Assignment On: Bio-preservation

Submitted by : Md Mohon Farazi

Student Number : 201922800080
Subject : Advanced Food Microbiology
Major : Food Science and Engineering
Degree : Masters Student
Submitted to : Nor Ainy Mahyudin

December 16, 2019

Table of Contents:

Title Page
Introduction
What is Bio-preservation?
Biological methods for food preservation
Natural antimicrobials for food bio
preservation
What is Bacteriocins?
Common bacteriocins and their possible
uses in food industry.
What is Probiotic?

Characteristics of probiotics

Probiotics health benefits

Food applications of probiotics

Probiotics application challenges

Conclusion

Referencing and Plagiarism

Introduction

The foods of animal origin are highly perishable due to high nutritional content, moisture
and neutral pH. These foods require proper preservation to maintain quality and safety.
Failing which leads to human illnesses and disease outbreaks. These food borne
illnesses are serious and costly public health concern worldwide. So to maintain the
quality and safety of foods various measures are generally adopted in food industry i.e.
good manufacturing practices, good hygienic practices tec. But preservation of food by
a suitable means is the key of food quality and safety. There are number of preservation
techniques started from low temperature preservation like refrigeration, freezing etc.
and thermal preservation techniques like pasteurization, sterilization and preservation
using certain chemicals. Now a day modern preservation techniques like bio-
preservation, irradiation and hurdle technologies are also common. In traditional
preservation techniques food alters their status and loss some nutrients. So the modern
techniques are more suitable to achieve the safety and quality of foods. It is also
relevant in today world due to globalization of food market, introduction of novel foods,
innovations of new technologies, demand of minimally processed products with ready–
to-eat quality and longer shelf life. Among all the preservation techniques adopted now
bio-preservation is more reliable from ‘farm to fork’ concept. It is capable to enhance
shelf life with great quality, hygienic status with minimal nutritional and organoleptic
losses.

Bio-preservation

Bio-preservation is a technique of food preservation in which antimicrobial potential of

naturally occurring organisms and their metabolites are exploited. It is capable to
harmonize and rationale the necessary safety standards with traditional means of
preservation and modern demand of the food safety and quality. The bio-preservation
techniques of various foods are mainly relying on the quality of biological antimicrobial
systems such as lactic acid bacteria (LAB) and/or their bacteriocins, bacteriophages
and bacteriophage-encoded enzymes. They are widely used in food industry to get a
typical texture and flavour of the food products. They are helpful in maintaining the
quality and safety of foods and are common bio-preservatives in industrialized world.

Biological methods for food preservation

The use of micro-biota and/or antimicrobials in shelf life enhancement of foods is a new
branch of science. The fermentation is a typical example of this process in which
microbes are grown naturally or by addition. Fermentation process produces numbers
of beneficial products with the bacteria which helps in reduction of food spoilage and
renders the food free from pathogenic microorganisms and metabolites. This organism
used for the purpose is LAB and their compounds like organic acids which are capable
to exert antimicrobial properties as well as imparts unique flavor and texture to the food
products. In industrialized world about 60 percent food items are subjected to
fermentation to assure homogeneity, quality, and safety of products. They prefer to use
native micro-biota under controlled environment to get typical texture and flavor for a
specific product. The major use of micro-biota in food industry is in dairy industry but
they can also be exploited in meat and vegetable products. Bio-preservatives eligible for
use in food must fall under the category of generally recognized as safe with no
pathogenic and toxic effect on food. Biological agents used in food may be categorized
as starter cultures and protective cultures. Starter cultures are generally used group of
microorganisms which are used to initiate the fermentation and helps in production of
certain compounds responsible for typical texture and flavoring compounds in
fermented products. The protective culture is mainly used to control the antimicrobial
activity and reduces the survival and growth of pathogenic micro-organisms in foods.
However, combination of both starter and protective culture is generally preferred for
use in food industry.
Natural antimicrobials for food bio-preservation

Antimicrobials Source Food bio-preservation

Decrease the pH of surrounding environment,
creating a selective barrier against non-
Main end products of acidophiles. Lactic acid exerts antimicrobial
Organic acids
fermentation. effect by disruption of the cytoplasmic
membrane and interference with membrane
potential.

Heterofermentative
LAB produces Creation of an anaerobic environment and
CO2 CO2 as a byproduct antagonistic effects specifically against aerobic
of sugar bacteria and produce carbonic acid.
fermentation.

Antibacterial activity against Listeria,

Diacetyl (2,3- LAB as a by-product
Salmonella, Escherichia coli, Yersinia, and
butanedione) of metabolic activity
Aeromonas.
Antimicrobial
substances Produces by LAB in

derived from presence of oxygen Antibacterial effect through oxidative damage

bacterial cell Hydrogen peroxide and action of of proteins and increase of membrane

metabolism flavoprotein oxidases permeability.

or NADH peroxidase.

Low molecular-
weight antimicrobial
Antimicrobial activity against bacteria as well
compound produced
Reuterin as yeasts and molds by inhibiting DNA
by Lactobacillus
synthesis.
reuteri and some
other LAB.

Reutericyclin acts as a proton ionophore and

dissipation of the proton motive force against
Reutericyclin- gram-positive bacteria
Reutericyclin producing strains of including Lactobacillus spp., Bacillus
LAB. subtilis, Bacillus cereus, Enterococcus
faecalis, Staphylococcus aureus and Listeria
innocua.

Antifungal Natamycin (pimaricin) Produced Broad-spectrum antifungal bio-preservative for

 Antimicrobials Source Food bio-preservation
foods and beverages by binding irreversibly to
the cell membrane of fungi and causes
from Streptomyces
compounds membrane hyper permeability leading to rapid
natalensis
leakage of essential ions and peptides and
ultimately cell lysis.

Antimicrobials Naturally found as

from Animal part of defense Generally recognized as safe (GRAS) for direct
Lysozyme
Sources system of living addition to foods.
organisms.

Antimicrobial activity due to its iron-binding

capacity and polycationic nature against a wide
Natural protein found range of bacteria including food borne
Lactoferrin in milk and other pathogens like Carnobacterium, Listeria
secretions. monocytogenes, Escherichia coli, Klebsiella
and viruses (Lönnerdal, 2011; Gyawali and
Ibrahim, 2014).

Antimicrobial system
Lactoperoxidase that originated from Effective against gram-negative bacteria.
milk.

Produced by
Ovotransferrin hydrolysis of natural Inhibits bacterial growth due to iron deprivation.
proteins.

Broad antimicrobial activity against gram-

Naturally present in
positive bacteria, gram-negative bacteria, and
spermatic cells of
Protamine fungi. Used as preservative in wide variety of
fish, birds and
foods ranging from confection items to fruits
mammals.
and rice.

Present in myeloid
Antimicrobial activity against several food
cells and mucosal
borne bacteria such as L.
Pleurocidin tissues of many
monocytogenes and E. coliO157:H7, and
vertebrates and
pathogenic fungi.
Invertebrates.

Chitosan Polycationic Considered as safe food additive and has

biopolymer naturally antibacterial activity against both Gram-positive
present in and Gram-negative bacteria such
 Antimicrobials Source Food bio-preservation
as Staphylococcus aureus, Listeria
monocytogenes, Bacillus cereus, E.
exoskeletons of coli, Shigella dysenteria, and Salmonella
crustaceans and typhimurium (Gyawali and Ibrahim, 2014).
arthropods. Used in biodegradable edible coatings, singly
or dosed with other antimicrobial substances
(Elsabee et al., 2013).

Essential oils such as The concentration of 0.05–0.1% of essential

Plant material like
saponins, flavonoids, oils has demonstrated activity against
Antimicrobials flowers, buds, seeds,
carvacrol, thymol, pathogens, such as Salmonella
derived from leaves, twigs, bark,
citral, eugenol, linalool, typhimurium, E. coli O157:H7, L.
plants herbs, wood, fruits
terpenes, and their monocytogenes, B. cereus and S. aureus in
and roots.
precursors food systems.

Table 1: Natural antimicrobials for food bio-preservation

Bacteriocins

Bacteriocins are bacterial ribosomally synthesized peptides or proteins with

antimicrobial activity. Initially bacteriocins are mainly described to the colicins which are
relatively large proteins of up to 80 kDa and were primarily obtained from E. coli. They
were capable to kill very closely related bacteria upon binding to the inner membrane or
other cytosolic targets.
Common bacteriocins and their possible uses in food industry

Bacteriocin Food application

Dairy Nisin Clostridium butulinum in cheese, L.

industry monocytogenes in cheeses such as
Camembert, Ricotta and Manchego

Pediocin AcH Effective against milk and Cheddar

and Munster cheeses against L.
monocytogenes, S. aureus and E.
coli O157:H7 lacticin against
undesirable LAB. L.
monocytogenes and B. cereus in
Cheddar, Cottage cheese and
yogurt and enterocin AS-48
against B. cereus, S. aureus and L.
monocytogenes in milk and
Manchego cheese.

Meat Nisin, Enterocin AS-48, Enterocins Quite effective against L.

industry A and B, Sakacin, Leucocin A and monocytogenes and other
especially Pediocin PA-l/AcH alone pathogens.
or in combination with several
physicochemical treatments like
modified atmosphere packaging,
high hydrostatic pressure (HHP),
heat and chemical preservatives

Bacteriocinogenic LAB Bio-protective cultures to protect

 Bacteriocin Food application

pathogens in food processing.

Pediocin PA-l/AcH It is more suitable for use in meat

and meat products than nisin but P.
acidilactici is not an indigenous
meat strain.

Vegetable Nisin In tinned vegetables and fruit juices.

products

Pediocin PA-1/AcH In salad and fruit juices.

Enterocin AS-48 Effective against B. cereus in rice

and vegetables and against
pathogens such as E.
coli O157:H7, S. aureusand the
spoilage bacterium Alicyclobacillus
acidoterrestris.

Fish Combination of nisin and Microgard Gram-negative microorganisms

products generally encountered in fresh
chilled salmon and L.
monocytogenes in frozen thawed
salmon.

Bacteriocins culture In inhibition of L. monocytogenes in

containing Carnobacterium
 Bacteriocin Food application

divergens culture in combination rainbow trout.

with lactic acid, sodium chloride,
and/or nisin.

Table 2: Common bacteriocins and their possible uses in food industry

Probiotic
Probiotics are live microbes that can be formulated into many different types of
products, including foods, drugs, and dietary supplements. Probiotic is a relatively new
word that is used to name the bacteria associated with the beneficial effects for the
humans and animals. The term probiotic means ‘‘for life’’ and it was defined by an
Expert Committee as ‘‘live microorganisms which upon ingestion in certain numbers
exert health benefits beyond inherent general nutrition’’.

Characteristics of probiotics
Characteristics of probiotics will determine their ability to survive the upper digestive
tract and to colonize in the intestinal lumen and colon for an undefined time period.
Probiotics are safe for human consumption and no reports have found on any
harmfulness or production of any specific toxins by these strains. In addition, some
probiotics could produce antimicrobial substances like bacteriocins. Therefore, the
potential health benefit will depend on the characteristic profile of the probiotics. Some
probiotic strains can reduce intestinal transit time, improve the quality of migrating motor
complexes [9], and temporarily increase the rate of mitosis in enterocytes.

Probiotics health benefits

Human health
Several studies have documented probiotic effects on a variety of gastrointestinal and
extraintestinal disorders, including prevention and alleviation symptoms of traveler’s
diarrhea and antibiotic associated diarrhea, inflammatory bowel disease, lactose
intolerance, protection against intestinal infections, and irritable bowel syndrome. Some
probiotics have also been investigated in relation to reducing prevalence of atopic
eczema later in life, vaginal infections, and immune enhancement, contributing to the
inactivation of pathogens in the gut, rheumatoid arthritis, improving the immune
response of in healthy elderly people and liver cirrhosis. Some probiotic preparations
have been used to prevent diarrhea caused by antibiotics, or as part of the treatment for
antibiotic-related dysbiosis. Although there is some clinical evidence for the role of
probiotics in lowering cholesterol but the results are conflicting. Probiotics have a
promising inhibitory effect on oral pathogens especially in childhood but this may not
necessarily lead to improved oral health. Antigenotoxicity, antimutagenicity and
anticarcinogenicity are important potential functional properties of probiotics, which have
been reported recently. Observational data suggest that consumption of fermented dairy
products is associated with a lower prevalence of colon cancer, which is suggested that
probiotics are capable of decreasing the risk of cancer by inhibition of carcinogens and
pro-carcinogens, inhibition of bacteria capable of converting pro-carcinogens to
carcinogens.
Animal health

Probiotics which are traditional idea in the human food have been extended to animals
by developing fortified feed with intestinal micro biota to benefit the animals. The micro
flora in the gastrointestinal tracts of animals plays a key role in normal digestive
processes and in maintaining the animal’s health. Probiotics can beneficially improve
the intestinal microbial balance in host animal. Commercial probiotics for animal use are
claimed to improve animal performance by increasing daily gain and feed efficiency in
feedlot cattle, enhance milk production in dairy cows, and improve health and
performance of young calves and in improving growth performance of chickens.
Probiotics can attach the mucosal wall, adjust to immune responses, and compete the
pathogenic bacteria for attachment to mucus. Probiotics provide the animal with
additional source of nutrients and digestive enzymes. They can stimulate synthesis
vitamins of the B-group and enhancement of growth of nonpathogenic facultative
anaerobic and gram positive bacteria by producing inhibitory compounds like volatile
fatty acids and hydrogen peroxide that inhibit the growth of harmful bacteria enhancing
the host’s resistance to enteric pathogens . Finally, probiotics can inhibit pathogens by
competition for a colonization sites or nutritional sources and production of toxic
compounds, or stimulation of the immune system.

Plant health

The more beneficial the bacteria and fungi are, the more “fertile” the soil is. These
microorganisms break down organic matter in the soil into small, usable parts that
plants can uptake through their roots. The healthier the soil, the lower need for synthetic
herb or pesticides and fertilizers. The concept that certain microorganisms ‘probiotics’
may confer direct benefits to the plant acting as bio-control agents for plants. The plant
probiotic bacteria have been isolated and commercially developed for use in the
biological control of plant diseases or bio-fertilization. These microorganisms have
fulfilled important functions for plant as they antagonize various plant pathogens, induce
immunity, or promote growth. The interaction between bacteria and fungi with their host
plants has shown their ability to promote plant growth and to suppress plant pathogens
in several studies.

Food applications of probiotics

The presence of probiotics in commercial food products has been claimed for certain
health benefits. This has led to industries focusing on different applications of probiotics
in food products and creating a new generation of ‘probiotic health’ foods.

Dairy-based probiotic foods

Probiotics can be found in a wide variety of commercial dairy products including sour
and fresh milk, yogurt, cheese, etc.

Drinkable fresh milk and fermented milks

Among probiotics carrier food products, dairy drinks were the first commercialized
products that are still consumed in larger quantities than other probiotic beverages.
Among the probiotic bacteria used in the manufacture of dairy beverages, L.
rhamnosus GG is the most widely used. 

Yogurt- Yogurt is one of the original sources of probiotics and continues to remain a
popular probiotic product today. Yogurt is known for its nutritional value and health
benefits. Yogurt is produced using a culture of L. delbrueckii subsp. bulgaricus and
Streptococcus salivarius subsp. thermophilus bacteria.

 Cheese- Yogurt and milk are the most common vehicles of probiotics among dairy
products. However, alternative carriers such as cheese seem to be well suited.

Other dairy based products- Other dairy products including quark, chocolate
mousse, frozen fermented dairy desserts, sour cream, and ice cream can be good
vehicles of probiotics.

 
Probiotics application challenges

From a technological standpoint, Champagne has listed many challenges in the

development of a probiotic food product including: strain selection, inoculation, growth
and survival during processing, viability and functionality during storage, assessment
the viable counts of the probiotic strains particularly when multiple probiotic strains are
added and when there are also starter cultures added, and the effects on sensory
properties.

Conclusion

Overall bio-preservation of food by bacteriocins, bacteriophages and endolysins has

promising role in food processing, preservation and food safety. Bacteriocins and
endolysins are believes to have more suitability for DNA shuffling and protein
engineering to generate highly potent variants with expanded activity spectrum.
Genetically modified bacteriophages may also be helpful in bio-preservation but their
safety issues must be addressed properly before selection as bio-preservative agent.
The uses of probiotics and their applications have shown tremendous increase in the
last two decades. Probiotics can turn many health benefits to the human, animals, and
plants. Applications of probiotics hold many challenges. In addition to the viability and
sensory acceptance, it must be kept in mind that strain selection, processing, and
inoculation of starter cultures must be considered. Probiotics industry also faces
challenges when claiming the health benefits. It cannot be assumed that simply adding
a given number of probiotic bacteria to a food product will transfer health to the subject.
Indeed, it has been shown that viability of probiotics throughout the storage period in
addition to the recovery levels in the gastrointestinal tract are important factors.
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