Block 1

Introduction to Food
UNIT 1 INTRODUCTION TO FOOD Microbiology
MICROBIOLOGY
Structure
1.0 Objectives
1.1 Introduction
1.2 The Science of Microbiology
1.3 Food Microbiology - its Origins and Scope
1.4 Importance of Micro-organisms in Foods
1.5 Classification and Nomenclature of Micro-organisms
1.6 Micro-organisms in Food
1.6.1 Bacteria
1.6.2 Molds
1.6.3 Yeasts
1.6.4 Viruses
1.6.5 Parasitic Organism
1.7 Important Micro-organisms in Food

1.7.1 Important Mold Genera
1.7.2 Important Yeast Genera
1.7.3 Important Viruses
1.7.4 Important Bacterial Genera
1.8 Normal Microflora of some Common Foods

1.9 Let Us Sum Up
1.10 Key Words
1.11 Answers to Check Your Progress Exercises
1.12 Suggested Reading
1.0 OBJECTIVES
After reading the Unit, we shall be able to:
z explain the applied aspects of microbiology;
z comprehend the need and scope of food microbiology;
z enumerate the important genera of micro-organisms associated with food;
and
z specify normal microflora of some common foods.
1.1 INTRODUCTION
Micro-organisms are living entities of microscopic size and include bacteria,
viruses, yeasts and molds (together designated as fungi), algae, and protozoa.
While bacteria are classified as prokaryotes (cells without definite nuclei), the
fungi, algae, and protozoa are eukaryotes (cells with nuclei); viruses do not
have regular cell structures and are classified separately. Micro-organisms are 7
Fundamentals of Food present everywhere on earth, which includes humans, animals, plants and
Microbiology
other living creatures, soil, water, and atmosphere, and they can multiply
everywhere except in the atmosphere. Together, their numbers far exceed all
other living cells on this planet. They were the first living cells to inhabit the
earth over 3 billion years ago; and since then they have played important
roles, many of which are beneficial to the other living systems.
Among the micro-organisms, some molds, yeasts, bacteria, and viruses have
both desirable and undesirable roles in our food. In this unit, the scope of
food microbiology, importance of microbes in food and predominant micro-
organisms associated with food have been discussed.
1.2 THE SCIENCE OF MICROBIOLOGY

Microbiology is the branch of the biological sciences that deals with micro-
organisms, i.e. bacteria, fungi, some algae, protozoa, viruses, viroids and
prions. Most micro-organisms have the following characteristics:-
1) They are generally too small to be seen with the unaided human eye, and
some form of microscopy is required for the study of their structure.
2) Cells or other structures are relatively simple and less specialized than
those of higher plants and animals.
3) They are handled and cultured in the laboratory in ways that are generally
quite similar.
Microbiology has developed into a science that can be studied from a number
of perspectives. A specialist study can be made of each of the individual
groups giving rise to the following disciplines:
• Bacteriology - the study of bacteria;
• Mycology - the study of fungi;
• Protozoology - the study of protozoa;
• Phycology (algology) - the study of algae;
• Virology - the study of viruses.
Micro-organisms can also be studied from the applied viewpoint, i.e. the
relationship between micro-organisms, the environment and human activity.
This again gives rise to a number of areas of specialist study:
• Medical microbiology includes some aspects of pathology (the study of
diseases), immunology (how the immune system operates to prevent
invasion by micro-organisms) and epidemiology (how diseases are
distributed and spread).
• Agricultural microbiology: The study of micro-organisms for crop/
plant health and related areas.
• Industrial microbiology / biotechnology: The study of the use of Micro-
organisms in large scale industrial processes.
• Food microbiology: The study of the role that micro-organisms play in
8 food spoilage, food production, food preservation and food-borne disease.
None of these areas of specialist study can operate in isolation, e.g. food Introduction to Food
Microbiology
microbiology encompasses various aspects of industrial microbiology and
biotechnology in the manufacture of fermented food and the production of
single-cell protein. A study of food-borne disease involves aspects of medical
microbiology and agricultural microbiology.
Specialist knowledge needs to be underpinned by an understanding of
fundamental principles. The food microbiologist, for example, needs to have
an understanding of microbial structure; the classification and identification of
micro-organisms; how micro-organisms grow; the factors that influence growth
and how growth can be controlled; death of micro-organisms; nutrition of
Micro-organisms and how they are cultured in the laboratory.
1.3 FOOD MICROBIOLOGY - ITS ORIGIN AND

SCOPE
Although processes of food spoilage and methods of food preservation and
food fermentation have been recognized since ancient times, it was not until
the 1800s that the relationship between foods and micro-organisms was
established. In 1837 Schwann proposed that the yeast which appeared during
alcoholic fermentation was a microscopic plant, and between 1857 and 1876
Pasteur showed that micro-organisms were responsible for the chemical
changes that take place in foods and beverages.
Their observations laid the foundation for the development of food microbiology
as we know it today. Soon after these early discoveries were made, knowledge
about the role that micro-organisms play in food preservation, food spoilage
and food preservation, food spoilage and food poisoning accelerated rapidly
until food microbiology gradually emerged as a discipline in its own right.
Food microbiology is now a highly developed area of knowledge with the
main areas of interest highlighted in Fig. 1.1.
Food Laboratory
fermentations management
Water Food
quality hygiene
Topics of major interest

to the food microbiologist
Food Food-borne
preservation disease
Food Quality
spoilage control
Fig. 1.1: Topics of major interest to the food microbiologist 9

Fundamentals of Food Not all groups of micro-organisms are of equal interest to the food
Microbiology
microbiologist. Bacteria come very much on top of the list with molds and
yeasts also of considerable importance and viruses less so. The associations
that these organisms have with the manufacture and consumption of foods are
summarized in Fig. 1.2.
Food-borne disease Food-borne disease
Food spoilage
Food spoilage
Bacteria Moulds Production of single cell protein
Food fermentations
Food fermentations
Production of food additives and enzymes Production of enzymes
Food spoilage Food-borne disease
Yeasts Food fermentations Viruses Identification of food

poisoning bacteria
Production of food additives and enzymes

Failure of dairy fermentations
Fig. 1.2: Various groups of micro-organisms and their associations with food
Protozoa and algae have minimum direct impact on the production, processing
and consumption of food. Food-borne disease can be caused by some protozoa
and others belonging to this group are important in the treatment of wastes.
Algae are used to produce alginates; some have the potential for use in the
production of single-cell protein and some marine species produce toxins that
might enter our food along with sea foods.
1.4 IMPORTANCE OF MICRO-ORGANISMS IN

FOODS
Since 1900 A.D. our understanding of the importance of micro-organisms in

food has increased greatly. Their role in food can be either desirable (food
bioprocessing) or undesirable (food borne diseases and food spoilage), which
is briefly discussed here.
1.4.1 Food-borne Diseases
Many pathogenic micro-organisms (bacteria, molds and viruses) can

contaminate foods during various stages of their handling, between production
and consumption. Consumption of these foods can cause food borne diseases.
Food borne diseases can be fatal and may also cause large economic losses.
Foods of animal origin are associated, more with food borne diseases than
foods of plant origin. Mass production of food, introduction of new technologies
10 in the processing and storage of food, changes in food consumption patterns,
and increased import of food from other countries have increased the chances Introduction to Food
Microbiology
of large outbreaks as well as the introduction of new pathogens. Effective
intervention technologies are being developed and implemented to ensure the
safety of consumers against food borne diseases. New methods are also
being developed to effectively and rapidly identify the pathogens in contaminated
foods.
1.4.2 Food Spoilage

Except for sterile foods, all foods harbor micro-organisms. Food spoilage
stems from the growth of these micro-organisms in food or is due to the
action of microbial enzymes. New marketing trends, consumers’ desire for
foods that are not overly processed and preserved, extended shelf life, and
chances of temperature abuse between production and consumption of foods
have greatly increased the chances of food spoilage and, in some instances,
with new types of micro-organisms. The major concerns are the economic
loss and wastage of food. New concepts are being studied to reduce
contamination as well as control the growth of spoilage microbes in foods.
1.4.3 Food Bioprocessing

Many food-grade micro-organisms are used to produce different types of
fermented foods using raw materials from animal and plant sources.
Consumption of these foods has increased greatly over the last 15 to 20 years
and is expected to increase further in the future. There have been great
changes in the production and availability of these micro-organisms (starter
cultures) to meet the large demand. In addition, novel and better strains are
being developed by using genetic engineering techniques.
1.4.4 Food Additives

Microbial enzymes are also being used to produce food and food additives.
By employing genetic recombination techniques, and using diverse microbial
sources enzymes of higher purity & activity are obtained. Many types of
additives from microbial sources are being developed and used in food. Some
of these include single-cell proteins, essential amino acids, colour compounds,
flavour compounds, stabilizers and organic acids.
1.4.5 Food Biopreservation

Antimicrobial metabolites (e.g. bacteriocins and organic acids like acetic,
propionic and lactic acids) of desirable Micro-organisms are being developed
and used in foods in place of preservatives of non-food (chemical) origin to
control pathogenic and spoilage micro-organisms in food. Economic production
of these antimicrobial compounds and their effectiveness in food systems have
generated wide interest.
1.4.6 Probiotics
Consumption of foods containing live cells of bacteria and that have apparent
health benefits has generated interest among consumers. The role of these
bacteria for health and bacterial efficacy benefits is being critically investigated.
11
Fundamentals of Food
Microbiology Check Your Progress Exercise 1
Note: a) Use the space below for your answer.
b) Compare your answers with those given at the end of the unit.
1) What are Micro-organisms?
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2) What is Food Microbiology?
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3) What is a probiotic?
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1.5 CLASSIFICATION AND NOMENCLATURE

OF MICRO-ORGANISMS
Living cellular organisms, on the basis of phylogenetic and evolutionary
relationships, are grouped into five kingdoms in which bacteria belong to
prokaryote (before nucleus), while the eukaryotic (with nucleus) molds and
yeasts are grouped under fungi. Viruses are not considered as living cells and
are not included in this classification system.
For the classification of yeasts, molds, and bacteria, several ranks are used
after the kingdom. These are divisions, classes, orders, families, genera (singular,
genus), and species. The basic taxonomic group is the species. Several species
with similar characteristics form a genus.
A family is made up of several genera, and the same procedure is followed in
the hierarchy. Ranks above species, genus, and family are seldom used in
food microbiology. Among bacteria, a species is regarded as a collection of
strains having many common features. A strain is the descendent of a single
colony (single cell). Among the strains in a species, one is assigned as the
type strain; it is used as a reference strain while comparing the characteristics
of an unknown isolate.
The basic taxonomic group in bacteria, yeasts, and molds is the species, and
each species is given a name. The name has two parts (binomial name); the
first part is the genus name and the second part is the specific epithet (adjective).
Both parts are Latinized; when written, they are italicised (or underlined) with
the first letter of the genus written in a capital letter and species name in small
12 letters. For e.g. Bacillus subtilis (genus is Bacillus and species is subtilis)
Introduction to Food
Microbiology
1.6 MICRO-ORGANISMS IN FOOD
The Micro-organisms most common to food are bacteria and fungi. The fungi,
which are less common than bacteria, consist of two major types of Micro-
organisms, viz. molds and yeasts. Apart from these, food may contain viruses
and other parasites such as protozoans, worms etc.
1.6.1 Bacteria
Bacteria are unicellular Micro-organisms that are approximately one micro
meter (10-3mm) in diameter with variations in morphology from short and
Fig. 1.3: Types of bacterial cells and their groupings 13

Fundamentals of Food elongated rods (bacilli), spherical or ovoid forms(cocci), vibrio (comma shaped)
Microbiology
and even spiral in shape (Refer Fig. 1.3). Cocci (meaning “berry”) are sphere
shaped bacteria. Individual bacteria closely combine in various forms according
to genera. Some sphere-shaped bacteria occur in clusters similar to a bunch
of grapes (i.e. staphylococci). Other bacteria (rod shaped or sphere shaped)
are linked together to form chains (i.e. streptococci in case of cocci chain).
Certain genera of sphere-shaped bacteria are found together in pairs
(diplococci i.e. Pneumococci) or as a group of four (Square or cubical packets
formation; i.e. Sarcinia), while other genera appear as an individual bacterium.
Other bacteria (in majority) are rod shaped and possess flagella and are
motile.
Bacteria produce various pigments which range from shades of yellow to
dark pigments such as brown or black. Certain bacteria have pigmentation of
intermediate colors such as red, pink, orange, blue, green, or purple. These
bacteria cause food discoloration, especially, among foods with unstable color
pigments such as meat. Some bacteria also cause discoloration by slime
formation.
1.6.2 Molds
Molds are multicellular micro-organisms with mycelial (filamentous) morphology.
These microbes are also characterized by their display of a variety of colors
and are generally recognized by their mildewy or fuzzy, cotton like appearance.
Molds can develop numerous tiny spores that are found in the air and can be
spread by air currents. These spores can produce new mold growth if they
are transferred to a location that has conditions conducive to germination.
Molds generally withstand greater fluctuation in pH than bacteria and yeasts
and can frequently tolerate more temperature fluctuation. Although molds thrive
best at or near a pH of 7.0, a pH range of 2.0 to 8.0 can be tolerated, even
though an acid to neutral pH is preferred. Molds thrive better at ambient
temperature than in a colder environment, even though growth can occur
below 0°C. Although mold growth is optimal at a water activity (Aw) of
approximately 0.85, growth can and does occur below 0.80. At an Aw of
0.90 or higher, bacteria and yeasts grow more effectively and normally utilize
available nutrients for growth at the expense of molds. When the Aw goes
below 0.90, molds grow more effectively. That is why foodstuffs, such as
pastries, cheeses, and nuts, that are low in moisture content are more likely to
spoil from mold growth.
1.6.3 Yeasts
Yeasts are generally unicellular and differ from bacteria in their large cell size
and morphology, and because they produce buds during the process of
reproduction by division. Like molds, yeasts can be spread through the air, or
other means, and alight on the surface of foodstuffs. Yeast colonies are generally
moist or slimy in appearance and creamy white colored. Yeasts prefer an Aw
of 0.90 - 0.94, but can grow below 0.90. These micro-organisms grow best
in the intermediate acid range, pH from 4.0 to 4.5. Food that is highly
contaminated with yeasts will frequently have a slightly fruity odour.
1.6.4 Viruses
Viruses are 10- 450 nm in size; cannot reproduce without a living host; attack
14 only susceptible host cell lines; infect plants, animals, and bacteria; and have
the capacity to produce specific diseases in specific hosts. Transmission occurs Introduction to Food
Microbiology
in foods, water and air. Viruses that infect bacteria are called bacteriophages.
Viruses are included in the order Virales.
Viruses are too small to be visualized with an ordinary compound microscope.
Only after the electron microscope was developed, the direct observation of
viruses was possible. Viruses consist of a DNA or RNA core surrounded by
a protein coat. Because they lack all the apparatus for normal cellular
metabolism, they must utilize the cellular machinery of the host cell in order to
grow and divide. Once they invade a host cell, however, viruses can multiply
very rapidly.
1.6.5 Parasitic Organisms
A number of parasitic worms can also be transmitted by food to cause diseases
in humans.
Cestodes are flatworms that inhabit the intestinal tract, heart, and lungs of
animals. Beef, swine, dogs and other canine species, bears, and fish can all
harbour tapeworms and flatworms, which can be transmitted to and can infect
humans.
Trematodes are non segmented flatworms that possess a mouth and oral
sucker and depend on a snail as an intermediate host before infecting humans
by being ingested in drinking water or aquatic plants. Intestinal flukes, pyriform
worms from fish, sheep and Chinese liver flukes, and oriental lung flukes are
all examples of food-transmitted parasites.
Nematodes or true roundworms also can be transmitted from animals to
humans. Eggs carried in excrement from roaches and dung beetles ingested
by cattle, sheep and hogs contaminate humans. Trichinosis is an inflammation
of the muscle tissue caused by ingesting the worm Trichinella spiralis. Pork
is the most common vector. Capillary worms, whipworms, and pinworms are
other examples of nematode parasites.
Protozoa are microscopic single-celled animals, which can be taken in with
food or water to cause human illness. Entamoeba histolytica, Toxoplasma
gondii, Balantidium coli, and Giardia lamblia are the most common food
borne protozoan parasites.
Check Your Progress Exercise 2

1) What is Systematics?
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2) What are the two most common Micro-organisms found in food?
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Fundamentals of Food 3) Classify bacteria on the basis of their morphology.
Microbiology
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4) What is a bacteriophage?
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1.7 IMPORTANT MICRO-ORGANISMS IN FOOD

1.7.1 Important Mold Genera
Molds are important in food because they can grow in conditions in which
many bacteria cannot, such as low pH, low water activity (a w ), and high
osmotic pressure. They are important spoilage micro-organisms. Many strains
also produce mycotoxins and have been implicated in food borne intoxication.
Many are used in food bioprocessing. Finally, many are used to produce food
additives and enzymes. Some of the most common genera of molds found in
food are listed here.
1 ) Aspergillus : They are widely distributed and contain many species that
are important in food. They have septate hyphae and produce a sexual
spores (black color) or conidia. Many are xerophilic (able to grow in
low Aw) and can grow in grains, causing spoilage. They are also involved
in spoilage of foods such as jams, cured ham, nuts, and fruits and
vegetables (rot). Some species/strains produce mycotoxin (e.g.,
Aspergillus flavus produces aflatoxin). Many species/strains are also
used in food and food additive processing. Aspergillus oryzae is used to
hydrolyze starch by alpha-amylase in the production of sake. Aspergillus
niger is used to process citric acid from sucrose and to produce enzymes
like-galactosidase.
2 ) Alternaria: They are also septate and form dark-brown colored many
celled conidia on the conidiophere. They cause rot in tomatoes and
rancid flavor in dairy products. Species: Alternaria tenuis.
3 ) Geotrichum: The hyphae are septate and form rectangular asexual
arthrospores (oidia). They grow forming a yeast like, cottony, creamy
colony. They establish easily in equipment and often grow on dairy
products (also known as dairy mold). Species: Geotrichum candidum.
4 ) Mucor : They are widely distributed. They have nonseptate hyphae and
produce sporangiophores. They produce cottony colonies. Some species
are used in food fermentation and production of enzymes. They cause
spoilage of vegetables. Species: Mucor rouxii.
5 ) Penicillium: They are widely distributed and contain many species.
They have septate hyphae and form conidiophore on a blue-green, brush-
16 like conidia head. Some species are used in food production, such as
Penicillium roquefortii and Penicillium camembertii in cheese. Many Introduction to Food
Microbiology
species cause fungal rot in fruits and vegetables.
6) Rhizopus: The hyphae are aseptate and form sporangiophores in
sporangium. They are involved in the spoilage of many fruits and
vegetables. Rhizopus stolonifer is the common black bread mold.
Botrytis Cladosporium Rhizopus Alternaria
Fusarium Asporgillus Pencillium
Fig. 1.4: Common mold species found in foods
1.7.2 Important Yeast Genera

Yeasts are important in food due to their ability to cause spoilage. Many are
also used in food bioprocessing. Some are used to produce food additives.
Several important genera are briefly described below.
1 ) Saccharomyces: Cells are round, oval, or elongated. It is the most

important genus and contains heterogeneous groups. Saccharomyces
cerevisiae variants are used in baking for leavening of bread and in
alcoholic fermentation. They are also involved in spoilage of food with
the production of alcohol and CO2.
2 ) Pichia: They are oval to cylindrical cells and form pellicle in beer, wine,
and brine to cause spoilage. Some are also used in oriental food
fermentation. Species: Pichia membranaefaciens.
3 ) Rhodotorula: They are pigment (red, pink or yellow) forming yeasts

and can cause discoloration of foods, such as in meat, fish, and sauerkraut.
Species Rhodotorula glutinis.
17
Fundamentals of Food 4 ) Torulopsis: They have spherical to oval structure. They cause spoilage
Microbiology
of milk due to the ability to ferment lactose (Torulopsis sphaerica).
They also spoil fruit juice concentrates and acid foods.
5 ) Candida: Many spoil foods with high acid, salt, and sugar and form
pellicle on the surface of liquids. Some can cause rancidity in butter and
dairy products (Candida lipolytica).
6) Zygosaccharomyces: Involved in spoilage of foods, containing high sugar/
salt levels ex. honey, sirups, molasses, soy sauce. (Zygosaccharomyces
nussbaumeri). These yeasts are termed osmophilic, because they can
grow in high concentrations of solutes.
1.7.3 Important Viruses

Viruses are important in food for three reasons. Some are able to cause
enteric disease and thus, if present in a food, can cause food borne diseases.
Hepatitis A and Norwalk viruses have been implicated in food borne outbreaks.
Several other enteric viruses, such as Poliovirus, Echovirus, and Coxsackievirus,
have the potential of causing food borne diseases. In some countries where
the level of sanitation is not very high, they can contaminate foods and cause
disease.
Some bacterial viruses (bacteriophages) are used in the identification of species/
strains by a process called tranduction (e.g., in Escherichia, coli, Lactococcus
lactis).
Finally, some bacteriophages can be very important due to their ability to
cause fermentation failure. Many lactic acid bacteria, used as starter cultures
in food fermentation, are sensitive to different bacteriophages. These phages
can infect and destroy starter culture bacteria, causing product failure. Among
the lactic acid bacteria, bacteriophages have been isolated for many species
in genera Lactococcus, Streptococcus, Leuconostoc, and Lactobacills.
Methods are being studied to genetically engineer lactic acid start cultures so
that they become resistant to multiple bacteriophages.
Table 1.1: Human Intestinal Viruses with High Potential as Food
Contaminants
Types of Viruses Example
Picornaviruses Polioviruses
Coxsackievirus A
Coxsackievirus B
Echovirus
Enterovirus
Reoviruses Reovirus
Rotavirus
Parvoviruses Human gastrointestional viruses
Papovaviruses Human BK and JC viruses
Adenoviruses Human adenoviruses
18
1.7.4 Important Bacterial Genera Introduction to Food
Microbiology
Bacterial classification is rapidly changing. In the following Table 1.2, only
those species and genera currently approved and listed in Bergey’s Manual have
been used.
Table 1.2 Genera of Bacteria Important in Food
Group Family Genera
Spiral and curved bacteria Spirallaceae Campylobacter
Gram-negative aerobic Pseudomonadaceae Pseudomonas, Altermonas,

rods and cocci Gluconobacter, Xanthomonas
Halobacteriaceae Halobacterium, Halococcus
Genera of uncertain affinity Alcaligenes, Acetobacter,

Brucella
Gram-negative facultative Enterobacteriaceae Escherichia, Citrobacter,

anaerobic rods Salmonella, Shigella,
Klebsiella, Enterobacter,
Serratia, Proteus, Yersinia,
Erwinia
Vibrionaceae Vibrio, Aeromonas
Genera of uncertain affinity Flavobacterium,

Chromobacterium
Gram-negative diplococci Neisseriaceae Moraxella, Acinetobacter

and diplococcobacilli Micrococcaceae Micrococcus, Staphylococcus
Gram-positive cocci Streptococcaceae Streptococcus, Leuconostoc,
Pediococcus, Lactococcus,
Enterococcus
Peptococcaceae Sarcina
Endospore forming rods Bacillaceae Clostridium, Bacillus

and cocci
Gram-positive Lactobacillaceae Lactobacillus

asporogenous rod of
regular shape
Genera of uncertain affinity Listeria
Non spore-forming
rods of irregular shape Coryneform bacteria Arthrobacter,
Brevibacterium,
Propionibacterium
Rickettsia Rickettsiaceae Coxiella
1.7.4.1 Common Bacterial Groups in Foods

Among the Micro-organisms found in foods, bacteria constitute a major
important group. This is not only because many different species can be
present in foods, but is also due to their rapid growth rate, ability to utilize
food nutrients, and their ability to grow under a wide range of temperatures,
aerobiosis, pH, and water activity, as well as to survive under adverse situations,
such as survival of spores at high temperature. For convenience, bacteria
important in foods have been arbitrarily divided into several groups on the 19
Fundamentals of Food basis of similarities in certain characteristics. This grouping does not have any
Microbiology
taxonomic significance. Some of these groups and their importance in foods
are listed here.
1) Lactic Acid Bacteria
Those bacteria that produce relatively large quantities of lactic acid from
carbohydrates. Include species mainly from genera Lactococcus,
Leuconostoc, Pediococcus, Lactobacillus and Streptococcus
thermophilus.
2) Acetic Acid Bacteria
Those bacteria that produce acetic acid, such as Acetobacter aceti.
3) Propionic Acid Bacteria
Those bacteria that produce propionic acid and are used in dairy
fermentation. Include species such as Propionibacterium freudenreichii.
4) Butyric Acid Bacteria
Those bacteria that produce butyric acid in relatively large amounts.
Some Clostridium spp., such as Clostridium butyricum.
5) Proteolytic Bacteria
Those bacteria that are capable of hydrolyzing proteins due to production
of extracellular proteinases. Species in genera Micrococcus,
Staphylocccus, Bacillus, Clostridium, Pseudomonas, Alteromonas,
Flavobacerium, and Alcaligenes; some in Enterobacteriaceae and
Brevibacterium are also included in this group.
6) Lipolytic Bacteria
Able to hydrolyze triglycerides due to production of extracellular lipases.
Species in genera Micrococcus, Staphylococcus, Serration,
Pseudomonas, Alteromonas, Alcaligenes and Flavobacterium are
included in this group.
7) Saccharolytic Bacteria
Able to hydrolyze complex carbohydrates. Include some species in genera
Bacillus, Clostridium, Aeromonas, Pseudomonas, and Enterobacter.
8) Thermophillic Bacteria
Able to grow at 500C and above. Include some species from genera
Bacillus, Clostridium, Pediococcus, Streptococcus, and Lactobacillus.
9) Psychrotrophic Bacteria
Able to grow at refrigerated temperature (<50C). Include some species
of Pseudomonas, Alteromonas, Alcaligenes, Flavobacterium, Serratia,
Bacillus, Clostridium, Lactobacillus, Leuconostoc, Listeria, Yersinia
and Aeromonas.
10) Thermoduric Bacteria
Able to survive pasteurization temperature. Include some species of
Micrococcus, Enterococcus, Lactobacillus, Pediococcus, Bacillus
20
(spores) and Clostridium (spores).
11) Halotolerant Bacteria Introduction to Food
Microbiology
Able to survive high salt concentrations (>10%). Include some species of
Bacillus, Micrococcus, Staphylococcus, Pediococcus, Vibrio
Streptococcus, Clostridium and Corynebacterium.
12) Aciduric Bacteria
Able to survive at low pH (below 4.0). Include some species of
Lactobacillus, Pediococcus, Lactococcus, Enterococcus and
Streptococcus.
13) Osmophilic Bacteria
Can grow at a relatively higher osmotic pressure (environment) than
other bacteria. Some species from genera Staphylococcus, Leuconostoc,
and Lactobacillus are included in this group. They are much less
osmophilic than yeasts and molds.
14) Gas-producing Bacteria
Produce gas (CO2, H 2, H 2S) during metabolism of nutrients. Include
spices from genera Leuconostoc, Lactobacillus, Brevibacterium and
Escherichia.
15) Slime Producers
Produce slime due to synthesis of polysaccharides. Include some species
or strains of Xanthomonas, Leuconostoc, Alcaligenes, Enterobacter,
Lactococcus, and Lactobacillus.
16) Spore formers
Ability to produce spore. Include Bacillus, Clostridium and
Desulfotomaculum spp. They are again divided into aerobic, anaerobic,
flat sour thermophilic and sulfide-producing spore formers.
17) Aerobes
Require oxygen for growth and multiplication. Species of Pseudomonas,
Bacillus, and Flavobacterium are included in this group.
18) Anaerobes
Cannot grow in the presence of oxygen. Include species of Clostridium.
19) Facultative Anaerobes
Able to grow both in the presence and absence of oxygen. Lactobacillus,
Pediococcus, Leuconostoc, enteric pathogens, some species of Bacillus,
Serratia, and coliforms are included in this group.
20) Coliforms
Include mainly species from Escherichia, Enterobacter, Citrobacter,
and Klebsiella, and used as index of sanitation.
21) Fecal Coliforms
Include mainly Escherichia coli. Also used as index of sanitation. 21
Fundamentals of Food 22) Enteric Pathogens
Microbiology
Include pathogenic Salmonella, Shigella, Campylobacter, Yersinia,
Escherichia, Vibrio, Listeria, Hepatitis A, and others that can cause
gastrointestinal infection.
1.8 NORMAL MICRO FLORA OF SOME

COMMON FOODS
Under normal conditions a food generally harbours only a few types of Micro-
organisms. They constitute those that are naturally present in raw foods (which
provide the ecological niche) and those that get in from outside sources to
which the foods are exposed from the time of production until consumption.
The predominant type(s) will be the ones for which the optimum growth
condition is present. The normal microflora of different food groups are listed
below.
1.8.1 Meat
The carcass of a healthy animal slaughtered for meat and held in a refrigerated
room is likely to have only nominal surface contamination while the inner
tissues are sterile. Fresh meat cut from the chilled carcass has its surface
contaminated with micro organisms characteristic of the environment and the
implements (saws or knives) used to cut the meat. Each new surface of meat,
resulting from a new cut, adds more micro organisms to the exposed tissue.
Among the most common species of bacteria occurring on fresh meats are
Pseudomonads, Staphylococci, Micrococci, Enterococci and Coliforms. The
low temperature at which fresh meats are held favors the growth of psychrophilic
Micro-organisms.
1.8.2 Poultry
Freshly dressed eviscerated poultry have a bacterial flora on their surface
(skin) that originates from the bacteria normally present on the live birds and
from the manipulations during killing, defeathering, and evisceration. Under
good sanitary conditions the bacterial count has been reported to be from
100 to 1000 bacteria per square centimeter of skin surface, whereas under
less sanitary conditions the count may increase 100-fold or more.
Pseudomonads constitute the major contaminants on the skin of freshly dressed
poultry.
1.8.3 Eggs
The interior of a freshly laid egg is usually free of micro organisms; its
subsequent microbial content is determined by the sanitary conditions under
which it is held, as well as the conditions of storage, i.e. temperature and
humidity. Micro-organisms particularly bacteria and molds, may enter the egg
through cracks in the shells or penetrate the shells when the “bloom” (thin
protein coat) covering the shell deteriorates. The type of micro organisms
involved reflect those present in the environment.
1.8.4 Fruits and Vegetables
They are normally susceptible to infection by bacteria, fungi, and viruses.
22 Microbial invasion of plant tissue can occur during various stages of fruit and
vegetable development, and the likelihood of spoilage increases. A second Introduction to Food
Microbiology
factor contributing to the microbial contamination of fruits and vegetables
pertains to their post-harvest handling. Mechanical handling is likely to produce
breaks in the tissue which facilitates invasion by micro organisms. The pH of
fruits is relatively acid i.e. ranging from 2.3 (for lemons) to 5.2 (for bananas).
This restricts bacterial growth but does not retard fungal growth. The pH
range for vegetables is slightly higher pH 5.0 to 7.0 and hence they are more
susceptible than fruits to bacterial attack.
1.8.5 Shellfish and Finfish

The microbial flora of freshly caught oysters, clams, fish, and other aquatic
specimens is very largely a reflection of the microbial quality of the water
from where they are harvested. Of particular significance is whether the water
is sewage-polluted, in which case the aquatic food is potentially capable of
transmitting various pathogenic Micro-organisms. Shellfish that grow in
contaminated water can concentrate viruses and may be the source of Hepatitis
infection. For example, raw oysters and clams from polluted waters have
caused numerous epidemics in various parts of the world.
1.8.6 Milk
Milk is an excellent growth medium for all of the common spoilage organisms,
including molds and yeasts. Fresh, non pasteurized milk generally contains
varying numbers of Micro-organisms, depending on the care employed in
milking, cleaning, and handling of milk utensils. Raw milk held at refrigerator
temperatures for several days invariably shows the presence of several or all
bacteria of the following genera: Enterococcus, Lactococcus, Streptococcus,
Leuconostoc, Lactobacillus, Microbacterium, Propionibacterium,
Micrococcus, Coliforms, Proteus, Pseudomonas, Bacillus, and others. Those
unable to grow at the usual low temperature of storage tend to be present in
very low numbers. The pasteurization process eliminates all but thermoduric
strains, primarily, Streptococci and Lactobacilli, and spore formers of the
genus Bacillus (and clostridia if present in raw milk). The spoilage of pasteurized
milk is caused by the growth of heat-resistant Streptococci utilizing lactose to
produce lactic acid, which depresses the pH to a point (about pH 4.5) where
curdling takes place.

1) Give the scientific names of the following :
i) Baker’s Yeast .......................................................
ii) Bread Mold .......................................................
iii) Cocci in bunches .......................................................
iv) Aerobic spore former .......................................................
v) Anaerobic spore former .......................................................
23
Fundamentals of Food 2) Define coliforms.
Microbiology
............................................................................................................
............................................................................................................
............................................................................................................
3) What is the main contaminant in the chilled and frozen meats?
............................................................................................................
............................................................................................................
............................................................................................................
1.9 LET US SUM UP

This unit outlines briefly the definition and applied aspects of science of
microbiology. It highlights the scope of food microbiology. The contents of the
unit emphasize on the types and importance of Micro-organisms in foods. The
types of Micro-organisms (i.e. bacteria, fungi, molds and yeasts), viruses and
parasitic organisms are discussed in terms of occurrence, morphology and
their significance in food. Their role in food spoilage, food production, food
preservation and food borne diseases has been discussed. Since micro-
organisms are associated with most foods we eat, the normal micro flora of
some common food groups is also mentioned. Thus, the entire unit deals with
predominant Micro-organisms in food and highlights their harmful as well as
beneficial role in foods.
1.10 KEY WORDS

Aerobe : An organism that grows in the presence of atmospheric
oxygen.
Bacillus : A rod-shaped bacterium.
Bacteriocin : A protein produced by a bacterial strain that kills other
closely related strains.
Binary Fission : Asexual mode of reproduction in which a cell or an
organism separates into two identical cells.
Binomial system : The nomenclature system in which an organism is given
two names; the first is the capitalized generic name, and
the second is the uncapitalized specific epithet.
Endospore : An extremly heat- and chemical-resistant, dormant,
thick-walled spore (resting structure) that develops within
bacterial cell.
Halophile : A microorganism that requires high levels of sodium
chloride for growth.
Hepatitis : Any infection that results in inflammation of the liver.
Also refers to liver inflammation as such.
24
Microbiology : The study of organisms that are usually too small to be Introduction to Food
Microbiology
seen with the naked eye. Special techniques are required
to isolate and grow them.
Prion : An infectious particle that is the cause of slow diseases
like scrapie in sheep and goat; it has a protein
component, but no nucleic acid has yet been detected.
Prokaryotic Cells : Cells that lack a true, membrane-enclosed nucleus;
bacteria are prokaryotic and have their genetic material
located in a nucleoid.
Taxon : A group into which related organisms are put together
(classified).
Taxonomy : The science of biological classification; it consists of
three parts: classification, nomenclature, and
identification.
Vibrio : A rod-shaped bacterial cell that is curved to form a
comma or an incomplete spiral.
Virion : A complete virus particle that represents the extracellular
phase of the virus life cycle; at the simplest, it consists
of a protein capsid surrounding a single nucleic acid
molecule.
Viroid : An infectious agent of plants that is a single-stranded
RNA not associated with any-protein; the RNA does
not code for any protein and is not translated.
1.11 ANSWERS TO CHECK YOUR PROGRESS

EXERCISES
Your answer should include the following points.
1) Micro-organisms are living entities of microscopic size which cannot be
seen by naked eye and include bacteria, viruses, yeasts and molds
(together designated as fungi), algae, and protozoa.
2) Food microbiology is the branch of microbiology that studies the role of
Micro-organisms in food spoilage, food production and food borne
diseases.
3) Probiotic is the oral administration of living Micro-organisms to promote
the health and growth of an animal or human.
1) The scientific study of organisms with the ultimate objective being to
characterize and arrange them in an orderly manner is known as
systematics.
25
Fundamentals of Food 2) The two most common Micro-organisms in foods are bacteria and fungi.
Microbiology
3) On basis of morphology bacteria can be classified as:

a) Bacilli: rod-shaped
b) Cocci: spherical
c) Vibrio: comma-shaped
d) Spirillum: spiral shaped
4) Bacteriophage is a virus capable of eating host bacteria.
1) i) Saccharomyces cerevisiae
ii) Rhizopus stolonifer
iii) Staphylococcous aureus
iv) Bacillus spp.
v) Clostridium spp.
2) Coliforms are a group of gram-negative, nonsporing, facultative rods that
ferment lactose into acid along with gas formation.
3) Pseudomonads
1.12 SUGGESTED READING

Banwart, G.J. (1979), Basic Food Microbiology, AVI Publishing Co. Inc.,
Westport, Connecticut.
Frazier, W.C. and Westoff, D.C. (1996), Food Microbiology, Tata McGraw
Hill Publishing Co. Ltd., New Delhi.
Pelczar, M. Jr.; Chan, E.C.S. and Kreig, N.R. (1993), Microbiology, Tata
McGraw Hill Inc., New York.
Garbutt, J. (1998), Essentials of Food Microbiology, Arnold International
Student’s Edition, London.
26
Food Contamination
UNIT 2 FOOD CONTAMINATION AND and Spoilage
SPOILAGE
Structure
2.0 Objectives
2.1 Introduction
2.2 Food Contamination

2.2.1 Contamination of Living Plants and Animals
2.2.2 Sources of Contamination
2.3 Food Spoilage

2.3.1 Types of Spoilage
2.3.2 Classification of Foods on the Basis of Stability
2.4 Role of Micro-organisms

2.4.1 Micro-organisms Involved in Spoilage
2.4.2 Growth of Micro-organisms
2.5 Factors Affecting Spoilage

2.5.1 Extrinsic Factors
2.5.2 Intrinsic Factors
2.5.3 Interaction Among Growth Factors
2.6 Deteriorative Effect of Micro-organisms

2.6.1 Physical Changes
2.6.2 Chemical Changes
2.7 Different Types of Spoilage

2.8 Common Methods of Food Preservation
2.9 Let Us Sum Up
2.10 Key Words
2.0 OBJECTIVES
After reading this Unit, we shall be able to:
• identify sources of food contamination;
• specify food spoilage – its types, causative agents and changes
associated with it;
• enumerate factors affecting the rate of spoilage; and
• explain principles and methods of food preservation.
27
Microbiology 2.1 INTRODUCTION
Most foods are excellent media for rapid growth of micro-organisms. There is
abundant organic matter in foods, their water content usually sufficient, and
the pH is either neutral or slightly acidic.
Foods consumed by man and animals are ideal ecosystems in which bacteria
and fungi can multiply. The mere presence of micro-organisms in foods in
small numbers however, need not be harmful, but their unrestricted growth
may render the food unfit for consumption and can result in spoilage or
deterioration. There are many opportunities for food to become contaminated
as it is produced and prepared. Many food borne microbes are present in
healthy animals (usually in their intestines) raised for food. Meat and poultry
carcasses can become contaminated during slaughter by contact with small
amounts of intestinal contents. Similarly, fresh fruits and vegetables can be
contaminated if they are washed or irrigated with water that is contaminated
with animal manure or human sewage. Some types of Salmonella can infect
a hen’s ovary so that the internal contents of a normal looking egg can be
contaminated with Salmonella even before the shell in formed. Oysters and
other filter feeding shellfish can concentrate Vibrio bacteria that are naturally
present in sea water, or other microbes that are present in human sewage
dumped into the sea.
Later in food processing, other food borne microbes can be introduced from
infected humans who handle the food, or by cross contamination from some
other raw agricultural products. For example, Shigella bacteria, hepatitis A
virus and Norwalk virus can be introduced by the unwashed hands of food
handlers who are themselves infected. In the kitchen, microbes can be
transferred from one food to another food by using the same knife, cutting
board or other utensil to prepare both without washing the surface or utensil
in between. A food that is fully cooked can become recontaminated if it
touches other raw foods or drippings from raw foods that might contain
pathogens microbes responsible for spoilage.
The way in which food is handled after it is contaminated can also make a
difference in whether or not an outbreak occurs. Many bacteri need to
multiply to a large number before enough are present in food to cause disease.
Given warm moist conditions and an ample supply of nutrients, a bacterium
that reproduces by dividing itself every half hour can produce 17 million
progeny in 12 hours. As a result, lightly contaminated food left out overnight
can be highly infectious by the next day. If the food is refrigerated promptly,
the bacteria multiply at a slower rate. In general, refrigeration or freezing
prevents virtually all bacteria from growing and multiplying but generally
preserves them in a state of suspended animation. This general rule has a few
surprising exceptions. Two food borne bacteria, Listeria monocytogenes
and Yersinia enterocolitica can actually grow at refrigerator temperatures.
2.2 FOOD CONTAMINATION

Food contamination is the introduction or occurrence of a contaminant in
food. A contaminant is any biological or chemical agent, foreign matter, or
28 other substance unintentionally added to food that may compromise food
safety or suitability. Among these contaminants are biological, chemical or Food Contamination
and Spoilage
physical agents in, or condition of, food with the potential to cause an adverse
health effect.
The contamination of food by chemicals is a worldwide public health concern
and is a leading cause of trade problems internationally. Contamination may
occur through environmental pollution, as in the case of toxic heavy metals,
Poly Chlorinated Biphenyl (PCBs) and dioxins, or through the intentional use
of chemicals, such as pesticides, animal drugs and other agrochemicals. Food
additives and contaminants resulting from food manufacturing and processing
can also adversely affect health. When foods are contaminated with unsafe
levels of pathogens, chemical contaminants, or metals, they can pose substantial
health risk to consumers and place severe economic burden on individual
communities or nation.
Cross-contamination of food is a common factor in the cause of food borne
diseases. Cross-contamination is the contamination of a food product from
another contaminated source. Foods can become contaminated by micro-
organisms (bacteria and viruses) from many different sources during the food
preparation and storage process. There are three main ways cross-
contamination can occur:
• Food to food
• People to food
• Equipment to food
2.2.1 Contamination of living plants and animals
The internal tissues of healthy plants and animals are essentially sterile including
in the case of animals body fluids such as blood. Plants have a natural micro
flora associated with the surfaces of root, stem, leaves, flowers and fruits.
Invasion of healthy tissues and subsequent growth of micro-organisms is
prevented by:
• Outer mechanical barriers, e.g. epidermis with an outer waxy layer, and
outer corky layers;
• Internal chemical constituents that are anti-microbial, e.g. tannins, organic
acids and essential oils;
• Inert cell walls welded into tissues that are difficult to penetrate;
• Active cells with intact membranes.
Plant materials are harvested in the living state and, as long as the mechanical
barriers remain intact, can remain in storage at low temperature for several
months without spoilage.
Animals have a natural micro flora associated with the skin, the gut content
and external openings, e.g. the mouth. Lymph nodes and liver may also be
contaminated with invading micro-organisms. Invasion of healthy tissues and
subsequent growth of micro-organisms is prevented by:
• Epithelial barriers e.g. stratified skin epithelium (epidermis) and intestinal
mucosa;
• The immune system consisting of the lymphatic system, white blood
corpuscles and antibodies; 29
Fundamentals of Food • Active cells with intact membranes;
Microbiology
• Presence of natural antimicrobials, e.g. lysozyme in tears, saliva and egg
white;
• Voiding mechanisms such as vomiting.
Once an animal or plant is dead the activity of the majority of factors (defense
mechanisms) that prevent microbial invasion of tissues by micro-organisms
ceases and invasion is only temporarily hindered by mechanical barriers such
as stratified epithelium or plant epidermis. Cell membranes are no longer
active and leak cell contents, providing nutrients for microbial growth.
2.2.2 Sources of Food Contamination
It helps to understand at which point our food might become contaminated, as
this will provide us with a better impetus for taking personal responsibility to
reduce the potential for further contamination. There are five main events that
can cause food contamination:
i) Food production: The use of chemicals, fertilizers, manures etc. all have
the potential to contaminate food as it is being grown.
ii) Environmental factors: Bacteria, parasites, fungal sporus etc. travel in
the wind, float in the water, hitch lifts with dust and reside snugly in the
soil. They are a part of nature’s web of life and will always be a possible
source of contamination if not dealt with appropriately as part of a
consistent and dedicated approach to food hygiene.
iii) Food processing: Whether in a large factory or in your own kitchen,
food processing can be a major source of contamination. Areas used for
processing need to be kept scrupulously clean or cross-contamination
can easily occur, especially with meat products (natural bacteria residing
in the intestines of animals are a major source of cross-contamination
when mishandled).
Aerosols
Air
Spoiled Water
foods
Processing Soil
Equipment Dust
Raw material
with natural
micro flora
Diseased plants Sewage
and animals
Packing Humans
Materials
Pests
Faeces
30 Fig. 2.1: Sources of contamination of food
iv) Food storage: Food that is stored incorrectly, for instance an uncooked Food Contamination
and Spoilage
chicken thigh resting next to a bunch of grapes, can be a source of
transferring bacteria and other contaminants from one food to another.
v) Food preparation: A great deal of food contamination occurs during the
preparation stage. A sick person can pass on germs, ranging from flu to
gastroenteritis. A chopping board used for meat that is not washed and
then used for vegetables is another source of possible contamination.
Unwashed hands, dirty kitchen spaces, insects and rodents in the kitchen
etc. are all possible sources of food contamination.
2.3 FOOD SPOILAGE

Spoilage of food may be due to chemical or biological causes; the latter
include action of inherent enzymes, growth of micro-organisms, invasion by
insects, and contamination with trichinae and worms. About one-fourth of the
world’s food supply is lost through the action of micro-organisms alone.
The “spoilage” concept includes concepts about edibility, means the food is
unfit to eat or fit to eat. Spoilage is decomposition. Many foods may not be
decomposed, but harbor certain kinds of bacteria, or their toxins, in number
or amounts which make the food poisonous and thus unfit for human
consumption.
The criteria for assurance in foods suitable for consumption are:
• The desired stage of development or maturity of the food.
• Freedom from pollution at any stage in the production and subsequent

handling of the food.
• Freedom from objectionable, chemical and physical changes resulting

from action of food enzymes; activity of microbes, insects, rodents, invasion
of parasites; and damage from pressure, freezing, heating, drying, and the
like.
• Freedom from micro-organisms and parasites causing food borne illnesses.
Enzymatic and microbial activities are undesirable when they are unwanted or
uncontrolled. An example is the souring of milk; if unwanted, it is spoilage, yet
the same process is purposely used in the production of certain cheeses and
other fermented products made from milk.
2.3.1 Types of Spoilage

The food may become unacceptable due to the following factors:
1) Growth and activities of micro-organisms principally bacteria, yeasts and

moulds (This is by far the most important and common cause of
food spoilage).
2) Activities of food enzymes (enzymatic browning is a common example).
3) Infestation by insects, parasites and rodents.
4) Chemical changes in a food (i.e. not catalyzed by enzymes of the tissues 31

Fundamentals of Food or of micro-organisms). For example the chemical oxidation of fats
Microbiology
producing rancidity as well as non-enzymatic browning reactions in foods
like Maillard Browning.
5) Physical changes or damages such as those caused by freezing (freezer
burn), by drying (caking) etc.
6) Presence of foreign bodies.
2.3.2 Classification of Foods on the Basis of Stability
Foods are frequently classified on the basis of their stability as nonperishable,
semi perishable, and perishable. An example of the first group is sugar. Few
foods are truly nonperishable. Hermetically sealed, heat-processed, and
sterilized (canned) foods are usually listed among the nonperishable items.
For all intents and purposes, they belong there. However, canned food may
become perishable under certain circumstances, when, by accident, there is a
chance for recontamination following processing because of faulty seams of
the cans, or through rusting or other such damage so that the can is no longer
hermetically sealed.
Classified as semi perishables are usually the dry goods, such as flour, dry
legumes, baked goods, hard cheeses, dried fruits and vegetables, and even
waxed vegetables. Frozen foods, though basically perishable, may be classified
as semi perishables provided they are freezer-stored properly.
The majority of our food materials must be classified as perishables. This

group includes meat, poultry, fish, milk, eggs, many fruits and vegetables, and
all cooked or “made” food items, except the dry and very acid ones.
2.4 ROLE OF MICRO-ORGANISMS

Microbial spoilage of foods is the beginning of the complex natural process of
decay that under natural circumstances leads to recycling of the elements
present in the animal or plant tissues in the natural environment.
2.4.1 Micro-organisms involved in Spoilage

Micro-organisms which may cause food to spoil include molds, yeasts, and
bacteria. The contamination with molds, as a rule, is easily detected because
of the presence of furry hyphae or threadlike structures which, in many
instances, are colored. They often contribute a musty odor and flavor to the
food they invade. Some molds, because of toxins they produce, are not
altogether harmless. Semi moist foods or foods with low water activity having
been partially dehydrated, and where the remaining water is sufficiently bound
to hold the growth of bacteria are ideal for contamination by molds and
yeasts.
Yeasts are unicellular organisms of small sizes which multiply by budding. In

general, sugars are the best food source for energy for yeast; Carbon dioxide
and alcohol are the end products of the fermentation mediated by yeats.
Spoilage due to yeast may usually be recognized by the presence of bubbles
and an alcoholic smell and taste.
32
Bacteria spoil food in many ways and it is not always possible to recognize Food Contamination
and Spoilage
the spoilage by sight, smell, or taste. Unfortunately, some of the bacteria that
are important from a public health point of view may multiply to dangerously
high numbers in food without changing the appearance, odor, or taste of the
food. Disease-producing food has usually no decomposed appearance, but is
certainly unfit for human consumption, and must be considered to be spoiled.
It is an important fact that almost any food will spoil if it is moist and not kept
frozen. Spoilage must be expected within a wide temperature range. The
various types of micro-organisms as well as the genera, species, and strains
vary in their temperature and food requirements. Thus the bacterial flora of a
spoiled food item will vary greatly.
The origin of micro-organisms also varies. The micro-organisms may include
the original flora of the particular food, as well as contaminants added during
handling, processing, transporting, storing, preparing, and serving.
2.4.2 Growth of Micro-organisms

The multiplication of spoilage organisms on or in the food materials depends
on many factors – the type of organism involved, its ability to gain nourishment
from the food, competition from other micro-organisms, initial load, and
environmental conditions. Micro-organisms grow rapidly; we call it logarithmic
growth. The time a bacterium takes to multiply (double its number) is known
as its generation time.
Resting Phase
Logof Numbers of Bacteria
Growth
(Log)
Lag Death Phase

Phase
Time
Fig. 2.1: A typical bacterial growth curve
Four distinct phases occur in the growth curve: lag; log or growth phase;
stationary phase and death phase (Fig. 2.1). Bacteria need about four hours
to adapt to a new environment before they begin rapid growth. In handling
food, this means we have less than four hours to make a decision to either
cool the food, heat it or eat it.
As micro-organisms grow, they tend to form colonies. These colonies are
made up of millions of individual cells. Once a colony forms, the food available
to each cell is limited and excretions from these millions of cells become toxic
to a microbe. This is the stationary phase. Some of the cells now begin to die.
If we can control bacterial growth, we can control the major cause of food
spoilage.
33
Fundamentals of Food The multiplication of spoilage organisms on or in the food materials depends
Microbiology
on many factors – the type of organism involved, its ability to gain nourishment
from the food, competition from other micro-organisms, initial load, and
environmental conditions.

1) What are spoilage bacteria?
.............................................................................................................
.............................................................................................................
.............................................................................................................
2) Classify foods on the basis of their stability.
.............................................................................................................
.............................................................................................................
.............................................................................................................
3) What are the types of food spoilage?
.............................................................................................................
.............................................................................................................
.............................................................................................................
4) What causes food spoilage?
.............................................................................................................
.............................................................................................................
.............................................................................................................
2.5 FACTORS AFFECTING SPOILAGE
Food is a chemically complex matrix, and predicting whether, or how fast,

micro-organisms will grow in any given food is difficult. Most foods contain
sufficient nutrients to support microbial growth. Several factors encourage,
prevent or limit the growth of micro-organisms in foods; the most important
are water activity aw, pH and temperature.
Factors affecting microbial growth are divided into two groups -intrinsic and
extrinsic parameters (Table 2.1). These factors affect the growth of micro-
organisms on foods. When spoilage of a food occurs under a given set of
circumstances, not all of the different types of organisms contaminating a food
are associated with the spoilage process. In fact, the spoilage flora always is
34
dominated by just a few and sometimes only one organism. Components of Food Contamination
and Spoilage
the micro flora compete with one another for the available nutrients and the
organism(s) with fastest growth under a particular set of circumstances will
become dominant and give rise to the spoilage symptoms.
Table 2.1: Intrinsic and Extrinsic Parameters Affecting Microbial Growth
Intrinsic parameters Extrinsic parameters
Water activity, humectant identity Temperature
Oxygen availability Relative humidity
pH, acidity, acidulant identity Atmospheric composition
Buffering capacity Packaging
Available nutrients
Natural antimicrobial substances
Presence and identity of natural microbial flora
Colloidal form
The component of the micro flora which becomes dominant is determined by

a complex interaction between the components of the contaminating micro
flora (implicit factors), the storage environment (extrinsic factors) and the
physico-chemical properties of the food (intrinsic factors).
All of the factors that influence the growth of micro-organisms have been
dealt with as under. Time is included because, under any given set of
circumstances, spoilage takes a finite period to occur and equates with the
storage life of a product.
A knowledge of the intrinsic and extrinsic parameters should enable you to

decide which broad group of organisms is likely to spoil a particular type of
food, i.e. whether the food is likely to be spoiled by bacteria, yeasts or
moulds. For example, foods that have a high water activity and a pH above
5.0 are likely to be spoiled by bacteria simply because under these conditions
bacteria grow the fastest. Foods with pH below 4.2 are likely to be spoiled
by yeasts and moulds even when the water activity is high.
2.5.1 Extrinsic Factors

Extrinsic factors relate to the environmental factors that affect the growth rate
of micro-organisms. They are as follows:
A. Temperature
Microbes have an optimum temperature as well as minimum and maximum

temperatures for growth. Therefore, the environmental temperature determines
not only the proliferation rate but also the genera of micro-organisms that will 35
Fundamentals of Food thrive and the extent of microbial activity that occurs. For example, a change
Microbiology
of only a few degrees in temperature may favor the growth of entirely different
organisms and result in a different type of food spoilage and food poisoning.
These characteristics have been responsible for the use of temperature as a
method of controlling microbial activity.
The optimal temperature for the proliferation of most micro-organisms is from

150 to 400C. However, many genera of microbes are capable of growth from
00 to 150C and other even micro-organisms will grow at subzero temperatures.
Still other genera will grow at temperatures up to and exceeding 1000C.
Microbes classified according to temperature of optimal growth include:
• Thermophiles (high-temperature-loving micro-organisms), with growth

optima at temperatures above 450C (e.g., Bacillus stearothermophilus,
Bacillus coagulans, and Lactobacillus thermophilus).
• Mesophiles (medium-temperature-loving micro-organisms), with growth

optima between 200 and 450C (e.g., most Bactobacilli and Staphylococci).
• Psychrotrophs (cold-temperature-tolerant micro-organisms), which

tolerate and thrive at temperatures below 200C (e.g., Pseudomonas and
Acinetobacter).
Bacteria, molds and yeasts each have some genera with temperature optima
in the range characteristic of thermophiles, mesophiles, and psychrotrophs.
Molds and yeasts tend to be less thermophilic than bacteria. As temperature
approaches 00C, fewer micro-organisms can thrive and their proliferation is
slower. As temperature falls below approximately 50C, proliferation of spoilage
micro-organisms is retarded as the growth of nearly all pathogens ceases.
B. Oxygen Availability
As with temperature, the availability of oxygen determines which micro-
organisms will be active. Some micro-organisms have an absolute requirement
for oxygen, whereas others grow in total absence of oxygen. Yet other micro-
organisms can grow either with or without available oxygen. Micro-organisms
that require free oxygen are called aerobic micro-organisms (e.g.,
Pseudomonas spp.) and those that thrive in the absence of oxygen are called
anaerobic micro-organisms (e.g., Clostridium spp.). Micro-organisms that
can grow with or without the presence of free oxygen are called facultative
micro-organisms (e.g., Lactobacillus spp.).
C. Relative Humidity
This extrinsic factor affects microbial growth and can be affected by

temperature. All micro-organisms have high requirements for water to support
their growth and activity. A high relative humidity can cause moisture
condensation on food, equipment, walls, and ceilings. Condensation causes
moist surfaces, which are conducive to microbial growth and spoilage. Also,
microbial growth is inhibited by a low relative humidity. Micro-organisms
bacteria require the highest relative humidity of the various. Optimal relative
humidity for bacteria is 92% or higher, whereas yeasts need 90% or higher
and for molds, the value of relative humidity is 85-90%.
36
2.5.2 Intrinsic Factors Food Contamination
and Spoilage
Intrinsic factors that affect the rate of proliferation relate more to the
characteristics of the substrates (foodstuff or debris) that support or affect
growth of micro-organisms. These major intrinsic factors are:
A. Water Activity (a w )
Water is required by micro-organisms, and a reduction of water availability
constitutes a method of food preservation through reduction of microbial
proliferation. It is important to recognize that it is not the total amount of
moisture present that determines the limit of microbial growth, but the amount
of moisture which is readily available for metabolic activity of microbes.
The unit of measurement for water requirement of microorganism is
usually expressed as water activity (a w ). Water activity is defined as the
vapor pressure of the subject solution divided by the vapor pressure of the
pure solvent: aw = p/p0, where p is the vapor pressure of the solution and po is
the vapor pressure of pure water. The approximate optimal aw for the growth
of many micro-organisms is 0.99, and most microbes require an aw higher
than 0.91 for growth. The relationship between relative humidity (RH) and aw
is RH = aw × 100. Therefore an aw of 0.95 is equivalent to an RH of 95%.
Generally, bacteria have the highest water activity requirements of the micro-
organisms. Molds normally have the lowest aw requirements, with yeasts being
intermediate. Most spoilage bacteria do not grow at an aw below 0.91, but
molds and yeasts can grow at an aw of 0.80 or lower. Molds and yeasts are
more likely to grow in partially dehydrated surfaces (including food), whereas
bacterial growth is retarded.
Approximate Minimum (aw) Values for Growth
ORGANISMS GROUPS WATER ACTIVITY
MOST SPOILAGE BACTERIA 0.90
MOST SPOILAGE YEASTS 0.88
MOST SPOILAGE MOLDS 0.80
B. pH
The pH for optimal growth of most micro-organisms is near neutrality (7.0).

Yeasts can grow in an acid environment, but grow best in an intermediate acid
(4.0-4.5) range. Molds tolerate a wider range of pH (2.0-8.0), although their
growth is generally greater with an acid pH. Molds can thrive in a medium
that is too acid for either bacteria or yeasts. Bacterial growth is usually
favoured by near-neutral pH values. However, acidophilic (acid-loving) bacteria
will grow on food or debris down to a pH value of approximately 5.2. Below
pH 5.2, microbial growth is dramatically reduced when compared from growth
in the normal pH range (Fig. 2.2).
37
Microbiology
Fig. 2.2: pH and growth of micro-organisms
C. Oxidation – Reduction Potential
The oxidation – reduction (redox) potential is an indication of the oxidizing

and reducing power of the substrate. To attain optimal growth, some micro-
organisms require reduced conditions while others need oxidized conditions.
Thus, the importance of the oxidation-reduction potential is apparent. Aerobic
micro-organisms grow more readily under a high oxidation – reduction potential
(oxidizing reactivity). A low potential (reducing reactivity) favors the growth
of anaerobes. Facultative micro-organisms are capable of growth under either
condition. Micro-organisms can alter the oxidation-reduction potential of food
to the extent that the activity of other micro-organisms is restricted. For
example, anaerobes can decrease the oxidation – reduction potential to such
a low level that the growth of aerobes can be inhibited.
D. Nutrient Requirements
In addition to water and oxygen (except for anaerobes), micro-organisms

have other nutrient requirements. Most microbes need external sources of
nitrogen, energy (i.e., carbohydrates, proteins, or lipids), minerals, and vitamins
to support their growth. Nitrogen is normally obtained from amino acids and
other nonprotein nitrogen sources; however, some micro-organisms utilize
peptides and proteins. Molds are the most effective in the utilization of proteins,
complex carbohydrates, and lipids because they contain enzymes capable of
hydrolyzing these molecules into less complex components. Many bacteria
have a similar capability, but most yeasts require the simple forms of these
compounds. Minerals (micronutrients) are needed by all micro-organisms, but
requirements for vitamins vary. Molds and some bacteria can synthesize enough
B vitamins to fulfill their needs, while other micro-organisms require a ready-
made supply.
E. Inhibitory Substances
Microbial proliferation can be affected by the presence or absence of inhibitory

substances. Substances or agents that inhibit microbial activity are called
38
bacteriostats, and those which destroy micro-organisms are called bactericides. Food Contamination
and Spoilage
Some bacteriostatic substances are added during food processing (i.e., nitrites).
Most bactericides are utilized as a method of decontaminating foodstuffs or
as a sanitizer for cleaned equipment, utensils, and rooms.
2.5.3 Interaction Between Growth Factors

The effects that factors such as temperature, oxygen, pH, and aw have on
microbial activity may be dependent on each other. Micro-organisms generally
become more sensitive to oxygen availability, pH, and aw at temperatures near
growth minima or maxima. For example, bacteria may require a higher pH,
aw, and minimum temperature for growth under anaerobic conditions than
when aerobic conditions prevail. Micro-organisms that grow at lower
temperatures are usually aerobic and generally have a high aw requirement.
Lowering aw by adding salt or excluding oxygen from foods such as meat
which have been held at a refrigerated temperature dramatically reduces the
rate of microbial spoilage. Normally, some microbial growth occurs when any
one of the factors that control the growth rate is at a limiting level. Yet, if more
than one factor becomes limiting, microbial growth is drastically curtailed or
completely stopped.
2.6 DETERIORATIVE EFFECTS OF MICRO-

ORGANISMS
Food is considered spoiled when it becomes unfit for human consumption.
Spoilage is usually equated with the decomposition and putrefaction that result
from activity of micro-organisms. Some of the more common physical and
chemical changes due to the micro-organisms have been described under:
2.6.1 Physical Changes

The physical changes caused by micro-organisms usually are more apparent
than the chemical changes. Microbial spoilage usually results in an obvious
change in physical characteristics such as color, body, thickening, odor, and
flavor degradation. Food spoilage is normally classified as being either aerobic
or anaerobic, depending upon the spoilage conditions, including whether the
principal microorganism causing the spoilage were bacteria, molds, or yeasts.
Aerobic spoilage by bacteria and yeasts usually results in slime formation;

undesirable odors and flavors (taints); color changes; and rancid, tallowy, or
chalky flavors from the breakdown of lipids. Slime formation by certain species
of bacteria or yeasts depends upon environmental conditions, especially those
of temperature and aw. Colour changes can be pigment oxidation resulting in a
gray, brown, or green color.
Physical deterioration through aerobic spoilage by molds results in sticky

surface of many foods. A filamentous appearance frequently referred to as
“whiskers” can also occur as a result of physical deterioration. Discoloration
from molds can give surface colorations, such as creamy, black, or green.
Deterioration from molds can affect appearance of lipids of foods in a way
similar to that of bacteria and yeasts and will produce musty odors and
39
alcohol flavors.
Fundamentals of Food Aerobic spoilage of foods from molds is normally limited to the food surface,
Microbiology
where oxygen is available. Therefore, molded surfaces of foods such as meats
and cheeses can be trimmed off and the remainder is generally acceptable for
consumption. This is especially true for aged meats and cheeses. When these
surface molds are trimmed, surfaces underneath usually have limited microbial
growth. However, if extensive bacterial growth occurs on the surface,
penetration inside the food surface usually follows and toxins may be present.
Anaerobic spoilage occurs within the interior of food products or in sealed

containers, where oxygen is either absent or present in limited quantities.
Spoilage is caused by facultative and anaerobic bacteria and is expressed
through souring, putrefaction, or taint. Souring occurs from the accumulation
of organic acids during the bacterial enzymatic degradation of complex
molecules (carbohydrate). Also, proteolysis without putrefaction may
contribute to souring. Souring can be accompanied by the production of
various gases. Examples of souring are milk, round sour or ham sour, and
bone sour in meat. These meat sours, or taints, are caused by anaerobic
bacteria that may originally have been present in lymph nodes or bone joints,
or which might have gained entrance along the bones during storage and
processing.
2.6.2 Chemical Changes

Through the activity of endogenous hydrolytic enzymes that are present in
foodstuffs (and the action of enzymes that micro-organisms produce), proteins,
lipids, carbohydrates, and other complex molecules are degraded to smaller
and simpler compounds. Initially, the endogenous enzymes are responsible
for the degradation of complex molecules. As microbial load and activity
increase, degradation subsequently occurs. These enzymes hydrolyze the
complex molecules into simpler compounds that are subsequently utilized as
nutrient sources for supporting microbial growth and activity. Oxygen availability
determines the end products of microbial action. Availability of oxygen permits
hydrolysis of proteins into end products such as simple peptides and amino
acids. Under anaerobic conditions, proteins may be degraded to a variety of
sulfur-containing compounds, which are odorous and generally obnoxious.
The end products of non-protein nitrogenous compounds usually include
ammonia.
Other chemical changes include action of lipases secreted by micro-organisms

which hydrolyze triglycerides and phospholipids into glycerol and fatty acids.
Phospholipids are also hydrolyzed into nitrogenous bases and phosphorus.
Lipid oxidation is also accelerated by extensive lipolysis.
Most micro-organisms prefer carbohydrates to other compounds as a source

of energy. When available, carbohydrates are more readily utilized for energy.
Utilization of carbohydrates by micro-organisms results in a variety of end
products such as alcohols and organic acids. In many foods such as sausage
products and cultured diary products, microbial fermentation of sugar that has
been added yields organic acid (e.g., lactic acid) which contribute to their
distinct and unique flavors.
40
Food Contamination
2.7 DIFFERENT TYPES OF FOOD SPOILAGE and Spoilage
Food undergoes different types of spoilage depending on its composition as

shown in Table 2.2.
Table 2.2: Types of Food Spoilage
Food Type of Spoilage Microorganisms Involved
Bread Moldy Rhizopus nigricans

Penicillium spp
Aspergillus niger
Ropy Bacillus subtilis
Maple sap and Syrups Ropy Enterobacter aerogenes
Yeasty Saccharomyces spp

Zygosaccharomyces spp
Pink Micrococcus roseus

Moldy Aspergillus spp
Fresh fruits and Soft rot Rhizopus, Erwinia

vegetables Grey mold rot Botrytis
Black mold rot Aspergillus niger
Pickles, sauerkraut Film yeasts, Rhodotorula

Pink yeasts
Fresh meat Putrefaction Alcaligenes

Clostridium
Proteus vulgaris
Pseudomonas fluorescens
Cured meat Moldy Aspergillus

Rhizopus
Pencillium
Souring Pseudomonas
Micrococcus
Greening, slime Lactobacillus
Leuconostoc
Fish Discoloration Pseudomonas

Colourless rot Alcaligenes
Flavobacterium
Eggs Green rot Pseudomonas fluorescens

Colourless rot Pseudomonas alcaligenes
Black rot roteus
Concentrated Off flavour Lactobacillus

Orange juice Leuconostoc
Acetobacter
Poultry Slime, odour Pseudomonas

Alcaligenes
41
Microbiology Check Your Progress Exercise 2
1) How do bacteria spoil food?
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2) Define water activity.
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3) What is a growth curve?
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4) What is temperature danger zone?
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2.8 COMMON METHODS OF FOOD

PRESERVATION
Principles of Food Preservation include the following:
i) Prevention or delay of microbial decomposition.
z By keeping microbes out (asepsis).
z By removal of microbes (e.g. filtration).
z By reducing the rate of microbial growth (e.g. by low temperature,
drying, anaerobic conditions and chemical inhibitors).
z By killing microbes (e.g. by heat or radiation).
ii) Prevention or delay of self-decomposition of food
z By inactivation of food enzymes (e.g. blanching).
z By prevention of chemical reactions (e.g. by using antioxidants).
42
Table 2.3 Methods of food preservation Food Contamination
and Spoilage
Operation Intended effect
Cleaning, washing Reduces microbial load
Cold storage (below 8OC) Prevents the growth of most pathogenic

bacteria; slows the growth of spoilage microbes
Freezing (below – 10OC) Prevents growth of all microbes
Pasteurizing (60-80OC) Kills most non-sporing bacteria, yeast and

molds
Blanching (95-110OC) Kills surface vegetative bacteria, yeast and

molds
Canning (above 100OC) ‘Commercially sterilizes’ food; kills all

pathogenic bacteria
Drying Stops growth of all microbes when a w <0.60
Salting Stops growth of most microbes
Syruping (sugars) Halts growth when a w < 0.70
Acidifying Halts growth of most bacteria (effects depend

on acid type)
aw denotes water activity
Food preservation aims at preventing the microbial spoilage of food products

and the growth of the food borne pathogens. Thus, the two principal goals of
food preservation methods are: (i) increasing the shelf life of the food and
(ii) ensuring the safety for human consumption. There are a variety of food
preservation methods. Table 2.4 consolidates the methods and details are
given here.
2.8.1 Heat
Heat kills micro-organisms by inactivating their proteins and by changing the
physical and chemical properties of their proteins. The most common use of
heat is in the process of canning. The food product is washed, sorted, and
graded and then subjected to steam heat for three to five minutes. This last
process called blanching, destroys many enzymes in the food product and
prevents further cellular metabolism.
The food is then peeled and cored, and diseased portions are removed. For
canning, containers are evacuated and placed in a pressurized steam sterilizer,
similar to an autoclave at 121°C. This removes especially Bacillus and
Clostridium spores.
If canning is defective, foods may become contaminated by anaerobic; bacteria
which produce gas. These are species of Clostridium, and coliform bacteria
(a group of Gram-negative nonspore-forming rods which ferment lactose to
acid and gas at 32°C in 48 hours). 43
Microbiology
2.8.2 Low Temperature
Exposure of micro-organisms to low temperatures reduces their rates of growth
and reproduction. This principle is used in refrigeration and freezing. Microbes
are however not killed. In refrigerators at 5°C, food remains unspoiled. In a
freezer at -5°C the crystals formed tear, shred and thereby may kill micro-
organisms.
However, some are able to survive. Salmonella spp. and Streptococci survive
freezing. For these types rapid thawing and cooking is necessary. Deep freezing
at -60°C forms smaller crystals. It reduces biochemical activities of microbes.
Blanching of fruits and vegetables, by scalding with hot water or steam prior
to deep freezing, inactivate plant enzymes that may produce toughness, change
in colour etc. A brief scalding prior to freezing also reduces the number of
micro-organisms on the food surface by up to 99% enhances the colour of
green vegetables.
2.8.3 Drying or Desiccation
Water from foods is removed in different ways. It may be done by spray
dryer which expels a fine mist of liquid such as coffee into a barrel cylinder
containing hot air. A heated drum may be used onto which liquids like soup
may be poured. Another example is a belt heater that exposes liquids like
milk to a steam of hot air that evaporates water and produces dried milk
solids.
A common process of freeze drying or lyophilization is used these days. The
food is deep frozen, after which the water is drawn off by a vacuum pump in
a machine. The dry product is then sealed in foil and is reconstituted with
water. This method is very useful for storing, transporting and preserving
bacterial cultures.
2.8.4 Osmotic Pressure
The principle of osmosis is applied in the age old traditional method of
adding Salt or Sugar to pickles, preserves and jams. Foods are preserved by
adding salts and sugars to them. These solutes remove the water out of
microbial cells causing them to shrink, thus stopping their metabolism. Jams,
jellies, fruit syrups, honey etc. are preserved by using high sugar concentration.
Fish, meat, beef and vegetable products are preserved with salt mediated
process.
2.8.5 Chemical Preservatives
The most commonly used preservatives are the acids, such as sorbic acid,
benzoic acid and propionic acid. These check mainly the growth of yeasts
and molds. Sorbic acid is used for preservation of syrups, salads, jellies and
some cakes. Benzoic acid is used for beverages, margarine, apple cider etc.
Propionic acid is an ingredient of bread and bakery products. Sulphur dioxide,
as gas or liquid is also used for dried fruits, molasses and juice concentrates.
Ethylene oxide is used for spices, nuts and dried fruits.
2.8.6 Radiation
Ultra Violet (UV) radiation is used in meat storage facilities which reduce
surface contamination, on meat products. Gamma rays are also used for some
44
meat products.
2.8.7 Anaerobiosis Food Contamination
and Spoilage
Packaging of food products under anaerobic conditions - anaerobiosis is
effective in preventing aerobic spoilage process. Vacuum packing in an airtight
container is used to eliminate air.
2.8.8 Controlled Atmosphere

Atmospheres containing 10% CO2 are used to preserve stored food products
such as apples and pears. This checks fungal growth. Ozone can also be
added.
2.8.9 Other Methods

These are asepsis i.e. washing utensils that come in contact with food and
filtration and centrifugation to remove microbes. Filtration is used for fruit
juices and other drinks. Bacteriological filters are used in industries.
2.9 LET US SUM UP

Food spoilage is caused by microbial growth in foodstuffs, chemical reactions
within the food itself or between the food and its environment, or the presence
of foreign material in the food. Food spoilage is a complex process and
Table 2.4: Preservation Methods and Effect on Micro-organisms
Effect on Preservation Factor Method of Achievement
Microorganisms
Reduction or Low temperature Chilled and frozen storage

inhibition of
growth
Low water activity Drying, curing and conserving
Restriction of nutrient Compartmentalization in water-in-

availability oil emulsions
Lowered Oxygen Vacuum and nitrogen packaging
Raised carbon dioxide Modified atmosphere packaging
Acidification Addition of acids; fermentation
Alcoholic fermentation Brewing; vinification; fortification
Use of preservatives Addition of preservatives:

inorganic (sulphite, nitrite);
organic (propionate, sorbate,
benzoate, parabens); bacteriocins
(nisin, natamycin)
Inactivation of Heating Pasteurization and sterilization

micro-organisms
Irradiating Ionizing irradiation
Pressurizing Application of high hydrostatic

pressure
45
Fundamentals of Food excessive amounts of foods are lost due to microbial spoilage even with
Microbiology
modern day preservation techniques. Despite the heterogeneity in raw materials
and processing conditions, the micro flora that develops during storage and
spoilage of foods can be predicted based on knowledge of the origin of the
food, the substrate base and a few central preservation parameters such as
temperature, atmosphere, aw and pH. Based on such knowledge, more detailed
sensory, chemical and microbiological analysis can be carried out on the
individual products to determine the actual specific spoilage organism.
Generally, foods carry a variety of organisms of which, most are saprophytic.
Their presence cannot be avoided since these are mostly from the environment
in which the food is prepared or processed. Also, their complete elimination is
difficult. However, it is possible to reduce their number or decrease their
activities by altering the environmental conditions. A knowledge of the factors
that either favour or inhibit their growth is therefore, essential in understanding
the principles of food spoilage and preservation.

1) What is food preservation?
.............................................................................................................
.............................................................................................................
.............................................................................................................
2) Food irradiation is ...........................................
3) What are the principles of food preservation?
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2.10 KEY WORDS

Additives : Natural and man-made substances added to a food
for an intended purpose (such as preservatives and
colours) or unintentionally (such as pesticides and
lubricants)
Adulteration : Deliberate contamination of foods with materials of
low quality.
Aerobic : Requires oxygen
Antimicrobials : Preservatives that protect food by slowing the
growth of bacteria, molds and yeasts.
Antioxidants : Preservatives that protect by preventing food
molecules from combining with oxygen (air).
Biological Hazard : Danger posed to food safety by the contamination
of food with pathogenic micro-organisms or
46 naturally occurring toxins.
Chemical Hazard : Danger posed to food safety by the contamination Food Contamination
and Spoilage
of food by chemical substances, such as pesticides,
detergents, additives, and toxic metals.
Exponential Phase : The period in which the cells of a defined bacterial

population are growing and dividing continuously.
Food Acids : Citric acid, tartaric acid, or malic acid used for
adjusting the pH value in food.
Food Contact Surface : Any surface of equipment, utensils, containers,

wrappings that come in direct contact with food.
Food Preservatives : Prevent spoilage either by slowing the growth of

organisms that live on food or by protecting the
food from oxygen.
Food Safety : Protecting the food supply from microbial, chemical

(i.e. rancidity, browning) and physical (i.e. drying
out, infestation) hazards or contamination that may
occur during all stages of food production and
handling-growing, harvesting, processing,
transporting, preparing, distributing and storing. The
goal of food safety monitoring is to keep food
wholesome.
Food Safety Hazards : Include all microbiological, chemical, and foreign

materials that, if consumed, could cause injury or
harm.
pH : Measure of the acidity or alkalinity of a solution,

defined as the - log10 of the hydrogen ion
concentration
Physical Hazard : Particles or fragments of items not supposed to be

in foods.
Spoilage Organisms : Micro-organisms naturally found within a food

source that cause food spoilage.
Spore : Inactive or dormant state of some rod-shaped

bacteria.
Stationary Phase : The stationary phase occurs at the maximum

population density, the point at which the maximum
number of bacterial cells can exist in an environment.
This typically represents the carrying capacity of
the environment.
Vegetative Cell : The vegetative cell state is the form in which an

organism is able to grow and divide continuously,
given favourable conditions. Unlike endospores,
vegetative cells are relatively poor at surviving
environmental stresses such as high temperature and
drying. 47
Microbiology 2.11 ANSWERS TO CHECK YOUR PROGRESS
EXERCISES
1) Spoilage bacteria are micro-organisms that cause food to deteriorate and
develop unpleasant odors, tastes, and textures.
2) Foods may also be classified on the basis of stability:
• Perishable foods such as meat and fish.
• Semi perishable foods such as potatoes.
• Stable foods such as cereals, flour and sugar.
(Any stable or semistable food becomes perishable food under moist
condition )
3) Types of spoilage in food:
Slime: Food becomes slimy when there are many bacteria
that they touch one another farming continuous film of
murcilage
Souring: Foods may go sour when the micro-organisms produce
acids, e.g. sour milk.
Discolouration: Food may become discoloured from microbiological
growth. Some molds have colored spores, e.g. black
mould on bread, or blue and green mold on citrus fruit.
Sometimes meat becomes green due to the growth of
micro-organisms
Gas: Bacteria often produces gas as a by product which
affects food, e.g.
- Meat becomes spongy in texture

- Packages, cans or vacuum packs will swell
Odour: Rotten smells develop from the breakdown of
components
4) Food spoilage can be caused by a combination of various factors such as
light, oxygen, heat, humidity and/or all kinds of micro-organisms.
1) There are different spoilage bacteria and each reproduces at specific
temperatures. Some can grow at the low temperatures in the refrigerator
or freezer. Others grow well at room temperature and in the “Danger
Zone.” Bacteria will grow anywhere if they have access to nutrients and
water. Under the right conditions, spoilage bacteria reproduce rapidly
and the populations can grow very large. In some cases, they can double
their numbers in as little as 20 minutes. The large number of micro-
organisms and their waste products cause objectionable changes in odor,
48 taste, and texture hence food spoilage.
2) Qualitatively, water activity / aw is a measure of unbound, free water in a Food Contamination
and Spoilage
system, available to support biological and chemical reactions. Water
activity affects micro-organisms survival and reproduction, enzymative
and chemical reactions. The water activity of a substance is quantitatively
equal to the vapor pressure of the substance divided by the vapor pressure
of pure water (both measured at the same temperature). Measurements
range from 0.00 (dry) to 1.00 (pure water).
3) Growth curve is a graph displaying the behavior of a bacterial population
over time.
4) Temperature danger zone are the temperatures between 5° and 60°C
(41° and 140°F) at which bacteria grow best.
1) Food preservation is the process used to slow or stop the progress of
spoilage by using heat treatment, sugar, salt, acid or preservatives.
2) Process of exposing food to radiation (rays of energy)
3) Asepsis is keeping out the micro-organisms.
4) Principles of Food Preservation are:
a) Prevention or delay of microbial decomposition.
• By keeping microbes out (asepsis);
• By removal of microbes (e.g. filtration);
• By reducing the rate of microbial growth (e.g. by low
temperature, drying, anaerobic conditions and chemical
inhibitors); and
• By killing microbes (e.g. by heat or radiation).
b) Prevention or delay of self-decomposition of food
• By inactivation of food enzymes (e.g. blanching);
• By prevention of chemical reactions (e.g. by using antioxidants).

Banwart, G.J. (1979). Basic Food Microbiology, AVI Publishing Co. Inc.,
Frazier, W.C. and Westoff, D.C. (1996). Food Microbiology, Tata McGraw
Pelczar, M. Jr.O, Chan, E.C.S. and Kreig, N.R. (1993). Microbiology, Tata
Garbutt, J. (1998). Essentials of Food Microbiology, Arnold International
49
Microbiology UNIT 3 FOOD-BORNE DISEASES
Structure
3.0 Objectives
3.1 Introduction
3.2 What is a Disease?
3.3 How Do Micro-organisms Cause Disease?
3.4 Food-borne Diseases and the Agents
3.4.1 Diseases by Bacteria
3.4.2 Diseases by Molds
3.4.3. Diseases by Viruses
3.4.4. Diseases by Parasites
3.4.5 Diseases by Natural toxins
3.4.6 Diseases by Prions
3.5 Types of Food-borne Diseases

3.6 Common Food-borne Pathogens and their Symptoms
3.7 Factors Responsible for Food-borne Diseases
3.8 Emerging Food-borne Pathogens
3.8.1 Examples of Emerging Food-borne Pathogens
3.8.2 Antimicrobial Resistance and Food-borne Pathogens
3.9 Let Us Sum Up

3.10 Key Words
3.0 OBJECTIVES
z specify food-borne diseases: their types and causative agents;
z enumerate common food-borne diseases and their symptoms; and
z state emerging food-borne pathogens and reasons for their emergence.
3.1 INTRODUCTION
Food-borne disease is caused by consuming contaminated foods or beverages.
Many different disease-causing microbes, or pathogens, can contaminate foods,
so there are many different Food-borne infections. In addition, poisonous
chemicals, or other harmful substances can cause Food-borne diseases if they are
present in food.
50
More than 250 different Food-borne diseases have been described. Most of Food Borne Diseases
these diseases are infections, caused by a variety of bacteria, viruses, and parasites
that can be Food-borne. Other diseases are poisonings, caused by harmful toxins
or chemicals that have been produced in or have contaminated the food, for
example, mercury in fish /sea foods, mycotoxins in peanuts, wheat etc. These
different diseases have many different symptoms, so there is no one “syndrome”
that is Food-borne illness. However, the microbe or toxin enters the body through
the gastrointestinal tract, and often causes the first symptoms there. Nausea,
vomiting, abdominal cramps and diarrhea are common symptoms in many Food-
borne diseases.
Many microbes can spread in more than one way, so we cannot always know
that a disease is Food-borne. The distinction matters, because public health
authorities need to know how a particular disease is spreading to take the
appropriate steps to stop it. For example, Escherichia coli O157:H7 infections
can spread through contaminated food, contaminated drinking water, contaminated
swimming water, and from toddler to toddler at a day care center. Depending on
which means of spread caused a case, the measures to stop other cases from
occurring could range from removing contaminated food from stores, chlorinating
a swimming pool, or closing a child day care center.
3.2 WHAT IS DISEASE?

The term disease is applied to any harmful change in the tissues and/or metabolism
of a plant, animal or human that produces the symptoms of illness. Micro-organisms
(bacteria, yeasts, moulds, viruses and protozoa) that cause diseases are known as
pathogens.
3.3 HOW DO MICRO-ORGANISMS CAUSE

DISEASE?
Soon after birth the external surfaces and cavities of our bodies are colonized by
large numbers of different types of micro-organisms that originate from other
humans and the environment in general. These organisms constitute our natural
permanent (native) micro flora. Most of the organisms are bacteria but some
yeasts also occur. This natural resident micro flora is symbiotic, i.e. it lives in
mutual harmony with our body tissues, and is essential for our well being.
Here are two examples of importance of our permanent micro flora:
• The permanent micro flora is essential in combating invasion of the body by
potential pathogens by competing for space and nutrients, and sometimes
producing antibiotics. The presence of a variety of strains of Escherichia
coli in the colon, for example, helps to prevent enteric pathogens such as
Salmonella spp. from becoming established. Laboratory animals that are
reared under sterile conditions and without a natural resident micro flora are
exceptionally prone to diseases caused by organisms that are not even normally
considered pathogens.
• Bacteria in the colon synthesize vitamin K and contribute significantly to our
requirement for this vitamin.
Our bodies are constantly being exposed and infected with organisms that
are not part of this permanent micro flora. Most of these organisms are 51
Fundamentals of Food harmless and transient. Others are pathogens and have the ability to invade
Microbiology
our tissues, or produce toxins, or both.
Toxins are chemical substances produced by microorganisms that are harmful to
human tissues and physiology. Many, but not all, of the toxins produced by micro-
organisms are proteins. Sometimes toxins are secreted into the environment in
which the micro organism is growing, for example, the enterotoxin produced by
Staphylococcus aureus can be secreted into food. Toxins of this type can come
into contact with or enter the human body and cause disease in the absence of the
organism.
3.4 FOOD-BORNE DISEASES AND THE AGENTS

Food-borne disease is simply disease that results from the ingestion of food.
Food-borne diseases (FBD) are defined by the World Health Organization as
“diseases of infectious or toxic nature caused by, or thought to be caused by the
consumption of food or water”. More than 250 FBDs have been described.
Symptoms vary widely, depending on the etiological agents. Diarrohea and vomiting
are the most common. In many countries, national health care organizations record
FBD outbreaks, defined as the occurrence of two or more cases of a similar
illness resulting from the ingestion of a common food.
Agents that can be responsible for food-borne disease are:
1) micro-organisms;
2) parasites;
3) chemicals;
4) naturally occurring plant toxicants;
5) naturally occurring fish toxicants;
6) metabolic disorders;
7) foods that. give rise to allergies;
8) radioactive materials.
Common food poisioning agents are given in the Table 3.1.
Micro-organisms are by far the most important agents of food-borne disease,
with bacteria causing the major bulk of food-borne disease outbreaks. Viruses
are also an important source of food-borne disease with food-borne transmission
of infective protozoa far less common, particularly in developed countries. Some
moulds produce substances that are toxic to man (mycotoxins) but their importance
in food-borne disease production is currently not known. A few algae produce
toxins that are associated with shellfish poisoning. Yeasts are very rarely associated
with food-borne disease (apart from the ability of Saccharomyces cerevisiae to
produce alcohol). The one documented example is associated with a yeast that
infects the surface of sun-dried fish in South America. The organism can cause a
skin infection in anyone handling the fish. Prions that cause degenerative diseases
of the nervous system, e.g. the agent causing bovine spongiform encephalopathy
52 (BSE) in cattle, may possibly be transmitted to man via infected offal.
Table 3.1: Common Food Poisoning Agents Food Borne Diseases
Agent Example
Bacteria Salmonella, Staphylococcus aureus, Clostridium

perfringens, C.botulinum, Bacillus cereus
Viruses Hepatitis A virus, Parvovirus, Norwalk virus
Molds Claviceps purpurea, Aspergillus flavus (foodborne

aflatoxins)
Marine protozoa Gonyaulax tamareusis (paralytic crustacean

poisoning). Gambierdiscus toxicus (poisoning with
ciguatera). Prorocentrum lima and some species
of Dinophys (diarrheic shellfish poisoning)
Parasites Trichinella spiralis, Taenia solium, Giardia

(protozoa, amoebas) lamblia, Cryptosporidium, Entamoeba histolytica
Chemical substances Heavy metals (lead, zinc, copper, cadmium, mercury,

arsenic, antimony), pesticides, herbicides, fungicides,
substances used for cleaning and disinfections.
Toxic plants Germinated potatoes, apricot core, red bean

(Phaseolus vulgaris)
Toxic animals Scombrotoxin from fish insufficiently cooked –

herring – (histamine)
3.4.1 Diseases by Bacteria

Toxins are particularly important in production of bacterial diseases. Bacterial
toxins are classified into two types, exotoxins and endotoxins. Exotoxins have
the following characteristics:
z generally proteins synthesized by metabolic activity;
z not structural components of the cell;
z secreted into the cell environment.
z produced by Gram-positive and Gram-negative organisms;
Endotoxins have the following characteristics:
z lipopolysaccharides;
z toxic components of the cell wall released when the cell dies and breaks
down;
z produced by Gram-negative organisms.
3.4.2 Diseases by Molds
Mushroom poisoning is caused by the consumption of raw or cooked fruiting
bodies (mushrooms, toadstools) of a number of species of higher fungi
(Basidiomycetis). The term toadstool (from the German Todesstuhl, death’s stool)
is commonly given to poisonous mushrooms, but for individuals who are not
53
Fundamentals of Food experts in mushroom identification there are generally no easily recognizable
Microbiology
differences between poisonous and non-poisonous species. The toxins involved in
mushroom poisoning are produced naturally by the fungi themselves, and each
individual specimen of a toxic species should be considered equally poisonous.
Mycotoxins are low molecular weight natural products (i.e., small molecules)
produced as secondary metabolites by filamentous fungi. These metabolites constitute
a toxigenically and chemically heterogeneous assemblage that is grouped together
only because the members can cause disease and death in human beings and
other vertebrates. Not surprisingly, many mycotoxins display overlapping toxicities
to invertebrates, plants, and microorganisms. Unfortunately, mycotoxins can also
be incredibly toxic to humans causing a variety of responses including cold/flu-like
symptoms, sore throats, headaches, nose bleeds, fatigue, diarrohea, dermatitis,
and immune suppression. Some mycotoxins may also be carcinogenic and
teratogenic. Molds that have been known to potentially produce toxins. Some of
these molds are Acremonium, Alternaria, Aspergillus, Chaetomium,
Cladosporium, Fusarium, Penicillium, and Stachybotrys. Even though these
molds may potentially produce mycotoxins, they will not do so unless specific
environmental conditions exist.
Common types of mycotoxins
• Aflatoxin. This mycotoxin is primarily produced by Aspergillus species. It
is one of the most potent carcinogens known to man and has been linked to
a wide array of human health problems.
• Ochratoxin. This mycotoxin is primarily produced by species of Penicillium
and Aspergillus. It can be damaging to the kidney/liver, and it is a suspected
carcinogen. There is also evidence supporting its role in impairing immune
system function.
• Tricothecene. This toxin is produced by Stachybotrys spp and Fusarium
spp and has even been indicated as a potential agent for use as a biological
weapon. One of the more deadly mycotoxins, if it is ingested in large amounts
it can severely damage the entire digestive tract and cause rapid death due to
internal haemorrhaging.
3.4.3 Diseases by Viruses

Unlike bacteria, viruses invade host cells, take over host cell metabolism and
induce the cell to produce new virus particles. Disease symptoms are caused by
the destruction of host cells and secondary effects resulting from host cell destruction.
Several viruses like Hepatitis A, Norwalk and Norwalk like viruses, Poliovirus
and Echovirus may cause Food-borne disease in consumers of virus-contaminated
water and foods. Some of the other viruses that have also been associated with
food are: Astrovirus, Calcivirus, Enteric Adenovirus, Parvovirus and
Rotavirus. These enteric viruses replicate in the intestine of infected individuals
and are transmitted by faecal-oral route. They must survive the acidic environment
of stomach, the alkaline conditions and digestive enzymes of the small intestine,
and the conditions encountered between the hosts. Such selective pressures result
in a stability that allows virtually any food to serve as a vehicle for transmission.
However, the most common types of food-borne viral diseases are Hepatitis A
(infectious hepatitis) and acute viral gastroenteritis.
54
The Hepatitis A virus (HAV) is one of more than 70 members of enterovirus Food Borne Diseases
group of Picornaviridae family. HAV is presumed to replicate initially in the
gastrointestinal tract and then spread primarily to liver, where it infects hepatocytes
and Kupffer cells. Food-borne viral gastroenteritis, on the other hand, is usually a
mild disease with various degrees of nausea, diarrhea, malaise, abdominal pain,
muscle pain, headache, and low-grade fever. Ice, water, ice cream, milk, pastries,
salads, sandwiches, shellfish, and other foods consumed raw or subjected to
additional handling after cooking are major food vehicles for virus transmission.
The highly pathogenic Avian Influenza H5N1 virus is a fast re emerging pathogen.
It has killed millions of poultry in a number of countries throughout Asia, Europe
and Africa. Although Avian Influenza viruses are essentially animal diseases, the
highly pathogenic H5N1 is able to infect and kill humans. Health experts are
concerned that the co-existence of human flu viruses and avian flu viruses (especially
H5N1) will provide an opportunity for genetic material to be exchanged between
species-specific viruses, possibly creating a new virulent influenza strain that is
easily transmissible and lethal to humans.
3.4.4 Diseases by Parasites
Numerous parasites can be transmitted by food including many protozoa and
helminths. The most common Food-borne parasites are protozoa such as
Cryptosporidium spp., Giardia intestinalis, Cyclospora cayetanensis,
Toxoplasma gondii, and Entamoeba histolytica; roundworms such as
Trichinella spiralis and Anisakis spp.; and tapeworms such as Diphylobothrium
spp. and Taenia spp.
Many of these organisms can also be transmitted by water, soil, or person-to-
person contact. Occasionally in the developed countries, but often in developing
countries, a wide variety of helminthic roundworms, tapeworms, and flukes are
transmitted in foods such as
• undercooked fish, crabs, and mollusks;
• undercooked meat; raw aquatic plants such as watercress;
• raw vegetables that have been contaminated by human or animal faces;
• foods contaminated by food service workers with poor hygiene or working
in unsanitary facilities.
Symptoms of Food-borne parasitic infections vary greatly depending on the type
of parasite. Protozoa such as Cryptosporidium spp., Giardia intestinalis and
Cyclospora cayetanensis most commonly cause diarrohea and other gastrointestinal
symptoms. Helminthic infections can cause abdominal pain, diarrhoea, muscle
pain, cough, skin lesions, malnutrition, weight loss, neurological and many other
symptoms depending on the particular organism and burden of infection. Treatment
is available for most of the Food-borne parasitic organisms.
3.4.5 Diseases by Natural Toxins

1) Ciguatera poisoning
2) Shellfish toxins (PSP, DSP, NSP, ASP)
3) Scombroid poisoning
4) Pyrrolizidine alkaloids 55
Fundamentals of Food 5) Phytohaemagglutinin (Red kidney bean poisoning)
Microbiology
6) Grayanotoxin (Honey intoxication)

7) Gempylotoxin (Gastrointestinal illness from consumption of Escolar and
Oilfish)
8) Tetrodotoxin (Pufferfish)
The most commonly occurring natural toxins are :
• Ciguatera Fish Poisoning
Ciguatera is a form of human poisoning caused by the consumption of
subtropical and tropical marine finfish which have accumulated naturally
occurring toxins through their diet. The toxins are known to originate from
several dinoflagellate (algae) species that are common to ciguatera endemic
regions in the lower latitudes. Manifestations of ciguatera in humans usually
involves a combination of gastrointestinal, neurological, and cardiovascular
disorders. Symptoms defined within these general categories vary with the
geographic origin of toxic fish.
• Shellfish Poisoning
Shellfish poisoning is caused by a group of toxins elaborated by planktonic
algae (dinoflagellates, Gonyaulax catenella and G. tamareusis in most cases)
upon which the shellfish feed. The toxins are accumulated and sometimes
metabolized by the shellfish. The 20 toxins responsible for paralytic shellfish
poisonings (PSP) are all derivatives of saxitoxin. Diarrheic shellfish poisoning
(DSP) is presumably caused by a group of high molecular weight polyethers,
including okadaic acid, the dinophysis toxins, the pectenotoxins, and
yessotoxin. Neurotoxic shellfish poisoning (NSP) is the result of exposure to
a group of polyethers called brevetoxins. Amnesic shellfish poisoning (ASP)
is caused by the unusual amino acid, domoic acid, as the contaminant of
shellfish.
Types of Shellfish Poisoning
• Paralytic Shellfish Poisoning (PSP)
• Diarrheic Shellfish Poisoning (DSP)
• Neurotoxic Shellfish Poisoning (NSP)
• Amnesic Shellfish Poisoning (ASP)
• Scombroid Poisoning (also called Histamine Poisoning)

Scombroid poisoning is caused by the ingestion of foods that contain high levels
of histamine and possibly other vasoactive amines and compounds. Histamine and
other amines are formed by the growth of certain bacteria (Proteins morganii,
Klebsiella pneumoniac) and the subsequent action of their decarboxylase enzymes
on histidine and other amino acids in food, either during the production of a
product such as Swiss cheese or by spoilage of foods such as fishery products,
particularly tuna or mahi mahi. However, any food that contains the appropriate
amino acids and is subjected to certain bacterial contamination and growth may
56 lead to scombroid poisoning when ingested.
3.4.6 Diseases by Prions Food Borne Diseases
Prions are normal proteins of animal tissues that can misfold and become infectious:
they are not cellular organisms or viruses. Prions are associated with a group of
diseases called Transmissible Spongiform Encephalopathies (TSEs). In humans,
the illness suspected of being Food-borne is variant Creutzfeldt-Jakob disease
(vCJD). The human disease vCJD and the cattle disease, bovine spongiform
encephalopathy (BSE), also known as “mad cow” disease, appear to be caused
by the same agent.

1) What are pathogens?
............................................................................................................
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2) Define FBD.
............................................................................................................
............................................................................................................
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3) Mycotoxins are ..................... metabolites.
4) What is an enterotoxin?
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3.5 TYPES OF FOOD-BORNE DISEASES (FBDs)

Types of food poisoning include the following :
• Intoxications
• Infections.
• Toxi-infections
Table 3.2 enlists the food-borne diseases and causative pathogens
3.5.1 Intoxications
Intoxications involve food poisoning in which the organism grows in the food and
releases a toxin from the cells. When the toxin is ingested along with the food, the
toxin gives rise to the food poisoning syndrome (signs and symptoms that indicate 57
Fundamentals of Food a particular disease). The presence of the organism in the food is irrelevant to
Microbiology
disease production. It is the toxin that gives rise to the disease. Bacterial toxins
that produce intoxications are exotoxins that are either enterotoxins affecting the
gut, as in the disease caused by Staphylococcus aureus, or neurotoxins, as in
the disease caused by Clostridium botulinum, the toxin in this case affecting the
nervous system.
Mycotoxicosis (diseases produced by the ingestion of food containing mycotoxin
produced by moulds) and the diseases produced by algal toxins that find their
way into shellfish can also be considered intoxications. Generally, intoxications
have short incubation periods (time from ingestion of the food to the appearance
of symptoms).
3.5.2 Infections
Infections involve food poisoning caused by the ingestion of live organisms when,
typically, the organisms grow in the gastrointestinal tract to produce the disease.
Most food poisoning caused by micro-organisms falls into this category, for
example, food poisoning caused by Salmonella spp (salmonellosis). Enteritis
associated with food poisoning infections is due to the production of exotoxins or
endotoxins that act as enterotoxins.
3.5.3 Toxi Infections
In some types of food poisoning, e.g. Clostridium perfringens, live cells need to
be ingested for the disease to occur but the organism does not grow and reproduce
in the gut. Vegetative cells sporulate after ingestion, and an enterotoxin is released
when the spore mother cells break down releasing the toxin. Because living cells
also need to be ingested to cause this type of food poisoning, it can be considered
as a food-borne toxi-infection.
Table 3.2: Microbial Food-borne Diseases and Causative Pathogens
Type of disease Causative microorganism Microbial group Major symptom(s)

type
Intoxication
Staph. poisoning Staphylococcus aureus Bacteria, Gm +* Gastric

strains
Botulism Clostridium botulinum Bacteria, Gm + Non gastric

strains
Mycotoxin Mycotoxins producing mold Molds Non gastric

poisoning strains, e.g., Aspergillus
flavus
Infection
Salmonellosis Over 2000 Salmonella Bacteria, Gm - * Gastric

serovars (except S. typhi
and S. paratyphi)
Campylobacter Campylobacter jejuni and C. Bacteria, Gm - Gastric

enteritis coli strains
Yersiniosis Pathogenic strains of Bacteria, Gm - Gastric

Yersinia enterocolitica
58
Food Borne Diseases
Type of disease Causative microorganism Microbial group Major symptom
(s) type
Intoxication Intoxication Intoxication
Enterohemorrhagic Escherichia coli Bacteria, Gm - Gastric and

E. coli colitis O157:H7 nongastric
Nonhaemorrhegic Shiga-like toxin (verotoxin) Bacteria, Gm - Gastric

E. coli colitis producing
E. coli strains like
E. coli 026:H11
Listeriosis Listeria monocytogenes Bacteria, Gm + Gastric and

(Pathogenic strains) nongastric
Shigellosis Four Shigella species e.g., Bacteria, Gm - Gastric

S. dysenterie
Vibrio Pathogenic strains of Bacteria, Gm - Gastric

parahaemolyticus V. parahaemolyticus
gastroenteritis
Vibrio vulnificus Vibrio vulnificus strains Bacteria, Gm - Gastric and

infection nongastric
Brucellosis Brucella abortus Bacteria, Gm - Gastric and

nongastric
Viral infections Pathogenic enteric viruses, Viruses Gastric and

e.g., Hepatitis A. virus nongastric
Toxi infection
Clostridium Clostridium perfringens Bacteria, Gm+ Gastric

perfringens strains
gastroenteritis
Bacillus cereus Bacillus cereus strains Bacteria, Gm + Gastric

gastroenteritis
E. coli Enteropathogenic and Bacteria, Gm - Gastric

gastroenteritis enterotoxigenic Escherichia
coli strains
Cholera Pathogenic strains of Vibrio Bacteria, Gm - Gastric

cholerae
Gastroenteritis by Opportunist Pathogens
Aeromonas Aeromonas hydrophila Bacteria, Gm - Gastric

hydrophila strains
gastroenteritis
Plesiomonas Plesiomonas shigelloidies Bacteria, Gm - Gastric

shigelloides strains
gastroenteritis
Gm +* , Gm -* : Gram-positive and – negative, respectively

59
Microbiology 3.6 COMMON FOOD-BORNE PATHOGENS AND
THEIR SYMPTOMS
Symptoms will vary depending on the type of pathogen, and may include diarrhoea,
vomiting, nausea, abdominal pain, and fever. Other symptoms may include
headache, jaundice, and numbness. Symptoms can take between a few hours to
a few days to develop and may last for a few days, depending on the type of
pathogen.
Some common pathogens and their features:
Pathogen Microscopic Incubation Symptoms Associated foods *

image of period (time
pathogen between eating
and onset of
symptoms)
Bacillus cereus 1 - 6 hours Sudden onset Improperly

toxin (vomiting) of severe refrigerated
nausea and cooked rice
vomiting
Bacillus cereus 6 - 24 hours Abdominal Meats, stews,

toxin cramps, gravy, vanilla
(diarrhoea) nausea and sauce
watery
diarrhoea
Campylobacter 2 - 5 days Fever, nausea, Raw and

abdominal undercooked
cramps and poultry,
diarrhoea unpasteurised milk
(sometimes and contaminated
bloody) water
Clostridium 6 - 24 hours Abdominal Meats, poultry,

perfringens cramps, gravy, dried or
toxin watery precooked foods
diarrhoea and
nausea
Escherichia 2 - 10 days Diarrhoea Improperly cooked

coli (STEC) more (often beef,
commonly 3 - 4 bloody), unpasteurised milk
days abdominal and juice, sprouts
cramps and contaminated
water
Hepatitis A 2 - 7 weeks Jaundice, Raw or poorly

fatigue, cooked seafood
anorexia, harvested from
nausea contaminated
waters, ready-to-
eat foods handled
by an infected
food handler
Listeria 3 days - Meningitis, Soft cheeses,

monocytogenes 10 weeks sepsis, fever unpasteurised
milk, ready-to-eat
deli meats
60
Food Borne Diseases
Pathogen Microscopic Incubation Symptoms Associated foods *
image of period (time
pathogen between eating
and onset of
symptoms)
Norovirus 24 - 48 hours Fever, nausea, Poorly cooked
vomiting, shellfish, ready-to-
abdominal eat foods touched
cramps, by an infected
diarrhoea and worker
headache
Salmonella 6 – 72 hours, Headache, Undercooked

usually 12-36 fever, poultry, raw egg
hours abdominal deserts and
cramps, mayonnaise,
diarrhoea, sprouts, tahini
vomiting and
nausea
Staphylococus 0.5 – 8 hours Sudden onset Cream deserts and

aureus toxin of vomiting pastries, potato
and abdominal salad
cramps
Vibrio 4–30 hours, Nausea, Undercooked or

parahae- usually 12-24 vomiting, raw seafood.
molyticus hours abdominal
cramps and
watery
diarrhoea
(*The foods shown in the table have previosly been found to be the source of
the pathogens listed. This does not mean that these foods are always unsafe
to eat or that such pathogens are always present, however the foods should
be purchased from a reputed supplier and handled with care. )

1) Give an example of the following:
a) Waterborne virus
b) Pathogen associated with poultry
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2) What are the common Food-borne diseases?
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61
Fundamentals of Food 3) Classify the FBDs.
Microbiology
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3.7 FACTORS RESPONSIBLE FOR FBD

Several factors have developed during the past 20 years that make the
emergence of Food-borne diseases a reality. Before discussing these new risk
factors, it would be good to review the five basic factors that must be present
in order for a Food-borne disease to occur. When reading about the newly
developed factors that predispose society to Food-borne diseases, keep these
basic factors in mind.
1) Presence of pathogenic organisms or toxins in the food consumed
2) Source of contamination
3) Medium for the growth and maintenance of the pathogen or toxin
4) Proper environmental conditions to maintain or allow for replication of
the pathogen
5) Consumption of a significant quantity of the contaminated or infectious
food, i.e. acquire an infectious dose.
Table 3.3: Factors Contributing to Outbreaks of Food-borne Disease
(Various Sources).
Contributing Factors Percentage
A. Factors relating to microbial growth
Storage at ambient (room) temperature 43
Improper cooling 32
Preparation too far in advance of serving 41
Improper warm holding 12
Use of leftovers 5
Improper thawing and subsequent storage 4
Extra large quantities prepared 22
B. Factors relating to microbial survival
Improper reheating 17
Inadequate cooking 13
C. Factors relating to contamination
Food workers 12
Contaminated processed non-canned foods 19
Contaminated raw foods 7
Cross-contamination 11
Inadequate cleaning of equipment 7
Unsafe source 5
Contaminated canned foods 2
62 (Percentages exceed a total of 100 since multiple factors often contribute to Food-borne illness.)
Food Borne Diseases
3.8 EMERGING FOOD-BORNE DISEASES
The basic definition of an emerging or re-emerging infectious disease is a
disease whose incidence has increased in a defined time period and
location. If the disease was unknown in the location before, the disease is
considered to be emerging. However, if the disease had been present at the
location in the past and was considered eradicated or controlled, the disease is
considered to be re-emerging. Diseases considered to be emerging or re-
emerging include avian influenza, West Nile virus, bovine tuberculosis in wildlife
etc.
Recognition of an emerging disease can occur because the disease is present in
the population for the first time, because the disease has been detected for the
first time, or because links between an infectious agent and a chronic disease or a
syndrome have only recently been identified. Many of these emerging diseases
are zoonotic, and rely on animal populations as reservoirs of infection.
Most emerging infections are caused by pathogens already present in the
environment, brought out of obscurity or given a selective advantage by changing
conditions and afforded an opportunity to infect new host populations. These
changes include ecological changes, such as those due to human activities or to
anomalies in climate; demographic changes and behavior; travel and commerce;
technology and industry; microbial adaptation and change; and breakdown of
public health measures. Many factors precipitate emergence by placing humans
or animals in contact with a natural reservoir or host for an infection unfamiliar but
already present (often a zoonotic or arthropod-borne infection), either by increasing
proximity or, often, also by changing conditions so as to favor an increased
population of the microbe or its natural host. New Food-borne disease threats
occur for a number of reasons:
1) Globalization of the food supply: A large outbreak of cyclosporiasis occurred
in North America in 1996-97 linked to contaminated raspberries imported
from South America.
2) Inadvertant introduction of pathogens into new geographic areas:
Vibrio cholerae was introduced into waters off the coast of southern United
States when a cargo ship discharged contaminated ballast water in 1991.
3) Travellers, refugees, and immigrants exposed to unfamiliar Food-borne
hazards while abroad: International travellers may become infected by Food-
borne pathogens that are uncommon in their countries. It is estimated that
about 90% of all cases of Salmonellosis in Sweden are imported.
4) Changes in microorganisms: Changes in microbial populations can lead to
the evolution of new pathogens, development of new virulent strains in old
pathogens, development of antibiotic resistance that might make a disease
more difficult to treat, or to changes in the ability to survive in adverse
environmental conditions.
5) Change in the human population: The population of highly susceptible
persons expanding world-wide because of ageing, malnutrition, HIV infections
and other underlying medical conditions. Age is an important factor in
susceptibility to Food-borne infections because those at the extremes of age
63
Fundamentals of Food have either not developed or have partially lost protection from infection.
Microbiology
Particularly for the elderly, Food-borne infections are likely to invade their
blood stream and lead to severe illness with high mortality rates. People with
a weakened immune system also become infected with Food-borne pathogens
at lower doses which may not produce an adverse reaction in healthier
persons. Seriously ill persons, suffering, for example, from cancer or AIDS,
are more likely to succumb to infections with Salmonella, Campylobacter,
Listeria, Toxoplasma, Cryptosporidium, and other Food-borne pathogens.
In developing countries reduced immunity due to poor nutritional status render
people, particularly infants and children, more susceptible to Food-borne
infections.
6) Changes in lifestyle: Greater numbers of people go out and eat meals
prepared in restaurants, canteens, fast food outlets, and by street food vendors.
In many countries, the boom in food service establishments is not matched
by effective food safety education and control. Unhygienic preparation of
food provides ample opportunities for contamination, growth, or survival of
Food-borne pathogens.
3.8.1 Examples of Emerging Food-borne Diseases

• Some Food-borne diseases are well recognized, but are considered emerging
because they have recently become more common. For example, outbreaks
of Salmonellosis have been reported for decades, but within the past 25
years the disease has increased in incidence on many continents. In the
Western hemisphere and in Europe, Salmonella serotype enteritidis (SE)
has become the predominant strain. Investigations of SE outbreaks indicate
that its emergence is largely related to consumption of poultry or eggs.
• While cholera has devastated much of Asia and Africa for years, its
introduction for the first time in almost a century on the South American
continent in 1991 makes it another example of an infectious disease that is
both well-recognized and emerging. While cholera is often waterborne, many
foods also transmit infection. In Latin America, ice and raw or under processed
seafood are important epidemiological pathways for cholera transmission.
• Other Food-borne pathogens are considered emerging because they are
new microorganisms or because the role of food in their transmission has
been recognized only recently. Infection with Escherichia coli serotype
O157:H7 (E. coli) was first described in 1982. Subsequently, it has emerged
rapidly as a major cause of bloody diarrhoea and acute renal failure. The
infection is sometimes fatal, particularly in children. Outbreaks of infection,
generally associated with beef, have been reported in Australia, Canada,
Japan, United States, in various European countries, and in southern Africa.
Outbreaks have also implicated alfalfa sprouts, unpasteurized fruit juice, lettuce,
game meat and cheese curd.
In 1996, an outbreak of Escherichia coli O157:H7 in Japan affected over
6,300 school children and resulted in 2 deaths. This is the largest outbreak
ever recorded for this pathogen.
• Listeria monocytogenes (Lm) is considered emerging because the role of
food in its transmission has only recently been recognized. In pregnant women,
64 infections with Lm can cause abortion and stillbirth, and in infants and persons
with a weakened immune system it may lead to septicemia (blood poisoning) Food Borne Diseases
and meningitis. The disease is most often associated with consumption of
foods such as soft cheese and processed meat products that are kept
refrigerated for a long time because Lm can grow at low temperatures.
Outbreaks of listeriosis have been reported from many countries, including
Australia, Switzerland, France and the United States. Two recent outbreaks
of Listeria monocytogenes in France in 2000 and in the USA in 1999 were
caused by contaminated pork tongue and hot dogs respectively.
• Food-borne trematodes are also emerging as a serious public health problem,
especially in South-east Asia but also in Latin America, in part due to a
combination of increased aquaculture production, often under unsanitary
conditions, and of consumption of raw and lightly processed fresh water fish
and fishery products. Food-borne trematodes can cause acute liver disease,
and may lead to liver cancer. An estimated 40 million people world wide are
affected.
• Bovine Spongiform Encephalopathy (BSE), a fatal, transmissible, neuro-
degenerative disease of cattle, was first discovered in the United Kingdom in
1985. The cause of the disease was traced to an agent related to scrapie in
sheep, which contaminated recycled bovine carcasses used to make meat
and bone meal additives for cattle feed. Recycling of the BSE agent led to a
distributed common source epidemic of more than 180,000 diseased animals
in the UK alone. The agent affects the brain and spinal cord of cattle udder
and lesions are characterized by sponge-like changes visible in a microscope.
At this time, 19 countries have reported endemic BSE cases and the disease
is no longer confined to the European Community: a case of BSE has been
reported in the cattle herd of Japan.
In human populations, exposure to the BSE agent (probably in contaminated
bovine-based food products) has been strongly linked to the appearance in 1996
of a new transmissible spongiform encephalopathy of humans called variant
Creutzfeldt-Jakob Disease (vCJD).
LIST OF EMERGING FOOD- AND WATERBORNE PATHOGENS
BACTERIA
• Diarrheagenic E.coli ( E. coli 0157:H7)

• Pathogenic Vibrios
• Shigella species
• Salmonella serotype enteritidis (SE)
• Listeria monocytogenes
• Campylobacter jejuni
• Yersinia enterocolitica
VIRUSES
• Caliciviruses
• Hepatitis A
65
Fundamentals of Food • Chikungunya virus
Microbiology
• Severe acute respiratory syndrome-associated coronavirus (SARS-CoV )

• Avian Influenza
PROTOZOA
• Cryptosporidium parvum
• Cyclospora cayatanensis
• Giardia lamblia
• Entamoeba histolytica
• Toxoplasma
• Microsporidia
PRIONS
3.8.2 Anti-Microbial Resistance and Food-Borne Pathogens

Bacteria become resistant to antibiotics as a consequence of antibiotic use in
humans and animals. Antibiotics used in humans can lead to resistant bacteria that
can be spread in communities. For example, there are significant increases in
resistant pneumococcus (a cause of pneumonia), gonococcus (the cause of
gonorrhea), and Mycobacterium tuberculosis (the cause of tuberculosis). In
addition, many pathogens that are spread among patients in hospitals are highly
resistant, and these antibiotic- resistant infections can be life-threatening and
untreatable with currently available antibiotics.
The use of antibiotics in animals similarly, leads to resistant bacteria in animals.

Bacteria from healthy animals can contaminate food and cause human illness.
When the bacteria from animals are resistant to antibiotics, the resulting human
infection may be more difficult to treat. There is a clear relationship between the
use of antibiotics in animals and the appearance of resistant human infections. One
particular class of antibiotics, fluoro-quinolones, has been particularly important in
the treatment of severe infections of humans. Resistance to fluoro-quinolones has
emerged in some human pathogens.. Therefore, collaborative effort, designed
specifically to detect the emergence of antibiotic resistance in Food-borne
pathogens, is an important component of the efforts to improve surveillance for
new and emerging pathogens.

1) What is an enterging pathogen?
............................................................................................................
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66
2) What is Mad Cow disease? Food Borne Diseases
............................................................................................................
............................................................................................................
............................................................................................................
3) Name few emerging Food-borne pathogens.
............................................................................................................
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3.9 LET US SUM UP

Food-borne disease generally results from consuming food or drink contaminated
with pathogenic bacteria, viruses or toxins. It often produces symptoms such as
nausea, vomiting, stomach pain, diarrohea, or fever. The symptoms are often the
same as for gastro-type illnesses caused by other sources (other people carrying
the bacteria, unhygienic surfaces, etc). This usually makes it hard to separate
illness caused by food from the other sources of illness.
Bacteria are found in many places, including in human and animal faeces. When
these potentially harmful pathogens enter the food supply they can cause Food-
borne diseases. Food-borne diseases can occur from any fresh or processed
foods consumed in a range of settings, such as homes, restaurants, large catering
establishments, schools and institutions. Most cases of Food-borne diseases can
be avoided through good hygiene and good food handling practices.
Most Food-borne diseases are caused by pathogenic bacteria or viruses in food.
Other less common Food-borne diseases occur from accidental chemical poisoning
and natural contaminants. The most common types of Food-borne diseases are:
• Bacterial e.g., Salmonella, Campylobacter, Ecolib, Shigella and Listeria;
• Viral eg Neurovirus, Rotavirus and Hepatitis A;
• Intoxication caused by toxins produced by pathogens such as Staphylococcus
aureus, Bacillus cereus and Clostridium perfringens.
Symptoms will vary depending on the type of pathogen, and may include diarrhoea,
vomiting, nausea, abdominal pain, and fever. Other symptoms may include
headache, jaundice, and numbness. Symptoms can take between a few hours to
a few days to develop and may last for a few days, depending on the type of
pathogen.
New Food-borne pathogens like E. coli will continue to be identified, and other
known but rare Food-borne pathogens like Salmonella enteritidis (SE) may re-
emerge as important public health problems. Infections may arise from changes in
the microbes, changes in the industrial technology that underlies food production
and processing, changes in our choices concerning the foods that we eat, how
they are prepared, and where we eat them, and changes in the demographics of
the population. The increasing number of elderly and chronically ill are at particular
risk for severe illness caused by Food-borne pathogens. 67
Microbiology 3.10 KEY WORDS
Aflatoxins : Dangerous toxins (carcinogen) produced by moulds of
the Aspergillus species (A. flavus), found in cereals,
oilseeds and nuts when incorrectly dried and stored.
Biological hazard : Danger posed to food safety by the contamination of
food with pathogenic microorganisms or naturally
occurring toxins.
Chemical hazard : Danger posed to food safety by the contamination of
food by chemical substances, such as pesticides,
detergents, additives, and toxic heavy metals.
Coliforms : Bacteria (primarily E. coli and Enterobacter
aerogenes) used as an indicator of the sanitary quality
of food. High levels of coliforms indicate the presence
of fecal contamination in food and water.
Enteric (entero-) : Relating to the intestine.
Enteropathogenic : Causing illness in the intestinal tract.
Exotoxin : Potent toxic substance (Protein) formed and released
extracellularly by species of certain bacteria.
Food safety : Protecting the food supply from microbial, chemical
(i.e. rancidity, browning) and physical (i.e. drying out,
infestation) hazards or contamination that may occur
during all stages of food production and handling-
growing, harvesting, processing, transporting, preparing,
distributing and storing. The goal of food safety
monitoring is to keep food wholesome.
Food Safety Hazards : Include all microbiological, chemical, and foreign
materials that, if consumed, could cause injury or harm.
Food-borne : Illness caused by ingestion of food containing a toxin
Intoxication (metabolic byproduct) that was formed and excreted
into the food as a result of pathogenic microbial growth
(i.e. Clostridium botulinium, Staphyloccccus aureus.)
Food-borne toxin : Disease that results from eating a food containing a
mediated infection large number of disease-causing microorganisms.
Microbial load : Total number of living microorganisms in a given volume
or mass of microbiological media or food.
Natural toxins : Naturally occurring substance (e.g., produced in some
cases by disease-causing microorganisms) which is
poisonous to certain other living organisms.
Norwalk virus : Virus that contaminates raw oysters/shellfish, water and
ice, salads, frosting, person-to-person contact.
Pathogenic : Capable of causing disease; harmful; any disease-causing
68 agent.
Physical hazard : Particles or fragments of items not supposed to be in Food Borne Diseases
foods.
Toxin : Poison produced by a living microorganism.
Virulence : Pathogenic or poisonous potential of bacteria, fungi, or
other agents.
Virus : Infectious microorganisms that reproduce only in living
cells. They cause diseases such as mumps and Hepatitis
A and can be transmitted through food.
Zoonotic : Microorganism normally found in or on animals.
3.11 ANSWERS TO CHECK YOUR PROGRESS

EXERCISES
1) Food-borne outbreak is an incident in which two or more people
experience the same illness after eating the same food.
2) Food-borne disease is the illness which result from ingestion of
contaminating microbial pathogens (i.e., bacteria, mold, viruses), chemicals,
parasites, viruses or from naturally occurring toxins or poisons.
3) Fungal
4) Enterotoxins are the proteins produced by bacteria that are either ingested
as pre-formed toxins or are produced by a pathogen that has colonized
the gastro-intestinal tract.
1) Hepatitis A and Campylobacter
2) Examples of diseases transmissible through food include Amoebiasis,
Botulism, Cholera, Cryptosporidiosis, Giardiasis, Listeriosis, Salmonella
infections, Shigellosis, Trichinosis, Typhoid fever.
3) Food-borne diseases can be classified as : Intoxications, Infections and
Toxi infections
1) Infection that has newly appeared in [a] population or has existed but is
rapidly increasing in incidence or geographic range.
2) Mad cow disease (also known as Bovine spongiform encephalopathy
(BSE). It is a rare, chronic degenerative disease affecting the brain and
central nervous system of cattle. Cattle with BSE lose their coordination,
develop abnormal posture and experience changes in behavior. Clinical
symptoms take 4-5 years to develop, followed by death in a period of
69
Fundamentals of Food several weeks to months unless the affected animal is destroyed sooner.
Microbiology
3) Diarrhoeagenic E.coli (E.coli 0157:H7), Salmonella serotype enteritidis

(SE), Listeria monocytogenes, Campylobacter jejuni and Yersinia
enterocolitica .

Cole, R.J. (Ed.) (1986). Modern Methods in the Analysis and Structural
Elucidation of Mycotoxins. Academic Press, Orlando, FL.
Frazier, W.C. and Westoff, D.C. (1996). Food Microbiology, Tata McGraw
Pelczar, M. Jr., Chan, E.C.S. and Kreig, N.R. (1993). Microbiology, Tata
Garbutt, J. (1998). Essentials of Food Microbiology, Arnold International
70
Beneficial Role of
UNIT 4 BENEFICIAL ROLE OF Micro-organisms
MICRO-ORGANISMS
Structure
4.0 Objectives
4.1 Introduction
4.2 Fermentation
4.3 Fermented Foods and their Importance
4.4 Food Fermentation-Science and Technology
4.5 Types of Food Fermentations
4.5.1 Acid Food Fermentation
4.5.2 Yeast food fermentation
4.5.3 Solid State Fermentation
4.6 Common Examples of Food Fermentation

4.6.1 Oriental and Indigenous Fermented Foods
4.6.2 Fermented Vegetable Foods
4.6.3 Fermented Soya Bean Products
4.6.4 Fermented Dairy Products
4.6.5 Economically Important Fermented Foods
4.7 Fermented foods as Functional Foods

4.7.1 Probiotics
4.7.2 Prebiotics
4.7.3 Synbiotics
4.7.4 Use of Probiotics
4.7.5 Health Benefits of Probiotics
4.8 Let Us Sum Up

4.9 Key Words
4.0 OBJECTIVES

z describe beneficial role of microorganisms;
z explain fermentation – its science and technology;
z comprehend the needs and types of food fermentations;
z state common examples of different types of food fermentations; and
z attribute fermented foods as functional foods.
71
Microbiology 4.1 INTRODUCTION
Food microbiology can be divided into three focus areas; beneficial microorganisms,
spoilage microorganisms, and disease causing microorganisms. Beneficial
microorganisms are those used in food fermentation to produce products such as
cheese, fermented meat (pepperoni), fermented vegetables (pickles), fermented
dairy products (yoghurt), and ethnic fermented products such as sauerkraut, idli
and kimchi. In fermented products (produced by natural or controlled fermentation),
microorganisms metabolize complex substrates to produce enzymes, flavor
compounds, acids, and antimicrobial agents to improve product shelf-life and to
prevent growth of pathogens and to provide product attributes. Micro organisms
with their enzymes, also breakdown indigestible compounds to make the product
more palatable and easy to digest. In addition, the beneficial microorganisms also
serve as probiotics to impart direct health benefit by modulating the immune
system to provide protection against chronic metabolic diseases, bacterial infection,
atherosclerosis, and allergic responses. Examples of beneficial microorganisms
are Lactobacillus acidophilus, Lactobacillus arabinosus, Lactobacillus lactis,
and Pediococcus cerevisiae. Food spoilage microorganisms are those which
upon growth in a food, produce undesirable flavour (odour), texture and
appearance, and make food unsuitable for human consumption. Sometimes
uncontrolled growth of many of the beneficial microorganisms can also cause
spoilage. Food spoilage is a serious issue in developing countries because of
inadequate processing and refrigeration facilities. Examples of food spoilage
microorganisms are Brochotrix, Lactobacillus, Bacillus, Pseudomonas spp.
and some molds. The micro-environment created in a spoilt food generally
discourages the growth of the pathogenic microorganisms, which are considered
poor competitors.
4.2 FERMENTATION
The uniqueness of several microorganisms and their often unpredictable nature
and biosynthetic capabilities, in a specific set of environmental conditions, have
made them ideal candidates, in attempts to solve difficult problems in life sciences
and other fields. Microorganisms have been used in various ways over the past
many decades, to advance medical technology, human and animal health, food
processing, food safety and quality, genetic engineering, environmental protection
and agricultural biotechnology. The use of beneficial microorganisms in the food
sector has been a long tradition, namely lactic acid bacteria and yeasts in
fermentation processes; the former are widely used in the manufacture of fermented
food and are among the best studied microorganisms. The fermentations may be
by yeasts, bacteria, molds or combination of these organisms. Detailed knowledge
of a number of physiological traits has opened novel potential applications for
these organisms in the food industry, while other traits might be specifically, beneficial
for human health.
The food and beverage industry exploits non-pathogenic microorganisms for the
production of fermented foods. These foods are prepared from raw materials and
acquire their characteristic properties by a process that involves microorganisms.
In certain cases the endogenous enzymes of the wild micro flora nature to the raw
material may play a decisive role. It is believed that fermented foods originated
72
from the Orient and date back to the prehistoric times. Originally, these were Beneficial Role of
Micro-organisms
fermented “spontaneously” by autochthonous strains found in the raw materials
or the environment; This was the start of traditional biotechnology. The most
important were cheese, yoghurt, wine, vinegar, beer, bread and the traditional
fungal fermentations used in Asia and Africa, for the production of food.
In the biochemical sense, the term fermentation refers to a metabolic process in

which organic compounds (particularly carbohydrates) are broken down to release
energy without the involvement of a terminal electron acceptor such as oxygen.
Partial oxidation of the substrate occurs so that only a relatively small amount of
ATP energy is released compared to the energy generated if a terminal electron
acceptor is involved. Partial oxidation of a carbohydrate can give rise to a variety
of organic compounds such as alcohols organic acids and acetone. The compounds
produced by micro-organisms vary from organism to organism and are produced
through different metabolic pathways.
The term fermentation can also be applied to any industrial process that produces
a material that is useful to humans and if the process depends on the activity of
one or more micro-organisms (consortia). These processes, known as industrial
fermentations, are usually carried out on a large scale and in vessels in which the
organisms are normally grown in liquid media. Some industrial fermentations are
fermentation in the biochemical sense but the majority of microorganisms involved
are aerobic and use oxygen (as terminal electron acceptor) and thereby metabolize
carbohydrates completely.
A vast range of materials are produced by industrial fermentations. These include
1) Organic chemicals used as fuels, food additives, antibiotics and enzymes for
use in the food and other industries. Vinegar is an example of a food additive
produced by an industrial fermentation.
2) Organisms are produced on a large scale for the extraction of protein (single
cell protein) that can be used as a part of the human diet. Quorn is an
example of a single cell protein, It is produced from the fungus Fusarium
graminearum. This mycoprotein, purified from the fungus, is currently available
for use as a food and is incorporated into a range of dishes that appear on
supermarket shelves. Meatless dishes with a high protein content made from
Quorn are particularly appealing to vegetarians.
3) Yeast cells produced for use in industries such as the baking industry, which
relies on the mass production of large amounts of baker’s yeast.
4) Foods produced on a large scale as a result of the activities of micro-

organism, e.g. cheese, yogurt and bread.
5) Production of alcoholic beverages, e.g. beer and wines.
6) Cellular extracts used as food additives, e.g., yeast extracts from yeast cells
produced as a by product of the brewing industry.
7) Industrial fermentations are now often considered under the heading of

biotechnology, i.e. technology that uses living organisms and their products in
the manufacturing and service industries.
73
Microbiology 4.3 FERMENTED FOODS AND THEIR
IMPORTANCE
Fermented foods are those foods produced by the modification of a raw
material of either animal or vegetable origin by the activities of micro-
organisms. Bacteria, yeast and moulds can be used to produce a diverse range
of products, that differ in flavour, texture and stability from the original raw
material. The production of many fermented foods involves organisms that are
biochemically fermentative. Lactic acid bacteria that ferment carbohydrates to
produce lactic acid are particularly important, but yeasts also play a major role in
some food fermentations, fermenting carbohydrates to produce ethanol and other
organic chemicals. Moulds that do not ferment carbohydrates, also play an essential
role in some food fermentations, for example, the production of cheeses (blue
cheeses) and oriental foods (soy sauce). Fermented foods are an extremely valuable
addition to the human diet for a whole variety of reasons:
1) Increase in variety: Fermented foods increase the variety of foods that are
available, adding to our diet a group of highly nutritious products with unique
characteristics. There are, for example, about 1000 different types of cheeses.
2) Use of ingredients: Fermented foods form an important ingredient for a
wide variety of dishes and are often used to impart special flavours, e.g.
pepperoni in pizzas, yoghurt in curries, cheeses in a whole range of dishes,
including soups, and soy sauce in stir-fry dishes.
3) Improvement in nutritional quality: The fermentation process may improve
the nutritional quality of a raw material. Here are some examples:
• Tempeh fermentation raises the vitamin B12 content of the original
soybean .
• Tapioca fermentation doubles the protein content of cassava and increases
the level of essential amino acids.
• The presence of yeasts in a fermented food will increase the vitamin B
content.
• Antinutritional factors such as phytates, glucosinolates and lectins may
be removed by the fermentation process.
• Fermentation may lead to an increase in the bio-availability of minerals.
These improvements in the nutritional value of raw material will have little effect in
the balanced diets of Western populations. However, for populations that subsist
on diets consisting largely of polished rice, maize or other starches, such as in
Africa and Asia, the contribution that fermented foods make to the intake of B
group vitamins and proteins is highly significant.
4) Preservation: Fermentation often preserves a raw material, improving safety
with regard to food-borne pathogens and increasing shelf-life; compare the
shelf-life of raw milk (only a few days) with the shelf-life of yoghurt (several
weeks).
5) Health benefits: Some fermented foods are said to have definite health
74 benefits, although the scientific evidence for this is limited. Reports suggest
that fermented milk products such as yoghurt can reduce serum cholesterol Beneficial Role of
Micro-organisms
levels and help avoid cancers, particularly, those associated with the colon.
‘Bio’ yoghurts (AB and ABT yoghurts ) are said to have a restorative effect
on a normal micro flora, assisting recovery of a normal balanced flora after
oral antibiotic therapy.
6) Improved digestibility: Some fermented foods are more easily digested
than the original raw material. People who cannot digest lactose properly
(show lactose intolerance) can often consume some types of fermented dairy
products (particularly yoghurts) without harmful effects. Lactose intolerance
is due to the absence of the enzyme galactosidase in digestive juices, which
converts lactose to glucose and galactose. Ingestion of dairy products leaves
unabsorbed lactose in the gut, which is fermented by the normal gut flora
giving flatulence, abdominal pain and diarrhoea. The fermentation of milk
converts the difficult to digest lactose to the more easily digested lactate, and
the galactosidase in live starter culture organisms appears to assist in the
digestion of any residual lactose. Legumes, e.g. soybean, contain
oligosaccharides such as stachyose which are fermented in the gut to yield
gas and the associated socially embarrassing flatus. The oligosaccharides are
broken down to readily digestible monosaccharides and disacccharides during
fermentation of legumes by moulds, thus removing the problem.
Food Safety
- antimicrobials
- bacteriocin
- cyclic
dipeptides
Organoleptic
Nutritional
- texture
- vitamins Starter - aroma
- biogenic amine Cultures - flavour
production
Health Technological
- nutraceuticals - phase resistance
- bioactive peptides - lactose-negative
- accelerated ripening
Fig. 4.1: Functional properties of starter cultures for food fermentation
7) Detoxification of raw materials : The fermentation process may remove

toxic chemicals present in the raw material. Cassava fermentation, for example,
removes a cyanogenic (cyanide producing) glycoside; cassava is toxic if
eaten raw.
75
Microbiology 4.4 FOOD FERMENTATION - SCIENCE AND
TECHNOLOGY
The science of fermentation is known as zymology. Food fermentation involves all
those fermentation processes where either the ultimate product is used directly as
a food, as an additive to food or is a basic ingredient to the food or the by-
product formed during fermentation, food waste utilization, their disposal or proper
management. As a science the food fermentation, has an element of biological
sciences especially the microbiology, genetics and biochemistry, as a technology
the food technology, chemical engineering along with integral component of sciences
involved in food toxicity, acceptability and food nutrition. For any process to
make fermented food, essential components are raw materials, micro-
organism(starter cultures), fermentation vessel and associated controls, processing,
recovery and packing systems, but from fermentation technology point of view,
the fermenter and micro-organism involved assumes prime importance. Since the
end product would serve as a food, it is essential to evaluate it from nutritional
toxicology and sensory quality point of view and thus, the knowledge of these
aspects would form an integral part of food fermentation technology, The
technological dimensions of food fermentation towards the application of engineering
sciences in designing the fermenters and associated controls for optimum
fermentation and product recovery are immense. As the product on the commercial
scale would be marketed, which can occur only if it is economical and therefore
the economics of such products and the associated marketing aspects cannot be
ignored. Similarly, transfer of the knowledge to the scale of technology operation
required is of paramount importance. The scale up operations, harvesting,
biocatalysis of micro-organisms, product recovery, effluent treatment etc. are of
concern to fermentation technologists. Food fermentation technology could be
organized based either on the product or the system used to make the fermented
foods or products derived from it similar to food biotechnology (refer Fig.4.2)
Raw material
Microbial cells
or enzymes
Pretreatment
Medium nutrients
Sterilization
Biocatalyst Substrate and nutrients
(immobilized or free)
Bioreactor
Purification
Products Waste Utilization
Processing treatment
(Thermal, Chemical,
Freezing, Drying etc.)
Packaging By-products
Useful products
Labelling
Marketing
Fig 4.2: A generalized scheme of various operations employed in food
76
fermentation technology
Beneficial Role of
Check Your Progress Exercise 1 Micro-organisms

1) What is fermentation?
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2) What is the purpose of fermentation?
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3) Define zymology.
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4.5 TYPES OF FOOD FERMENTATIONS

A number of different types of food fermentation can be recognized.
4.5.1 Acid Food Fermentation

These include acid fermented dairy products, e.g. cheese, butter, yoghurt and
kefir; acid fermented vegetable products, e.g. sauerkraut, olives and various pickles;
acid fermented meat products, e.g. the semi-dry fermented meats such as cerevelat
and the dry fermented meats such as salami and pepperoni; sour dough breads.
The common feature of all these products is the use of lactic acid bacteria to
carry out the basic fermentation process. Modern production systems usually
involves the use of starter cultures. An exception is the fermentation of sauerkraut
for which the process depends on lactic acid bacteria, that are natural inhabitants
of the surface of cabbage leaves. Sometimes sugar is added to raw material to
allow the lactic acid bacteria to produce sufficient acid for a successful fermentation.
This is the case with fermented meats in which the sugar content of the raw
material is very low. Salt may be added to suppress the growth of the normal
spoilage micro flora and allow the lactic acid bacteria to dominate, e.g. sauerkraut,
pickles and fermented meats. The raw materials may be pasteurized to eliminate
pathogens and suppress natural contaminants that compete with the lactic acid
bacteria, used in the starter culture. 77
Microbiology
4.5.2 Yeast Fermentation
Yeasts are important in food fermentation because of their ability to produce
carbon dioxide and ethanol. Carbon dioxide is the important metabolic product in
the manufacture of leavened bread whereas ethanol is metabolized in the production
of beer, wine and spirits.
4.5.3 Solid State Fermentation
Solid state fermentation involves the use of a solid substrate into which the
fermenting organism is inoculated. The organisms used are often molds. Examples
are the ‘koji’ process and the second stage of tempeh fermentation.
4.6 COMMON EXAMPLES OF FOOD

FERMENTATIONS
A very wide range of inumerable products of the food industry, such as sour
cream, yoghurt, cheese, fermented meat, bread and other bakery products,
alcoholic beverages, vinegar, fermented vegetables and pickles, etc., are produced
through microbial fermentation processes. The efficiency of the strains of the
organisms used, and the processes are being continuously improved to market
quality products at more reasonable costs.
4.6.1 Oriental and Indigenous Fermented Foods
A large number of fermented foods can be grouped under the heading of oriental
and indigenous fermented foods. Fermented foods of this type are produced in
Asia and Africa and are often associated with specific countries or areas. Most of
the products are unknown in the West but frequently have major nutritional role in
the diets of the local population. Lactic acid bacteria are involved in some of the
fermentation but yeast and moulds are often the main organisms responsible.
Many are solid state fermentations or involve fermentations of more than one
type. Some of the products are manufactured on large scale but many are carried
out on a cottage industry or household basis. The major groups of these products
with example/s are given below:
Table 4.1: Oriental and Indigenous Fermented Foods
S. Product Country/ Substrate Micro- Nature and Use
No. Region Organism(s)
Involved
1. KANIMA Nepal, Sikkim, Soyabeans — Solid snack food

& Darjeeling
2. KHAMAN India Bengal gram — Solid, cake like

breakfast food
3. NATTO Northern Soyabeans Bacillus natto Solid cake, as a meat

Japan substitute
4. PAPADAM India Black gram Saccharomyces Solid, crisp,

sp. condiment
5. SOY SAUCE Japan, China, Soybeans & Lactic Acid Liquid drink,
Philippines & Wheat Bacteria, seasoning agent for
Oriental A. oryzae, meat, fish and
countries A. sojae, cereals.
Lactobacillus sp.,
Saccharomyces
78 rouxii
S. Product Country/ Micro- Nature And Use Beneficial Role of
Substrate
Organism(s) Micro-organisms
No. Region
Involved
6. TEMPEH Indonesia Soyabeans Rhizopus sp. Fried in oil, roasted,

and nearby as a meat substitute
regions
7. WARIES India Black gram Candida sp. Spongy, spicy

flour Saccharomyces condiment
sp.
8. JALEBIES India, Nepal Wheat flour Saccharomyces Syrup filled

& Pakistan bayanus confectionary
9. KANJI India Rice & Hansenula Sour liquid added to

Carrots anomala vegetables
10. MISO Japan, China Rice & Aspergillus Paste, soup base
soyabeans oryzae
Torulopsis
etchellsii
Lactobacillus sp.
Saccharomyces
rouxii
11. NAN India Unbleached —- Solid snack food/

Pakistan wheat flour bread
Afghanistan
Iran
12. DHOKLA India Bengal gram —- Spongy condiment

& wheat
13. DOSAI India Black gram & Yeasts Spongy breakfast

rice food
14. IDLI India Black gram & L. mesenteroides Spongy, moist

rice T. candida breakfast food
Trichosporon
pullulans
15. TORANI India Rice H. anomala Seasoning for

C. guilliermondii vegetables
C. tropicalis
G. candidum
4.6.2 Fermented Vegetable Foods

a) Sauerkraut
Sauerkraut is fermented fresh cabbage product. It is popular in USSR and Europe.
The main organism involved in the fermentation of this pickle is lactic acid bacteria,
Leuconostoc mesenteroides followed by Lactobacillus plantarum.
b) Cucumber pickle
Cucumber pickle is a fermentation product of fresh cucumbers. Several lactic acid
bacteria are involved in preparation of this pickle. Lactobacillus plantarum is
the most important organism required for fermentation of cucumber pickle.
4.6.3 Fermented Soyabean Products
a) Tempeh
Tempeh is a highly popular soyabean preparation in Indonesia. The chief organism
in this preparation is the mold Rhizopus oligosporus. The boiled soybean seeds 79
Fundamentals of Food are mashed and wrapped in banana leaves or kept in boxes or hollow tubes. It is
Microbiology
inoculated with spores of tempeh fungus by addition of a portion of previous
batch and allowed to ferment for about 20 hours at a temperature 32oC until there
is a good growing mycelium but little sporulation. It is then sliced and prepared as
per the taste such as roasting or frying. The taste of the tempeh is considered to
be bland but it is highly nutritious.
b) Soya sauce
Soya sauce is a very popular preparation of Japan, which has received wide
acceptance world over. This is prepared by inoculating Aspergillus oryzae (A.
Soyae) in a mixture of soaked and steamed soy bean with roasted wheat in the
ratio of 2:1. The mixture is incubated at 25-300C for a period of 3 to 5 days.
Subsequently, it is subjected to various processing steps using bacterium
Lactobacillus delbruckii and the yeast Saccharomyces rouxii. After 3 months,
the final product is filtered, pasteurized and bottled for use.
c) Miso
It is made from fermented soyabeans and is a thick paste-like substance. Miso is
brownish in color and tastes extremely salty and tangy on its own. Saccharomyces
rouxii and Torulopsis are the yeasts and Pediococcus halophilus and Streptococcus
faecalis are the bacteria, principally involved in Miso fermentation. While the most
common use of miso is in Japanese-style miso soup recipes, miso also adds a
unique burst of flavour to salad dressings, sauces and marinades, baked tofu, or
vegetable dishes.
4.6.4 Fermented Dairy Products

The fermented dairy products assume greater importance in the human diets as
invariably most of the diet consist of milk products especially the cheese, butter,
yoghurt, curd etc. There are a number of fermented dairy products as shown
below:
Table 4.2: Types of Fermented Dairy Products
Name Country of Milk types, Micro flora
origin conditions
Dahi India Milk (Cow/ L. lactis subsp. lactis, S. salivarius

(Dadhi) Persia Buffalo) subsp. thermophilus, L. delbrueckii
subsp. bulgaricus, plantarum, lactose
fermenting yeast, Mixed culture
Srikhand India Milk (Cow/ S. salivarious subsp.thermophilus, L.

(chakka) Buffalo) delbrueckii subsp. bulgaricus.
Lassi India Milk (Cow/ S. salivarius subsp. thermophilus, L.

Buffalo) delbrueckii subsp.bulgaricus.
Cultured butter Scandinevian Milk (Cow/ L. lactis subsp. lactis, L. lactic subsp.
milk and European Buffalo) diacetylactis, Leuconostoc
countries dextranicum subsp. citrovorum
Acidophilus milk Australia Cow’s milk L. acidophilus
Yoghurt Middle Asia, Cow’s milk S, salivarius subsp.thermophilus, L.

(bio-ghurt) Balkans Goat’s or delbrueckii subsp. bulgaricus,
mixed milk Micrococcus and other lactic acid,
80 bacteria, cocci, yeasts, molds
Beneficial Role of
Name Country of Milk types, Micro flora Micro-organisms
origin conditions
Kefir Caucasus Sheep’s, Cow’s, L lactis subsp. lactis, Leuconostoc

goat’s or mixed spp., L.delbrueckii subsp.
milk, caucassiucu, Saccharomyces kefir,
fermentation in Torula kefir, micrococci, spore
skin bag or in forming bacilli
wooden barrels
Kumiss Asiatic Mare’s, camel’s L. delbrueckii subsp. bulgaricus,

Steppes or ass’s milk Torula kumiss, Saccharomyces
fermentation lactis, micrococci, spore forming
in skin bag bacilli lactis, micrococci, spore
forming bacilli
Leben, Lebanon Goat’s or L. lactis subsp. lactis, S. salivarius

Labneh Arab sheep’s milk, subsp. thermophilus, L. delbrueckii
countries Fermentation in subsp. bulgaricus, lactose
skin bag/ fermenting yeasts
earthenware
4.6.5 Economically Important Fermentation Products

The growth of micro-organisms or other cells results in a wide range of products.
Each culture operation has one or few set objectives. The process has to be
monitored carefully and continuously, to maintain the precise conditions needed
and recover optimum levels of products. Accordingly, fermentation processes
aim at one or more of the following:
a) production of cells (biomass) such as yeasts;
b) extraction of metabolic products such amino acids, proteins (including
enzymes), vitamins, alcohol, etc., for human and/or animal consumption or
industrial use such as fertilizer production;
c) modification of compounds (through the mediation of elicitors or through
bio transformation); and
d) production of recombinant products.
A. Microbial Biomass
Microbial biomass is produced commercially as single cell protein (SCP) using
such unicellular algae as species of Chlorella or Spirulina for human or animal
consumption, or viable yeast cells needed for the baking industry, which was also
used as human feed at one time. Bacterial biomass is used as animal feed. The
biomass of Fusarium graminearum is also produced for a similar use.
B. Microbial Metabolites
i) Primary metabolites
During the log or exponential phase organisms produce a variety of substances
that are essential for their growth, such as nucleotides, nucleic acids, amino acids,
proteins, carbohydrates, lipids, etc., or by-products of energy yielding metabolism
such as ethanol, acetone, butanol, etc. This phase is described as the trop-
phase, and the products are usually called primary metabolites. Commercial
examples of such products are given in Table 4.3.
81
Fundamentals of Food Table: 4.3: Examples of Commercially Produced Primary Metabolites
Microbiology
Primary Metabolite Organism Significance

Ethanol Saccharomyces cerevisiae alcoholic beverages
Kluyzveromyces fragilis
Citric acid Aspergillus niger food industry
Acetone and Clostridium
butanol acetobutyricum solvents
Lysine Corynebacterium nutritional additive
Glutamic acid glutamacium flavour enhancer
Riboflavin Ashbya gossipii nutritional
Eremothecium ashbyi
Vitamin B12 Pseudomonas nutritional
denitrificans
Propionibacterium
shermanii
Dextran Leuconostoc mesenteroides industrial
Xanthan gum Xanthomonas campestris industrial
ii) Secondary metabolites

Organisms produce a number of products, in addition to the primary metabolites.
The microbial growth phase, during which products that have no obvious role in
metabolism of the synthesizing culture organisms are produced, is called the
idiophase, and these products are called secondary metabolites. In reality, the
distinction between the primary and secondary metabolites is not a straight jacket
situation. Many secondary metabolites are produced from intermediates and end
products of secondary metabolism. Some like the Enterobacteriaceae do
not undergo secondary metabolism. Examples of secondary metabolites are
given in Table 4.4.
Table 4.4: Examples of Commercially Produced Secondary Metabolites
Metabolite Species Significance
Penicillin Penicillium chrysogenum antibiotic
Erythromycin Streptomyces erythreus antibiotic
Streptomycin Streptomyces griseus antibiotic
Cephalosporin Cephalosporium acrimonium antibiotic
Griseofulvin Penicillium griseofulvin antifungal antibiotic
Cyclosporin A Tolypocladium inflatum immunosuppressant
82
iii) Production of Enzymes Beneficial Role of
Micro-organisms
Industrial production of enzymes is needed for the commercial production of food
and beverages. Enzymes are also used in clinical or industrial analysis and now
they are even added to washing powders (cellulase, protease, lipase). Enzymes
may be produced by microbial, plant or animal cultures. Even plant and animal
enzymes can be produced by microbial fermentation through techniques of genetic
manipulations. While most enzymes are produced in the tropophase, some like
the amylases (by Bacillus stearothermophilus) are produced in the idiophase,
and hence are secondary metabolites. Examples of enzymes produced through
fermentation processes are given in Table 4.5.
Table 4.5: Examples of Commercially Produced Enzymes
Organism Enzyme
Aspergillus oryzae Amylases
Aspergillus niger Glucoamylase
Trichoderma reesii Cellulase
Saccharomyces cerevisiae Invertase
Kluyzveromyces fragilis Lactase
Saccharomycopsis lipolytica Lipase
Aspergillus species Pectinases and proteases
Bacillus species Proteases
Mucor pusillus Microbial rennet
Mucor meihei Microbial rennet

1) What are the various types of fermentations?
............................................................................................................
............................................................................................................
............................................................................................................
2) Give one example of each fermented food type-
Oriental, Indian, Caucasus and Lebanon.
............................................................................................................
............................................................................................................
............................................................................................................
83
Fundamentals of Food 3) What are the major groups of commercially important fermentations?
Microbiology
............................................................................................................
............................................................................................................
............................................................................................................
............................................................................................................
4.7 FERMENTED FOODS AS FUNCTIONAL

FOODS
Dr. Elias Metchnikoff, a Nobel Laureate, around 100 years ago proposed that
“Lactic acid bacteria can render a great service in the fight against intestinal
putrefaction” and might “ postpone and ameliorate old age”. This concept
was developed further through the decades, and today, the emerging probiotics,
prebiotics and synbiotics era is a subject of scientific debate and intense research.
The use of probiotics, prebiotics, and synbiotics is a promising area for the
development of functional foods.
Functional foods are generally characterized as foods similar in appearance to

conventional foods, consumed as part of a usual diet, and providing health-related
benefits beyond meeting basic nutritional needs. A food can be considered
naturally ‘functional’ if it contains a food component that affects one or
more targeted functions in a beneficial way. Dairy foods can be included in
the functional food category because of their content of calcium, specific health-
enhancing proteins, conjugated linoleic acid, sphingolipids, butyric acid, and probiotic
cultures.
Dairy foods appear to be the preferred medium for introducing probiotic bacteria
such as human-derived species of lactic acid bacteria (e.g., L. acidophilus,
L. casei, L. gasseri, L. rhamnosus, L. reuteri, Bifidobacterium bifidum,
B. breve, B. infantis and B. longum). Lactobacillus spp. (naturally found in the
human small intestine) and various Bifidobacterium spp. (a major organism in the
human large intestine) are the most commonly used probiotic cultures.
4.7.1 Probiotics
The term Probiotic means “for life”, is derived from the Greek language. It is now
defined as “live microorganisms which when administered in adequate amounts
confer a health benefit on the host”. The most common probiotic organisms
clinically useful include members of the bacteria such as Lactobacillus and
Bifidobacteria species and some selected strains of Streptococcus, Lactococcus
and members of yeast and molds such as Saccharomyces, Aspergillus, Acanthosis
and Candida species.
4.7.2 Prebiotics
The term Prebiotic is defined as “short chained carbohydrates that are indigestible
by human enzymes in the GIT (Gastro-intestinal tract) and selectively stimulate the
growth and activity of specific species of bacteria in the gut, usually bifidobacteria
84 and lactobacilli, with benefits to health”. The most commonly used prebiotics in
supplements are Fructo-oligosaccharides (FOS). Bifidobacteria, due to the Beneficial Role of
Micro-organisms
presence of beta-fructofuranosidase enzyme are liable to break down and utilize
FOS. This helps in stimulation of bifidobacterium growth in the GIT. FOS exhibits
nutritional properties on colonic pH and stool bulking. It also increases bioavailability
of essential minerals and decreases serum triglycerides.
The other types of prebiotic substrates include-
• Xylitol, Sorbitol, Mannitol
• Disaccharides-Lactulose, Lactilol
• Oligosaccharides-Raffinose, Soybean, Palatinose, Isomaltose, Lactosucrose
• Polysaccharides-In-ulin, resistant starch
4.7.3 Synbiotics
The term synbiotic is used when a product contains both probiotics and prebiotics.
Since the word alludes to synergism, this term should be reserved for products in
which the prebiotic compound selectively favors the probiotic compound, e.g.,
FOS in combination with strains such as Bifidobacterium B. infantis, B. longum
etc. Combining probiotics with prebiotics could improve the survival of the bacteria
crossing the upper part of the GIT, thus enhancing their effects in the large bowel.
Moreover, the local and the systemic beneficial effects of probiotics and prebiotics
might be additive or even synergistic.
4.7.4 Use of Probiotics

As per definition of probiotics the live microorganisms that are administered have
to be in adequate amounts to confer a health benefit to the host. Clinical bodies
have used probiotic levels of 1 billion to 10 billion or above.
Dosage
According to Earl Mindell, an internationally recognized expert on nutrition, healthy

person should take 2-5 billion CFUs (colony stimulating units) of probiotics a day
and those with GI conditions can take up to 10 billion CFUs per day. In acute
infectious diarrhoea, lactobacillus is most effective at a dose of 10 billion CFUs
during the first 48 hours, which translates to 5 billion CFUs per day. For prescription
probiotics, the current daily intake recommended is 5-10 billion CFUs per day.
Capsules and Sachets of Probiotic plus Prebiotic combination (Pro-wel) and
Probiotic alone (Darolac) are commercially available. Benefits offered by Probiotic
and Prebiotic combination formula are :-
• Maximum Colony Forming Units (CFUs): ensure complete action.
• Fructo-oligosaccharide (FOS): offers nutrition to the probiotics and normal

intestinal flora.
• Acid-resistant cells: reach intestine in full force.
• Freezed-dried and nitrogen-flushed cells: offers excellent stability.
• Vegetable capsules: ensure universal appeal.

85
Microbiology
4.7.5 Health Benefits of Probiotics
Few well designed, well conducted human clinical trials of probiotics have been
conducted over the past 30 years. Only in recent years, has the importance of
choosing probiotic strains of demonstrated efficacy been recognized. With respect
to health benefits of probiotics, research studies indicate the following:
1) Improved intestinal health.
2) Modulation of the immune response and immune function .
3) Reduced risk of cancer.
4) Reduced risk of heart disease.
5) Improved tolerance to milk.
6) Reduce food allergies.
7) Reduce ulcers by decreasing the growth of ulcer-inducing bacteria (i.e.,

Helicobacter pylori).
Beneficial
modulation
of intestinal
Alleviation microbial
o f lactose flora
Lowering
intolerance biomakers
symptoms (harmful
faecal
enzymes)
Treatment
of viral and Established Immune
antibiotic Effects modulation
associated Immune
diarrhoea modulation
Treatment
of infant Alleviation
gastro- of atopic
enteritis dermatitis
Positive symptoms
effects on in children
bladder and
cervical
cancer
86 Fig. 4.3: Beneficial health effects claimed for probiotics

Beneficial Role of
Check Your Progress Exercise 3 Micro-organisms

1) What is a probiotic?
............................................................................................................
............................................................................................................
............................................................................................................
2) What are the ideal properties of a probiotic?
............................................................................................................
............................................................................................................
............................................................................................................
3) Define a prebiotic.
............................................................................................................
............................................................................................................
............................................................................................................
4.8 LET US SUM UP

Fermentation is a relatively efficient, low energy preservation process which
increases the shelf life and decreases the need for refrigeration or other forms of
food preservation technologies. It is therefore a highly appropriate technique for
use in developing countries and remote areas where access to sophisticated
equipment is limited. Fermented foods are popular throughout the world and in
some regions, make a significant contribution to the diet of millions of individuals.
In Asia the preparation of fermented foods is a widespread tradition. The fermented
products supply protein, minerals and other nutrients that add variety and nutritional
fortification to otherwise starchy and bland diets. For instance Soy sauce is
consumed throughout the world and is a fundamental ingredient in diets from
Indonesia to Japan. Over one billion litres are produced each year in Japan alone.
Although fermentation of foods has been in use for thousands of year, it is likely
that the underlying microbial and enzymatic processes responsible for the
transformations were largely unknown. It is only recently that there has been a
development in the understanding of these processes and their adaptation for
commercialization. There is tremendous scope and potential for the use of micro-
organisms towards meeting the growing world demand for food, through efficient
utilization of available natural food and feed stocks and the transformation of
waste materials. Genetic improvement of the organism is fundamental to the success
of fermentation technology. Mutation and recombination are the two ways to
meet this end.
The alarming increase in inappropriate use of antibiotics and enhanced bacterial
resistance, along with renewed interest in ecological methods to prevent infections
makes probiotics, prebiotics and synbiotics a very interesting field for research.
87
Fundamentals of Food Evidence is beginning to accumulate, describing their beneficial effects in a variety
Microbiology
of GI (Gastro Intestinal) and non-GI disorders. They offer dietary means to
support the balance of the intestinal flora. They may be used to counteract local
immunological dysfunction, to stabilize the intestinal barrier function, to prevent
infectious succession of pathogenic microorganisms and to influence intestinal
metabolism. Looking ahead, this field holds immense promise for the future in
delivering novel therapies in different fields.
4.9 KEY WORDS

Carbohydrates : Organic compounds that consist of carbon, hydrogen and
oxygen. They vary from simple sugars containing from three
to seven carbon atoms to very complex polymers.
CFUs : Colony forming units.
Enzyme : A protein that acts as a catalyst. Every chemical reaction in
living organisms is facilitated by an enzyme.
Fermentation : Enzymatic breakdown (catabolism) of carbohydrates
generally in the absence of oxygen or changes in food
caused by intentional growth of bacteria, yeast or mold.
Fructo-oligosac- : A type of prebiotic/probiotic found in Jerusalem artichokes,
charides (FOS) shallots and onion powder which may improve
gastrointestinal health.
Food : Those substances that are eaten or otherwise taken in the
body to: sustain psychological and physiological life support
growth and repair and replacement of tissues; and provide
energy and nutrition. In essence, the sum of all the processes
concerned with the growth, maintenance and repair of the
body and/or its organs and systems.
Food processing : Using food as a raw material and changing it in some way
to make a food product.
Gastro intestinal : Pertaining to the stomach, small and large intestines, colon,
rectum, liver, pancreas, and gallbladder.
Yeast : Single cell organism, which as it grows converts
carbohydrates by fermentation into alcohol and carbon
dioxide.
4.10 ANSWERS TO CHECK YOU PROGRESS

EXERCISES
Your answer should include the following points:
1) In the biochemical sense, the term fermentation refers to a metabolic process
in which organic compounds (particularly carbohydrates) are broken down
to release energy without the involvement of a terminal electron acceptor
88 such as oxygen.
2) Food fermentation serves five main purposes: Beneficial Role of
Micro-organisms
• Enrichment of the diet through development of a diversity of flavors,
aromas, and textures in food substrates.
• Preservation of substantial amounts of food through lactic acid, alcohol,
acetic acid and alkaline fermentations.
• Biological enrichment of food substrates with protein, essential amino
acids, essential fatty acids, and vitamins.
• Detoxification during food-fermentation processing.
• A decrease in cooking times and fuel requirements.
3) The science of fermentation is known as zymology.
1) The various types of food fermentations are acid, yeast and solid state
fermentation.
2) An example of each fermented food type-
• Oriental- Tempeh
• Indian- Idli
• Caucasus-Kefir
• Lebanon- Leben
3) The major groups of commercially important fermentations are:
• Microbial cells or biomass as the product, e.g. bakers yeast, lactobacillus
etc.
• Microbial enzymes: catalase, amylase, protease, pectinase, glucose isomerase,
cellulase, hemicellulase, lipase, lactase, streptokinase, etc.
• Microbial metabolites : Primary metabolites – ethanol, citric acid, glutamic
acid, lysine, vitamins, polysaccharides etc. Secondary metabolites: all antibiotic
fermentation
1) Probiotics are dietary supplements containing potentially beneficial bacteria
or yeasts. According to the currently adopted definition by FAO/WHO,
probiotics are: ‘Live microorganisms which when administered in adequate
amounts confer a health benefit on the host’.
2) Probiotics supplements are considered ideal, if they fulfill the following
important characteristics: normal gut inhabitant, nonpathogenic, genetically
stable, resistant to acid, capable of survival, proliferation and metabolic activity
at the target site, high stability, viability at high populations and able to exert
clinically documented health benefits
89
Fundamentals of Food 3) Prebiotics are a category of functional food, defined as non-digestible food
Microbiology
ingredients that beneficially affect the host by selectively stimulating the growth
and/ or activity of one or a limited number of bacteria in the colon, and thus
improve host health.

Frazier, W.C. and Westoff, D.C. (1996). Food Microbiology, Tata McGraw Hill
Publishing Co. Ltd., New Delhi.
Pelczar, M. Jr., Chan, E.C.S. and Kreig, N.R. (1993). Microbiology, Tata McGraw
Hill Inc., New York.
Garbutt, J. (1998). Essentials of Food Microbiology, Arnold International Student’s
Edition, London.
90

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Introduction to Food

UNIT 1 INTRODUCTION TO FOOD Microbiology

1.7 Important Micro-organisms in Food

1.8 Normal Microflora of some Common Foods

1.2 THE SCIENCE OF MICROBIOLOGY

1.3 FOOD MICROBIOLOGY - ITS ORIGIN AND

Topics of major interest

Fig. 1.1: Topics of major interest to the food microbiologist 9

Food-borne disease Food-borne disease

Bacteria Moulds Production of single cell protein

Production of food additives and enzymes Production of enzymes

Food spoilage Food-borne disease

Yeasts Food fermentations Viruses Identification of food

Production of food additives and enzymes

1.4 IMPORTANCE OF MICRO-ORGANISMS IN

Since 1900 A.D. our understanding of the importance of micro-organisms in

1.4.1 Food-borne Diseases

Many pathogenic micro-organisms (bacteria, molds and viruses) can

1.4.2 Food Spoilage

1.4.3 Food Bioprocessing

1.4.4 Food Additives

1.4.5 Food Biopreservation

1.5 CLASSIFICATION AND NOMENCLATURE

Fig. 1.3: Types of bacterial cells and their groupings 13

Check Your Progress Exercise 2

1.7 IMPORTANT MICRO-ORGANISMS IN FOOD

Botrytis Cladosporium Rhizopus Alternaria

Fusarium Asporgillus Pencillium

Fig. 1.4: Common mold species found in foods

1.7.2 Important Yeast Genera

1 ) Saccharomyces: Cells are round, oval, or elongated. It is the most

3 ) Rhodotorula: They are pigment (red, pink or yellow) forming yeasts

1.7.3 Important Viruses

Spiral and curved bacteria Spirallaceae Campylobacter

Gram-negative aerobic Pseudomonadaceae Pseudomonas, Altermonas,

Halobacteriaceae Halobacterium, Halococcus

Genera of uncertain affinity Alcaligenes, Acetobacter,

Gram-negative facultative Enterobacteriaceae Escherichia, Citrobacter,

Vibrionaceae Vibrio, Aeromonas

Genera of uncertain affinity Flavobacterium,

Gram-negative diplococci Neisseriaceae Moraxella, Acinetobacter

Endospore forming rods Bacillaceae Clostridium, Bacillus

Gram-positive Lactobacillaceae Lactobacillus

Genera of uncertain affinity Listeria

Rickettsia Rickettsiaceae Coxiella

1.7.4.1 Common Bacterial Groups in Foods

1.8 NORMAL MICRO FLORA OF SOME

1.8.5 Shellfish and Finfish

Check Your Progress Exercise 3

1.9 LET US SUM UP

1.10 KEY WORDS

1.11 ANSWERS TO CHECK YOUR PROGRESS

3) On basis of morphology bacteria can be classified as:

1.12 SUGGESTED READING

2.2 Food Contamination

2.3 Food Spoilage

2.4 Role of Micro-organisms

2.5 Factors Affecting Spoilage

2.6 Deteriorative Effect of Micro-organisms

2.7 Different Types of Spoilage

2.2 FOOD CONTAMINATION

2.3 FOOD SPOILAGE

The criteria for assurance in foods suitable for consumption are: