lack of power supply or power failures
5. Lack of computer knowledge
6. Poor internet connection for computers
7. Lack of computer training programmers
8. Maintenance and repair of software and hardware
problems
9. Lack of trained personnel in this field
10. Increased risk for establishment and maintenance
11. Language barriers in using internet
12. Lack of organizational and institutional infrastructure
facility
education and literacy programs, health, business and commerce (e-commerce),

political development and civic engagement (e-government), tourism, food safety,
etc.
The reasons for reluctance towards the adaption of information technology by
marginal farmers are detailed in Table 16.1.
16.3.1 ICT for Agriculture in Rural India
Information and communication technologies have become so common that its

usage is no more considered as an advanced technology. During the last decade,
there is a tremendous surge in the integration of information technology in the life-
style of people. This results in the opening of new opportunities and practices in
technologies related to agriculture, education, health and society. But the challenge
is to provide them with the basic requirements for information technology (IT)-
based systems. Over the past 20 years, the use of ICT has made an incredible change
in the face of business as well as governance. ICT has started its functioning in the
education field and the role of ICT in education is becoming more and more relevant
as it is a student-centred learning approach. The major tools in the agricultural sec-
tor include mobiles, telephones and other telecommunication, personal computers,
devices, etc. ICT tools can be effectively used for agricultural development in the
following ways as illustrated in Fig. 16.1.
Key government initiatives in India to promote the use of ICT in rural areas
include National e-Governance Plan in Agriculture (NeGP-A), various Touch
Screen Kiosks, Krishi Vigyan Kendras (KVKs), Kisan Call Centres (KCC), Agri-
Clinics, Common Service Centers, mKisan, Kisan TV and various other applica-
tions. In order to exploit the advances in ICT, Internet extension and broadband
technology are now given the main priority. Various government projects and
Awareness
and
Education
Online
Weather
farmer
forecasting
communities
ICT
Agrifinance Pricing and

and market
Agriclinics information
Dissemination
of government
schemes
Fig. 16.1 Advantages of ICT
corporate initiatives are working to fulfill the requirements of Internet and broad-
band facility in rural India. Some of them are pointed here:
• Golden quadrilateral project to create kiosks.

• Project Saksham-Microsoft India’s project to set-up kiosks in about 200,000 vil-
lages around India.
• Hindustan Unilever Limited (HUL) project i-Shakti for 7500 villages.
• VICTERS (Virtual Class Technology on Edusat for Rural Schools).
• Hughesnet Fusion centres project by Hughes for setting 1000 rural ICT kiosks.
• Kisan Soochana Kendras to put up over 3000 kiosks in Uttaranchal by an NGO
Jai Kisan.
• State Bank of India set up over 5000 kiosks to encourage rural banking habits.
• Around 200 e-government projects in various states under state governments to
enhance the connectivity of farmers in rural areas.
Some research projects are going on in order to improve the Indian agriculture
and rural development which includes e-Sagu, pest prediction systems, etc. With the
main emphasis of government lies in giving services in single-point centre, private
agencies and the civil society are also realizing the role of ICTs in continuing incor-
porated services to the citizens. There are some ICTs for social and economic
development, named as Integrated Citizen Service Schemes, such as e-Seva,

Akshaya, Gyandoot, Akashganga, TARAhaat, Bhoomi, etc. (https://www.icfa.org.
in/assets/doc/reports/ICT_in_Agriculture.pdf).
16.3.2 Integrated Citizen Service Centre
Integrated citizen service centres are one-stop centres which can provide services of
various government offices. The use of this centres elliminates the need of people to
visit diverse government offices and interacting with various employees for any
help. These are constructed on the public-private partnership (PPP) model in which
the government helps the citizens by giving services through a private vendor who
put up kiosks for serving the needy and is paid in return on basis of the transaction-
cost model. Integrated Citizen Service Centers are established in the states of
Andhra Pradesh (e-Seva), Tamilnadu (SARI), Kerala (Akshaya), Maharashtra
(SETU), Gujarat (Swagat), West Bengal, Himachal Pradesh (LokMitra) and
Karnataka (Bhoomi and Rural Digital Services) (https://iesgeneralstudies.com/ict-
applications-and-government-schemes/; https://www.manage.gov.in/
studymaterial/e-gov-E.pdf), which are briefed below:
16.3.2.1 e-Seva in Andhra Pradesh

e-Seva is the one among the first initiatives in India in the Integrated Citizen Services
field which has already been implemented in Hyderabad, Andhra Pradesh. Other
projects such as Computer-aided Administration of Registration Department
(CARD), Vijayawada Online Information Centre (VOICE), e-Procurement System,
etc. are implemented in the state as a part of e-Governance.
16.3.2.2 Akshaya in Kerala

Akshaya Project was implemented by the state Kerala IT Mission. The main
achievement of this project was to establish a network of 6000 information centres.
These centres help at least one common member from each family of Kerala to get
access to basic IT literacy. It also offers services such as data entry sources, data
publishing, computer training and Internet telephony to the needy; develops public
delivery of services to accelerate transactions such as certificates issues, tax collec-
tions and licenses. The Akshaya centres act as agri-business centres by ensuring
agriculture-related information and services to the farmers which offers various
agriculture-related inputs.
16.3.2.3 Setu in Maharashtra

Setu Suvidha Kendra (electronic bridge) was recognized in 25 district headquarters
and 225 taluka in Maharashtra which provides information related to collector
office in each place with the help of several wide area networks. Similar projects
have been set up in Gujarat such as Mahiti Shakti in Panchmahal district, Gyan
Ganga Project and Swagat (Online Grievance Redressal System).
16.3.2.4 SARI in Tamil Nadu

Sustainable Access in Rural India (SARI), a pilot project, was implemented at the
outset of Melur taluk in Madurai district during 2003–2004. The main motto of this
project was to promote the use of Internet among villagers for communal uplift-
ment, prosperity enhancement and job creation and to create connectivity among
rural people with the help of touch screen sources through public-private-
partnerships at a low cost. Later on, the project has been set up in ten more districts
and renamed as RASI (Rural Access Services through the Internet) (http://www.
tnagrisnet.tn.gov.in/).
16.3.2.5 LokMitra in Himachal Pradesh

LokMitra, a pilot project, was initiated by the state government of Himachal
Pradesh. The project has made Intranet connection throughout the entire district by
using 25 citizen information booths at rural area and servers at district
headquarters.
16.3.2.6 Bhoomi in Karnataka

To control the land registry and to maintain the land records, Government of India
and Government of Karnataka jointly headed and funded a project named ‘Bhoomi’
to digitalize the paper-based records to create a software means to manage changes
to the land registry in the state. The project was intended to eradicate the long-term
problem of ineffectiveness and sleaze in handling and preservation of land records
at block-level offices. The project was developed and implemented by the National
Informatics Centre (NIC). The main features of this project are:
• Development of printing software for maintaining land records at any time.

• Online updating of record certificates needed for farmers.
• For the authentication of users.
–– Bio-Login metrics system through fingerprints helps in order to shun users
imitation and database system hacks.
• The software can be used for generation of reports on soil, land-holding size,
types of crops grown and many more to construct knowledgeable policy
decision.
As far as farmers are concerned, they can quickly get their land records easily
from kiosks which makes them secure about their property. Further, these land
records can help them in getting credits/loans and can be used in legal matters. For
administrators, the software helps ease in maintenance and updating of speedy and
trouble-free access to the documents and land records, for study and monitoring of
government lands. This helps different financial institutions to offer farm credit by
accessing information on the financial status of each farmer as well as general infor-
mation regarding their land. Bhoomi and Rural Digital Services are initiatives taken
up by the state’s rural areas in Karnataka.
Banglore One (B1) which is another initiative of the government aims to extend
the interaction between citizen and government, funded of which has been mainly
done by the Ministry of Communications and Information Technology (MOCIT)

and the United Nations Development Programme (UNDP) funded ICT develop-
ment project which are being implemented by the National Institute for Smart
Government (NISG). (https://bagalkot.nic.in/en/service/land-records/; https://e-
governnsc.weebly.com/the-bhoomi-project.html; http://nisg.org/projects/project_
bangaloreone/bangaloreone.html; http://www.bangaloreone.gov.in/public/
aboutbone.aspx).
16.3.2.7 Gyandoot in Madhya Pradesh

Gyandoot, a government-to-citizen Intranet-based service portal, launched in
January 2000 in the Dhar district of Madhya Pradesh. This project was initiated with
21 telecentres located in councils of five district blocks in separate villages. The
project gradually reached over 600 villages, covering 9 of 13 development blocks in
the Dhar district. Gyandoot is an innovative, unique and creative e-governance con-
cept. Gyandoot has been developed to inform the front end of public services
throughout the region. Gyandoot has developed the idea of rural telecentres in India
and the concept of the project has been replicated in India by another initiative for
the growth of information and communication technologies. The project aimed at
linking villagers and government with IT through data kiosks. The kiosks provide
access to a wide range of public facilities, such as complaint registration and appli-
cation for certificate and loan issuance. There are also data available on the prices of
agricultural plants in various industries. In the early years of its execution, Gyandoot
was regarded as very successful and Stockholm information technology challenge
award for public service and democracy was given in year 2000 to this project
(http://documents.worldbank.org/curated/en/621851468267614396/Gyandoot-Project-
ICT-initiative-in-the-district-of-Dhar-Madhya-Pradesh).
16.3.2.8 Technology and Action for Rural Advancement (TARAhaat)

TARAhaat is a development alternative (DA) enterprise, a pioneering sustainable
development company founded in 1983, acknowledged by the ministry of science
and technology as a scientific research institution, not for profit but for research,
development and section organization. With two decades of experience and over
400 experts, the DA group had a deep effect on creating sustainable livelihoods,
particularly in innovation and applying suitable methods and distributing them in
rural India through micro-enterprises. TARAhaat provides the villagers with count-
less data resources and possibilities depending on the market.
Initially, the project was focused on the main concerns of the villagers such as
commodity prices, health facilities, land records, local development programmes,
business opportunities, jobs, etc. But these days farmers are able to shop farm inputs
such as seeds, machinery, spare parts and household items such as bicycles, scooters
and refrigerators. TARAhaat is designed specially to address the needs of its par-
ticular customers. The data, analysis and communication structures of TARAhaat.
com are vigilantly designed so as to function smoothly in response to the require-
ments of its users, making it a well participatory and responsive network. TARAhaat
is successful in literacy, language, financial liquidity and levels of understanding.
Useful information and user-friendly access to TARAhaat.com is critical to the suc-

cess of TARAhaat. TARAhaat works in collaboration with the premier institutions
in each field within the country like All India Institute of Medical Sciences (http://
www.tarahaat.com/ICT.aspx).
16.3.2.9 Information Village Research Project in Pondicherry

M.S. Swaminathan Research Foundation (MSSRF) has implemented the informa-
tion village research project for the Pondicherry fishermen. ICT is used as the way
of transferring services to these fishermen. Computer-equipped village centre is
connected to the Internet and the weather forecast of the Indian meteorological
offices are made accessible to the farmers by using loudspeakers and very high fre-
quency (VHF) radios help to conclude the low and high tide frequencies to the
fishermen before sailing off to the sea (https://www.mssrf.org/; https://www.mssrf.
org/mssrfoldsite/?q=iec).
16.3.2.10 Akashganga
Akashganga (Shree Kamdhenu Electronics Pvt. Ltd.—SKEPL) is targeted to
improve the efficiency of milk cooperative societies, transparency and quality of
milk collection process and provide a complete automated solution to rural milk
cooperative societies from milk quality monitoring to milk accounting, thus improv-
ing timely milk collection that results in higher earnings to farmers. Under the proj-
ect, a computerized system has installed instead of a manual time-consuming
process for collection of milk and quality testing. The automated process involves
the distribution of a unique token to each farmer in which the quantity and quality
of the poured milk is mentioned upon which the farmer will be paid for milk. The
system includes electronically milk weighing, fat testing, capturing exclusive ID
followed by the printing of payslip and payment settlement. The initiative aid timely
milk collection, fetching higher salary and payments for the farmers (http://akash-
ganga.in/profile.htm; https://egov.eletsonline.com/2005/08/akashganga/).
16.3.2.11 e-Choupal
The project was initiated by Swagatika Barika which offers substitute marketing
channel, information on weather, agricultural activities, input sales, profits, etc. The
information kiosk system, with Internet access managed by trained sanchalak, is
placed in a village. Through ITC (Indian Tobacco Company) E-choupal, ten million
farmers in 10,000 villages were benefited by using computerized mandis(markets)
for agricultural products. Traditionally, mandis are the main market for the rural
manufacturers and mostly controlled by the middlemen who make profit by block-
ing the price and market information. But this computerized Mandis minimizes the
role of middlemen and set up the direct link with the rural manufacturers. Each
choupal has one coordinator known as ‘Sanchalak’. Due to the absence of middle-
man, the rural manufacturers earns high profit through good market exposure.
(https://www.itcportal.com/businesses/agri-business/e-choupal.aspx).
16.3.2.12 e-Sagu
The e-Sagu is a web-based agricultural cost-effective expert counsel dissemination
system in order to distribute expert agricultural knowledge to the farmers to improve
their agricultural productivity. The aim of pest prediction systems is to make an
understanding of pest population and its dynamics to develop a prediction model by
applying the data available on pest surveillance. e-Sagu offers personalized expert
advice about each and every aspect of farming-related information in a timely man-
ner to the doorstep of a farmer. Digital photographs and text information about the
farm is sent to the expert person and he analyses the scenario. When expert advice
is ready, it has delivered to the farmer involved in the overall process in the same or
subsequent days (http://agriculture.iiit.ac.in/esagu2012/overview.php).
16.3.2.13 Other Projects
Agmarknet
Agmarknet helps to make available the daily market price and information of about
300 commodities in 2000 varieties in different eight local languages. Information
on prices, arrival, grades, standards, packaging, etc. are collected and circulated
among different markets in the country by networking major agricultural markets.
e-Krishi
The communication network formed under e-Krishi is utilized to inform farmers
regarding price information, arrivals, weather forecast and issue disaster alert. The
motive of the project is to enable farmers to take decisions on the sale of products
and fetch transparency in the functioning of the State Agricultural Marketing Board
in Madhya Pradesh.
Warana
Warana project provides information on agriculture and other fields to the beneficia-
ries in the local language. Crops information, market fares, employment-related
schemes, educational opportunities, etc. are provided through information kiosks
placed in the village to the farmers through kiosk operators.
IFFCO Kisan Sanchar Limited (IKSL)

The project helps the farmers by providing the required information or customized
solutions of problems on their mobile phones in local language through voice calls.
The farmers can also get expert advice on the precise subject through special
‘phone-in’ programmes using helpline numbers.
Pravara
The project aims to connect a hundred villages in Ahmednagar by an enlightening
rural population about the information on a government scheme, agricultural mar-
keting, healthcare, education, agro-processing and economic development. It has
contributed in the improvement of life quality of rural people.
iKisan
iKisan is a one-stop solution for farmers facilitating information on crops, crop
management techniques, fertilizers, pesticides, market updates, weather forecasts,
etc.
Earik
This is a single-window approach towards agricultural information and technology
in northeastern India. It offers expert advice on manufacture fortification and mar-
keting of agricultural materials especially plants.
Digital Mandi
Digital Mandi is an electronic trading platform for agricultural commodities through
the eradication of geographical barriers and temporal constraints and the removal of
crunchy cases to benefit the farmers and traders from communication technology.
Digital mandi is influenced by the vision of media laboratories and Asia sustainable
village by using fresh technologies which are culturally suitable (http://digitalmandi.
iitk.ac.in).
aAQUA
An online multilingual system for solving the farmer’s problem by providing
answers to their query by experts in related fields within days.
Mahindra Kisan Mitra

The scheme offers information related to agriculture (market prices, weather
updates, crop advisories and agri-related news) as well as other sections (insurance,
Mandi database, loans, cold storage and warehouses). It also provides information
on the website about the success stories of farmers which makes others get
motivated.
Haryali Kisan Bazar (HKB)

HKB has established different centres within country to provide useful information
to the farmers with alternatives to issues under one roof. The centre provides infor-
mation on crops, modern technologies, weather forecast, market prices, agri-inputs,
financial services, farm-output services, round-the-clock expert advice and tailored
services based on the farmer database kept under the scheme (http://www.dcmshri-
ram.com/haryali-kisaan-bazaar).
AGRISNET
This provides an infrastructure network that facilitates connectivity among agricul-
tural offices, agricultural extension services and agribusiness activities to enhance
the rural development.
Digital Green
Under this scheme, the agricultural-related information of any local relevance is
transmitted through digital video. The system comprises a digital video database
prepared for farmers with expert help. The recordings of the same are shown to
individuals or small groups using laptops, digital versatile disc (DVD) player, tele-
vision and to communities through village cable network (https://www.digitalgreen.
org/).
Fisher Friend Mobile Advisory

The scheme provides important information on wind speed/direction, government
schemes, wave height, market prices, fishing zones, etc. through mobile phone for
fishermen in the local language.
KCC (Kisan Call Centres)

The project provides customized information to farmers on agriculture and allied
fields using telecommunication using toll-free numbers.
Reuters Market Light

Reuters Market Light provides customized information in local languages across 13
states through Short Message Service (SMS) in a mobile phone as per the prefer-
ences and needs of farmers on 440 crops and varieties, more than 1400 markets and
2800 locations (https://en.wikipedia.org/wiki/Reuters_Market_Light).
e-Agri Kiosk
This programme is initiated by the National Bank For Agriculture and Rural
Development (NABARD) and India and Central Agricultural University among
tribal farmers of Arunachal Pradesh for technology transfer using touch screen
kiosk.
MSSRF Friends Foreign Missionary Association Fisher Friend

The programme consists of a Binary Runtime Environment for Wireless (BREW)-
based application offering a mobile handset with a graphical interface, an iconic
menu and programmable shortcut keys. Despite the security and weather informa-
tion, fishing areas and market places were made available in the local languages
(https://iesgeneralstudies.com/ict-applications-and-government-schemes/).
16.4 Entrepreneurship Development in Dairying in India
A person who begins a new venture for deriving profit under conditions of uncer-
tainty is called an entrepreneur. He/she possesses certain unique characteristics like
risk-bearing, self-confidence, achievement, motivation, optimism, information
seeking, innovative, networking, leadership, long-term involvement, personal drive
and energy, etc. (Mondal and Ray 2007). There is no age, sex, caste or class dis-
crimination in becoming an entrepreneur. Anyone who wants to make an identity of
their own can start a new venture. Bahal (2008) stated entrepreneurship as a process
change which involves three behavioural conducts as given below:
1 . Identifying and evaluating an opportunity.

2. Facilitating the production of new goods and services by proper management of
the transformed or new organization.
3. Successful use of new idea (i.e. innovation) for creating value.
Our country is witnessing the start-up revolution through the implementation of

various schemes which promotes entrepreneurship. By attracting youth to become
an entrepreneur, the unemployment level can be reduced. In the dairy sector, the
government has initiated the Dairy Entrepreneurship Development Scheme (DEDS)
to encourage entrepreneurship.
16.4.1 Dairy Entrepreneurship Development Scheme (DEDS)
A model scheme titled ‘Venture Capital Scheme for Dairy and Poultry’ was launched
by the central government in the year 2005–2006 with the main aim of extending
support for setting up of small dairy farms and other components in the dairy entre-
preneurship. After the midway evaluation of this scheme, certain recommendations
were made to improve its performance. Taking into account the representations
received from different stakeholders of dairying and the recommendations of the
evaluation study, the Department of Animal Husbandry, Dairying and Fisheries
(DAHDF) revised this scheme with a new name as Dairy Entrepreneurship
Development Scheme (DEDS) which has came into operation from 1 September,
2010 with the following objectives:
1 . Self-employment generation and provision of infrastructure for the dairy sector.

2. Building modern dairy farms and infrastructure for clean milk production.
3. Good breeding stock development and conservation by encouraging the heifer
calf rearing.
4. Structural changes of the unorganized sector to equip initial milk processing at
the village level.
5. Traditional technology upgradation for commercial-scale milk handling.
6. Milk value addition by processing and production of milk products.
This scheme covers all the activities like improvement of milk production, pro-
curement, preservation, transportation, processing and distribution of milk. The
beneficiaries of this scheme can be individual entrepreneurs, farmers, groups of
organized and unorganized sectors, nongovernmental organizations (NGOs), com-
panies, etc. The groups of the organized sectors include self-help groups (SHGs),
dairy cooperatives, milk unions, milk federations, etc. Every individual can avail
assistance once for each component under the scheme. The scheme provides assis-
tance to more than one member of a family if they set up separate units at different
locations. The minimum distance flanked by the two farms, in this case, should be
500 m.
This centrally sponsored scheme was implemented through the National Bank
for Agriculture and Rural Development (NABARD) by providing back-end capital
subsidy for bankable projects. The subsidy wise ceiling of the parts has to be subject
to indicative costs rising from moment to time from NABARD.
16.4.1.1 Components Which Can Be Funded

• Establishment of small dairy units with crossbreed cows/indigenous descript
milch cows/graded buffaloes up to ten animals (minimum unit size is two ani-
mals and maximum unit size ten animals). Indicative unit cost is Rs. 7 lakh for
ten animals per unit.
• Heifer calves crossbreed/indigenous cattle/graded buffaloes rearing facility up to
20 calves. Indicative unit cost is Rs. 9.70 lakh for 20 calves per unit (minimum
unit size 5 calves and maximum 20 calves).
• Vermicompost of a milch animal per unit with indicative unit cost Rs. 25,200/−.
• Milking machines/milk testers/bulk milk cooling units (up to 5000 L capacity)
purchase having an indicative unit cost of Rs. 20 lakh.
• Purchase unit cost of Rs 13.2 lakhs for dairy processing equipment of indigenous
milk products.
• Dairy products transportation facilities with indicative unit cost of Rs. 26.5 lakh.
• Milk and milk products cold storage facility with an indicative unit cost of Rs. 33
lakh.
• Establishment of private veterinary clinics with an indicative unit cost of Rs. 2.6
lakh for mobile clinic and Rs. 2 lakh for stationary clinic.
• Dairy marketing with an indicative unit price of Rs. 3 lakh for outlets/parlours.
This scheme promotes self-employment opportunities in the dairy sector by

credit-linked assistance for the projects sanctioned by the suitable financial institu-
tions like state cooperative banks, commercial banks, regional rural and urban
banks, etc.
16.4.2 Technology Business Incubator in Dairying
Technology business incubator (TBI) is a body that offers all the support and types
of equipment needed to help entrepreneurs such as technology/prototype develop-
ment support, infrastructure support, financial aid, business consultancy assistance,
marketing assistance and any other support needed to set up a company. The tech-
nology incubation concept for start-up companies helps in reducing the failure rate.
A business incubator’s primary objective is to promote economic development by
enhancing the survival and growth of enterprises from the idea level to autonomous
productive company. The TBI provides an initial space for entrepreneurs with a sup-
portive environment, generates high-skilled employment and develops technology-
driven new enterprises for the promotion of national development. The objectives of
TBI are as follows:
1. Awareness creation among graduating students and unemployed graduates on

entrepreneurship and self-employment.
2. Assist the entrepreneurs in the identification and evaluation of technologies and
know-how.
3. Undertake research and develop educational materials regarding the evolution of
different innovations into viable enterprises.
4. Disseminate information and research findings on the management of innova-
tion, incubation and entrepreneurship.
5. Spread data and study results on innovation management, incubation and

entrepreneurship.
6. Help the entrepreneur to carry out their survey of feasibility, project evaluation,
market research and financial study.
7. Help the current units and entrepreneurs to enhance their technical, economic,
marketing and leadership abilities by training their staff.
At the ICAR-National Dairy Research Institute (ICAR-NDRI), Karnal Haryana,

a TBI operates the dairy industry to support knowledge-based and innovation-driven
enterprises to the nation. The TBI is an initiative of the Department of Science and
Technology (DST)-Government of India and promoted by Society for Innovation
and Entrepreneurship in Dairying (SINED), for reducing the time lag between tech-
nology development and commercialization. This TBI works for:
• Conducting awareness camps for entrepreneurship.

• Development programmes for entrepreneurship.
• Provides specialized services to the region’s current small and medium
enterprises.
• Different training programmes including short-term courses.
• Start-up incubation and mentoring.
A panel of specialists deal with prospective entrepreneur’s apps for TBI affilia-
tion. After assessing the business idea and innovation content, viability and develop-
ment prospectus, promoter and market accessibility, the panel picks up the
incubation enterprise. The TBI helps to train prospective entrepreneurs through
continuous nurturing and interactive workshops to encourage entrepreneurship and
bring them through various phases such as the phase of development, the phase of
incubation, the phase of execution and the starting phase.
16.5 G
overnment Initiatives in Animal Husbandry and Dairy
Sector in India
16.5.1 National Livestock Mission (NLM)
From 2014 to 2015, the national livestock mission has begun. The task is intended
to cover all operations necessary to guarantee the quantitative and qualitative
enhancement of livestock production systems and capacity building of all stake-

holders. The task includes various aspects to enhance the productivity of livestock.
The mission is developed with the aim of sustainable livestock industry growth and
with the aim of enhancing the accessibility of quality feed and fodder. The NLM
consist of following four sub-missions:
16.5.1.1 Sub-mission on Livestock Development

Risk management and insurance have been introduced in all the district of the nation
as a component of this sub-mission instead of 300 districts that have chosen previ-
ously. All animals are now covered by the system, including indigenous/crossbreed
milk cattle, pack animals (horses, donkeys, mules, camels, ponies and buffaloes)
and other animals (goats, sheep, pigs, rabbits and yaks) rather than just milk cattle
and buffaloes that were covered previously. The subsidy advantage has been
increased and is limited to five cattle per unit per household beneficiary instead of
just two milk animals per household. One unit of cattle is regarded as equivalent to
ten livestock in the case of goat, sheep, pigs and rabbit.
16.5.1.2 S ub-mission on Skill Development, Technology Transfer

and Extension
The extension equipment for livestock operations at the field level is very weak. As
a consequence, farmers are unable to adapt the study institutions’ technique. For
this, new technique and procedures need to be linked between all stockholders.
Accordingly, a structured approach to livestock expansion has been incorporated
into the NLM to tackle the various livestock owner requirements.
16.5.1.3 Sub-mission of Fodder and Feed Development

It deals with non-forest wasteland/grassland/non-arable land fodder manufacturing,
forest land, coarse grain cultivation and dual-purpose plants, fodder seed procure-
ment and allocation, etc.
16.5.1.4 Sub-mission on Pig Development in Northeastern Region

A sub-mission on pig development was initiated for the first time under NLM in
which the Indian government would support the state piggy farms and the importa-
tion of germplasm.
16.5.2 National Dairy Plan Phase-1 (NDP-1)
National Dairy Plan Phase-1 (NDP-1) is also known as Mission Milk. This pro-
gramme was started in April 2012 at the National Dairy Development Board
(Anand, Gujarat) for a period up to 2018–2019. This programme is a multi-state
initiative covering 18 major milk-producing states, namely, Uttarakhand, Kerala,
Maharashtra, Karnataka, Andhra Pradesh, Telangana, Odisha, Bihar, Tamil Nadu,
Haryana, Madhya Pradesh, Gujarat, Chhattisgarh, Uttar Pradesh, Punjab, Rajasthan,
West Bengal and Jharkhand. All the states together account for over 90% of the
country’s milk production (National Dairy Development Board 2019). This pro-
gramme is systematically intended with the following objectives:
• To assist boost the milk productivity and thus boost the milk output to satisfy the
quickly increasing demand for milk.
• To help ensure higher access for rural milk manufacturers to the organized milk
processing industry.
By the adoption of technical and logical processes, these objectives would be

materialized. Appropriate regulatory and policy measures along with the provision
of technical inputs will be pursued. This project was implemented by the National
Dairy Development Board through end implementing agencies located in the differ-
ent states. This project is largely financed with a loan from the International
Development Association of World Bank. The budget provision for the scheme dur-
ing 12th 5-year plan was of Rs. 1400 crores.
16.5.2.1 Components of NDP-1
Productivity Enhancement
Improved animal nutrition is the main expected outcomes from the proposed inter-
ventions under this component. Improved artificial insemination (AI) design rates,
reduced feeding costs per kg of milk production, increased milk production by
increasing productivity per animal and reduced methane released per kg of animal
covered under Ration Balancing Programme (RBP). This constituent focuses on
enhancing bovine productivity through a methodical approach related to the animal
breeding and nutrition.
Milk Procurement Systems for Collection, Weighing, Testing the Quality

of Milk and Making Payment to Milk Producers at the Village Level
This includes investment in milk collection infrastructure at village levels such as
milk cans, bulk milk coolers for a village cluster, associated weighing and testing
facilities and linked IT facilities. This aims at establishing village-based milk pro-
curement schemes for the fair and transparent collection of milk and ensuring their
timely payments. A rise in the number of extra villages covered and more milk
producers organized into dairy cooperative societies and milk producer institutions
are the primary anticipated outcomes of the measures suggested under this
initiative.
16.5.2.2 Project Management and Learning

The main purpose of this is to achieve outcome of effective harmonization of project
activities between different agencies for the end implementations, preparing and
implementing annual plans in good time, regular review and reporting of advance-
ment and outcomes of the project and extensive and functional information system
for the project management. It also promotes the creation of abilities and under-
standing of staff engaged in the project execution and develops a capacity building
that extends beyond the life of the project. The project provides funding for the
effective collection and spreading of data for the continuous operations and man-
agement of matters related to breeding, nutrition and milk procurement systems.
16.5.3 National Programme for Bovine Breeding and Dairy

Development (NPBB and DD)
During the 12th 5-year plan, this Programme was launched in 2014 with a budget of
Rs. 1800 crores for implementation (Vikaspedia 2019). This programme consists of
two components with the subsequent objectives:
(a) National Programme for Bovine Breeding (NPBB)

• To facilitate and understand artificial insemination techniques to farmers
doorstep.
• To gather good breedable female cattle through AI or natural facility by
using germplasm of high genetic virtues.
• To provide high worth breeding input services to significant indigenous
breeds so as to prevent the extinction and destruction of species.
• To preserve, expand and propagate certain native bovine breeds of high
socio-economic significance.
(b) National Programme for Dairy Development (NPDD)
• Action to build and reinforce infrastructures and facilities for the production
of superiority milk including cold chain supply chain, helping and linking
the farmer to the consumer.
• To develop and strengthen procurement infrastructure, collection, process-
ing and distribution of milk.
• To create scheduled training facilities for dairy farmers.
• To support dairy cooperative societies/manufacturers at the village level.
• To enhance milk production by facilitating technical input facility such as
cattle feeds, mineral mixture, etc.
• To help in the upbringing of underdeveloping milk federations/unions.
16.6 Gender Mainstreaming in Dairying
Dairying plays a critical role in the livelihood of farmers. Men and women partici-
pate in dairying, but the activities performed and the benefits gained by them vary
in many places. Even though women contribute highly to the growth of the dairy
sector, they remain as invisible farmers or nonpaid family labours. In dairying, 80%
of the activities are done by the women in rural households, but the major decision-
makers are men. So without solving the gender issues in dairying, we cannot achieve
the desired growth. Gender analysis is essential for maintaining gender equality.
Gender refers to the socially and mentally developed roles and various responsi-
bilities of men and women. It differs from sex, as sex determines the biological
differences between them. Sex is determined by birth, but gender defines the
acquired roles which can be changed over time and between cultures. Gender equal-
ity means favouring men and women equally to develop their personal abilities
without any limitations. The fairness and justice in the distribution of the benefits to
men and women are known as gender equity. Gender mainstreaming involves the
development of strategies, policies and interventions at all levels for the equal ben-
efits of men and women. Both men and women should get equal access to the
resources around them. For that, they should be sensitized regarding the inequalities
existing and the better ways available to solve them.
16.6.1 Gender Issues in Dairying
There are different gender issues in dairy farming also. Normally at rural household
levels, the dairy activities are divided between men and women. Women perform
mainly cleaning of the shed, feeding animals, milking, washing of milk vessels, etc.
Men handles the marketing part mostly. The control over the resources mostly lies
with men. Many times, sharing of the benefits does not take place judiciously. The
time utilized by women and the labour involved in dairying may be equal or high as
that of men but still faces the problem of invisibility as a dairy farmer. The access to
credits by women is another issue. As women don’t have much access to resources,
they find difficulty in getting credit. Women receive normally fewer training or
exposure visits compared to their counterparts. Normally the participation of women
in the government organized extension programs is low due to their other assigned
roles at home or due to some social or cultural barriers. Women rarely go to market-
places for purchasing the inputs for dairying or for selling the milk. They always
prefer to sell the milk or milk products in the neighbourhood or to some local trad-
ers. This prevents them from getting good remuneration from their dairy farming.
Due to less education and fewer negotiation skills, they lie below the men in har-
nessing the benefits. All these gender issues can be solved with proper
interventions.
16.6.2 Dimensions of Women Empowerment
The women empowerment is a widely acknowledged concept for the development

of every sector. The empowerment book of World Bank defines empowerment as
‘the extension of liberty of selection and action to form one’s life’. For empowering
women they should be involved in the change process to improve gender equality
rather than being its recipients. Women should be motivated and enabled enough to
set their goals and achieve them without getting afraid of any obstacles on their
path. By empowering women, they take ownership and control of their own choices.
There are different dimensions for women empowerment which includes the eco-
nomic, cultural, social, political, legal and psychological empowerment.
Economical empowerment means women get financial independence. They gain

ownership of assets and control over income. Social and cultural empowerment
gives greater visibility to women in social platforms. Through this, they will be free
from domestic and sexual acts of violence. The political empowerment ensures the
presence of women in the political system. The legal dimension of empowerment
includes expanding the knowledge of women regarding legal rights and how to get
benefit or protection by using the judicial system. Psychological empowerment
makes women happy and feels self-worth. By understanding of all these dimen-
sions, policies have to be developed to increase gender equality (Ponnusamy and
Sharma 2015).
16.6.3 Role of Self-Help Groups (SHG) in Women Empowerment
Self-help groups are a voluntary association of poor people for solving their prob-
lems. The number of people involved can be 10–20. The members of SHG should
be from the same socio-economic background and work through self-help and
mutual support. The formation of women SHGs can greatly contribute to their
socio-economic development. The SHGs inculcate the habit of saving small amounts
of money among the women and also increase their access to credit.
Nowadays, rural dairy sector also has many SHGs that are functioning which are
involved in dairy farming, processing and marketing. The formation of SHGs has
been carried out as an intervention by many organizations as a tool for the rural
development. This helps the women to work together by the creation of a common
fund. The SHGs easily get credit from the financial institutions which help them to
invest in some income-generating activities. An example of SHGs working in the
dairy sector is ‘Anmol Dughd Mahila Samitee’ at Amritpurkalan village in
Gharaunda block of Karnal district. This group was constituted by 15 women who
undergone a 3-month training programme on the production of value-added dairy
products at the ICAR-National Dairy Research Institute, Karnal.
16.6.4 Strategies for Women Empowerment in Dairying
The empowerment of women in dairying can be achieved by adopting different

measures starting from the grassroot level. Bringing some extension programs spe-
cifically for women and arranged by women extension officers can make differ-
ences. By targeting women as the audience for improving their knowledge will
contribute to the empowerment. The specific training programmes can be arranged
on scientific dairy farming and production of value-added milk products. Establishing
custom hiring centres of newly developed technologies at village level for women
farmers will be helpful. By connecting them to markets by creating linkages can
help to gain more profits. Special types of equipment should be designed to reduce
the drudgery of farm women. Village level sensitization is required to change the
attitude of male and female farmers regarding their roles and responsibilities in
performing dairy farming. Formation of SHGs and institutional frameworks for col-
lective action of women can improve their decision-making ability and help them
raise their voice. Gender policy is essential for improving the livelihood of farm
women. Negotiation skills and market literacy should be improved by capacity
building programs. The gender issues should be studied thoroughly to plan specific
programs which cater the gender needs. A proper monitoring system should be
developed at the village level for ensuring the effectiveness of different measure
undertaken to bring gender equality. Adequate attention is required in sensitizing
various departmental officials for providing equal opportunities for men and women
to perform better in the dairy sector.
16.7 Conclusions
Agriculture and allied activities play a vital role in the existence of a human being.
A portion of the food products produced is being wasted due to improper manage-
ment and handling. Proper food handling can reduce this problem by establishing
and implementing measures for quick marketing of food products to the consumers.
In case of the dairy industry, milk is very much prone to spoilage in short time,
which needs more attention. Thus, technologies are improved nowadays to handle
these problems and an extension person can educate as well as solve the problems
of farmers. The extension provides support to farmers to face new challenges in
farming such as transformation in global food and agricultural system, marketing,
the importance of food safety in agricultural farming, etc. Information and commu-
nication technology helps farmers make use of advanced technologies in the field of
agriculture and dairying. Entrepreneurship development programmes can strengthen
the backbone of farming by attracting persons to agriculture by giving in-hand train-
ing. There were different projects implemented in the dairy and animal husbandry
sector in India for improving the communication among farmers, entrepreneurship
development and women empowerment.
References
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initiative-in-the-district-of-Dhar-Madhya-Pradesh. Accessed 12 Jul 2019
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https://www.mssrf.org/mssrfoldsite/?q=iec. Accessed 21 Jul 2019
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Mondal S, Ray GL (2007) Textbook on rural development entrepreneurship and communication
skills. Kalyani Publishers, Ludhiana
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National Dairy Research Institute, Karnal
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pedia.in/agriculture/policies-and-schemes/livestock-poultry related/ national-project-for-cat-
tle-buffalo-breeding. Accessed 10 Jul 2019

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Jagrani

ISBN 978-981-15-2607-7 ISBN 978-981-15-2608-4 (eBook)

© Springer Nature Singapore Pte Ltd. 2020

range of readers including dairy scientists, dairy engineers, dairy technologists,

Lincoln, NE, USA Jagrani Minj

1 Basic Facts About Dairy Processing and Technologies�������������������������� 1

About the Editors

Jagrani Minj, Ph.D. (Dairy Microbiology), is currently a Postdoctoral Research

Aparna Sudhakaran V, Ph.D. (Dairy Microbiology), is an Assistant Professor of

Anuradha Kumari, Ph.D. (Dairy Chemistry), is an Assistant Professor of Dairy

Santosh Anand, Ph.D. (Dairy Microbiology) Dairy Microbiology Department,

M. Misha Madhavan, Ph.D. (Agricultural Extension) ICAR-National Research

Veterinary and Animal Sciences University, Thrissur, Kerala, India

FDV Flow Diversion Valve

IC50 Inhibitory Concentration

MST Milli Second Technology

RFID Radio Frequency Identification

USA United States of America

Aparna Sudhakaran V (*)

© Springer Nature Singapore Pte Ltd. 2020 1

1.2 Overview of the Global Dairy Industry

1.3 The Impact of Dairy Processing

1.4 Classification of Milk Products

Codex Alimentarius defines a milk product as a ‘product obtained by any processing

Dried milk Fermented

Fig. 1.1 Basic classification of milk products

1.5 Major Dairy Processing Area and Applied Technologies

1.5.1 Liquid Milk Processing

Table 1.1 Liquid milk Liquid milk

Table 1.2 Fat-rich dairy Cream

Table 1.3 Classification of cream based on fat content and processing methods

1.5.2 Fat-Rich Dairy Product Processing Technologies

Table 1.5 Frozen dairy Ice cream

1.5.3 Frozen Dairy Product Processing Technologies

1.5.4 Fermented Dairy Product Processing Technologies

Fermentation of milk by means of specific microorganisms which coagulate the

Table 1.6 Fermented dairy Fermented milk

antibacterial activity (Namaei et al. 2015; Taha et al. 2017; Mohammadi-Gouraji

Table 1.7 Fermented milk available in different countries

1.5.5 Enzyme Coagulated Dairy Products

Cheese is basically the protein coagulation obtained by coagulating wholly or

1.5.6 Dehydrated Dairy Products

Table 1.8 Classification of Enzyme coagulated milk products

Table 1.9 Dehydrated milk Evaporated milk

Table 1.10 Classification of evaporated milk

Table 1.11 Classification of condensed milk

1.5.7 By-Products in Dairy Industry

Table 1.12 Types of milk powder

Fig. 1.2 By-products of the Dairy Industry

Table 1.13 The processing technologies employed in by-product utilization

1.5.8 Specialized Dairy Products

1.5.8.1 Functional Foods

Table 1.14 Classifications of functional foods

several diseases like hypertension, coronary vascular diseases, obesity, osteoporo-

1.5.8.2 Designer Milk

1.5.8.3 Organic Milk

1.6 I mportance of Advanced and Innovative Technologies

Application of advanced and innovative approaches in the dairy processing sector is

FDA (2017) Guide to inspections of dairy product manufacturers. https://www.fda.gov/

© Springer Nature Singapore Pte Ltd. 2020 25

c­ hapter, the outlines of non-thermal techniques used in dairy processing like

2.2 Ultrasound Technology

Ultrasound is an emerging technology with immense potential in food processing

2.2.1.1 Microbial Inactivation

Lincoln, NE, USA Jagrani Minj

1 Basic Facts About Dairy Processing and Technologies�� 1

1.2 Overview of the Global Dairy Industry

1.3 The Impact of Dairy Processing

1.4 Classification of Milk Products

1.5 Major Dairy Processing Area and Applied Technologies

1.5.1 Liquid Milk Processing

1.5.2 Fat-Rich Dairy Product Processing Technologies

1.5.3 Frozen Dairy Product Processing Technologies

1.5.4 Fermented Dairy Product Processing Technologies

1.5.5 Enzyme Coagulated Dairy Products

1.5.6 Dehydrated Dairy Products

1.5.7 By-Products in Dairy Industry

1.5.8 Specialized Dairy Products

1.5.8.1 Functional Foods

1.5.8.2 Designer Milk

1.5.8.3 Organic Milk

1.6 I mportance of Advanced and Innovative Technologies

c hapter, the outlines of non-thermal techniques used in dairy processing like

2.2 Ultrasound Technology

2.2.1.1 Microbial Inactivation

2.2.1.2 Ultrasound Applications for Dairy Products

2.3 High Pressure Processing

2.3.1.1 Microbial Inactivation

2.3.1.2 Effects of HPP on Milk Components

2.3.1.3 High Pressure Processing of Dairy Products

2.4 Cold Plasma Processing

2.5 Supercritical Carbon Dioxide Technology

2.6.1.1 Microbial Inactivation

2.6.1.2 Irradiation of Dairy Products

2.7 Pulsed Electric Field Processing

3.2 The Beginning

3.3 Engineering the Dairy

3.4 Milk Processing

3.5 The Cold Chain

3.6 Farm Automation

3.8 Engineering in the Dairy Plant

3.9 Pointers to the Future

4.2 Advances in Farm-Level Interventions

4.3 Engineering in Milk Testing

4.4 Advances in Dairy Processing

4.4.1 Pasteurization and Sterilization

4.4.2 Cream Separation, Standardization, and Homogenization

4.4.3 Evaporation and Drying

4.4.4 Butter and Cheese

4.4.5 Ice Cream

4.6 Cleaning and Sanitation

4.7 Dairy Effluent Treatment

4.8 Storage and Distribution

5.2 Importance of Fortification in Milk and Milk Products

5.3 omponents Used for the Fortification of Milk

5.3.6 Phytochemicals or Plant-Based Ingredients

5.3.7 Fruit/Fruit Pulp/Fruit Purees

5.3.8 Short-Chain Fatty Acids Such as Omega-3 Fatty Acids