Mob: 9473826765 [Link].

in
Copyright Disclaimer

Copyright © 2022

All rights reserved. This book or any portion thereof may not be reproduced or used in any
manner whatsoever without the express written permission of the publisher except for the
use of brief quotation in a book review.

Published by:
Agriculture & GK Publishing House
Lucknow – 226101
[Link]
E-mail: contact@[Link]
Mobile Number – 9473826765

Disclaimer: Despite having taken all possible precautions during editing/proofing to make
this book free from error/s, but it may possible that some may be left. Reader’s discretion is
required.

Agriculture & GK Page 1

Mob: 9473826765 [Link]

INDEX
[Link] Title Name Page No
Food Microbiology, Hygiene & Sanitation
1. Microorganism 3 to 26
2. Sources of Microorganisms in Food 27 to 46
3. Food Spoilage 47 to 65
4. Food Hazard & Hygiene 66 to 78
5. Food Contamination & Disease 79 to 98
6. Food Safety Management System 99 to 122

Agriculture & GK Page 2

Mob: 9473826765 [Link]

MICROBIOLOGY

Microorganisms:
Microorganisms or microbes are microscopic organisms that exist as unicellular,
multicellular, or cell clusters. Microorganism are widespread in nature and are beneficial to
life, but some can cause serious harm.

What Is Microbiology?
Microbiology is the study of living organisms that are so small or the study of
microscopic organisms, such as bacteria, viruses, archaea, fungi and protozoa.

As a tribute to his work and study in the field of microbiology Anton van Leeuwenhoek
is considered the Father of Microbiology. He was a Dutch microscopist and biologist born in
Delft, Netherland.

Louis Pasteur (1822-1895) was a French biologist who is often regarded as the father
of modern microbiology because of his many contributions to science.

Microorganisms 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 present everywhere on earth, which

includes humans, animals, plants and 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.

BIOLOGICAL ENTITIES STUDIES BY MICROBIOLOGIST

Cellular Acellular
Fungi e.g. Yeast Molds Virus composed of Protein and nucleic acid
Protista e.g. Algae, Protozoa Viroid composed of RNA
Bacteria e.g. Escherichia coli Satellites Composed of Nucleic Acid often RNA
Archaea e.g. Methanogens Prions Composed of Protein

Basic Concept of Microbiology:

Agriculture & GK Page 3

Mob: 9473826765 [Link]
Microbiology is the branch of the biological sciences that deals with microorganisms, i.
e. bacteria, fungi, some algae, protozoa, viruses, viroid and prions.

Most micro-organisms have the following characteristics:

➢ 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.
➢ Cells or other structures are relatively simple and less specialized than those of higher
plants and animals.
➢ 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.

Louis Pasteur is considered as father of Bacteriology. Louis Pasteur was a French

chemist and microbiologist who was one of the most important founders of medical
microbiology. He pioneered the study of molecular heterogeneity; discovered that
microorganisms cause fermentation and disease; Initiated the process of pasteurization and
developed vaccines against anthrax and rabies.

Heinrich Anton de Bary is known as the Father of mycology and Father of Plant
pathology (Phytopathology). Heinrich was a German botanist, microbiologist, and
mycologist and researched for roles of fungi and their role in causing disease.

Antony van Leeuwenhoek - Father of Protozoology

F.E. Fritsch (1935) divided algae into following eleven classes in his book "Structure
and Reproduction of the Algae", mainly on the basis of pigmentation, reserve food and
flagellation, thallus structure, modes of reproduction and life cycles and he is known as
father all algae.

Martinus Beijerinck is often called the Father of Virology.

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:

Agriculture & GK Page 4

Mob: 9473826765 [Link]
Medical microbiology:
It 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 Microorganisms in large scale industrial processes.

Food microbiology:
The study of the role that micro-organisms play in food spoilage, food production,
food preservation and food-borne disease.

Food Microbiology and Its Origin:

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 below.

Major Area of Food Microbiology:

➢ Food fermentations
➢ Laboratory management
➢ Food hygiene
➢ Food-borne disease
➢ Quality control
➢ Food spoilage
➢ Food preservation
➢ Water quality

Not all groups of micro-organisms are of equal interest to the food microbiologist.
Bacteria come very much on top of the list with molds and yeasts also of considerable

Agriculture & GK Page 5

Mob: 9473826765 [Link]
importance and viruses less so. The associations that these organisms have with the
manufacture and consumption of foods are summarized.

Bacteria:

• Food-borne disease
• Food spoilage
• Food fermentations
• Production of food additives and enzymes

Molds:

• Food-borne disease
• Food fermentations
• Food spoilage
• Production of single cell protein
• Production of enzymes

Viruses:

• Food-borne disease
• Identification of food poisoning bacteria
• Failure of dairy fermentations

Yeast:

• Food spoilage
• Food fermentations
• Production of food additives and enzymes

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.

Role 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.

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.

Agriculture & GK Page 6

Mob: 9473826765 [Link]
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 in the processing and storage of food,
changes in food consumption patterns and increased import of food from other countries
have increased the chances 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.

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.

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
microorganisms (starter cultures) to meet the large demand. In addition, novel and better
strains are being developed by using genetic engineering techniques.

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, color compounds, flavor compounds, stabilizers and organic acids.

Food Bio-preservation:
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 nonfood (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.

Probiotics:

Agriculture & GK Page 7

Mob: 9473826765 [Link]
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.

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 italicized (or underlined) with the first letter of the genus written in a
capital letter and species name in small letters. For e. g. Bacillus subtilis (genus is Bacillus
and species is subtilis).

NOMENCLATURE
The basic taxonomic group in bacteria, yeasts, and moulds 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 italicized (or underlined), with the first letter of the genus written in a capital letter (e.g.,
Saccharomyces cerevisiae, Penicillium roqueforti, and Lactobacillus acidophilus).

A bacterial species can be divided into several subspecies (subsp. or ssp.) if the
members show minor but consistent differences in characteristics. Under such conditions, a
trinomial epithet (subspecific epithet) is used (e.g., Lactococcus lactis ssp. lactis or
Lactococcus lactis ssp. cremoris). In some instances, ranks below subspecies are used to
differentiate strains recognized by specific characters (e.g., serovar, antigenic reaction;
biovar, producing a specific metabolite; and phagovar, sensitive to a specific phage). Such

Agriculture & GK Page 8

Mob: 9473826765 [Link]
ranks have no taxonomic importance but can be practically useful (e.g., Lactococcus lactis
ssp. lactis biovar diacetilactis is a Lactococcus lactis ssp.

Actis strain that produces diacetyl, an important flavour compound in some

fermented dairy products. Each strain of a species should be identified with a specific strain
number, which can be alphabetic or numeric or a mixture of both (e.g., Pediococcus
acidilactici LB923). At the family level, bacterial names are used as plural adjectives in
feminine gender and agree with the suffix “aceae” (e.g., Enterobacteriaceae). The species
and strains in a genus can be represented collectively, either using “spp.” after genus (e.g.,
Lactobacillus spp.) or plural forms of the genus (e.g., lactobacilli for Lactobacillus; lactococci
for Lactococcus; leuconostocs for Leuconostoc, or salmonellae for Salmonella).

The scientific names of bacteria are given according to the specifications of the
International Code of Nomenclature of Bacteria. The International Committee on Systematic
Bacteriology of the International Union of Microbiological Association examines the validity
of each name and then publishes the approved lists of bacterial names from time to time. In
the Bergey's Manual of Systematic Bacteriology, only the first letter is used (e.g., Listeria
monocytogenes and then L. monocytogenes).

MORPHOLOGY AND STRUCTURE OF MICROORGANISMS IN FOODS

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 Microorganisms, viz. moulds
and yeasts. Apart from these, food may contain viruses and other parasites such as
protozoans, worms etc. Micro-organisms will be discussed in three arbitrary groups,
normally used by food microbiologists: moulds, yeasts and bacteria.

BACTERIA
Many of us know bacteria only as “germs,” invisible to naked eyes that can invade our
bodies and make us sick. Few know that many bacteria not only coexist with us all the time,
but help us do an amazing array of useful things like make vitamins, break down garbage,
and even maintain our atmosphere. These are unicellular microorganisms that are classed
as plants. A bacterial cell is about 1µm in length and somewhat smaller in diameter. Bacteria
are classified according to their shape. Cocci are spherical, bacilli are cylindrical and spirilla
are spiral. Bacterial spores are more heat resistant than yeast or mould spores to most
processing conditions. Bacteria, with a few exceptions cannot grow in acid media in which
yeasts and moulds thrive. They multiply by ‘binary fission’. When a bacterium becomes
mature it divides into two, these two become four and so on. Bacteria can be found virtually
everywhere. They are in the air, the soil, and water, plants and animals, including us. A
single teaspoon of topsoil contains about a billion bacterial cells (and about 120,000 fungal
cells and some 25,000 algal cells). Microbes that dwell in these extreme habitats are aptly
called extremophiles. The growth of bacteria is very rapid and depends upon the nature of
the food material, moisture, temperature and air.

Agriculture & GK Page 9

Mob: 9473826765 [Link]
Some bacteria do not grow in air but temperature plays a major role in their growth,
the optimum being generally 37°C for bacteria pathogenic to humans. Bacteria are very
sensitive to acids and are destroyed in their presence even at temperature of boiling water.
Hence, most fruits being acidic can be easily sterilized at 100°C whereas vegetables being
non-acidic require a higher temperature of 116°C. A bacterium’s genetic information is
contained in a single DNA molecule suspended in a jelly-like substance called cytoplasm. In
most cases, this and other cell parts are surrounded by a flexible cytoplasmic membrane
that is itself surrounded by a tough, rigid cell wall. A few species, such as the mycoplasmas,
don’t have cell walls. Even though bacteria have only one cell each, they come in a wide
range of shapes, sizes, and colours.

The important groups of bacteria are:

a. Bacillus: rod-shaped.
b. Coccus: spherical.
c. Coccobacillus: oval-shaped.
d. Aerobes: require atmospheric oxygen for growth, e.g., Acetobacter aceti. 67 TSPSC
Food Safety Officer Book (Swa Education)
e. Facultative anaerobes: can grow with or without atmospheric oxygen.
f. Obligate anaerobes: do not grow in atmospheric oxygen.
g. Mesophiles: require a temperature below 38°C for growth.
h. Obligate thermophiles: grow between 38°C and 82°C.
i. Facultative thermophiles: grow over a wide range of temperatures covered by
mesophiles and obligate thermophiles and below.
j. Psychrotrophs: grow fairly well at refrigeration temperatures and some can even
grow slowly at temperatures below freezing.

Some bacteria have natural colours. Certain species contain pigments, such as various
chlorophylls, that make them naturally green, yellow, orange, or brown. Colonies of millions
of bacteria may appear pink, yellowish, or white.

Important Food Spoilage Bacteria

Group Genus
Acetic Acetobacter and Gluconobacter
Lactic Lactobacillus, Leuconostoc, Pediococuus, Sreptococcus
Butyric Clostridium
Propionic Propionibacterium
Proteolytic Bacillus, Pseudomonas, Clostridium, Proteus etc.

Some useful bacteria the following bacteria are of great importance in the food
processing industry. Acetobacter sp. These bacteria, also known as “vinegar bacteria”, cause
significant spoilage in the wine industry but are necessary for vinegar production. The
important species are Acetobacter aceti, A. orleansis and [Link] .They are very
small, usually non-motile and generally do not form spores. These bacteria are aerobes and

Agriculture & GK Page 10

Mob: 9473826765 [Link]
in the presence of oxygen convert ethyl alcohol to acetic acid. These bacteria can be easily
destroyed by heating to 65°C. Lactobacillus sp. Different organisms of this group, also
known as “lactic acid bacteria”, have different properties but all of them produce lactic acid
from carbohydrates. The important species include Lactobacillus plantarum, Pediococcus
cerevisiae, Leuconostoc mesenteroides, Streptococcus faecalis and Lactobacillus brevis.
These bacteria cause “lactic souring” and spoil wines, which can be easily prevented by
maintaining a sulphur dioxide concentration of 0.007 per cent in wine.

YEASTS
Yeasts are unicellular fungi which are widely distributed in nature. They are somewhat
larger than bacteria. The cell length is about 10µm and the diameter is about a third of this.
Most yeasts are spherical or ellipsoidal. Yeasts that multiply by means of ‘budding’ are
known as ‘true yeasts.

Yeasts grow luxuriously at a moderate temperature in a solution of sugar in plenty of

water. Under suitable conditions the sugar is converted into alcohol and carbon dioxide is
evolved. This is the reason that carbon dioxide is evolved from food materials spoiled by
yeasts and pushes out corks from bottles with great force. Most of them do not develop in
media containing more than 66% sugar or 0.5% acetic acid.

Boiling destroys the yeast cells and spores completely. Some of the yeasts which grow
on fruits are Saccharomyces, Candida and Brettanomyces. Pseudo-yeasts these are like true
yeasts but do not form spores. All the members of this group are particularly unsuitable for
fermentation purposes as they produce off-flavours and cloudiness.

Yeasts causing food spoilage

Yeast Product Spoilage

Saccharomyces Low sugar products
Candida High-acid foods, salty foods, butter
Brettanomyces Beers, wines
Zygosaccharomyces Honey, syrups, molasses, wines, soy
(osmophillic) sauce
Pichia Wines
Hansenula Beers
Torulopsis Milk products, fruit juices, acid foods
Rhodotorula Meat, sauerkraut

MOULDS
Moulds are multicellular micro-organisms with mycelial (filamentous) morphology.
They are nonmotile, filamentous, and branched. The cell wall is composed of cellulose,
chitin, or both. A mould (thallus) is composed of large numbers of filaments called hyphae.
An aggregate of hyphae is called mycelium.

Agriculture & GK Page 11

Mob: 9473826765 [Link]
A hypha can be non-septate, septate-uninucleate, or septate-multinucleate. A hypha
can be vegetative or reproductive. The reproductive hypha usually extends in the air and
form exospores, either free (conidia) or in a sack (sporangium). Shape, size, and colour of
spores are used for taxonomic classification. These microbes are also characterized by their
display of a variety of colours and are generally recognized by their mildewy or fuzzy, cotton
like appearance.

Moulds can develop numerous tiny spores that are found in the air and can be spread
by air currents. These spores can produce new mould growth if they are transferred to a
location that has conditions conducive to germination. Moulds generally withstand greater
fluctuation in pH than bacteria and yeasts and can frequently tolerate more temperature
fluctuation. Although moulds 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.

Moulds thrive better at ambient temperature than in a colder environment, even

though growth can occur below 0°C. Although mould 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 moulds. When the Aw goes below 0.90, moulds 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 mould growth.

The principles parts of a mould are a web-like structure known as mycelium and the
spore. The mycelium is often white and cottony and penetrates into the attacked foodstuff.
After fixing itself the mould produces viable spores which resist the favourable conditions
after the dispersal and germinate when they get favourable conditions. They thrive best in
closed, damp and dark situations with an adequate supply or warm, moist air but require
less free moisture than yeasts and bacteria. They prefer sugar containing substances and
may spoil jams, jellies and other sugar-based products. Acid medium favours their growth
and, therefore, they grow well in pickles, juices etc. this is the main reason that fruit and
fruit products are attacked by moulds which not only consume nutrients present in the food
thereby lowering its food value but also spoil the flavour, texture and appearance of the
product. Moulds are sensitive to heat; boiling quickly destroys moulds and their spores.

The most important moulds are:

a. Penicillium sp. (Blue moulds)
b. Aspergillus sp. (Black moulds)
c. Mucor sp. (Gray moulds)
d. Byssochlamys fulva

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 moulds, 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

Agriculture & GK Page 12

Mob: 9473826765 [Link]
coloured. 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.

VIRUSES
Viruses are 10- 450 nm in size; cannot reproduce without a living host; attack only
susceptible host cell lines; infect plants, animals, and bacteria; and have 15 Introduction to
Food Microbiology the capacity to produce specific diseases in specific hosts. Transmission
occurs 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.

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.

Classification of Microorganisms:
A. On basis of temperature for growth Microorganisms can be classified into:

• Hermophillic:
Microbes who require high temperature for their growth and survival (optimum
temperature=45-65⁰C).

Agriculture & GK Page 13

Mob: 9473826765 [Link]

• Thermoduric:
Microbes which do not grow at high temperatures but can survive in it.

• Mesophillic:
Microorganisms which require optimum temperature of 20-50ºC for growth and
multiplication.

• Psychrophillic:
Microorganisms requiring less than 20ºC as optimal temperature for growth.

• Psychroduric:
Microorganisms which do not grow at low temperature but can survive.

B. On basis of oxygen requirement for growth:

• Obligate Aerobes:
Require oxygen for growth and multiplication e. g. moulds.

• Obligate Anaerobes:
Strictly grow only in absence of oxygen.

• Facultative:
Microorganisms than can grow in both presence and absence of oxygen e. g. yeasts.

• Microaerophillic:
Organisms which are able to grow at very low oxidation-reduction potential.

C. On basis of requirement of water activity:

In general, bacteria require more moisture than yeasts and yeasts more than moulds.

The classification according to requirement of aw is as follows:

Group of microorganisms Minimal aw value
Bacteria 0.91
Yeast 0.88
Moulds 0.80
Halophillic bacteria 0.75
Xerophillic fungi 0.65
Osmophillic Yeasts 0.60

• Halophillic bacteria:
Bacteria which grow in high salt solutions

Agriculture & GK Page 14

Mob: 9473826765 [Link]
• Osmophillic Yeasts:
Yeasts which can grow best in high concentrations of sugar.

• Xerophillic Fungi:
Fungi which can grow in low water activity

D. On basis of nutrient degradation capacity:

• Proteolytic:
Microorganisms which are capable of protein degradation because of extracellular
proteinases produced.

• Lipolytic:
Microbes which catalyse the hydrolysis of fats to fatty acids and glycerol.

• Sacchrolytic:
These microorganisms hydrolyse disaccharides or polysaccharides to simpler sugars.

• Pectinolytic:
These microorganisms hydrolyse pectin.

E. On basis of staining: On basis of staining the bacteria can be classified as:

Gram positive:
Those bacteria that stain violet after Gram stain test. In these the cell wall is mostly
comprised of peptidoglycan layer.

Gram negative:
Those bacteria that do not stain violet after Gram stain test. Cell wall mainly
comprised of lipopolysaccharides.

CHARACTERISTICS (MORPHOLOGICAL, CULTURAL AND PHYSIOLOGICAL) OF

MICROORGANISMS BACTERIA

Morphological Characteristics:
One of the first step in the identification of bacteria in food is microscopic examination
to ascertain the shape, size, aggregation, structure and staining reactions of the bacteria
present.

The following characteristics may be of special significance:

Encapsulation:

Agriculture & GK Page 15

Mob: 9473826765 [Link]
The presence of capsules or slime may account for sliminess or ropiness of a food.
Most capsules are polysaccharides of dextrin, dextran or levan and they serve as a source of
reserve nutrients and increase the resistance of bacteria under adverse conditions.

Formation of Endospores:
Bacteria of genera Bacillus, Clostridium, Sporosarcina etc have the ability to form
endospores. Endospores are formed at an intracellular site and are resistant to heat,
ultraviolet light and desiccation. Lysis of the vegetative cell releases the free endospore,
which may remain dormant with no detectable metabolism for years. Sporulation usually
appears in the late logarithmic phase of growth, possibly because of nutrient depletion or
product accumulation. The acquisition of heat resistance is closely related to the formation
of dipicolinic acid and the Ca2+ uptake. Germination is favoured by conditions that are
favourable for growth.

Formation of Cell Aggregates:

It is characteristic of some bacteria to form long chains or of others to clump under
certain conditions. It is more difficult to kill all bacteria in intertwined chains or sizable
clumps than to destroy separate cells.

Cultural Characteristics:
Bacterial growth in and on foods often is extensive enough to make the food
unattractive in appearance or otherwise objectionable. Pigmented bacteria cause
discolouration on the surfaces of foods; films which may cover the surfaces of liquids;
growth may make surfaces slimy; or growth throughout the liquids may result in undesirable
cloudiness or sediment.

Physiological Characteristics:
Most bacteria may be placed into one of three groups based on their response to
gaseous oxygen. Aerobic bacteria thrive in the presence of oxygen and require it for their
continued growth and existence. Other bacteria are anaerobic, and cannot tolerate gaseous
oxygen, such as those bacteria which live in deep underwater sediments, or those which
cause bacterial food poisoning. The third group are the facultative anaerobes, which prefer
growing in the presence of oxygen, but can continue to grow without it. Bacteria may also
be classified both by the mode by which they obtain their energy. Classified by the source of
their energy, bacteria fall into two categories: heterotrophs and autotrophs.

Heterotrophs derive energy from breaking down complex organic compounds that
they must take in from the environment − this includes saprobic bacteria found in decaying
material, as well as those that rely on fermentation or respiration. The other group, the
autotrophs, fix carbon dioxide to make their own food source; this may be filled by light
energy (photoautotrophic), or by oxidation of nitrogen, sulphur, or other elements
(chemoautotrophic). While chemoautotrophs are uncommon, photoautotrophs are

Agriculture & GK Page 16

Mob: 9473826765 [Link]
common and quite diverse. They include the cyanobacteria, green sulphur bacteria, purple
sulphur bacteria, and purple non-sulphur bacteria. The sulphur bacteria are particularly
interesting, since they use hydrogen sulphide as hydrogen donor, instead of water like most
other photosynthetic organisms, including cyanobacteria.

Microbe is a term for tiny creatures that individually are too small to be seen with
the unaided eye. Microbes include bacteria, archaea, fungi and protists. You've probably
heard of bacteria and fungi before. Archaea are bacteria-like creatures that have some traits
not found in any true bacteria. Protists include primitive algae, amoebas, slime moulds and
protozoa. We can also include viruses as a major type of microbe, though there is a debate
as to whether viruses can be considered living creatures or not.

Important Bacterial Genera:

Bacterial classification is rapidly changing. In the following, only those species and
genera currently approved and listed in Bergey’s Manual have been used:

Group Family Genera

Spiral and curved bacteria Spirallaceae Campylobacter
Gram-negative aerobic rods Pseudomonadaceae Pseudomonas, Altermonas,
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,
Grampositive cocci Streptococcaceae Staphylococcus,
Streptococcus, Leuconostoc,
Pediococcus, Lactococcus,
Enterococcus
Peptococcaceae Sarcina
Endospore forming rods and Bacillaceae Clostridium, Bacillus
cocci
Gram-positive Lactobacillaceae Lactobacillus
asporogenous rod of regular Genera of uncertain affinity Listeria
shape
Nonspore-forming rods of Coryneform bacteria Arthrobacter,
irregular shape Brevibacterium,

Agriculture & GK Page 17

Mob: 9473826765 [Link]
Propionibacterium
Rickettsia Rickettsiaceae Coxiella

Common Bacterial Groups in Foods:

Among the Micro-organisms found in foods, bacteria constitute a major important
group. This grouping does not have any taxonomic significance. Some of these groups and
their importance in foods are listed here.

• 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.

• Acetic Acid Bacteria:

Those bacteria that produce acetic acid, such as Acetobacter aceti.

• Propionic Acid Bacteria:

Those bacteria that produce propionic acid and are used in dairy fermentation. Include
species such as Propionibacterium freudenreichii.

• Butyric Acid Bacteria:

Those bacteria that produce butyric acid in relatively large amounts. Some Clostridium
spp., such as Clostridium butyricum.

• 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.

• 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.

• Saccharolytic Bacteria:
Able to hydrolyze complex carbohydrates. Include some species in genera Bacillus,
Clostridium, Aeromonas, Pseudomonas, and Enterobacter.

• Thermophillic Bacteria:

Agriculture & GK Page 18

Mob: 9473826765 [Link]
Able to grow at 500 C and above. Include some species from genera Bacillus, Clostridium,
Pediococcus, Streptococcus, and Lactobacillus.

• Psychrotrophic Bacteria:
Able to grow at refrigerated temperature.

Halotolerant Bacteria:
Able to survive high salt concentrations (>10%). Include some species of Bacillus,
Micrococcus, Staphylococcus, Pediococcus, Vibrio Streptococcus, Clostridium and
Corynebacterium.

Aciduric Bacteria:
Able to survive at low pH (below 4.0). Include some species of Lactobacillus,
Pediococcus, Lactococcus, Enterococcus and Streptococcus.

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 moulds.

Gas-producing Bacteria:
Produce gas (CO2, H2, H2S) during metabolism of nutrients. Include spices from
genera Leuconostoc, Lactobacillus, Brevibacterium and Escherichia.

Slime Producers:
Produce slime due to synthesis of polysaccharides. Include some species or strains of
Xanthomonas, Leuconostoc, Alcaligenes, Enterobacter, Lactococcus, and Lactobacillus.

Spore formers:
Ability to produce spore. Include Bacillus, Clostridium and Desulfotomaculum spp.
They are again divided into aerobic, anaerobic, flat sour thermophilic and sulphide-
producing spore formers.

• Aerobes Require oxygen for growth and multiplication. Species of Pseudomonas, Bacillus,
and Flavobacterium are included in this group.

• Anaerobes Cannot grow in the presence of oxygen. Include species of Clostridium.

• 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.

Agriculture & GK Page 19

Mob: 9473826765 [Link]
• Coliforms Include mainly species from Escherichia, Enterobacter, Citrobacter, and
Klebsiella, and used as index of sanitation.

• Fecal Coliforms Include mainly Escherichia coli. Also used as index of sanitation.

• Enteric Pathogens Includes Salmonella, Shigella, Campylobacter, Yersinia, Escherichia,

Vibrio, Listeria, Hepatitis A, and others that can cause gastrointestinal infection.

MOULDS

• General Characteristics:
The term “mould” is a common one applied to certain multicellular, filamentous fungi
whose growth on foods usually is readily recognized by its fuzzy or cottony appearance.
The main part of the growth commonly appears white but may be coloured or dark or
smoky. Coloured spores are typical of mature mould of some kinds and give colour to
part or all of the growth. The thallus, or vegetative body, is characteristic of thallophytes,
which lack true roots, stems and leaves.

• Morphological Characteristics:
The morphology, i.e. the form and structure, of moulds, as judged by their macroscopic
and microscopic appearance, is used in their identification and classification.

• Hyphae and Mycelium:

The mould thallus consists of a mass of branched, intertwined filaments called hyphae
(singular hypha), and the whole mass of these hyphae are known as the mycelium.

• Reproductive Parts or Structures:

Moulds can grow from a transplanted piece of mycelium. Reproduction of moulds is
chiefly by means of asexual spores. Some moulds also form sexual spores. Culture
Characteristics The gross appearance of a mould growing on a food often is sufficient to
indicate its class or order. Some moulds are loose and fluffy; others are compact. Some
look velvety on the upper surface, some dry and powdery, and others wet or gelatinous.
Some moulds are restricted in size, while others seem limited only by the food or
container. Pigments in the mycelium – red, purple, yellow, brown, gray, black, etc. – are
characteristic, as are the pigments of mass of asexual spores; green, blue-green, yellow,
orange, pink, lavender, brown, gray, black, etc. Physiological Characteristics The
physiological characteristics of moulds will be reviewed only briefly here and will be
discussed in more detail subsequently.

• Moisture Requirements:

Agriculture & GK Page 20

Mob: 9473826765 [Link]
In general, most moulds require less available moisture than do most yeasts and bacteria.
It has been claimed that below 14 to 15 percent total moisture in flour or some dried
fruits will prevent or greatly delay mould growth.

• Temperature Requirements:
Most moulds would be considered mesophilic i.e. able to grow well at ordinary
temperature. The optimal temperature for most moulds is around 25 to 30°C, but some
grow well at 35 to 37°C or above, e.g. Aspergillus spp. And some at still higher
temperatures. A number of moulds are psychrotophic or psychroduric i.e. they grow
fairly well at temperatures of refrigeration, and some can grow slowly at temperatures
below freezing. Growth has been reported at as low as – 5 to 10°C. A few are
thermophilic; i.e. they have a high optimal temperature. Oxygen and pH Requirements
Moulds are aerobic; i.e. they require oxygen for growth; this is true at least for the
moulds growing on foods. Most moulds can grow over a wide range of hydrogen-ion
concentration (pH 2 to 8.5), but the majority are favoured by an acid pH.

• Food Requirements:
Moulds in general can utilize many kinds of foods, ranging from simple to complex. Most
of the common moulds possess a variety of hydrolytic enzymes, and some are grown for
their amylases, pectinases, proteinases, and lipases. Inhibitors: Compounds inhibitory to
other organisms are produced by some moulds, such as penicillin from Penicillium
chrysogenum and clavacin from Aspergillus clavatus. Certain chemical compounds are
mycostatic, inhibiting the growth of moulds (sorbic acid, propionates, and acetates are
examples), or are specifically fungicidal, killing moulds. Initiation of growth of moulds is
slow compared to that of bacteria or yeasts, so that when conditions are favourable for
all these organisms, moulds usually lose out in the competition. After mould growth is
under way, however, it may be very rapid.

Classification of Moulds and Moulds of Industrial Importance.

In the following only genera of industrial importance will be shortly overviewed.

• Genus Mucor:
(Mucor racemosus, Mucor rouxii). Mucors are involved in the spoilage of some foods and
in the manufacture of others e.g. oriental fermented foods.

• Genus Rhizopus
Rhizopus nigricans, sometimes called „bread mould”, is very common and is involved in
the spoilage of many foods such as berries, fruits, vegetables, bread, etc. Genus
Aspergillus. The members of this genus are very widespread. Many are involved in the
spoilage of foods and some are useful in preparation of fermented foods.

Agriculture & GK Page 21

Mob: 9473826765 [Link]
• Many groups and hundreds of aspergillus species are known. Aspergillus niger is the
leading species important for food microbiologists. Selected strains are used for
commercial production of citric and gluconic acids. Genus Penicillium. This is another
widespread genus important in foods. Penicillium expansum, a green spored species,
causes soft rot of fruits. Penicillium camemberti with grayish conidia, useful in the
ripening of Camembert cheese, and Penicillium roqueforti, used in ripening of blue
cheeses, are also well known, members of this genus.

• Genus Bothrytis:
The species Bothrytis cinerea causes the noble rot of grape in some wine producing areas
such as Tokay (Hungary).

• Genus Alternaria:
Moulds of this genus are common causes of the spoilage of foods. Alternaria citri,
Alternaria tenuis and Alternaria brassicae are the common species. Genus Neurospora
(Monilia). The species of this genus grow on various foods.

YEASTS

Like mould, the term “yeast” is commonly used but hard to define. As used here it
refers to those fungi which are generally not filamentous but unicellular and ovoid or
spheroid and which reproduce by budding or fission. Yeasts may be useful or harmful in
foods. Yeast fermentations are involved in the manufacture of foods such as bread, beer,
wines, vinegar, and surface ripened cheese, and yeasts are grown for enzymes and for food.
Yeasts are undesirable when they cause spoilage of sauerkraut, fruit juices, syrups,
molasses, honey, jellies, meats, wine, beer, and other foods.

Morphological Characteristics Form and structure:

The form of yeasts may be spherical to ovoid, lemon shaped, pear-shaped, cylindrical,
triangular, or even elongated into a false or true mycelium. They also differ in size.

Reproduction:
Most yeasts reproduce asexually by multilateral or polar budding, a process in which
some of the protoplasm bulges out the cell wall; the bulge grows in size and finally walls off
as a new yeast cell. A new species or yeasts reproduce by fission, and one reproduces by
combination of fission and budding. Sexual reproduction of “true” yeasts (Ascomycotina)
results in the production of ascospores, the yeast cell serving as the ascus. The ascospores
may differ in colour, in smoothness or roughness of their walls, and in their shape (round,
oval, reniform, bean or sickle-shaped, hemispherical, angular, fusiform, or needle-shaped).
“False” yeasts, which produce no ascospores or other sexual spores, belong to the Fungi
imperfecti. Cells of some yeasts become chlamydospores by formation of a thick wall about
the cell, for example, Candida, Rhodotorula, and Cryptococcus.

Agriculture & GK Page 22

Mob: 9473826765 [Link]

Cultural Characteristics For the most part, the appearance of massed yeast growth is
not useful in the identification of yeasts, although growth as a film on the surface of liquid
media suggests an oxidative or film yeasts, and production of a carotenoids pigment
indicates the genus Rhodotorula. However, the appearance of the growth is important
when it causes coloured spots on foods. Yeasts are oxidative, fermentative, or both. The
oxidative yeasts may grow as a film, pellicle, or scum on the surface of liquid and then are
termed film yeasts. Fermentative yeasts usually grow throughout the liquid and produce
carbon dioxide.

Physiological Characteristics:
Most common yeasts grow best with a plentiful supply of available moisture. But since
many yeasts grow in the presence of greater concentration of solutes (such as sugar or salt)
than most bacteria it can be concluded that these yeasts require less moisture than the
majority of bacteria.

Most yeast require more moisture than moulds, however, on the basis of water
activity or aw yeasts may be classified as ordinary if they do not grow in high concentrations
of solutes, i.e. in a low aw, and as osmophilic if they do. However, limits of aw for ordinary
yeasts tested thus far ranges from 0.88 to 0.94. The range of temperature for growth of
most yeasts is, in general, similar to that for moulds, with the optimum around 25°C to 30°C
and the maximum about 35°C to 47°C. Some kinds can grow at 0°C or less. The growth of
most yeasts if favoured by an acid reaction in the vicinity of pH 4 to 4.5, and they will not
grow well in an alkaline medium unless adapted to it.

Yeasts grow best under aerobic conditions, but the fermentative types can grow
anaerobically, although slowly. In general, sugars are the best source of energy for yeasts,
although oxidative yeasts, e.g., the film yeasts, oxidize organic acids and alcohol. Carbon
dioxide produced by bread yeasts accomplishes the leavening of bread, and alcohol made
by the fermentative yeasts is the main product in the manufacture of wines, beer, industrial
alcohol, and other products. The yeasts also aid in the production of flavours or “bouquet”
in wines. Nitrogenous foods utilized vary from simple compounds such as ammonia and
urea to amino acids and polypeptides.

In addition, yeasts require accessory growth factors. Microorganisms, namely,

bacteria, yeasts and moulds can be found in any environment. The eight environmental
sources of organisms to foods are: soil and water, plants and plant products, food utensils,
intestinal tracts of humans and animals, food handlers, animal feeds, animal hides, air and
dust. Although we see that the microorganisms are beneficial to the humans in many ways,
there are many microorganisms that are the causative agents for food borne diseases. e.g.
Staphylococcus aureus and Clostridium botulinum cause food borne intoxication whereas
Salmonella, E. coli, Campylobacter, Listeria, Yersinia, Bacillus etc cause food borne
infections. Moulds are responsible for causing food intoxication by production of

Agriculture & GK Page 23

Mob: 9473826765 [Link]
mycotoxins, which are lethal for the human body e.g. Aflatoxin produced by Aspergillus
flavus, patulin produced by Penicillium expansum, ochratoxins produced by Aspergillus
ochraceus etc.

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 discolouration of
foods, such as in meat, fish, and sauerkraut. Species Rhodotorula glutinis. Botrytis
Cladosporium Rhizopus Alternaria Fusarium Asporgillus Pencillium 18 Fundamentals of
Food Microbiology

4. Torulopsis:
They have spherical to oval structure. They cause spoilage 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.

VIRUSES

Agriculture & GK Page 24

Mob: 9473826765 [Link]
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 transduction (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, phage 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 Lactobacillus. Methods are being studied to genetically engineer lactic
acid start cultures so that they become resistant to multiple bacteriophages.

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

Human Intestinal Viruses with High Potential as Food Contaminants

SPORE

Spore is a small, single cell structure produced by certain bacteria, fungi, algae and
nonflowering plants. Spores are asexually or sexually made. They can resist harsh
environmental conditions, and they can survive under low nutrient conditions. Once the
favorable conditions happen, spores can become active and grow into a new organism.
Furthermore, some bacteria produce spores known as endospores that are dormant
structures developed from the bacterial cell.

A spore is a dormant, reproductive cell with a thick cell wall, which is highly resistant
to unfavorable environmental conditions. When the conditions are favorable, a spore gives
rise to a new individual of the same species. Spores do not fuse with other spores to
produce an individual, like gametes. Therefore, spore formation is a type of asexual
reproduction. Typically, plants, algae, fungi, and bacteria produce spores. A spore is very
similar to a plant seed but may contain some stored food compared to a seed. Plants that
undergo the alteration of generations produce spores as reproductive cells of the asexual
Agriculture & GK Page 25
Mob: 9473826765 [Link]
generation. Lower plants such as ferns, mosses, hornworts, and liverworts produce spores,
which act as seeds.

Microspores and megaspores are the two types of spores produced by both
angiosperms and gymnosperms. Microspores give rise to male gametophyte whereas
macrospores give rise to female gametophytes.

Spores are metabolically inactive at the most time, and they contain less water
content. They are resistant to disinfectants, chemicals, heat, radiation, etc. Some
endospores remain unharmed even after boiling.

VEGETATIVE CELL

The cell which produces spores and which is metabolically active is known as a
vegetative cell. Vegetative cell grows rather than producing spores. It contains a high
amount of water content, and it is not resistant to harsh environmental conditions. Unlike
spores, vegetative cells are susceptible to disinfectants, heat, chemicals, radiation, etc.

Vegetative cell is active and reproductive. When the environmental conditions are not
favorable, vegetative cell produces spores that are dormant structures. Moreover,
vegetative cells have high enzymatic activity.

Agriculture & GK Page 26

Mob: 9473826765 [Link]

SOURCES OF MICROORGANISMS IN FOOD

Microbes obtain nutrients from organic matter, some of which constitutes our food
items. In this process, microbes contribute immensely to the maintenance of our
environment through the nitrogen cycle and cycle of other elements, which has been shown
below in the following general reaction:

All organic matter (carbohydrates, proteins, lipids, etc).

↓
1 Energy + Inorganic compound (nitrates, sulfates, etc.)

Some of the microorganisms are desirable in production of certain foods like

fermented food products such as fermented meat sausages, ham, alcohol, beverages etc.,
whereas, other type of microorganisms bring about spoilage of foods. Some microbes that
are also known as 'pathogens' or more popularly as 'germ' may cause food poisoning
leading to gastroenteritis in man and animals. Therefore, it becomes relevant to know about
the different sources of microorganisms found in foods, since each genus of microorganisms
has its own particular nutritional requirements and has adopted itself to a particular
environment. These environmental food sources of organisms may be classified in following
eight categories:

A. Plants (Fruits and Vegetables):

The inside tissue of foods from plant sources are essentially sterile, except for a few
porous vegetables (e.g., radishes and onions) and leafy vegetables (e.g., cabbage and
Brussels sprouts).1– 5 Some plants produce natural antimicrobial metabolites that can limit
the presence of microorganisms. Fruits and vegetables harbour microorganisms on the
surface; their type and level vary with soil condition, type of fertilizers and water used, and
air quality. Moulds, yeasts, lactic acid bacteria, and bacteria from genera Pseudomonas,
Alcaligenes, Micrococcus, Erwinia, Bacillus, Clostridium, and Enterobacter can be expected
from this source.

B. Animals, Birds, Fish, and Shellfish:

Food animals and birds normally carry many types of indigenous microorganisms in
the digestive, respiratory, and urinogenital tracts, the teat canal in the udder, as well as in
the skin, hooves, hair, and feathers. Their numbers, depending on the specific organ, can be
very high (large intestinal contents can have as high as 1010 bacteria/g). Many, as carriers,
can harbour pathogens such as Salmonella serovars, pathogenic Escherichia coli,
Campylobacter jejuni, Yersinia enterocolitica, and Listeria monocytogenes without showing
symptoms. Laying birds have been suspected of asymptomatically carrying Salmonella
Enteritidis in the ovaries and contaminating the yolk during ovulation Fish and shellfish also
carry normal microflora in the scales, skin, and digestive tracts. Water quality, feeding
habits, and diseases can change the normal microbial types and level. Pathogens such as

Agriculture & GK Page 27

Mob: 9473826765 [Link]
Vibrio parahaemolyticus, Vib. vulnificus, and Vib. cholerae are of major concern from these
sources. Many spoilage and pathogenic microorganisms can get into foods of animal origin
(milk, egg, meat, and fishery products) during production and processing. Milk can be
contaminated with fecal materials on the udder surface, egg shells with fecal material
during laying, meat with the intestinal contents during slaughtering, and fish with intestinal
contents during processing. In addition to enteric pathogens from fecal materials, meat
from food animals and birds can be contaminated with several spoilage and pathogenic
microorganisms from skin, hair, and feathers, namely Staphylococcus aureus, Micrococcus
spp., Propionibacterium spp., Corynebacterium spp., and moulds and yeasts.

C. Air:
Microorganisms are present in dust and moisture droplets in the air. They do not grow
in dust, but are transient and variable, depending on the environment. Their level is
controlled by the degree of humidity, size and level of dust particles, temperature and air
velocity, and resistance of microorganisms to drying. Generally, dry air with low dust
content and higher temperature has a low microbial level. Spores of Bacillus spp.,
Clostridium spp., and moulds, and cells of some grampositive bacteria (e.g., Micrococcus
spp. and Sarcina spp.), as well as yeasts, can be predominantly present in air. If the
surroundings contain a source of pathogens (e.g., animal and poultry farms or a sewage-
treatment plant), different types of bacteria, including pathogens and viruses (including
bacteriophages), can be transmitted via the air. Microbial contamination of food from the
air can be reduced by removing the potential sources, controlling dust particles in the air
(using filtered air), using positive air pressure, reducing humidity level, and installing UV
light.

D. Soil:
Soil, especially the type used to grow agricultural produce and raise animals and birds,
contains several varieties of microorganisms. Because microorganisms can multiply in soil,
their numbers can be very high (billions/g). Many types of moulds, yeasts, and bacterial
genera (e.g., Enterobacter, Pseudomonas, Proteus, Micrococcus, Enterococcus, Bacillus, and
Clostridium) can enter foods from the soil. Soil contaminated with fecal materials can be the
source of enteric pathogenic bacteria and viruses in food. Sediments where fish and marine
foods are harvested can also be a source of microorganisms, including pathogens, in those
foods. Different types of parasites can also get in food from soil. Removal of soil (and
sediments) by washing and avoiding soil contamination can reduce microorganisms in foods
from this source.

E. Sewage:
Sewage, especially when used as fertilizer in crops, can contaminate food with
microorganisms, the most significant of which are different enteropathogenic bacteria and
viruses. This can be a major concern with organically grown food and many imported fruits
and vegetables, in which untreated sewage and manure might be used as fertilizer.
Pathogenic parasites can also get in food from sewage.
Agriculture & GK Page 28
Mob: 9473826765 [Link]

F. Water:
Water is used to produce, process, and, under certain conditions, store foods. It is
used for irrigation of crops, drinking by food animals and birds, raising fishery and marine
products, washing foods, processing (pasteurization, canning, and cooling of heated foods)
and storage of foods (e.g., fish on ice), washing and sanitation of equipment, and processing
and transportation facilities. Water is also used as an ingredient in many processed foods.
Wastewater can be recycled for irrigation. However, chlorine-treated potable water
(drinking water) should be used in processing, washing, sanitation, and as an ingredient.
Although potable water does not contain coliforms and pathogens (mainly enteric types), it
can contain other bacteria capable of causing food spoilage, such as Pseudomonas,
Alcaligenes, and Flavobacterium. Improperly treated water can contain pathogenic and
spoilage microorganisms. To overcome the problems, many food processors use water,
especially as an ingredient, that has a higher microbial quality than that of potable water.

G. Food:
Ingredients In prepared or fabricated foods, many ingredients or additives are
included in different quantities. Many of these ingredients can be the source of both
spoilage and pathogenic microorganisms. Various spices generally have very high
populations of mould and bacterial spores. Starch, sugar, and flour might have spores of
thermophilic bacteria. Pathogens have been isolated from dried coconut, egg, and
chocolate. The ingredients should be produced under sanitary conditions and given
antimicrobial treatments. In addition, setting up acceptable microbial specifications for the
ingredients will be important in reducing microorganisms in food from this source.

QUANTIFICATION OF MICROORGANISMS

Food serves as excellent substrate for the growth of different kinds of

microorganisms. Microorganisms enter into food and grow as contaminants. Growth of
microorganisms in food may spoil food quality and consumption of such food creates
hazardous health effects in human and animal. So is becomes necessary to examine the
microorganism in food.

Microbiological testing of food is the examination of the microscopic organisms in

food. These organisms could be single cell, multiple cell or without cell. Microbiology
includes various sub-disciplines like Virology, Mycology, Parasitology and Bacteriology.
Microorganisms play an important role in our surroundings as all humans, plants & animals
and probably they are the largest mass of living materials on our earth.

These diversified living organisms can grow in extreme conditions where no other
living organisms can survive. They can grow in a temperature like at boiling points and can
similarly can grow and survive in extreme freezing conditions like -20 degrees & -30 degrees.

Agriculture & GK Page 29

Mob: 9473826765 [Link]
Microbial pathogens, such as Bacteria & Archaea known as prokaryotes and various
types of Fungi, protozoa etc. which are called eukaryotes need to be taken care well as this
may be harmful to some extent.

As per the latest Food Safety and Standards Act, 2006, every food product has to be
tested & approved before it is put up for sale in the market.

There is a direct relationship between microorganisms and diseases as these harmful

living organisms are responsible for taking many lives with diseases like diphtheria,
pneumonia, typhoid, amoebiasis, botulism, cholera, dysentery etc.

The microorganisms can be found in various foods & beverages and they can be
harmful if they enter into a human body. Some of these microorganisms could prove to be
resistant to one or more types of antibiotics.

METHOD FOR MICROBIOLOGICAL EXAMINATION OF FOOD

A. CONVENTIONAL METHODS:
Standards Plate Count Methods
Direct Microscopic Examination
Dye Reduction Techniques
Most Probable Number Counts

B. RAPID METHODS FOR THE DETECTION OF SPECIFIC ORGANISMS AND

TOXINS
Immunological Methods
DNA/RNA Methodology

C. CHEMICAL METHODS
Limulus Lysate Method for Endotoxins
Adenosine Triphosphate Measurement
Radiometry

A. CONVENTIONAL METHODS

1. STANDARD PLATE COUNT

SPC can be performed by two methods:

(i) Spread plate
(ii) Pour plate

(i) Spread Plate Method:

Agriculture & GK Page 30
Mob: 9473826765 [Link]
In spread plate method pre poured and hardened agar plates with dry surfaces are
used. The food samples are serially diluted and 0.1 mm inoculum is taken using sterile
pipette and evenly distributed over the agar surface. The inoculum is distributed over the
agar surface with the help of bent glass rod.

The dispersed
cells develop into
isolated colonies called
Colony Forming Units
(CFU). Because the
number of colonies
should equal the
number of viable
organisms in the food
sample and spread
plates can be used to
count the microbial
population in food
sample.

Surface plating offers advantages in determining the number of heat-sensitive

psychrotrophsin a food product because the organisms do not come in contact with melted
agar. It is the method of choice when the features of a colony are important to its
presumptive identification and for most selective media. Strict aerobes are obviously
favoured by surface plating, but microaerophilic organisms tend to grow slower rate.

The disadvantages of surface plating are the problem of spreaders (especially when the
agar surface is not adequately dry prior to plating) and the crowding of colonies, which
makes enumeration more difficult.

(ii) Pour Plate Method:

The original food sample is diluted several times to reduce the microbial population
sufficiently to obtain separate colonies when plating. Then small volumes, around one ml of
several diluted samples mixed with liquid agar that has been cooled to about 45°C, and the
mixtures are poured immediately into sterile culture dishes.

After the agar has hardened, each cell is fixed in place

and forms an individual colony (CFU). Plates containing
between 30 and 300 colonies are counted.

The total number of colonies equals the number of

viable microorganisms in the diluted sample. Colonies

Agriculture & GK Page 31

Mob: 9473826765 [Link]
growing on the surface also can be used to inoculate fresh medium and prepare pure
cultures.

2. DIRECT MICROSCOPIC EXAMINATION

When examining foods, the possibility of detecting the presence of micro-organisms
by looking at a sample directly under the microscope should not be missed. A small amount
of material can be mounted and teased out in a drop of water on a slide, covered with a
cover slip, and examined, first with a low magnification, and then with a X45 objective. The
condenser should be set to optimize contrast even though that may result in some loss of
resolution.

Alternatively dark-field illumination or phase-contrast microscopy may be used. It is

usually relatively easy to see yeasts and moulds and with care and patience it is possible to
see bacteria in such a preparation.

The high refractive index of bacterial endospores

makes them particularly easy to see with phasecontrast
optics and, if the preparation is made as a hanging drop on
the cover glass mounted over a cavity slide, it should also
be possible to determine whether the bacteria are motile.

Since only a small sample of product is examined in this way, micro-organisms will not
be seen unless present in quite large numbers, usually at least 106 ml-1 . In the case of some
liquid commodities, such as milk, yoghurt, soups and fruit juices, it may be possible to
prepare and stain a heat-fixed smear. But the food constituents often interfere with the
heat fixing and care is needed to prevent the smear being washed away during staining.

It may be necessary to dilute the sample with a little water, although that will reduce
the concentration of micro-organisms further. The great advantage of such techniques is
their rapidity, although in their simplest forms they do not distinguish between live and
dead cells.

3. DYE REDUCTION TECHNIQUES

A group of tests which have been used for some time in the dairy industry depend on
the response of a number of redox dyes to the presence of metabolically active micro-
organisms. They are relatively simple and rapid to carry out at low cost. The redox dyes are
able to take up electrons from an active biological system and this results in a change of
colour.

Agriculture & GK Page 32

Mob: 9473826765 [Link]
Usually, oxidized form is coloured and the reduced form is colourless but the
triphenyltetrazolium salts are an important exception. There are three most widely used
redox dyes, methylene blue, resazurin and triphenyltetrazolium chloride.

Methylene blue is a dye which remains blue in its oxidized state and turn colourless on
its reduction state. The result of this test is expressed in terms of time required for the
colour of methylene blue to disappear at incubation temp. of 37°[Link] 1937, and until
relatively recently, the methylene blue test was a statutory test for grading the quality of
milk in England and Wales. Changes in the technology of handling bulk milk, especially
refrigeration, have made this test less reliable and it is no longer a statutory test because
results show little correlation with the numbers of psychrotrophic bacteria.

The reduction of resazurin takes place in two stages, from blue to pink to colourless,
there is a wider range of colour that can be scored using a comparator disc and the ten-
minute resazurin test is still useful for assessing the quality of raw milk at the farm or dairy
before it is bulked with other milk.

Triphenyltetrazolium salts and their derivatives are initially colourless and become
intensely coloured, and usually insoluble, after reduction to formazans.
Triphenyltetrazolium chloride itself is most widely used as a component of diagnostic and
selective agar media on which some bacterial colonies will become dark red to maroon as
formazan becomes precipitated within the colony.

The crystals of the formazan produced from 2-(p-iodo-phenyl)-3-( p– nitro-phenyl)-5-

phenyltetrazolium chloride (INT) are so intensely coloured that they are readily seen in
individual microbial cells under the microscope and their presence may be used to assess
the viability of cells. One possible development would be the incorporation of INT as part of
the staining procedure in the DEFT analysis.

4. MOST PROBABLE NUMBER COUNTS

Monitoring and detection of indicator and disease-causing microorganisms are a
major part of sanitary microbiology. A wide range of viral, bacterial and protozoan diseases
result from the contamination of water with human fecal wastes. Although many of these
pathogens can be detected directly, environmental microbiologists have generally used
indicator organisms as an index of possible water contamination by human pathogen

Coliforms are defined as gram-negative, non-sporing, facultatively anaerobic, rod-

shaped bacteria that ferment lactose with gas formation within 48 hours at 35°C.

THE FOLLOWING ARE AMONG THE SUGGESTED CRITERIA FOR INDICATOR

ORGANISMS

Agriculture & GK Page 33

Mob: 9473826765 [Link]
➢ The indicator bacterium should be suitable for the analysis of all types of water tap,
river, ground, impounded, recreational, estuary, sea, and waste.
➢ These organisms should be present wherever enteric pathogens are present.
➢ The indicator bacterium should survive longer than the hardiest enteric pathogen.
➢ The indicator bacterium should not reproduce in the contaminated water and produce
an inflated value.
➢ The assay procedure for the indicator should have great specificity; in other words,
other bacteria should not give positive results. In addition, the procedure should have
high sensitivity and detect low levels of the indicator.
➢ The testing method should be easy to perform.
➢ The indicator should be harmless to humans.
➢ The level of the indicator bacterium in contaminated water should have some direct
relationship to the degree of fecal pollution.

MPN test is the most widely used method to find out the potability of water by testing
the viable bacterial count. This method was introduced by Mc Crady in 1915. The original
test for coliforms that was used to meet this definition involved the presumptive,
confirmed, and completed tests. The presumptive step is carried out by means of tubes
inoculated with three different sample volumes to give an estimate of the most probable
number (MPN) of coliforms in the water.

The complete process, including the confirmed and completed tests, requires at least
4 days of incubations and transfers. So, water to be tested to find out presence of fecal
coliforms. These coliforms are derived from the intestine of warm-blooded animals, which
can grow at the more restrictive temperature of 44.5°C. Three serial aliquots or dilutions are
then planted into 9 or 15 tubes of appropriate medium for the three- or five-tube method,
respectively. Numbers of organisms in the original sample are determined by use of
standard MPN tables. It is not a precise method of analysis; the 95% confidence intervals for
a three-tube test range from 21 to 395.

B. RAPID METHODS FOR THE DETECTION OF SPECIFIC ORGANISMS AND

TOXINS MOLECULAR BIOLOGY TECHNIQUES

1. NUCLEIC ACID (DNA/RNA) METHODOLOGY

All biochemical, immunological and other characteristics used in the detection of
micro-organisms are governed directly or indirectly by the base sequences encoded in the
organism’s genome. The specificity of this information can now be mobilized to provide
methods capable of identifying genera, species or even strains within a species.

Nucleic acid probes can be designed which recognize and bind (hybridize) to specified
regions of either chromosomal or plasmid DNA or to RNA, and the region chosen to give the
desired level of specificity. Thus, for example, ribosomal RNA contains both conserved and
variable regions, the former being suitable for recognition at the genus level whereas the
latter may be considerably more specific.
Agriculture & GK Page 34
Mob: 9473826765 [Link]

Although RNA is a more labile molecule than DNA, there are many more copies of
ribosomal RNA in a cell than genomic DNA which should make methods based on this
molecule more sensitive.

The nucleic acids have to be released from the cells by some form of lysis and, in the
case of doublestranded DNA, it has also to be denatured, usually by heat treatment, to the
single-stranded form. The denatured nucleic acid is then adsorbed onto a membrane, fixed
to it by heat or alkali treatment, and the membrane is treated with some form of blocking
agent to prevent nonspecific binding of the probe.

After incubating with the labelled probe and washing off un-adsorbed probe, the
presence of the hybridization product is measured using the label attached to the probe. In
the earliest stages of the development of this methodology probes were directly labelled
with radioactive isotopes such as 32P or 35S and hybridization was detected by
autoradiography.

This is a very sensitive method but the routine use of radioactive compounds in a
food-associated environment is not usually acceptable. Probes can be labelled with an
enzyme and detected with a chromogenic substrate or they can be labelled with a small
molecular weight hapten for which an enzyme-linked monoclonal antibody is available.

Such probes are available for the enterotoxin gene of Staphylococcus aureus, the
haemolysin gene and rRNA of Listeria monocytogenes, 23S rRNA of Salmonella, as well as
several other systems. One interesting example is a ribosomal RNA probe to detect Listeria
monocytogenes which uses a chemiluminescent label.

The single-stranded DNA probe has a chemiluminescent molecule bound to it. When
the probe binds to its RNA target, the chemiluminescent molecule is protected from
degradation in a subsequent step so that successful hybridization is indicated by light
emission measured in a luminometer.

2. DNA AMPLIFICATION (PCR)

PCR has been used to detect Enterotoxigenic E. coli, Vibrio, Clostridium, etc. and this
method is an elegant technique to determine the pathogen by amplifying their DNA using
specific primer.

IMMUNOLOGIC METHODS

Because of the potential specificity of immunoassays using polyclonal or monoclonal

antibodies, there has been considerable effort devoted to developing their application in
food microbiology. Commercial immunoassay kits are now available for detecting a variety
of foodborne micro-organisms and their toxins, including mycotoxins.

Agriculture & GK Page 35

 Mob: 9473826765 [Link]
Raising antibodies to specific surface antigens of micro-organisms, or to
macromolecules such as staphylococcal or botulinum toxins, is relatively straightforward
and can be achieved directly. Mycotoxins, however, belong to a class of molecules known as
haptens which can bind to an appropriate antibody but are of relatively low molecular
weight and are not themselves immunogenic.

Serological reactions are effective method for detecting the pathogenic

microorganisms or their toxin. The most commonly used serological methods are discussed
below:

I. FLUORESCENT ANTIBODY
An antibody to a given antigen is made fluorescent by coupling it to a fluorescent
compound and when the antibody reacts with its antigen, the antigen-antibody complex
emits fluorescence and can be detected by the use of a fluorescence microscope. The
fluorescent markers used are rhodamine B, fluorescein isocyanate, and fluorescein
isothiocyanate with the last being the most widely used.

The fluorescent antibody (FA) technique can be carried out by use of either of two
basic methods. The direct method employs antigen and specific antibody to which is
coupled the fluorescent compound (antigen coated by specific antibody with fluorescent
label) with the indirect method, the homologous antibody is not coupled with the
fluorescent label, but instead an antibody to this antibody is prepared and coupled in the
indirect method, the labelled compound detects the presence of the homologous antibody;
in the direct method, it detects the presence of the antigen.

II. ENRICHMENT SEROLOGY

The use of Enrichment Serology (ES) is a more rapid method for recovering
salmonellae from foods than the conventional culture method. It is carried out in four steps:
pre-enrichment in a nonselective medium for 18 hours; selective enrichment in selenite-
cystine and/or tetrathionate broth for 24 hours; elective enrichment in M broth for either 6-
8 hours or 24 hours; and agglutination with polyvalent H antisera at 50°C for 1 hour.

The Oxoid Salmonella Rapid Test (OSRT) is a variation of ES. It consists of a culture
vessel containing two tubes, each of which contains dehydrated enrichment media in the
lower compartments and dehydrated selective media in the upper compartments.

The media are hydrated with sterile distilled water and a special salmonella elective
medium is added to the culture vessel along with a novobiocin antibiotic disk, followed by
1ml of pre-enrichment culture of sample. Following incubation at 41°C for 24 hours, media
in the upper compartment (selective media) of each tube are examined for colour change,
indicating the presence of salmonella.

III. SALMONELLA 1-2 TEST

Agriculture & GK Page 36
 Mob: 9473826765 [Link]
Salmonella 1-2 Test employs the use of a semisolid phase. The test is conducted in a
specially designed plastic device that has two chambers, one for selective broth and the
other for a nonselective motility medium. In addition to selective ingredients, the
nonselective medium contains the amino acid LSerine, which is selective for salmonellae.

Following inoculation of the selective medium chamber, the device is incubated,

during which time motile salmonellae move into the nonselective medium chamber. The
non-selective medium contains flagellar antibodies, and when the motile organisms enter
the antibody area, an immune band develops, indicating antigen-antibody reaction.

IV. RADIOIMMUNOASSAY
This technique consists of adding a radioactive label to an antigen, allowing the
labelled antigen to react with its specific antibody, and measuring the amount of antigen
that combined with the antibody by the use of a counter to measure radioactivity. Solid-
phase radioimmunoassay (RIA) refers to methods that employ solid materials or surfaces
onto which a monolayer of antibody molecules binds electrostatically.

The solid materials used include polypropylene, polystyrene and bromacetyl cellulose.
The ability of antibody-coated polymers to bind specifically with radioactive tracer antigens
is essential to the basic principle of solid-phase RIA. When the free-labelled antigen is
washed out, the radioactivity measurements are quantitative.

V. ELISA
The enzyme-linked immunosorbent assay (ELISA, enzyme immunoassay, or EIA) is an
immunological method similar to RIA but employing an enzyme coupled to either an antigen
or an antibody. A typical ELISA is performed with a solid-phase (polystyrene) coated with
antigen and incubated with antiserum. Following incubation and washing, an enzyme-
labelled preparation of anti-immunoglobulin is added. After gentle washing, the enzyme
remaining in the tube or microtiter well is assayed to determine the amount of specific
antibodies in the initial serum. A commonly used enzyme is horseradish peroxidase and its
presence is measured by the addition of peroxidase substrate.

The amount of enzyme present is ascertained by the colorimetric determination of

enzyme substrate. One variation of basic ELISA consists of a “sandwich” ELISA in which the
antigen is required to have at least two binding sites. The antigen reacts first with excess
solid-phase antibody, and following incubation and washing, the bound antigen is treated
with excess labelled antibody. The ELISA technique is used widely to detect and quantify
organisms and/ or their products in foods.

C. CHEMICAL METHODS

1. LIMULUS LYSATE FOR ENDOTOXINS

Agriculture & GK Page 37

Mob: 9473826765 [Link]
Endotoxins, a type of pyrogen, are natural compounds found in the outer cell
membrane of Gramnegative bacteria and can impact over 30 biological activities. Endotoxin
can lead to cell death by initiating complement activation. The Limulus amebocyte lysate
(LAL) test was commercially introduced in the 1970s. LAL is derived from the blood cells, or
amebocytes, of the horseshoe crab, Limulus polyphemus. Gram-negative bacteria are
characterized by their production of endotoxins, which consist of a lipopolysaccharides (LPS)
layer (outer membrane) of the cell envelope. The LPS is pyrogenic and responsible for some
of the symptoms that accompany infections caused by gramnegative bacteria. The Limulus
amoebocyte lysate (LAL) test employs a lysate protein obtained from the blood cells
(amoebocytes) of the horseshoe crab (Limulus Polyphemus).

The LAL test is performed by adding aliquots of food suspensions or other test
material to small quantities of lysate preparation, followed by incubation at 37° C for 1hr.
The presence of endotoxins causes gel formation of the lysate material.

2. ATP (ADENOSINE -TRIPHOSPHATE) MEASUREMENT

ATP is a molecule found in and around living cells, and as such it gives a direct
measure of biological concentration and health. ATP is quantified by measuring the light
produced through its reaction with the naturally occurring firefly enzyme luciferase using a
luminometer. The amount of light produced is directly proportional to the amount of ATP
present in the sample.

Adenosine-triphosphate (ATP) is the primary sources of energy in all living organism. It

disappears within 2 h after cell death, and the amount per cells is generally constant. The
simplest ways to measure ATP is by use of the firefly luciferin-luciferase system. In the
presence of ATP, luciferase emits light, which is measured with a liquid scintillation
spectrometer or a luminometer. The amount of light produced by firefly luciferase is directly
proportional to the amount of ATP added.

The application of the measurement as a rapid method for estimating microbial

numbers has been used in clinical microbiology.

FACTORS AFFECTING GROWTH OF MICROORGANISMS IN FOOD

In most cases, micro-organism utilizes food supply as a source of nutrient for their
growth. This course can result in deterioration (decay) of food. The organism not only
deteriorates the food but may also pose risks of disease to the human being on
consumption of such contaminated food. However, the growth of microorganisms in food
may be affected by several factors like physical, chemical and biological.

These factors can broadly divide into two categories i.e.

A. Intrinsic parameters or intrinsic factors

Agriculture & GK Page 38

 Mob: 9473826765 [Link]
B. Extrinsic parameters or extrinsic factors

A. INTRINSIC PARAMETERS
The natural environment inside the plant or animal tissues used as food may either
favour or discourage the microbial growth. This internal environment of the food tissues is a
result of the combination of several parameters. Since such tissue parameters are an
inherent part of food item, these are known as intrinsic parameters and have been listed
below:

I. Hydrogen-ion concentration pH
II. Water activity (aw)
III. Redox potential
IV. Nutrient contents
V. Antimicrobial constituents
VI. Biological structures.

I. Hydrogen-Ion Concentration pH:

The hydrogen ion concentration of the growth medium has a marked effect on the
growth of bacteria. This concentration is normally expressed in terms of pH, which is
defined as the negative logarithm of the hydrogen ion concentration.

The ultimate pH of meat has a significant bearing on the growth of microorganisms

present in [Link] producing excess of hydrogen ions (H') in solution are acidic while
excess of hydroxyl ions (OH-) are termed as basic or alkaline. Every microorganism has its
minimal, maximal and optimal requirement of pH for growth. Most bacteria grow optimally
at pH 7.0 but not so well below 4.0 or above 9.0. Most bacteria favour a pH near neutrality
or slightly on the alkaline side (6.8-7.5). The acidity and alkalinity (pH) of an environment
has a strong influence on the activity and stability of macromolecules such as enzymes.
These enzymes play an important role during growth of microorganisms and in their
metabolism. Thus, growth and metabolism of microorganisms are influenced by pH.

The pH ranges of some common food items and pH range of some common
food microflora

Food pH range Organism pH range

Citrus fruits 2-5 Moulds 0-11
Soft drinks 2.5-4 Yeasts 1.5-8.5
Beer 3.5-4.5 Lactic acid bacteria 3.2-10.5
Meat 5.5-6.2 Staphylococcus 4-9.8
aureus
Fish 6.5-7.3 Salmonella spp. 4.1-9
Egg white 8.6-9.6 Escherichia coli 4.3-9
Milk 6.5-7 Yersinia 4.5-9
enterocolitica
Flour 6.2-7.2 Clostridium 4.8-8.2
Agriculture & GK Page 39
 Mob: 9473826765 [Link]
botulinum
Vegetables 4.8-7 Clostridium 5.4-8.7
perfringens
Fermented shark 10-12 Bacillus cereus 4.7-9.3
Bananas 4.5 - 4.7 Listeria 4.39- 9.4
monocytogenes

pH minima and maxima of microorganisms also varies due to other important factors
like temperature, moisture content, salt concentration, redox potential etc.

For example, in the presence of 0.2 M NaCl, Alcaligenes faecalis can grow over a wider
pH range than in the absence of NacI or in the presence of 0.2 M sodium citrate. Yeasts and
moulds grow well in an acid environment of pH 3.5-4.5; moulds, whilst favouring acid
conditions, usually grow over and its Control a wide pH range (3.5-8.0).

II. Water Activity (aw):

We know that water is essential for growth and survival of most of the living forms
present on this planet. Similarly, the availability of moisture present in foods is an important
factor for microbial growth. The water requirement is best expressed in terms of 'available
water' or 'water activity (aw)'. It is defined as the ratio of the vapour pressure of the food to
the vapour pressure of pure water at the same temperature. The most rapid growth of
microorganisms occurs at aw in the range of 0.995- 0.980, and it is reduced as aw value goes
down. That's why some of the food preservation methods like freezing or drying cause
reduction in the aw of foods

Approximate aw values for growth of selected pathogens in

food
Organism aw values
Campylobacter spp. 0.98- 0.99
Clostridium botulinum type E 0.97
Salmonella spp. 0.94- 0.99
Bacillus cereus 0.93
Clostridium perfringens 0.943- 0.97
Approximate aw values of selected food categories
Animal Products Aw Values
fresh meat, poultry, fish 0.99 - 1.00
Eggs 0.97
Honey 0.75
cured meat 0.87 - 0.95
Plant Products Aw Values
fresh fruits, vegetables 0.97 - 1.00
Jam 0.75 - 0.80
Flour 0.67 - 0.87
Cereal 0.10 - 0.20
bread, white 0.94 - 0.97

Agriculture & GK Page 40

 Mob: 9473826765 [Link]
Jellies 0.82 - 0.94

III. Redox Potential:

The oxidation-reduction or redox potential of a substance is defined in terms of the
ratio of the total oxidizing (electron accepting) power to the total reducing (electron
donating) power of the substance. In effect, redox potential is a measurement of the ease
by which a substance gains or loses electrons. The redox potential (Eh) is measured in terms
of millivolts. A fully oxidized standard oxygen electrode will have an Eh of +810 mV at pH
7.0, 30 °C (86 °F), and under the same conditions, a completely reduced standard hydrogen
electrode will have an Eh of -420 mV. The Eh is dependent on the pH of the substrate;
normally the Eh is taken at pH 7.0 (Jay 2000, p 45-7). Examples of foodborne pathogens for
each of these classifications include Aeromonas hydrophila, Clostridium botulinum,
Escherichia coli O157:H7, and Campylobacter jejuni. The measurement of redox potential of
food is done rather easily, either for single or multicomponent foods. For multicomponent
foods, in addition to measurement of each component, the redox potential of the interface
areas and microenvironments should be considered. These values can be highly variable
depending on changes in the pH of the food, microbial growth, packaging, the partial
pressure of oxygen in the storage environment, and ingredients and composition (protein,
ascorbic acid, reducing sugars, oxidation level of cations, and so on). Another important
factor is the poising capacity of the food.

IV. Nutrient Content:

The microorganisms b present in foods also require certain items as their food in order
to grow and action normally. These nutrients include the following items: Water Source of
energy Source of nitrogen Vitamins and related growth factors Minerals. The other four
substances listed above are required least by moulds followed by yeasts, Gram-negative and
Gram-positive bacteria. As a source of energy, foodborne microorganisms may utilise
sugars, alcohols and amino acids. Some of the microbes have ability to first breakdown the
complex carbohydrates such as starches and cellulose into simple sugars for their utilization.
Only very few microbes that are capable of breaking down the fats (lipolytic microbes) can
use fats as sources of energy. The food based on their nutrient composition can be classified
as:
• foods for energy,
• foods for growth, and
• accessory food substances, or vitamins, which may be necessary for energy or growth.

Microorganisms may require low quantities of B-vitamins, which are present in most
of the natural foods. In general, Gram-positive bacteria cannot synthesize more quantities
of these compounds, therefore, need supplementation of B-vitamins. On the other hand,
Gram-negative bacteria and moulds can synthesize these compounds at their own, and
therefore, grow freely on foods which are low in B-vitamins such as fruits. This helps to
explain why fruits are spoiled frequently by moulds rather than bacteria.

Agriculture & GK Page 41

 Mob: 9473826765 [Link]
In general, the nutritional factors which encourage or support the microbial growth in
foods include high moisture, low salt (although Staphylococci can tolerate more salt than
other pathogenic bacteria), low acid, low fat, and moderate sugar. The microbial growth is
retarded by heat, cold and dehydration, high concentrations of salt, sugar, acid and fat.

V. Antimicrobial Constituents:
The stability of some foods against attack by microorganisms is due to the presence of
certain naturally occurring substances that have been shown to have antimicrobial activity.
Foods may contain a variety of substances, which may affect microbial growth and these
may occur naturally, be produced by microbial growth or added artificially. Some foods
intrinsically contain naturally-occurring antimicrobial compounds that convey some level of
microbiological stability to them. There are a number of plant-based antimicrobial
constituents, including many essential oils, tannins, glycosides, and resins, that can be found
in certain foods. Specific examples include eugenol in cloves, allicin in garlic, cinnamic
aldehyde and eugenol in cinnamon, allyl isothiocyanate in mustard, eugenol and thymol in
sage, and carvacrol (iso thymol) and thymol in oregano. It is also known that some types of
food processing result in the formation of antimicrobial compounds in the food. Some types
of fermentations can result in the natural production of antimicrobial substances, including
bacteriocins, antibiotics, and other related inhibitors.

VI. Biological Structures:

Let us understand the function of the natural covering of foods. Such coverings are
seen in the form of the outer covering of fruits, shell of nuts, hide of animals, and the shell
of eggs. These natural barriers provide excellent protection against the entry and
subsequent damage by spoilage organisms. The skin covering of fish and meats such as beef
and pork prevents the contamination and spoilage of these foods, partly because it tends to
dry out faster than freshly cut surfaces.

B. EXTRINSIC PARAMETERS
The conditions under which foods are stored greatly affect both the food stored and
their microorganisms. These storage conditions that make the external environment around
the food are referred to as extrinsic factors or parameters and include the following:

i. Temperature of storage
ii. Relative humidity (RH) in the storage environment
iii. Presence and concentration of gases in the storage environment.

i. Temperature of Storage:
Temperature is the most important factor that affects all the, chemical reactions
related to microbial growth in foods. Therefore, the temperature of food storage or growth
medium has a marked influence on the rate of growth of any microorganism. For any
organism, the temperature at which growth is most rapid is known as the 'optimum
temperature'. The 'maximum temperature' is the highest temperature at which the

Agriculture & GK Page 42

Mob: 9473826765 [Link]
organism will grow while the 'minimum temperature' is the lowest temperature at which
growth of the organisms can occur, which is usually below the optimum temperature. Based
on their optimum temperature for growth, the bacteria have been classified into three
groups namely the 'psychrophiles' with optimal temperature of 20-35⁰C, 'mesophiles' with
optimal temperature of 30-35⁰C, and 'thermophiles' with 45-70⁰C.

ii. Relative Humidity (RH):

The relative humidity of the storage environment is important both from the
standpoint of water activity (aw) within foods and the growth of microorganisms at the
surface. It is important that the food should be stored under conditions of RH that do not
allow the food to pick up moisture from the air.

iii. Presence and Concentration of Gases in the Environment:

When food is stored in an environment (e.g., a packaging film) that contains increased
amounts (up to about 10 per cent) of carbon-dioxide (CO2), it is referred to as"control1ed
atmosphere" or "modified atmosphere" storage. A gas mixture of 10 per cent CO2 five per
cent oxygen (O2) and 85 per cent nitrogen (N2) is found to be very effective for the
preservation of meat steaks.

Growth and Death Kinetics:

Typically, to understand and define the growth of a particular microbial isolate, cells
are placed in a liquid medium in which the nutrients and environmental conditions are
controlled. If the medium supplies all nutrients required for growth and environmental
parameters are optimal, the increase in numbers or bacterial mass can be measured as a
function of time to obtain a growth curve. Several distinct growth phases can be observed
within a growth curve. These include the lag phase, the exponential or log phase, the
stationary phase, and the death phase. Each of these phases represents a distinct period of
growth that is associated with typical physiological changes in the cell culture. As will be
seen in the following sections, the rates of growth associated with each phase are quite
different.

1) The Lag Phase:

The first phase observed under batch conditions is
the lag phase in which the growth rate is essentially zero.
When an inoculum is placed into fresh medium, growth
begins after a period of time called the lag phase. The lag
phase is defined to transition to the exponential phase
after the initial population has doubled ( Yates and
Smotzer, 2007 ). The lag phase is thought to be due to the physiological adaptation of the
cell to the culture conditions. The lag phase may also be due to low initial densities of
organisms that result in dilution of exoenzymes (enzymes released from the cell) and of
nutrients that leak from growing cells. Normally, such materials are shared by cells in close

Agriculture & GK Page 43

Mob: 9473826765 [Link]
proximity. But when cell density is low, these materials are diluted and not as easily taken
up. As a result, initiation of cell growth and division and the transition to exponential phase
may be slowed. The lag phase usually lasts from minutes to several hours. The length of the
lag phase can be controlled to some extent because it is dependent on the type of medium
as well as on the initial inoculum size.

2) The Exponential Phase:

The second phase of growth observed in a batch
system is the exponential phase. The exponential phase
is characterized by a period of the exponential
growth—the most rapid growth possible under the
conditions present in the batch system. During
exponential growth the rate of increase of cells in the
culture is proportional to the number of cells present at
any particular time.

There are several ways to express this concept both theoretically and mathematically.
One way is to imagine that during exponential growth the number of cells increases in the
geometric progression 20 , 2 1 , 2 2 , 2 3 until, after n divisions, the number of cells is 2n.

This can be expressed in a quantitative manner; for example, if the initial cell number
is X0 the number of cells after n doublings is 2n X0 . As can be seen from this example, if one
starts with a low number of cells exponential growth does not initially produce large
numbers of new cells. However, as cells accumulate after several generations, the number
of new cells with each division begins to increase explosively.

In the example just given, X0 was used to represent cell number. However, X0 can also
be used to represent cell mass, which is often more convenient to measure than cell
number Whether one expresses X0 in terms of cell number or in terms of cell mass, one can
mathematically describe cell growth during the exponential phase using the following
equation:

𝒅𝑿
= 𝝁𝑿 (𝑬𝒒. 𝟑. 𝟏)
𝒅𝒕
where X is the number or mass of cells (mass/volume), t is time, and is the specific
growth rate constant (1/time). The time it takes for a cell division to occur is called the
generation time or the doubling time. Equation 3.1 can be used to calculate the generation
time as well as the specific growth rate using data generated from a growth curve.

The generation time for a microorganism is calculated from the linear portion of a
semi log plot of growth versus time. The mathematical expression for this portion of the
growth curve is given by Eq. 3.1, which can be rearranged and solved as shown in Eqs. 3.2 to
3.6 to determine the generation time.

Agriculture & GK Page 44

Mob: 9473826765 [Link]
𝒅𝑿
= 𝝁𝑿 (𝑬𝒒. 𝟑. 𝟏)
𝒅𝒕
Rearrange:

𝒅𝑿
= 𝝁𝒅𝒕 (𝑬𝒒. 𝟑. 𝟐)
𝑿
Integrate:
𝑿 𝒕
𝒅𝑿
∫ = 𝝁 ∫ 𝒅𝒕 (𝑬𝒒. 𝟑. 𝟑)
𝑿
𝑿𝟎 𝟎

𝐈𝐧 𝑿 = 𝝁𝒕 + 𝐈𝐧 𝑿𝟎 𝐨𝐫 𝑿 = 𝑿𝟎 𝒆𝝁𝒕 (𝑬𝒒. 𝟑. 𝟒)

For 𝑿 to be doubled:

𝑿
= 𝟐 (𝑬𝒒. 𝟑. 𝟓)
𝑿𝟎

Therefore:

𝟐 = 𝒆𝝁𝒕 (𝑬𝒒. 𝟑. 𝟔)

Where 𝒕 = generation time.

3) The Stationary Phase:

The third phase of growth is the stationary phase. The stationary phase in a batch
culture can be defined as a state of no net growth, which can be expressed by the following
equation:

𝒅𝑿
= 𝟎 (𝑬𝒒. 𝟑. 𝟕)
𝒅𝒕
Although there is no net growth in stationary phase, cells still grow and divide. Growth
is simply balanced by an equal number of cells dying. There are several reasons why a batch
culture may reach stationary phase. One common reason is that the carbon and energy
source or an essential nutrient becomes completely used up. When a carbon source is used
up it does not necessarily mean that all growth stops. This is because dying cells can lyse and
provide a source of nutrients. Growth on dead cells is called endogenous metabolism.
Endogenous metabolism occurs throughout the growth cycle, but it can be best observed
during stationary phase when growth is measured in terms of oxygen uptake or evolution of
carbon dioxide.

Thus, in many growth curves the stationary phase actually shows a small amount of
growth. Again, this growth occurs after the substrate has been utilized and reflects the use

Agriculture & GK Page 45

Mob: 9473826765 [Link]
of dead cells as a source of carbon and energy. A second reason that stationary phase may
be observed is that waste products build up to a point where they begin to inhibit cell
growth or are toxic to cells. This generally occurs only in cultures with high cell density.
Regardless of the reason why cells enter stationary phase, growth in the stationary phase is
unbalanced because it is easier for the cells to synthesize some components than others. As
some components become more and more limiting, cells will still keep growing and dividing
as long as possible. As a result of this nutrient stress, stationary phase cells are generally
smaller and rounder than cells in the exponential phase.

4) The Death Phase:

The final phase of the growth curve is the death phase, which is characterized by a net
loss of culturable cells. Even in the death phase there may be individual cells that are
metabolizing and dividing, but more viable cells are lost than are gained so there is a net
loss of viable cells. The death phase is often exponential, although the rate of cell death is
usually slower than the rate of growth during the exponential phase. The death phase can
be described by the following equation:

𝒅𝑿
= −𝒌𝒅 𝑿 (𝑬𝒒. 𝟑. 𝟖)
𝒅𝒕
where k d is the specific death rate. It should be noted that the way in which cell
growth is measured can influence the shape of the growth curve.

Agriculture & GK Page 46

Mob: 9473826765 [Link]

FOOD SPOILAGE

Food spoilage is a metabolic process that causes foods to be undesirable or

unacceptable for human consumption due to changes in sensory characteristics.

➢ Spoilage of food is identified by off-colours, off-odours, softening of vegetables, fruits,

and slime production.
➢ Spoilage may arise from insect damage, physical damage, and indigenous enzyme
activity in food or by microorganisms (bacteria, viruses, fungi).
➢ The spoilage microbe’s common inhabitants are soil, water, or the intestinal tracts of
animals or they are dispersed through the air and water.
➢ Food spoilage by microorganisms depends upon intrinsic (pH, water activity, nutrient
content, oxidation-reduction potential, antimicrobial property) and extrinsic factors
(temperature, relative humidity, pressure).
➢ Different spoilage-causing microorganisms have different nutrients requirements.
➢ Microorganisms are the biological agents that cause foodborne diseases when
consumed however the microorganisms not only cause spoilage, some of them are
beneficial for food fermentation.

Bacteria in food:
➢ Food is most commonly spoiled by bacteria as it can grow in a wide variety of
conditions however bacteria are used for beneficial fermentations of pickles, milk
products, and some fermented vegetable products.
➢ Bacteria do not grow at a water activity level below 0.91 and require neutral pH
(6.5-7) to cause food spoilage (e.g. milk, meat, green vegetables, fruits, etc.).

Agriculture & GK Page 47

Mob: 9473826765 [Link]
➢ Some bacteria are capable of spore formation so they are highly heat resistant and
some are capable of producing heat-resistant toxins.
➢ The consumption of such spoiled food leads to foodborne illness.
➢ The most common bacteria that cause food spoilage are:
• Gram-positive bacteria such as Staphylococcus aureus, Bacillus spp, Clostridium
spp, Lactic acid bacteria Streptococcus spp, Brochothrix spp, Weissella spp,
Mycobacterium bovis, etc.
• Gram-negative bacteria such as Salmonella spp, Shigella, Vibrio spp, Escherichia
coli, Campylobacter jejuni, Yersinia enterocolitis, Brucella spp, Coxiella burnetii,
Aeromonas spp, Plesiomonas shigelloides, etc.
➢ These bacteria cause off-odours and off-flavors, discolourations, gas production, slime
production, and decreases in pH in food.

Fungi in food:
➢ Fungi is the most abundant group of microorganisms that plays important role in food
spoilage.
➢ Fungi are osmotrophic they obtain their nutrients by absorption.
➢ Fungi can be divided into mould and yeast.

Moulds:
➢ Moulds are the most common food spoilage-causing microorganisms.
➢ Moulds grow on the surface of food (they require free oxygen for growth) and in a
wide range of pH values (from 2 to 8.5), but the majority of them prefer acidic pH.
➢ Moulds can grow at very low water activity levels (0.7–0.8) on dried foods (e.g. grains,
beans, peanuts, and some spices).
➢ The most common food spoilage causing moulds are Mucor, Aspergillus spp, Rhizopus
spp, Penicillium spp, Alternaria spp, Bothrytis, Byssochlamys, Fusarium spp.
➢ This mould causes off-flavors, mycotoxins contamination, discolouration, rotting, and
is externally visible in the food.

Yeasts:
➢ Compared to bacteria and moulds, yeasts play a minor role in food spoilage.
➢ Yeasts can grow with or without oxygen and are used for beneficial fermentation in
bread and alcoholic drinks fermentation.
➢ They often spoil food that has high sugar or salt content (e.g. maple syrup, pickles,
jams, soy sauce, and sauerkraut.)
➢ Yeasts require a water activity level of 0.90–0.95 for growth and they can grow in a
wide range of pH (3 – 8) but in general, they prefer acidic pH (4.5-5.5).
➢ Most commonly food spoilage causing yeasts are Zygosaccharomyces spp,
Saccharomyces spp., Candida spp, Dekkera spp
➢ These yeasts cause a change in colour, a change in texture, an unpleasant odour, or an
undesirable taste in food.

Agriculture & GK Page 48

Mob: 9473826765 [Link]
Protozoa in Food:
➢ Protozoa are one-celled microorganisms without a rigid cell wall and the transmissible
form of these organisms is termed cysts.
➢ Protozoan parasites are highly associated with foodborne and water-borne outbreaks
of disease. The water and food act as a carrier for transmission of protozoan parasites
from one host to another.
➢ The most common foodborne parasites are Giardia lamblia, Entamoeba histolytica,
Cyclospora cayetanensis, Toxoplasma gondii, and Trichinella spiralis.

Algae in Food:
➢ Algae are primary producers which are a source of different nutrients and they are
usually of aquatic habitats.
➢ They contaminate the water source with their toxin and cause them to accumulate in
fish and marine life. The toxic may or may not be harmful to marine lives. When such
fish or other marine lives are consumed by humans, it leads to foodborne illness.
➢ The algae that cause poisoning are Gonyaulax catenella, Gonyaulax tamarensis,
Gambierdiscus toxicus, Ptychodiscus brevis, Microcystis aeruginosa, Blue-green Algae.

Viruses in Food:
➢ Viruses are obligate intracellular parasites that cause a wide range of diseases in
plants, animals, and humans.
➢ Viruses need specific living cells to replicate and therefore they cannot replicate in
food or water. The water and food act as a carrier for transmission of virusesfrom one
host to another.
➢ Foodborne viruses are quite stable outside the host and are acid-resistant.
➢ Some of the foodborne viruses are Norovirus, Hepatitis A virus (HAV), Hepatitis E virus
(HEV), Astrovirus (AstV), Rotavirus (RV), Coronavirus, Sapovirus (SaV).

Prions in Food:
➢ Prions are infectious disease-causing agents which are the normal protein of a brain
that gets misfolded that lacks genome resulting in a pathological, infectious
conformation.
➢ Once misfolded, it can induce other normally folded proteins to become misfolded.
➢ Prion diseases can affect both humans and animals. It can also get transferred from
animal to human through the consumption of infected meat and meat products.
➢ Some examples of prion disease are Bovine spongiform encephalopathies’ (BSE),
Scrapie, Chronic wasting disease (CWD), Creutzfeldt-Jacob Disease (CJD).

SPOILAGE OF MILK
The main carbohydrate in milk is lactose, the microorganisms producing lactose
hydrolyzing enzyme (lactase or β-galactosidase) have an advantage over those unable to

Agriculture & GK Page 49

Mob: 9473826765 [Link]
metabolize lactose. Milk fat can be hydrolyzed by microbial lipases to produce small
molecular volatile fatty acids (butyric, capric, and caproic acids).

Types of some microbial spoilage of milk

A. Raw Milk:
Spoilage Raw milk is an excellent medium for microorganisms due to high moisture,
nearly neutral pH, and rich in nutrients. Microbial spoilage of raw milk occurs from the
metabolism of lactose, nitrogenous compound (such as proteins, amino acids, ammonia,
urea, and others), unsaturated fatty acids, triglycerides, and minerals

Spoilage Spoilage bacteria Enzyme Metabolic product

Acid proteolysis Lactobacillus, Proteinases, lactase Peptides, amino
Micrococcus, B.
Cereus
Alcoholic flavor Yeasts Alcohol Ethanol
dehydrogenase
Bitter flavor Psychrotrophic Proteinase, Lipases Bitter peptides
bacteria, Bacillus
Fishiness Aeromonas Proteinase Fish flavors
hydrophila
Fruity flavor P. fragi, Esterase Ethyl esters, lactons
Staphylococcus,
Candida,
S. Cerevisiae, P.
notatum, C.
butyricum
Caramel and malty L. lactis var. Oxidase 3-Methylbutanol
flavor maltigenes
Putrefaction P. putrefaciens, Proteinase Peptones, amino
Clostridium acids
Rancid flavor Lipolytic bacteria Lipase Free fatty acids
Ropy texture LAB Polymerase Exopolysaccharide
Sour and acid flavor Lactococcus lactis Lactase Lactate
Sour flavor at 10 – 40 LAB, coliforms, Lactase Lactate, acetate
0C Enterococcus
Sour flavor at 37 – 50 L. thermophiles, S. Lactase Lactate, acetate
0C thermophilus

Organoleptic and Physical Changes:

Microorganisms cause different undesirable organoleptic and physical changes in raw
milk. Refrigeration of raw milk prevents growth of most mesophilic microorganisms, while
psychrotrophic microorganisms (such as species of Pseudomonas, Flavobacterium,
Micrococcus, Bacillus, Enterobacter, Aeromonas, and Alcaligenes) are able to grow.
Although psychrotrophic microorganisms present initially in low numbers, but extended
refrigeration allows their growth to high numbers. Organoleptic changes (such as malty,

Agriculture & GK Page 50

 Mob: 9473826765 [Link]
rancid, yeasty, bitter, fruity, or putrid off-flavors), pigments, and ropiness can associate with
growth of psychrotrophic microorganisms.

Different off-flavors are produced by microorganisms:

i. Sour or acid flavor is caused by Lactococcus lactis. Volatile fatty acids are produced by
coliforms and Clostridium.
ii. Bitter flavors result from proteolysis and lipolysis.
iii. Burn or caramel flavor is caused by Lactococcus lactis subsp. lactis var. maltigenes,
which resembles the cooked flavor of overheated milk.
iv. Miscellaneous flavors— barny flavor is caused by Enterobacter oxytocum, soapiness
with ammonia production by Proteus sapolactica, malty flavor by Micrococcus, fruit
flavor by Pseudomonas fragi, proteolytic flavor by Pseudomonas mucidolens, fishiness
by Aeromonas hydrophila, putrefaction by Clostridium and Pseudomonas
putrefaciens, and fruity and alcoholic flavors by yeasts. Lactones, which can give fruity
and coconut-like flavor to dairy products, can be produced by various microbial
growth (such as Candida and Saccharomyces cerevisiae) and molds (such as
Penicillium notatum and Cladosporium butyricum).

The spoilage bacteria in raw milk are mostly aerobic Gram-negative psychrotrophic
rods, such as Alcaligenes, Flavobacterium, Pseudomonas, and some coliforms. About 65–
70% of psychrotrophic microorganisms in raw milk is Pseudomonas spp. Psychrotrophic
spoilage microflora of milk is generally proteolytic, lipolytic, and phospholipolytic, but not
lactose hydrolyzing. Pseudomonas spp. (such as P. fluorescens, P. fragi, P. putida, P.
lundensis, and P. aeruginosa) outgrow other bacteria when milk is stored at 3–7 °C. They
can produce extracellular heat-stable lipases, proteinases, and phospholipases. Milk
products that may be affected by residual heat-stable enzymes include ultrahigh
temperature (UHT) milk, butter, cheese, and fluid milk products. Raw milk is rapidly cooled
after collection and is kept cold until pasteurization. But there is often sufficient time
between milk collection and consumption for psychrotrophic bacteria to grow. Generation
time of psychrotrophic Pseudomonas spp. in milk is 8–12 h at 3 °C and 5–10 h at 5 °C. These
growth rates are sufficient to cause spoilage within 5 days even if the milk initially contains
only one Pseudomonas per milliliter.

2 Souring:
Spoilage of milk and milk products results from growth of fermentative bacteria when
storage temperatures are sufficiently high for psychrotrophs. Genera of bacteria producing
acids in milk and milk products are Enterococcus, Lactobacillus, Lactococcus, Leuconostoc,
Pediococcus, and Streptococcus. Alkaline reaction occurs by P. fluorescens and Alcaligenes
viscolactis at chilling and refrigeration temperatures. Little acid formation takes place in milk
held at temperatures near freezing, but proteolysis may take place at this temperature. If
raw milk is not refrigerated and kept at room temperature, growth of mesophiles
predominates. These include species of Bacillus, Clostridium, Enterococcus, Lactobacillus,
Lactococcus, Micrococcus, Proteus, Pseudomonas, coliforms, and others. The unpleasant

Agriculture & GK Page 51

 Mob: 9473826765 [Link]
sour odor and taste of spoiled milk result from the production of small amounts of acetic
and propionic acids by lactic acid bacteria (LAB). Lactose hydrolyzing Lactococcus lactis
subsp. lactis will generally predominate to cause souring with enough acid production to
lower the pH and inhibit growth of other microorganisms. A malty flavor can also result
from growth of Lactococcus lactis subsp. lactis var. maltigenes. When coliforms,
Enterococcus, Lactobacillus, and Micrococcus grow in milk, they cause curdling, gas
formation, proteolysis, and lipolysis. At higher temperatures, from 37 to 50 °C,
Streptococcus thermophilus and Enterococcus faecalis grow to produce acid and then
growth can be followed by Lactobacillus (such as Lactobacillus bulgaricus). Some of
Lactobacillus can grow at temperatures above 50 °C, but produce less acid. The
pasteurization of milk kills some acid-forming bacteria, but heat-resistant thermoduric LAB
(such as S. thermophilus, Enterococcus, and Lactobacillus) can survive.

3. Proteolysis:
Psychrotrophic species of Alcaligenes, Flavobacterium, Bacillus, Micrococcus, and
Pseudomonas can grow at low temperatures and produce extracellular proteinases. Most of
proteinases in raw milk are produced by P. fragi and P. fluorescens. Proteinases hydrolyze
proteins (proteolysis). The hydrolysis of proteins at low temperatures produces bitter, fruity,
and putrid flavors due to release of peptides.

Types of proteolysis are:

i. acid proteolysis due to proteolysis and acid production at the same time,
ii. proteolysis with little acidity or alkalinity,
iii. sweet curdling, which is caused by rennin-like enzymes of bacteria,
iv. slow proteolysis by intracellular enzymes of bacteria after lyses of their cells, and
v. residual proteolytic activity of heat-stable proteinases. Acid proteolysis is caused by
Enterococcus faecalis var. liquefaciens, Macrococcus caseolyticus, and Bacillus cereus.

4. Lipolysis:
Psychrotrophic bacteria (such as Alcaligenes, Bacillus, Clostridium, Micrococcus,
Pseudomonas, Proteus, and Staphylococcus) produce extracellular lipases in milk and
hydrolyze milk fat. Lipaseproducing bacteria are Pseudomonas spp. P. fragi and S. aureus,
and these enzymes are fairly heat resistant. Lipases hydrolyze fat to fatty acids and glycerol.
Oxidation of the unsaturated fatty acid results in the formation of aldehydes, acids, and
ketones. They can cause odors and unwanted tastes in milk. Combined effects of oxidation
and hydrolysis of fat cause rancidity.

5. Ropiness:
Ropiness (sliminess) results with the formation of polymers from microbial and
nonmicrobial activities on polysaccharides. Most LAB (such as Lactobacillus spp.),
Alcaligenes spp. (such as A. viscolactis), and coliforms produce extracellular polymers that
increase the viscosity of milk and cause ropiness. The polymers produced from

Agriculture & GK Page 52

 Mob: 9473826765 [Link]
polysaccharides contain glucose and galactose with small amounts of mannose, rhamnose,
and pentose. The main sources of bacteria causing ropiness are water, manure, utensils, and
feed. Nonbacterial ropiness can also occur in milk.

This may be due to:

i. the presence of fibrin and leukocytes (from mastitis) in milk,
ii. the thickness of cream (at the top), and
iii. the films of casein or lactalbumin.

Bacterial ropiness is caused by slime capsular polymers that are located on the surface of
cells. Bacteria causing ropiness are A. viscolactis and Micrococcus freudenreichii (surface
ropiness); Enterobacter aerogenes (ropiness near the top); Enterobacter cloacae, E. coli,
Klebsiella oxytoca, and Bacillus (ropiness throughout the milk); and certain species of LAB
(such as S. thermophilus, L. bulgaricus, Lactobacillus casei, Lactobacillus. plantarum,
Lactococcus lactis subsp. hollandicus, Lactococcus lactis subsp. lactis, and Lactococcus lactis
subsp. cremoris).

6. Color Changes and Gas Formation:

Pigmented bacteria and molds can change the color of milk. Blue milk is caused by
Pseudomonas syncyanea and Geotrichum, deep-blue color by P. syncyanea growing
together with L. lactis; red milk by P. synxantha, Serratia marcescens, Brevibacterium
erythrogenes, Micrococcus roseus, and Flavobacterium spp.; and brown milk by P.
putrefaciens and P. fluorescens. The chief gas formers are coliforms in raw milk. Clostridium,
Bacillus, and thermoduric heterofermentative Lactobacillus can produce gas in heat-treated
milk.

PATHOGENS IN RAW MILK

Mycobacterium bovis, Mycobacterium tuberculosis, Brucella abortus, and Brucella
melitensis can cause milk-borne diseases. Raw milk is an important vehicle for Salmonella
enterica subsp. enterica serovars (such as S. Aanatum, S. Thompson, S. Heidelberg, S.
Enteritidis, and S. Newport) that cause human infection. Salmonella enterica subsp. enterica
ser. Dublin and Salmonella enterica subsp. enterica ser. Typhimurium most frequently
associate with cattle. These Salmonella serovars may show multiple resistances to
antibiotics. Campylobacteriosis with Campylobacter spp. can also associate with raw milk.
Raw milk contaminates with pathogenic E. coli through exposure to fecal material and can
cause foodborne outbreaks (such as E. coli O157:H7). The most probable source of S. aureus
in raw milk. milk is the infected bovine udder. S. aureus can multiply and produce
enterotoxin in raw milk. Raw milk may be contaminated with L. monocytogenes from
poorsilage conditions, fecal contamination, and improperly cleaned equipment. Raw milk
may be contaminated with Yersinia enterocolitica, Streptococcus agalactiae, other
hemolytic Streptococcus and Coxiella burnetii. Drinking milk containing bovine tubercle
bacilli causes alimentary tuberculosis, especially in infants. In regions where consumption of

Agriculture & GK Page 53

Mob: 9473826765 [Link]
raw milk continues, multiple outbreaks of brucellosis, salmonellosis, campylobacteriosis,
and enterohemorrhagic colitis (by E. coli O157: H7) can occur.

FLUID MILK PRODUCTS SPOILAGE

Types of bacterial spoilage in fluid milk products:

B. Pasteurized Milk Spoilage:

Pasteurization of milk kills most of the acid-forming bacteria, but not heatresistant
thermoduric bacteria (such as Bacillus, Clostridium, Corynebacterium, Enterococcus,
Lactobacillus, Micrococcus, and Streptococcus). These thermoduric microorganisms can
grow at low temperatures, since some species are also psychrotrophic. Coliforms,
Alcaligenes, Flavobacterium, Pseudomonas, and others can enter into milk as
postpasteurization contaminants. Pasteurized milk storage in refrigerator has a limited shelf
life, mainly due to growth of psychrotrophic contaminants. The most common fermentative
spoilage of fluid milk products is souring caused by thermoduric LAB. Lactic acid by itself has
a clean pleasant acid flavor and no odor. The unpleasant “sour” odor and taste of spoiled
milk result from formation of small amounts of acetic and propionic acid. Sour odor can
appear before an acid flavor when the microbial population reaches 106 cells per milliliter of
fluid milk products. Pasteurized milk spoils by the growth of heatresistant Streptococcus
utilizing lactose to reduce pH (curdling at 4.59). Lactobacillus continues fermentation and
may reduce the pH to 4.0 or below.

If molds present in milk, they begin to grow on the surface of soured milk and raise
the pH toward neutrality, thus allowing the growth of proteolytic bacteria (such as
Pseudomonas) and causing liquefaction of the milk curd. Pasteurization temperature
activates spore germination and spoilage by sporeforming Bacillus, and Clostridium chiefly
occurs in heated packed fluid milk with gas production. Milk can be coagulated by spore-
forming bacteria without significant acid or off-flavor formation. Microbial growth in milk
may become visible as bacterial colonies (buttons) at the bottom of the carton.

Milk products Type of spoilage Bacteria

Pasteurized milk Proteolysis Pseudomonas, Flavobacterium, Bacillus
Souring Acid proteolysis Strepococcus, Lactobacillus
Acid proteolysis Micrococcus, B. cereus, Clostridium
Sweet proteolysis Alcaligenes, Proteus, B. cereus
Malty flavor L. lacits, L. lacits. Lactis var. maltigenes,
Sour flavor Micrococcus
Repines Bacillus circulans
Blue color LAB, A. viscolactis
Gases, coagulation P. syncyanea, L. lactis
Buttons Bacillus, Clostridium
Microbial growth
UHT milk Off-tastes and off-odors, G stearothermophilus,
Physical changes G thermosaccharolyticum
Pink color, off-flavor, B sporothermodurans

Agriculture & GK Page 54

Mob: 9473826765 [Link]
Coagulation
Age gelation, increasing Milk and bacterial proteinases
Viscosity
Rancid flavors, odors Phosphlipases, lipases
Maillard products Heat treatment
Acidity Free fatty acids, other organic acids
Concentrated Acid proteolysis Bacillus
milk
Bitter flavor B licheniforms, C botulinum
Swelling, off-flavors C sporogenes, chemical reaction, overfilling.
Xerophilic mold.
Late sour B licheniformis, B macerans, B subtilis, G
stearothermophilus
Buttons, coagulation Wallemia sebi
Sweet coagulation B coagulans, B cereus, T stearothermophilus
Flat sour spoilage G stearothermophilus B licheniformis, B
macerans

Degradation of casein with microbial enzymes can cause bitter flavor. Spores of
psychrotrophic Bacillus spp. surviving at pasteurization can germinate and multiply at
lowtemperature storage to cause a spoilage known as “bitty.” They produce the enzyme
lectinase. Lectinase hydrolyzes phospholipids of the fat and causes aggregation of the fat
globules that adhere to the container surfaces. Production of rennin-like enzymes by the
psychrotrophs can cause sweet curdling of milk at high pH. Nonaseptic packaged
refrigerated fluid milk may be spoiled by growth of psychrotrophic B. cereus and Bacillus
polymyxa in the absence of more rapidly growing Gram-positive psychrotrophs (such as
Pseudomonas spp.). Psychrotrophic Bacillus circulans is the predominant spoilage bacterium
in aseptically packaged heat-treated milk. This bacterium produces acid from lactose, giving
sour flavor to milk. Bacillus mycoides is another spoilage psychrotrophic spore former in
milk.

C. UHT Milk Spoilage:

UHT processing is a method of milk preservation by which both the microorganisms
and enzymes are reduced to a commercially acceptable level (commercialsterility; not more
than one package (1 l) spoiled per 10,000 orless), which ensures consumer safety and
extended shelf life of milk. However, during heat treatment and storage period, a number of
changes can occur in chemical, physical, and microbiological characteristics of the product.
These changes may make the product unacceptable because of the development of off-
flavor, color, and gelatin. Contamination of spore-forming bacteria (such as Geobacillus
stearothermophilus, Bacillus subtilis, and Clostridium botulinum) is possible with milk during
UHT process. A large number of spores can occur at the dairy farm. Feed and milking
equipment can act as reservoirs or entry points for potentially high-heat-resistant spores
into raw milk. Lowering the spore numbers by good hygienic measures can probably further
reduce the contamination level of raw milk. All microbial vegetative cells and most bacterial
spores in milk are destroyed by UHT treatments. However, some heat-resistant endospores

Agriculture & GK Page 55

Mob: 9473826765 [Link]
(such as Bacillus sporothermodurans, G. stearothermophilus, and Thermoanaerobacterium
thermosaccharolyticum) survive and heat-resistant extracellular enzymes (proteinases,
lipases, and phospholipases) retain activities at the UHT processing of milk. These bacteria
are not pathogenic and cannot grow at the temperature below 35 °C. They can cause
offtastes and off-odors, and physical changes (curdling or coagulation) in milk. Bacterial
heat-resistant extracellular proteinases can cause spoilage of UHT milk by producing bitter
peptides. Occasionally, microbial spoilage can occur in UHTtreated milk, usually as a result
of contamination during filling operations. Members of Bacillus spp. (such as Bacillus badius,
B. cereus, Bacillus licheniformis, B. polymyxa, Bacillus subtilis, and G. stearothermophilus)
are common spoilage bacteria in such situations. B. sporothermodurans spoilage in milk can
occur with a pink color change, off-flavors, and coagulation, especially in containers with a
low-oxygen barrier (such as plastic bottles). Despite its rather poor growth characteristics in
UHT milk, UHT milk can be regarded as a new ecological source for B. sporothermodurans
because of the lack of competition from other microorganisms.

D. Spoilage of Concentrated Milk Products:

Concentrated milk products can be divided into three groups: (i) evaporated milk,
(ii)sweetened or unsweetened condensed milk, and (iii) concentrated milk. These products
are subjected to sufficient heat treatments to kill microorganisms. Evaporated milk is
prepared by heating milk at 93–100 °C and then evaporating it under partial vacuum (at 50
°C). Evaporated milk results with 7.5% milk fat and 25% total solids. It is packed in
hermetically sealed cans and they are commercially sterilized at 115– 118 °C for 14–18 min.
Unsweetened condensed milk is prepared by heating milk at 100–120 °C for 1– 3 min
followed by chilling to 70 °C to stabilize the proteins. It is then concentrated at 45–70 °C
under partial vacuum. After concentration, milk is homogenized, supplemented with
stabilizers, cooled to 14 °C and packed in hermetically sealed cans. Unsweetened condensed
milk results with 10–12% fat and 36% total solids. Sweetened condensed milk is prepared by
heating milk at a high temperatures (from 71 to 100 °C for 10–30 min) and then condensed
at 50–57 °C under vacuum. Condensed milk results with 8.5% fat, 28% total solids, and 42–
64% sugar (sucrose). Only thermophilic bacterial spores can survive high-temperature
treatment of concentrated products. Exposure of milk to heat treatment over 43 °C or
higher can activate germination of spores that subsequently grow. Under such conditions,
Bacillus spp. (such as B. coagulans) can cause coagulation of products. Swelling of can
containing concentrated milk is caused by gas-forming anaerobic spore formers (such as
Clostridium spp.), overfilling of the can, and action of acid constituents of concentrated
product on the iron of the can. Bitterness usually results from proteolysis by Bacillus and
Clostridium spp. Thermophilic bacteria are a problem only if the product is stored at
elevated temperatures (over 35 °C). Halophilic and xerophilic microorganisms (such as
Micrococcus and fungi) are able to grow due to their low water activity. Growth of
xerophilic fungi may cause blowing of cans and development of off-flavors. Wallemia sebi
forms small brownish “buttons” of mycelium and coagulates casein without gas production.
Poor hygienic conditions allow contamination of spoilage microorganisms. Due to low aw (
0.85), these products are susceptible to spoilage with gas production by postprocess

Agriculture & GK Page 56

Mob: 9473826765 [Link]
contaminant yeasts (such as Kluyveromyces marxianus, Debaryomyces hansenii, Candida
famata, Candida kefyr, Rhodotorula mucilaginosa, Yarrowia lipolytica, and Pichia). Yeasts are
able to produce proteolytic or lipolytic enzymes in condensed milk.

Water activity of concentrated milk products is about 0.85, and this is only borderline
for S. aureus. However, anaerobic conditions in the condensed milk prevent growth of S.
aureus and enterotoxin formation. Sweetened condensed milk may be produced with low-
sugar content and high aw and used, for example, for pastry and confectionery. In such
products, growth of S. aureus is possible if the temperature is abused. Spores of Clostridium
and Bacillus spp. may be present in these products, but their growth is prevented by low aw.
Growth and survival of pathogens in ultrafiltered milk is similar to that in milk, except that
Listeria monocytogenes can grow faster and achieve higher populations.

E. Dried Milk Spoilage:

In the case of low-heat milk powders, the heat treatments correspond to
pasteurization; in the case of medium-heat milk powders, temperatures of 85–95 °C for 20–
30 s are applied and temperatures above 120 °C for up to 30 s are used to obtain high-heat
milk powders. Due to the extremely low aw (0.30–0.40) of dried milk products, growth of
spoilage microorganisms is not possible. Condensation of water on packaging material may
allow growth of microorganisms and must be avoided. Extent of microbial destruction
during drying depends on the types of microorganisms, drying temperature, and retention
time of process. Thermophilic and thermoduric bacteria may survive the thermal treatments
of drying process. The flora of dried milk includes (i) thermoduric Micrococcus usually
associated with dairy equipment, (ii) thermoduric Streptococcus (mainly S. thermophilus)
that grows at temperatures between 3 and 15 °C, (iii) thermoduric species of Enterococcus
(such as E. bovis, E. durans, and E. faecalis), (iv) thermoduric species of Corynebacterium
coming from milk, (v) bacterial spores of mostly B. subtilis, and (vi) miscellaneous
contaminants, such as E. coli that indicates poor sanitation and postprocess contamination
from the human reservoir. Whole milk powder contains bacterial lipases and they can cause
rancidity as a potential problem in dried milk. Low-fat milk powders (such as nonfat dried
milk, whey, and whey protein concentrate) may contain residual lipases, which become
active when these products are used in fat-containing food formulations. Dried milk may be
rehydrated and consumed directly, but it is more commonly used as ingredients in a number
of products (such as bakery, yogurt, chocolate, confectionery, baby foods, ice cream, and
animal feeds). L. monocytogenes and Salmonella (such as S. enterica subsp. enterica var.
Typhimurium and S. enterica [Link] var. Agona) may survive insufficient heat
treatment of dried milk or contaminate during processing of milk powder. Outbreak of S.
aureus can occur due to preformed staphylococcal enterotoxin when the milk powder is
used as an ingredient. Enterobacter sakazakii may present in dried milk and can cause
neonatal meningitis with the consumption of contaminated infant formula. The product
must be protected against microbial contamination, from pasteurization to the packaging
operations.

Agriculture & GK Page 57

Mob: 9473826765 [Link]
F. Fermented Milk Products Spoilage:
Buttermilk, yogurt, and cheese are a few of the fermented milk products that are
generally produced by inoculating milk with specific starter culture. They differ in acidity,
water activity, susceptibility to spoilage, and storage stability.

Cheese Spoilage:
Cheese is a ready-to-eat food and easily contaminates with microorganisms from
processing and storage environments. Cheese undergoes spoilage with bacteria, yeasts, and
molds Factors determining the type of spoilage of cheese are processing conditions, water
activity, pH, salt, moisture ratio, temperature, characteristics of the lactic starter culture,
types and viability of microorganisms, and characteristics and quantities of residual
enzymes. The types of spoilage microorganisms and their effects depend on the
characteristics of cheese, survival during processing step, contamination during processing,
and microorganisms either on the surface or in the interior of cheese. Many bacteria,
especially psychrotroph, can contaminate with cheese during processing. Psychrotrophic
Gram- negative bacteria can readily spoil the cheese, and cause visual and physical defects,
such as green or yellow slime, fruity odors, and putrid off-flavors. Bacterial spoilage may
occur in fresh cheese having a sufficiently high pH (such as cottage cheese). Microorganisms
can contaminate curd from wash water. Microorganisms can produce gases during
processing, at the beginning of ripening, and during ripening periods.

Spoilage by LAB:
Some species of LAB produce unwanted flavor in cheese. The use of high ripening
temperatures (such as 15 °C rather than 8 °C) encourages growth of heterofermentative
Lactobacillus. Lactobacillus brevis and Lactobacillus casei subsp. pseudoplantarum can
produce gas in cheese. Lactobacillus casei subsp. casei produces a soft defect in cheese.
Some unripened soft cheese with vacuum packaging and storage atrefrigerated
temperature are spoiled by heterofermentative Leuconostoc spp. This is characterized by
gas and liquid accumulation in the package. Another common spoilage of ripened cheese is
the appearance of white crystalline deposits on the surface. These deposits do not affect
flavor, but reduce consumer acceptability. Lactobacillus spp. produce higher amount of
lactic acid during cheese ripening and this results in the formation of insoluble calcium
lactate crystals.

Spoilege Microorganisms Products

Early gas Coliforms, yeasts, Lactobacillus CO2, H2
Fruity Lactococcus, Pseudomonas, yeasts Ethanol, esters
Late gas Clostridium spp. CO2, H2
Phenolic flavor Lb. casei subsp. alactosus, Lb. casei Phenolic compounds
subsp. rhamnosus
Pink Lb. bulgaricus, leuconostoc, High redox
Propionbacterium, Fusrium,
Discoloration Culmorum, yeasts potential
Rancid Pseudomonas, Micrococcus, Free fatty acids

Agriculture & GK Page 58

Mob: 9473826765 [Link]
Serratia
Slime and off- Acinetobacter, Enterococcus, Polymeric
Bacillus, Alcaligenes,
Flavor Pseudomonas, Leuconostoc products
Soft spoilage Lb. casei subsp casei Organic acids
White crystals Heterofermentative Lactobacillus Excessive D - lactate
spp.
Ropiness Micrococcus, Pseudomonas, Polymers
Serratia, Leuconostoc
Holes ih curd Bacillus, Pseudomonas, Leuconostoc CO2, H2

Lactobacillus casei subsp. alactosus and Lactobacillus casei subsp. rhamnosus

associate with the development of phenolic flavor, similar to that of horse urine after 2–6
months of aging. Fruity offflavor in cheese is usually caused by LAB (usually Lactococcus
spp.) with production of esterase. The major esters contributing fruity flavor to cheese are
ethyl hexanate and ethyl butyrate.

1. Spoilage by Psychrotrophic Bacteria:

Psychrotrophic bacteria (such as Pseudomonas, Alcaligenes, Achromobacter, and
Flavobacterium) have primary importance in cheese spoilage since these bacteria produce
very active proteolytic and lipolytic enzymes. The presence of psychrotrophic bacteria in
cheese indicates postprocess contamination since these bacteria are sensitive to
pasteurization. These bacteria grow rapidly at normal refrigeration temperature (7 °C) and
produce lipases. Lipases can lipolyze fats in cheese. Lipolysis produces short-chain fatty
acids that combine with ethanol to form fruity esters. P. fluorescens, P. fragi, and P. putida
cause bitterness, putrefaction, rancid odor, liquefaction, gelatinization, and slime formation
on cheese. The most common bacterial spoilage is “slimy curd” caused by Alcaligenes spp.
(such as A. viscolactis). Alcaligenes metacaligenes causes flat and poor flavorspoilage in
cheese. Flavobacterium spp. produce yellow flavin pigments that discolor cheese.
Psychrotrophic Bacillus species cause bitterness and proteolytic changes, and the product is
dark pigmented. P. fluorescens causes discoloration because of the formation of water-
soluble fluorescent pigments. Other Pseudomonasspp. also causes darkening or yellowing of
the cheese surface. Yellow discoloration of cheese surface may be attributed to production
of flavin pigment by Flavobacterium spp.

Spoilage by Fungi Molds, especially from the genera Alternaria, Aspergillus,

Cladosporium, Fusarium, Geotrichum, Monilia, Mucor, Penicillium, and others, lead to
undesirable changes in cheese; Penicillium most frequently occurs on cheese. Penicillium,
Mucor, and other molds grow well on cheese and produce stale, yeasty, and musty flavors.
During the first days of ripening, yeasts and/or molds (such as Debaryomyces hansenii,
Geotrichum candidum, and Penicillium camemberti) colonize on the surface of cheese and
utilize lactate. This leads to deacidification of the cheese surface, enabling the growth of less
acid-tolerant bacteria (such as Arthrobacter arilaitensis, Brevibacterium aurantiacum,
Brevibacterium linens, and Corynebacterium casei). Development of visible molds on the

Agriculture & GK Page 59

Mob: 9473826765 [Link]
surface of cheese is often the first sign of spoilage, followed by the appearance of musty off-
taints and odors. Moldsfrom the raw materials can play a major role in spoilage. Some
molds can grow under low oxygen tension. Molds commonly present in vacuum-packaged
cheese include Penicillium spp. and Cladosporium spp. Cream cheese are susceptible to
spoilage by heat-resistant molds (such as Byssochlamys nivea). B. nivea grows in reduced
oxygen atmospheres, as well as in atmospheres containing 20, 40, and 60% CO2 with less
than 0.5% oxygen.

Once this mold presents in the milk supply, it can be difficult to eliminate during
normal processing of cheese. The D value of B. nivea ascospores in milk and cheese is 1.3–
2.4 s at 92 °C, respectively. Yeasts most frequently associated with cheese spoilage include
Candida, Pichia, Yarrowia lipolytica, G. candidum, K. marxianus, and D. hansenii. The low pH
and the nutritional profile of most cheese are favorable for the growth of yeasts. The
characteristic yeast spoilage is gassiness, odors, off-flavors, and slime formation. Surface
moisture, lactic acid, peptides, and amino acids favor rapid growth of yeasts on cheese.
Many yeasts produce alcohol and CO2, resulting in yeasty taste. Yeasts can produce large
amounts of gases(such as CO2) in vacuum- and modified atmosphere-packed cheese. This
can cause swelling of packages. Some proteolytic yeast produces sulfides, resulting in an egg
odor. Yeasts cause spoilage, especially in the fresh or soft cheese during storage. Yeasts also
grow on the surface of ripened cheese. If the surface becomes wet, yeasts can cause slime
formation. A “fermented yeasty” flavor in cheese results from growth of Candida spp.

Yeasts and molds can contaminate with cheese during processing from air, brine,
equipment, the environment (floors, walls, ventilation ducts, etc.), ingredients (such as fruit
concentrates, cereals, honey, chocolate and cocoa, nuts, or spices), and stabilizing agents
(such as thickeners). Molds and yeasts contamination can be prevented during ripening by
rigorous cleaning procedures, supplying sterilized air through filtration or ultraviolet
treatment of room. In countries where there is legal allowance, natamycin or sorbic acid
may be incorporated into the packaging to prevent cheese spoilage.

2. Butter:
Spoilage Butter contains around 15% water, 81% fat, 0.4% carbohydrates, 0.6%
proteins, and 2.5% ash, and can be salted or unsalted. Cream for butter production is
heated at 71 °C for 30 min or at 93 °C for a few seconds. The microbiological quality of
butter depends on the quality of cream and the sanitary conditions used in the processing.
The main source of microorganisms for butter is cream, milk, equipment, water, and
processing conditions. Buttermilk, cream, and butter can be spoiled by psychrotrophs,
coliforms, yeasts, and LAB. Bacteria can cause five types of spoilage in butter: First, the
surface taint (putrid), which is caused by Shewanella putrefaciens, P. putrefaciens, and
Flavobacterium spp. at chilling and refrigerating temperatures within 7–10 days. Second, the
rancidity of butter, which is caused by hydrolysis of butterfat with liberation of free fatty
acids. Rancidity and fruity odors are caused by the growth of P. fragi and P. fluorescens.
Third, flavor formations—Malty flavor (due to formation of 3-methylbutanol) is caused by
Lac. lactis subsp. lactis var. multigenes. Diacetyl can be reduced by diacetyl reductase

Agriculture & GK Page 60

Mob: 9473826765 [Link]
enzymes produced by Pseudomonas, Lactococcus, and coliforms in these products, leading
to a “green” or yogurt-like flavor from an imbalance of the diacetyl to acetaldehyde ratio.
Coliforms and Enterococcus can grow in water phase of butter and produce flavor defects.
Cheesiness is caused by Lactobacillus, rancidity by lipolytic bacteria and molds, barny flavor
by Enterobacter, yeasty flavor by dissolving metals in high-acid cream, unclean flavor by
coliforms, fishiness by A. hydrophila, and ester-like flavor by P. fragi. Chemical reactions
(oxidation of unsaturated fats, rancidity, and trimethylamine formation from lecithin) can
also produce flavors. Fourth, discoloration caused by growth of molds, yeasts, and bacteria
on the surface of butter. Dark, smoky, or greenish color on butter are caused by Alternaria
and Cladosporium, small black color by Stemphylium and P. nigrificans, green color by
Penicillium, bright reddish-pink color by Fusarium culmorum, and black color by Torula spp.
Fifth, the formation of excessive viscosity in buttermilk and sour cream with the growth of
encapsulated, slimeforming Lactococcus. Pathogenic bacteria can survive and grow in butter
and cream. The presence of staphylococcal enterotoxin A is possible in butter since
enterotoxin may be formed in the cream used to make the butter and is carried over into
the finished product. The growth and survival of S. aureus, L. monocytogenes, and C. jejuni
in butter and cream is possible due to the use of contaminated ingredients, poor hygiene,
low salt concentration (or inadequate salt dispersal), insufficient heat treatment of cream,
and temperature abuse. L. monocytogenes may grow slowly from 4 to 13 °C in the butter.
AFM1 can contaminate with butter. Most of the mycotoxins present in the cream would be
removed with the buttermilk and very little will remain in the butter.

3. Yogurt Spoilage:
Yogurt generally has a low pH (with about 1% lactic acid) and is not spoiled by most
bacteria. The acid also restricts the growth of food-poisoning bacteria. So, whereas milk is a
potential source of food poisoning and has a shelf life of only a few days, yogurt is safer and
can be kept for longer time under proper storage conditions. During storage of yogurt, the
yogurt bacteria can continue to produce acids and cause an objectionable sharp acid taste.
Bacillus is the predominant spoilage bacterium in yogurt, followed by Lactobacillus,
Streptococcus, and Actinomyces. In general, coliforms cannot survive at high acidity. The
presence of coliforms indicates the poor sanitary conditions during production of yogurt.
Yeasts (especially in fruit yogurt) may grow in the acid environment and produce CO2 as
well as yeasty and fruity off-flavors. Yeasts are major spoilage organisms of yogurt in which
the low pH provides a selective environment for their growth. Yogurt produced under
hygienic conditions with good manufacturing practices should contain no more than 10 cells
per gram. However, yogurt having initial yeast counts of >100 cells per gram tend to spoil
quickly. Yeasty and fermented off-flavors and gassy appearance are detectable when yeasts
grow up to 105 –106 cells per gram. Galactose results from lactose hydrolysis by the lactic
starter cultures that are fermented by galactose-positive species of yeasts (such as S.
cerevisiae and Hansenula anomala). The most common yeast spoilage is caused by S.
cerevisiae. The other yeasts causing spoilage are Trichosporon brassicae, Cryptococcus
curvatus, K. marxianus, Trichosporon cutaneum, D. hansenii, Pichia farinosa, Candida
blankie, and G. candidum. In a higher temperature, the growth of yeasts is stimulated,

Agriculture & GK Page 61

Mob: 9473826765 [Link]
leading to the gas production and aroma and flavor formation in the yogurt. Especially,
galactose-positive yeasts can cause blowing in the fruit yogurt packages. The most
commonly associated yeasts in yogurt are K. marxianus and S. cerevisiae. In addition,
Saccharomyces exiguous, Rhodotorula glutinis, Yarrowia lipolytica, and Debaryomyces
hansenii likely cause quality problems in yogurt stored at relatively higher temperatures
(such as 15– 20 °C). Some metabolites (such as acetic acid) produced by yogurt starter
bacteria can show inhibitory effect against yeasts. The major sources of yeasts in yogurt are
ingredients that are added into the product after heat treatment.

MICROBIAL SPOILAGE OF MEAT AND MEAT PRODUCTS

➢ Meat and its product are highly nutritious food that is widely consumed by people all
over the world.
➢ Meat can be obtained from various birds (chicken, turkey, ducks, etc.) or mammals
(pork, mutton, buffalo, sheep), and after slaughtering, carcasses and primary cuts are
processed to raw or processed food products.
➢ It is a nutritious, protein-rich food that is highly perishable and has a short shelf life.
➢ The biological and chemical nature of meat leadsto its deterioration from the time
ofslaughter until consumption.
➢ Meat and its products such as ham, sausages, cooked meat, dry meats, smoked meats,
vacuum-packed meat, minced meat, etc. are all susceptible to microbial spoilage.

Contamination Source and Causes:

Meat spoilage can be caused by natural processes, such as lipid oxidation or autolytic
enzymatic that occurs in the muscle after slaughtering. Several factors are responsible for
microbial contamination of meat such as:
1) Bacterial flora of animal.
2) Knives, utensils, hands, and clothing of the workers.
3) Pre-slaughter handling of livestock and post-slaughter handling of meat.
4) handling during slaughtering, evisceration, and processing
5) temperature controls during slaughtering,
6) processing and distribution
7) type of packaging used 8. Handling and storage

Spoilage of Fresh Meat:

Fresh meat is subjected to spoilage by its enzymes and microbial action.

The autolysis changes cause proteolytic action on muscle and connective tissue and
hydrolysis of fats.

The survival and growth of microorganisms are influenced by the composition of the
atmosphere surrounding the meat.

Fresh meat contains nutrients such as sugars, amino acids, vitamins, cofactors, etc and
it had pH (5.5-5.9) and Aw (0.85) values that influence the growth of microorganisms.
Agriculture & GK Page 62
Mob: 9473826765 [Link]
The most common bacteria isolated from fresh meat are bacteria of the genera
Acinetobacter, Pseudomonas, Brochothrix thermosphacta, Flavobacterium, Psychrobacter,
Moraxella, Staphylococci, Micrococci, lactic acid bacteria (LAB), and various genera of the
Enterobacteriaceae.

The microbial pathogens found in fresh meat are Salmonella, Campylobacter, [Link],
Listeria monocytogenes.

There are two types of spoilage of meat:

1. Spoilage under aerobic condition
2. Spoilage under anaerobic condition

The kind of defects caused by microorganisms on fresh meat

Condition Kind of defects Microorganisms

Surface slime Pseudomonas, Moraxella,
Streptococcus, Bacillus,
micrococcus
The red color of meat called Lactobacillus, Leuconostoc
“bloom” caused by the
Aerobic condition production of an oxidizing
compound
Oxidative rancidity Pseudomonas spp,
Archromobacter
Red spot Serratia marcescens,
Blue color Pseudomonas syncyanea
Greenish blue or brownish Chromobacterium lividum
black spot
Stickiness, whiskers, Green Mold
patches
Anaerobic condition Putrefaction Clostridium spp,
Alcaligennes, Proteus
Souring Lactic acid bacteria

Spoilage of Meat:
➢ Microbial growth, oxidation, and enzymatic autolysis are the three basic mechanisms
responsible for the spoilage of meat.
➢ The nutrient composition, high water content, and moderate pH of meat make it an
excellent medium for microbial growth.
➢ The normal flora of an animal’s lymph nodes contaminating meat is Staphylococcus,
Streptococcus, Clostridium, and Salmonella.
➢ Meat may contain different bacteria that include species of Acinetobacter,
Aeromonas, Alcaligenes, Alteromonas, Brochothrix, Carnobacterium, Escherichia,
Enterobacter, Enterococcus, Flavobacterium, Lactobacillus, Leuconostoc, Micrococcus,
Proteus, Pseudomonas, Sarcina, Serratia, and Streptococcus.

Agriculture & GK Page 63

Mob: 9473826765 [Link]
➢ Pathogenic microbial species contaminating meat are Salmonella enteric strains,
Yersinia enterocolitica, Campylobacter jejuni, Aeromonas hydrophila, Listeria
monocytogenes, and Escherichia coli.
➢ Mold species found in meat include Cladosporium, Sporotrichum, Geotrichum,
Penicillium, and Mucor while yeasts species include Candida spp., Cryptococcus spp.,
and Rhodotorula spp.
➢ The main defects observed in meat are off-odor, off-flavor, discoloration, and gas
production.

Spoilage of Refrigerated Meat:

➢ When fresh meat is refrigerated at 4 ± 1°C, they remain in good condition for 5-7 days
➢ Refrigerated temperature favors the growth of psychrophilic organisms in due course
of time.
➢ The contaminations occur during slicing and serving operations, from hands, slicing
machines, and other equipment.
➢ Inadequate hygiene can lead to meat contamination by spoilage and pathogenic
microorganisms
➢ The important bacterial genera associated with spoilage of refrigerated meat are
Acinetobacter, Moraxella, Pseudomonas, Aeromonas, Alcaligenes, and Micrococcus.
➢ The mold genera associated with spoilage of refrigerated meat are Alternaria,
Cladosporium, Geotrichum, Mucor, Monilia, Penicillium, Sporotrichum, and
Thamnidium; and yeast genera associated with spoilage of refrigerated meat are
Candida, Torulopsis, Debaryomyces, and Rhodotorula.
➢ Generally, Brochothrix thermosphacta and lactic acid bacteria are the bacteria that
cause spoilage of refrigerated meat.
➢ Pathogenic microorganisms found in refrigerated meats include C. botulinum type E,
Yersinia enterocolitica, enteropathogenic Escherichia coli, Listeria monocytogenes,
and Aeromonas hydrophila as they are capable of growing at temperatures below 5°C.

Spoilage of Cured Meat:

➢ Cured meat is the meats in which are preserved by aging, drying, canning, brining, or
smoking for enhancement of flavor and to extends its shelf life.
➢ Some examples of cured meats are sausage, bacon, salami, ham, canned meat, dry
spicy meat, meat pickles, kebab, meatballs, meat patty, etc.
➢ The cured meat has a long shelf-life compared to fresh and raw meat however they
are not immune to spoilage. The bacterial spoilage causing organisms in processed
and cured meats are lactic acid bacteria (such as Lactobacillus sake, Lactobacillus
curvatus, Leuconostoc gelidium, Leuconostoc carnosum, Leuconostoc mesenteroides),
Acinetobacter, Bacillus, Micrococcus, Serratia, and Staphylococcus.
➢ The spoilage causing mold found in cured meat includes Aspergillus, Penicillium,
Rhizopus, and Thamnidium

Agriculture & GK Page 64

Mob: 9473826765 [Link]
➢ The spoilage causing yeast found in cured meat includes Candida, Debaryomyces,
Torula, Torulopsis, and Trichosporon.
➢ Other spoilage causing microorganism found in cured meats includes Clostridium spp,
Hafnia spp, Weisella spp, Shewanella spp, Pseudomonas spp, Enterococcus spp, etc.
➢ The pathogenic microorganisms found in cured meat include Escherichia coli,
Salmonella, Staphylococcus aureus, Listeria monocytogenes, Clostridium botulinum,
and Toxoplasma gondii.
➢ Microbial growth in cured meat can result in slime formation, structural components
degradation, decrease in water holding capacity, off odors, and texture and changes in
appearances.

Agriculture & GK Page 65

Mob: 9473826765 [Link]

FOOD HAZARD & HYGIENE

INTRODUCTION

Food is a major determinant of health, nutritional status and productivity of the

population. It is, therefore, essential that the food we consume is wholesome and safe.
Unsafe food can lead to a large number of food-borne diseases. You may have seen reports
in the newspapers about health problems caused by contaminated or adulterated foods.
Globally, foodborne illness is a major problem of public health concern. In India, the
Ministry of Health and Family Welfare, in September 2010 stated that more than 300 million
episodes of acute diarrhoea occur every year in children less than five years of age. Food-
borne illness can not only result in mortality but can damage trade and tourism, lead to loss
of earnings, unemployment and litigation and thus can impede economic growth, and
therefore food safety and quality have gained worldwide significance.

FOOD QUALITY

The term food quality refers to attributes that influence a product’s value to
consumers. This includes both negative attributes such as spoilage, contamination,
adulteration, food safety hazards as well as positive attributes such as colour, flavour,
texture. It is therefore a holistic concept integrating factors such as nutritional traits,
sensorial properties (colour, texture, shape, appearance, taste, flavour, odour), social
considerations, safety. Safety is a preliminary attribute and precursor of quality. In order to
ensure that foods are safe and of good quality, across the world various governments and
international bodies have laid down food standards that manufacturers/suppliers are
expected to adhere to.

Thus, all food service providers (those involved at all stages of pre-preparation and
preparation/processing, packaging and service) should adhere to good manufacturing
practices and ensure food safety.

Salient points to be borne in mind are:

➢ Quality of raw materials and water.

➢ Cleanliness of the premises, personnel, equipment, food preparation and storage and
serving areas.
➢ Storage of food at appropriate temperature
➢ Food hygiene
➢ Good service practices.

SIGNIFICANCE OF FOOD SAFETY AND QUALITY

Agriculture & GK Page 66

Mob: 9473826765 [Link]
Food safety and quality are important at the home level, but are critical in large scale
food production and processing, and also where food is freshly prepared and served. In the
past, many foods were processed at home. Advancement in technology and processing,
larger per capita incomes and better purchasing power as well as increased consumer
demand have led to a variety of products of processed foods, food for health / functional
foods being manufactured. Safety of such foods needs to be assessed.

Quality of food stuff, raw as well as processed is of public health concern and must be
addressed. In the past decade, safety challenges faced globally as well as in India have
changed significantly and issues related to food quality and food safety have gained
tremendous importance.

A number of factors are responsible for this:

➢ with fast changing lifestyles and eating habits, more people are eating outside their
homes. In commercial settings, foods are prepared in bulk handled by many persons,
thus there are more chances of food getting contaminated. Further, food items are
prepared many hours in advance, and may spoil if not stored appropriately.

➢ There are many processed and packaged food, safety of these foods is important.

➢ Spices and condiments, oilseeds were processed at home in former times and purity
of these were not a concern. In today’s world, pre-packaged individual spices,
condiments, spice powders and mixes are in demand, especially in cities and metros.
Quality of even raw food stuff besides processed foods is of public health concern and
must be addressed.

➢ Logistics governing transport of bulk food is complex and there is a long gap between
processing and consumption. Thus, risk assessment and safety management during
mass production and mass distribution is critical.

➢ Microbial adaptations, antibiotic resistance, altered human susceptibility and

international traveling have all contributed to increasing incidence of food-borne
microbial diseases. Nearly half of all known food-borne pathogens have been
discovered during the past 25-30 years. There are still many foodborne illnesses of
unknown etiology. This is an issue of global public health concern and there is a need
to detect, identify and recognise emerging pathogens and establish active surveillance
networks, nationally and internationally.

➢ India is a signatory to the World Trade Organisation (WTO) non-tariff agreement,

which has provided greater access to world markets and opportunities to all countries
to enter international trade. In this scenario, it has become essential for every country
to protect the safety and quality of foods and also ensure that imported foods are of
good quality and safe to eat. Effective food standards and control systems are

Agriculture & GK Page 67

Mob: 9473826765 [Link]
required to protect food production within the country as well as to facilitate trade
with other nations. All food manufacturers are required to meet the given standards
of quality and safety, and need to have their products regularly tested.

➢ Pollution in atmosphere, soil and water, including use of pesticides in agriculture,

bring their share of contaminants. Also use of additives such as preservatives,
colourants, flavouring agents and other substances such as stabilisers makes the
analysis of food for various components, both nutrients and contaminants imperative
Owing to the above factors, there is a growing concern for safe, wholesome and
nutritious foods in a highly dynamic food business environment, which in turn has
greatly expanded the scope and has increased career opportunities in this sector.
Before learning about the various career options in this field, it will be worthwhile for
us to understand the basic concepts regarding food quality, food safety, risk
assessment, food standards and quality management systems.

FOOD SAFETY

Food safety means assurance that food will not cause any harm to the consumers. An
understanding of food safety is improved by defining two other concepts - toxicity and
hazard.

Toxicity:
Toxicity is the capacity of a substance to produce harm or injury of any kind under any
conditions.

Hazard:
Hazard is the relative probability that harm or injury will result when substance is not
used in a prescribed manner and quantity.

Hazards can be physical, chemical and biological causing harmful/ adverse effects on
the health of consumers.

Why food safety is important?

• Reduce food borne illness

• Protect and enhance the business reputation leading to increased profits to FBOs
• Enhance consumer confidence on domestic and international foods
• Reduce food wastage and associated costs

FOOD INFECTION
Food infection /Food Poisoning results from ingestion of live pathogenic organisms
which multiply in the body and cause disease. Salmonella is a classic example. This organism

Agriculture & GK Page 68

Mob: 9473826765 [Link]
exists in the intestinal tract of animals. Raw milk and eggs are also sources. Heat destroys
salmonella, however, inadequate cooking allows some organisms to survive. Often
Salmonella is spread through cross-contamination. This could happen when a cook cuts raw
meat/poultry on a chopping board and without cleaning uses it for another food which does
not involve any cooking, such as salad. Food may become infected by Salmonella if an
infected food handler does not wash hands with soap after using bathroom and before
touching food. Salmonella can reproduce very quickly and double their number every 20
minutes. The symptoms of Salmonella infection include diarrhea, fever and abdominal
cramps.

FOOD INTOXICATION
Food intoxication: Some bacteria produce harmful toxins which are present in food
even if pathogen has been killed. Organisms produce toxins when the food has not been hot
enough or cold enough. Toxins in food cannot be detected by smell, appearance or taste.
Hence foods which smell and appear good are not necessarily safe. One example of such an
organism is Staphylococcus aureus. Such organisms exist in air, dust, water. They are also
present in the nasal passage, throat and on skin, hair of 50 per cent of healthy individuals.
People who carry this organism, contaminate food if they touch these places on body while
food handling. Diarrhea is also one of the symptoms of this contamination. Parasites can
also cause infestation, e.g. worm infestation by tape worm in pork. In addition to this, food
can be infested by pests and insects.

Among the various hazards, biological hazards are

an important cause of food-borne illnesses. In spite of all
the efforts in the area of food safety, microbial food-borne
pathogens are still a serious concern and new pathogens
continue to emerge.

Factors that are important in the emergence of

pathogens include human host, animal hosts and their interactions with humans, the
pathogen itself, and the environment including how food is produced, processed, handled
and stored. For example, changes in host susceptibility due to malnutrition, age and other
conditions can allow for the emergence of new infections in vulnerable populations. Genetic
exchange or mutations in the organisms can create new strains with the potential to cause
disease. Exposure to new pathogens through changes in eating habits, climate, mass
production, food processing and increased globalisation of the food supply can allow
pathogens to emerge in new populations or new geographic areas. Examples are Norovirus,
Rotavirus, hepatitis E contributing to about 70 per cent of cases. New pathogens will
continue to evolve and there is a need to develop methods to isolate them, control them
and detect their presence in foods.

In the context of food safety, it is important to understand the terms contamination

and adulteration.

Agriculture & GK Page 69

Mob: 9473826765 [Link]
Contamination:
It is the presence of harmful, or objectionable foreign substances in food such as
chemicals, micro-organisms, diluents before/during or after processing or storage.

Adulteration:
It is intentional or accidental addition of impure or cheap or unnecessary ingredient(s),
to cheat, cheapen or falsify a preparation, that will alter the properties and composition and
diminish the quality of the food.

FOOD SAFETY RISK ANALYSIS

In order to manage safe food production, it is important to have a thorough
knowledge of food safety hazards and the associated risks which can enter at any stage in
the food supply chain.

International Agencies like Food & Agriculture Organisation (FAO) and World Health
Organisation (WHO) as well as CODEX Alimentarius Commission recommend Risk Analysis
Approach for managing the risk in the food production process.

Probability of risk is associated with the exposure to hazard. The likelihood of

occurrence of risk is when there is exposure to hazard and the rate of the likelihood of a
specific risk can be high, medium or low.

FOOD HAZARDS-THE BIGGEST THREAT TO FOOD SAFETY

Hazard is defined as a biological or chemical or physical agent in a food or condition of
the food with the potential to cause an adverse effect.

Biological hazard are living organisms, including microbiological organisms, bacteria,

viruses, fungi and parasites.

Chemical hazard are in two categories: naturally occurring poisonous substances and
chemicals or deleterious substances. The first group covers natural constituents of foods
that are not the result of environmental, agricultural, industrial or other contamination
examples being aflatoxins and shell fish poison. The other group covers poisonous chemicals
or deleterious substances which are intentionally or unintentionally added to food at some
point in the production chain, examples are pesticides and fungicides as well as lubricants
and cleaners.

Physical hazard is any physical material not normally found in food which causes
illness or injury. Physical hazards include glass, wood, stone, bone and metal.

1. PHYSICAL HAZARD
Physical hazard is any physical material not normally found in food, which causes
illness or injury and includes wood, stones, parts of pests, hair etc.

Agriculture & GK Page 70

Mob: 9473826765 [Link]

2. CHEMICAL HAZARDS
Chemical hazards are chemicals or
deleterious substances which may be
intentionally or un-intentionally added to foods.
This category of hazards includes pesticides,
chemical residues, toxic metals, polychlorinated
biphenyls, preservatives, food colours and other
additives.

3. BIOLOGICAL HAZARDS
Biological hazards are living
organisms and include
microbiological organisms. Those
micro-organisms which are
associated with food and cause
diseases are termed food-borne
pathogens. There are two types of
food-borne diseases from microbial
pathogens—infections and
poisoning.

Agriculture & GK Page 71

 Mob: 9473826765 [Link]

RISK ANALYSIS
Risk analysis plays an important role for a National Food Control System. It is a
powerful tool to carry out science based analysis and to achieve a sound and consistent
solution to food safety problems. It provides information on hazard in food to be linked
directly to data on the risk to human health and to improve food safety decision making
process.

It comprises of three stages:

i. Risk Assessment
ii. Risk Management
iii. Risk Communication and Stakeholder Environment

Risk analysis involves identifying the risk and weighing their likelihood and impact on health
and then establish system to reduce or mitigate the risks.

i. Risk Assessment:
Is a scientifically based process consisting of four steps:

Step 1 Hazard identification:

“Could this food or anything in it be harmful?” Risk assessors collect and review
scientific data and identify biological or chemical hazards in food.

Step 2 Hazard characterization:

“What effects do the hazards cause?” Risk assessors evaluate scientific data to
determine whether evidence is strong enough to demonstrate that a substance has the
potential to cause harm and the nature of the harm.

Step 3 Exposure assessment:

Agriculture & GK Page 72
Mob: 9473826765 [Link]
“Who may be harmed and at what level of exposure may be harmful?” Experts
estimate how much of the food or ingredient consumers in general population groups (e.g.
infants, children, adults) or subpopulations (e.g. vegetarians, vegans) are likely to be
exposed to under real-life conditions where both dose and duration are considered. The
exposure must be evaluated to determine if a hazard presents an actual risk (step 4). With
increased exposure, the risk also increases.

Step 4 Risk characterization:

“How likely is it that people will experience exposure at a level that can cause harm in
real life?” The level of exposure that can cause harm is compared to the actual level of
exposure that someone would experience in real life. If the exposure level is higher than
that which causes harm, there may be a safety concern for consumers in general or for
specific groups.

Risk assessment is carried out by the independent Scientific Panels and Scientific
Committee (of FSSAI) to provide scientific inputs on the risk and potential adverse effect of
the risk to the health of the consumers. These inputs are based on sound scientific
principles, data and evidence.

ii. Risk Management:

It is the process of weighing policy alternatives in consultation with all interested
parties, considering risk assessment and other factors relevant for the health protection of
consumers and if needed, selecting appropriate prevention and control measures. A range
of risk management options are available to FSSAI as risk managers for preventing or
reducing health risks associated with food. These options can be regulatory i. e. those
specified in the Regulations, such as end product standards or outcome based standards or
non-regulatory, such as industry codes of practice, guidelines or information campaigns.

iii. Risk Communication:

Is the interactive exchange of information and opinions throughout the risk analysis
process concerning hazards and risks, risk related factors and risk perceptions, among risk
assessors, risk managers, consumers, industry, the academic community and other
interested parties. FSSAI is responsible for risk communication, which is a two-way process
and involves sharing the information internally with risk analysis team and with external
stakeholders including general public in an open and transparent way including the
explanation of risk assessment findings and the basis of risk management decisions. Risk
communication is also important to bridge the gap which sometimes exists between the
scientific assessment and consumers' perceptions of risk.

FOOD HYGIENE

Agriculture & GK Page 73

 Mob: 9473826765 [Link]
Food is a potential source of infection and is liable to contamination by microbes, at
any point during its journey from the producer to the consumer. Food hygiene may be
defined as the sanitary science which aims to produce food which is safe for the consumer
and of good keeping quality. It covers a wide field and includes the rearing, feeding,
marketing and slaughter of animals as well as the sanitation procedures designed to prevent
bacteria of human origin reaching foodstuff. Food hygiene, in its widest sense, implies
hygiene in the production, handling, distribution and serving. WHO (1984) has defined Food
Hygiene or Food Safety as all conditions and measures that are necessary during the
production, processing, storage, distribution and preparation of food to ensure that it is
safe, sound, wholesome and fit for human consumption. The primary aim of food hygiene is
to prevent food poisoning and other food-borne illness. The objective of food control has
three aspects-economic, aesthetic and public health. The laws and regulations are intended
to protect the public against injury to health and fraud and deceit. The laws and regulations
restrain the sale of foods which are decomposed, adulterated, improperly preserved or
misbranded. They provide specifications for safe handling of such perishable and potentially
dangerous foods such as milk or milk products and meats. Methods and procedures have
been developed to minimize the danger of contamination of foods with poisonous
substances or with pathogenic microbes.

DIFFERENT BRANCHES OF FOOD HYGIENE INCLUDE

i. Milk hygiene
ii. Meat hygiene
iii. Fish hygiene
iv. Egg hygiene
v. Hygiene of vegetables and fruits
vi. Food-handlers hygiene
vii. Sanitation of eating place

i. Milk Hygiene:
Milk is an efficient vehicle for a great variety of disease agents. Milk gets
contaminated by various sources like udder, utensils, personel hygiene of the
handlers, storage environment, water etc., This may lead to various milk-borne
diseases that may affect the population. Requisites in the production of clean and safe
milk are:
a. Healthy and clean animal - Milk from a healthy udder contains only few
microorganisms.
b. Sanitary conditions of the dairy farm i.e., the premises where the animal is housed and
milked should be sanitary.
c. Containers and equipment should be sterile and kept covered.
d. Water supply should be bacteriologically safe.
e. Milk handlers must be free from communicable diseases and before milking they must
wash their hands and arms. Milking machines should be used as far as possible.9

Agriculture & GK Page 74

Mob: 9473826765 [Link]
f. Milk must be cooled immediately to 10°C after it is drawn to retard bacterial growth.
g. Milk should be properly pasteurized to increase shelf life.

ii. Meat Hygiene:

A number of diseases are transmitted through meat and meat-based foods since
animal tissues are important vehicles for transmission of various protozoan diseases
like Taeniasis, Trichinellosis and a number of bacterial infections. Meat inspection is a
very important process before being accepted or rejected. Two types of inspection, i.
e., Ante-mortem and post-mortem inspection are being carried out.

Ante-mortem rejection:
It is based on emaciation, exhaustion, pregnancy, sheep-pox, foot-rot actinomycosis,
brucellosis, febrile conditions, diarrhea and other diseases.

Post-mortem rejection:
It is based on Cysticercus bovis, liver fluke, abscesses, sarcocystis, hydatidosus,
septicaemia, parasitic and nodular infection of liver and lungs, tuberculosis and Cysticercus
cellulosae.

Good meat qualities:

The meat should be firm and elastic to touch, not be pale pink or deep purple color.

Slaughter house hygiene:

Hygiene of slaughter house is of paramount importance to prevent the contamination
of meat during the process of dressing. There is a Model Public Health Act (1955) in India,
which standardizes on the location, structure, disposal of wastes, water supply, examination
of animal, storage of meat, transportation of meat and miscellaneous other activities
connected with meat processing.

iii. Fish Hygiene:

Fishes are also important agents for disease transmission since they are perishable
foods. It is an intermediate host of tapeworm and consumption of spoilt fishes may
lead to fish poisoning. Fresh fish is in a state of stiffness or rigor mortis with bright red
gills, and clear and prominent eyes. Fishes also get contaminated through various
means during catching, transporting and processing. Prevention of contamination is of
utmost importance at each level of processing for increasing the shelf life and quality
of fish.

iv. Egg Hygiene:

Majority of freshly laid eggs are sterile. Shells of eggs become contaminated by faecal
matter from the hens or ducks. The microorganisms can penetrate the shell of the
egg, cross the various chemical barriers to reach the yolk leading to spoilage of the

Agriculture & GK Page 75

Mob: 9473826765 [Link]
egg. Eggs must be stored in a dry condition for preventing spoilage. Eggs can also be
pasteurized to increase the shelf life.

v. Vegetable and Fruit Hygiene:

Vegetable and fruits host many pathogens like bacterial, fungal, protozoan which can
enter the plant materials during or after their harvest. Standards have been laid down
for effective storage of the vegetable and fruits to prevent their spoilage and further
disease transmission.

vi. Hygiene for Food-Handlers:

Food sanitation rests directly upon the state of personal hygiene and habits of the
person working in food industries. They may transmit infections like diarrhea,
dysenteries, typhoid, enteroviruses, viral hepatitis, protozoan cysts, eggs of helminths,
staphylococcus/streptococcal infections. Simple rules to be followed in food handling
are given below:

Medical examination to be carried out at the time of employment, Persons with above
diseases or communicable diseases (TB) are not to be employed. Persons with wounds,
otitis media, skin infections, etc., should not be permitted to handle food or utensils. The
day-to-day health appraisal of food-handlers is important. Those who are ill should be
excluded from food handling. Any illness which occurs in a food-handler’s family, should be
at once notified. Education of food-handlers in matters of personal hygiene, food handling,
utensil/dishwashing and insect or rodent control is the best mean of promoting food
hygiene.

vii. Personal hygiene to be promoted:

a. Hands – scrubbed and washed with soap and water immediately after visiting lavatory
and as often as necessary at other times. Nails to be kept trimmed and free from dirt.
b. Hair – to provide covering to the head.
c. Overalls – clean white overalls to be worn by all food-handlers.
d. Habits – coughing and sneezing in the vicinity of food, licking the fingers before picking
up an article e) of foods, smoking on food premises are to be avoided.

viii. Hygiene in Public Eating Places:

The principles of food hygiene in public eating or drinking places are the same as in
any other type of food handling activity. The complexity of the problem is mainly due to the
ever increasing number of establishments which requires some supervision. At one time,
public eating was restricted to special occasions, to the traveler, patient, boarding schools,
etc., Due to increased urbanization, distance of working places, mobility and employment,
many resort to dining in public eating places.

Agriculture & GK Page 76

Mob: 9473826765 [Link]
The six minimum essentials for cleanliness and sanitation for public eating or drinking places
are as follows:

a. Avoid hand contact with food as far as possible.

b. Keep perishable food below 40°F or above 140°F.
c. Keep food protected from personal contact and contaminating insects, dust and
animals.
d. Discard all food and food products which are not in good quality.
e. Clean and disinfect equipment that comes into direct contact with food.
f. Keep premises presentable and in a sanitary manner at all times.

Sanitation of eating establishments is a challenging problem in India. The Model Public

Health Act, Govt. of India (1955) has suggested the following minimum standards for
restaurants and eating places in India.

a. Location:
Shall not be near any accumulation of filth or open drain, stable, manure pit and other
sources of nuisances.

Floors:
To be higher than the adjoining land, made with impervious material and easy to keep
clean.

b. Rooms:
Rooms where meals are served shall not be less than 100 sq. feet and shall provide
accommodation for a maximum of 10 persons. Walls up to 3 feet should be smooth
with rounded corners and should be impervious and easily washable. Lighting and
ventilation should be ample with natural and artificial lighting systems along with a
good circulation of air.

c. Kitchen:
It should be with ample floor space, window opening, proper flooring and ventilation.

d. Storage of Cooked Food:

Separate rooms to be provided for storing cooked foods. For long storage, control of
temperature is necessary.

e. Furniture:
Furniture should be reasonably strong and easy to keep clean and dry. (f). Disposal of
refuse Refuse to be collected in covered and impervious bins and disposed-off twice a day.
(g). Water Supply It should be an independent source, adequate, continuous and safe. (h).
Washing Facilities Cleaning of utensils or crockery to be done in hot water and followed by
disinfection.

Agriculture & GK Page 77

Mob: 9473826765 [Link]

f. Disposal of refuse:
Refuse to be collected in covered and impervious bins and disposed-off twice a day.

g. Water Supply:
It should be an independent source, adequate, continuous and safe.

h. Washing Facilities:
Cleaning of utensils or crockery to be done in hot water and followed by disinfection.

Agriculture & GK Page 78

Mob: 9473826765 [Link]

FOOD CONTAMINATION & DISEASE

Food contamination refers to the presence of harmful chemicals and microorganisms

in food, which can cause consumer illness. This article addresses the chemical
contamination of foods, as opposed to microbiological contamination, which can be found
under foodborne illness.

A food contamination has been defined as any substances not intentionally added to
food, which is present in such food as a result of the production, manufacture, processing,
preparation, treatment, packing, transport or storage of such food as a result of
environmental contamination.
Contaminants are substances either added to food or not intentionally added to food in the
process of their production (including operations carried out in crop husbandry, animal
husbandry and veterinary medicine), manufacture, processing, preparation, treatment,
packing, packaging transport or holding of articles of such food as a result of environmental
contamination called Crop Contaminants. Since contamination generally has a negative
impact on the quality of food and may imply a risk to human health, so measures to
minimise contaminants in foods up to the levels that can pose no risk to human health are
in place. The basic regulatory control of chemical contaminants in food are laid down in FSS
(Contaminants, Toxins and Residues) Regulation, 2011.

There are three categories of food contamination:

• Physical

Agriculture & GK Page 79

Mob: 9473826765 [Link]
• Chemical
• Microbial

(i) PHYSICAL CONTAMINANTS

Physical contaminants are any physical objects that can be present in food, and not
meant to be in the food. This can include hair, glass, and insects. Some physical
contaminants, such as hair, are not considered food safety concerns. While others, such as
glass, are considered food safety concerns.

Physical contaminants often enter the product through not following good
manufacturing practices (GMPs) which include wearing proper protective equipment, such
as hair nets. It also includes keeping the premises clean with well-maintained equipment
and having a pest control policy in place to prevent infestation with insects and pests.

(ii) CHEMICAL CONTAMINANTS

Chemical contaminants mean chemicals in food that are not meant to be in the food
products and their presence in food is unintentional and undesirable. These include
antibiotics residues, pesticides residues and cleaning agents. Many of these contaminants
have food safety and quality concerns. Chemical contaminants may be harmful to health at
certain levels. It is necessary to manage their levels in food and reduce dietary exposure of
consumers. Several measures are required to be in place to manage the risk from these
contaminants and reduce the levels at which they are present in food - including good
practices and regulatory controls.

Examples of chemical contaminants include the following:

a. Mycotoxins
b. Heavy metals - lead and mercury, Arsenic, Cadmium
c. Pesticides and dioxins
d. Antibiotics and veterinary drug residues

Mycotoxins:
Mycotoxins are toxic compounds that are naturally produced by certain types of
moulds (fungi). Moulds that can produce mycotoxins grow on numerous foodstuffs such as
cereals, dried fruits, nuts and spices. Mould growth can occur either before harvest or after
harvest, during storage, on/in the food itself often under warm, damp and humid
conditions. Most mycotoxins are chemically stable and survive food processing. Several
hundred different mycotoxins have been identified, but the most commonly observed
mycotoxins that present a concern to human health and livestock include aflatoxins,
ochratoxin A, patulin, fumonisins, zearalenone and nivalenol/deoxynivalenol. Mycotoxins
appear in the food chain as a result of mould infection of crops both before and after
harvest. Exposure to mycotoxins can happen either directly by eating infected food or
indirectly from animals that are fed contaminated feed, in particular from milk.

Agriculture & GK Page 80

Mob: 9473826765 [Link]

How to minimise the risk from Mycotoxins:

Moulds that produces mycotoxins can grow on a variety of different crops and
foodstuff and can penetrate deep into food and do not just grow on the surface. Moulds
usually do not grow in properly dried and stored foods, so efficient drying of commodities
and maintenance of the dry state, or proper storage, is an effective measure against mould
growth and the production of mycotoxins.

To minimize the health risk from mycotoxins, it is advisable to:

➢ Inspect whole grains (especially corn, sorghum, wheat, rice), dried figs and nuts such
as peanuts, pistachio, almond, walnut, coconut, hazelnuts etc. which are prone to
contamination with aflatoxins for evidence of mould, and discard any that look
mouldy, discoloured or shrivelled;
➢ Avoid damage of grains before and during drying and in storage, as damaged grain is
more prone to invasion of moulds and therefore mycotoxin contamination;
➢ Buy grains and nuts as fresh as possible;
➢ Make sure that foods are stored properly – kept free of insects, in dry and not too
warm conditions;
➢ Do not keep foods for extended periods of time before being used;

Aflatoxins are a class of mycotoxins produced primarily by Aspergillus Flavus and

Aspergillus parasitic group of fungi. The spores of these fungi grow on a suitable food
substrate under favourable conditions. Aflatoxins prevalence is aggravated in pre-harvest
crops by conditions of drought, floods, delayed harvest, pest infestation, inadequate drying
including improper postharvest handling and storage of crops and food.

The maximum limits for mycotoxins including aflatoxins are prescribed under FSS
(Contaminants, Toxins, and Residues) Regulations 2011. The table below contains the
maximum limits of the aflatoxins in various food conditions as per the regulation.

[Link]. Name of the Contaminants Article of Food Limit

µg/kg
1. Aflatoxin All articles of food 30
2. Aflatoxin M1 Milk 0.5
3. Patulin Apple juice & Apple juice ingredients in 50
other beverages
4. Ochratoxin A A Wheat, barley & rye 20

Heavy metal contaminants in food:

Heavy Metal contaminants in food crops is an issue of global concern that ultimately
results in toxicity and diseases in humans and animals through consumption of
contaminated soils and food crops. Metals such as arsenic, cadmium, lead and mercury that
are naturally occurring chemical compounds present at various levels in the environment, e.
g. soil, water and atmosphere can also occur as residues in food because of their presence in

Agriculture & GK Page 81

Mob: 9473826765 [Link]
the environment. The presence of these metals in foods is the result of human activities
such as farming, industry or from contamination during food processing and storage. People
can be exposed to these metals from the environment or by ingesting contaminated food or
water. Their accumulation in the body can lead to harmful effects over time. Due to the
adverse effects of these heavy metals on humans, FSSAI has established maximum levels for
these commonly occurring metal contaminants in foods. Additionally, these standards are in
a constant state of revision because of ongoing studies. The commonly occurring metal
contaminants found in food products are lead, copper, arsenic, tin, zinc, cadmium, mercury,
chromium, nickel and their limits in ppm across different food products is prescribed in FSS
(Contaminants, Toxins and Residues) Regulations, 2011.

(i) Metal contaminants of concern in human diet:

Concerns regarding toxicity of metals exist primarily due to their bioaccumulation in
the tissues of the body where they are taken up and stored faster than they are excreted.
Secondly, their presence in the body disrupts normal cellular processes, leading to toxicity in
a number of organs. Four heavy metals, namely lead, arsenic, cadmium and mercury, are of
particular concern in food. Ingestion of such metals from food and water is a major route of
exposure for the general population. In order to protect human health, it is necessary to
control the levels of these toxic metals in foodstuffs.

(ii) Exposure pathways of heavy metals:

Human exposure to metals occur through consumption of contaminated food stuffs,
sea foods, and drinking water, through inhalation of polluted air as dust fumes, or through
occupational exposure at workplace. These metals can be taken up through several routes.
The chain of contamination usually follows from industry, to the atmosphere, soil, water,
foods and then human. Some heavy metals such as lead, cadmium, mercury, arsenic can
enter the body through the gastrointestinal route that is, through the oral ingestion after
consuming food including fruits, vegetables, drinking water or other beverages. Some
metals can enter the body by inhalation while others such as lead can be absorbed even
through the skin.

(a) Lead:
Lead is a ubiquitous heavy metal with no physiological function. Exposure to lead
occurs mainly at occupational sites, production of lead-acid batteries or pipes, metal
recycling industries and foundries. Therefore, it may get into soil and flow into water bodies
which can be taken up by plants, and human exposure occurs via food or drinking water.
The water supplies get contaminated through lead pipes, where lead is used as a
construction or welding material. The contamination of food can occur through food
containers containing lead, e.g. storage in lead-soldered cans, ceramic vessels with lead
glazes and leaded crystal glass. The lead gets accumulated in fish and shellfish in addition to
offal (liver and kidney) of animals. It can settle on soil or be absorbed by plants grown for
fruits or vegetables. The amount of lead in a food product should not be high enough to
raise a person's blood lead level to a point of concern. Too control this, the regulatory
Agriculture & GK Page 82
Mob: 9473826765 [Link]
agencies like FDA have established a maximum daily intake for lead, called the Interim
Reference Level (IRL) at 3 Hg per day for children and 12.5 Hs per day for adults.

Toxicity of Lead:
Lead poisoning occurs mainly by ingestion of food or water contaminated with lead.
Lead affects almost every organ in the body. The nervous system is the most affected target
in lead toxicity. both in children and adults. Infants and young children may undergo
behavioral problems, learning deficits and lowered intellectual quotient. Long-time
exposure to lead has been reported to cause anaemia, damage to kidneys, reproductive and
immune systems. Severe damage to brain and kidneys, both in adults and children, were
found to be linked to exposure to high lead levels even resulting in death.

(b) Mercury:
Mercury is a toxic heavy metal present in the environment both in organic and
inorganic form. It is the only metal which is liquid at room temperature. Inhaled elemental
mercury is retained in lungs and gets distributed in the plasma and erythrocytes to almost
equal degree. It can pass the blood brain barrier and the placenta. Oxidation to mercuric
form, the toxicological relevant form is fast. Methylmercury is the most toxic form and is
found at significant levels in fish and seafood. The industrial release of inorganic mercury
into marine environments results in its uptake by marine microorganisms which then
convert the inorganic mercury, which is less toxic, into the more toxic methyl mercury. It is
accumulated in food chain due to its low rate of breakdown, reaching potentially toxic levels
in species at the top of the food chain which forms part of the human diet. The major
potential dietary sources of exposure to methylmercury are fish and shellfish.

Toxicity of Mercury:
Excessive exposure to mercury is associated with a wide spectrum of adverse health
effects including damage to the central nervous system and the kidneys. Organic forms of
mercury can cross the placental barrier between the mother and the unborn baby, and
epidemiological studies in exposed populations of humans and toxicological studies in
animals have shown that this can result in a range of neurological disturbances from
impaired learning to clear brain damage.

(c) Cadmium:
Cadmium has a high phyto-accumulation index because of its low adsorption
coefficient and high soilplant mobility and thereby may enter the food chain. Its uptake by
plants depends on soil factors such as pH, Cadmium: Zinc ratio etc. Zinc limits the cadmium
uptake and thus protects the food chain. However, rice has unusual capacity to exclude zinc
which helps transfer of high level of cadmium from soil to plant in bioavailable form. Highest
levels of cadmium are found in the offal (kidney and liver) of mammals and in mussels,
oysters and scallops. Certain wild mushrooms may also contain high levels. These foodstuffs
are however minor contributors to overall intake of cadmium.

Agriculture & GK Page 83

Mob: 9473826765 [Link]
Toxicity of Cadmium:
The principal toxic effect of cadmium is on kidneys. It is also involved in lung damage
and skeletal changes in occupationally exposed populations. Cadmium is relatively poorly
absorbed in the body, but once absorbed it is slowly excreted, like other metals, and
accumulates in the kidney causing renal damage. Its uptake in the intestine occurs in two
phases i.e. lumen to mucosa and mucosa to blood. It is also evident that these are
influenced by interaction with other metals and proteins. Iron deficiency increases cadmium
absorption.

(d) Arsenic:
People are exposed to elevated levels of inorganic arsenic through various sources.
The greatest threat to public health from arsenic originates from contaminated
groundwater. Rice plants take up more arsenic, approx. 10 folds, as compared to other
cereal crops. Arsenic-contaminated water used for irrigation and food preparation is the
source of the high arsenic content detected in cultivated grains and vegetables and in
cooked food. Terrestrial foods typically contain low levels of arsenic but arsenic in rice is
elevated when grown on arsenic-rich groundwater and soil, arsenical pesticides, phosphate
fertilizer, processing industries, and through pollution from mining activities.

Toxicity of Arsenic:
People affected with arsenicosis generally show symptoms of skin lesions like
pigmentation, keratosis or melanosis which could range anywhere between moderate to
severe. Symptoms of early acute toxicity include muscular pain, nausea, vomiting and rice-
watery stool diarrhea. Severe toxicity is known to cause kidney failure, seizures and finally
death due to shock.

REGULATORY STATUS

Regulations have been established in many countries for metal contaminants in foods.
Internationally, Codex Alimentarius Commission has set limits for metal contaminants in
foods under the "General standard for contaminants and toxins in food and feed (CXs 193-
1995)" which serves as the reference standard for international trade in food.

Provisions under Food Safety and Standards Act, 2006:

The maximum limits for metal contaminants are prescribed under Food Safety and
Standards (Contaminants, Toxins and Residues) Regulations, 2011 (FSSR). These Regulations
are available on FSSAI website ([Link]). The regulations establish maximum levels
(MLs) for various metal contaminants in a range of foodstuffs. Maximum limits of some of
the major food categories covered above are reproduced below:

Name of metal contaminant Article of food Parts per million by

weight

Agriculture & GK Page 84

Mob: 9473826765 [Link]
Brassica vegetable excluding 0.3
Kale, Leafy vegetables
(excluding spinach)
Lead Bulb vegetables, Fruiting 0.1
vegetables, cucurbits and Root
& tuber vegetables
Foods not specified 2.5
Arsenic Foods not specified 1.1
Rice Polished 0.4
Brassica, Bulb and fruiting 0.005
Cadmium vegetables, Cucurbits
Leafy vegetables 0.2
other foods 1.5
Fish 0.5
Predatory Fish (Tuna, Marlin, 1.0
Mercury Sword Fish, Elasmobranch
Non predatory fish, 0.5
Cephalopods, crustaceans,
molluscus
other Foods 1.0

Test methods for determination of metal contaminants in food:

Various analytical methods are employed in analysis of metal contaminants in food.
The analytical Plasma-Optical methods Emission include Atomic Absorption
Spectrophotometer, Inductively Coupled Spectrometer, Spectrophotometer, colorimetric
method, and titrimetric method. The detailed methodology for sample preparation,
extraction, and analysis is provided in the Manual of methods of analysis of foods- Metals.

Pesticides in foods:
Pesticide residues in food commodities and their entry into the food-chain has
become a major cause of concern all-over the world. Food safety has become crucial for all
involved in the value chain and consumers have to be assured that they are not exposed to
an unacceptable level of pesticide residues. The presence of the residues above the
permissible level is also a major bottleneck in the international trade of food commodities.

FSSAI regulates the pesticide residues detected in various food items through FSS
(Contaminants, Toxins and Residue) Regulations, 2011. The regulation covers more than 150
insecticides/pesticides whose tolerance limits have been prescribed under this regulation.
There is a harmonized monitoring of pesticide residues in the country which started under
central sector scheme called, “Monitoring of Pesticide Residues at National Level” (MPRNL)
in food commodities since 2005. Pesticide residues in the crops are monitored through the
use of maximum residue limits (MRL), which are based on the analysis of the quantity of a
given chemical remaining on food product samples. Regulating pesticides on the ground
revolves around this regulatory yardstick called the MRLs of the pesticides in a food
commodity.

Agriculture & GK Page 85

Mob: 9473826765 [Link]
Antibiotic and veterinary drug residues:
Antibiotics are naturally formed metabolites derived from fungi or bacteria or are
produced by modern biotechnology and chemical synthesis as well. Antibiotics are able to
kill microorganisms or inhibit their growth. In human and veterinary medicine, antibiotics
are therefore used as drugs for the treatment of bacterial diseases.

FSSAI has notified the FSS (Contaminants, toxins and Residues) Amendment
Regulations, 2018 specifying 'Tolerance Limits' of antibiotics and other veterinary drugs in
meat/meat products, poultry, fish and milk. Through this amendment, the existing
regulations have been further expanded to include new tolerance limits for 103 antibiotics
and veterinary drugs in meat/meat products (including poultry and fish) and milk. Among
the new provisions, tolerance limits for 76 antibiotics (these are either prohibited or not-
intended for use in food producing animals) have been specified at 0.01 mg/kg of the listed
food and largely reflects the level of detection of such antibiotics/drugs by existing method
of analysis. The amendment also includes revisions to the prohibited list of antibiotics and
veterinary drugs applicable to meat and meat products, and poultry and eggs, sea foods
including shrimps, prawns or any variety of fish and fishery products.

ORIGIN AND HEALTH RISKS OF ANTIBIOTIC RESIDUES IN FOOD

The use of antibiotics as drugs for the treatment of diseased animals is a matter of
animal welfare and therefore inevitable. As a result of application failures, such as
noncompliance with the statutory withdrawal period or use of prohibited antibiotics, misuse
of antibiotics as growth promoters, residues of antibiotics can occur in food of animal origin
such as meat, milk or eggs and this can pose health risk for consumers.

The major public health significances of antimicrobial residues include the

development of antimicrobial drug resistance, hypersensitivity reaction, carcinogenicity,
mutagenicity, teratogenicity, bone marrow depression, and disruption of normal intestinal
flora. Indiscriminate use of antimicrobials in aquaculture is also resulting in occurrence of
residues in aquaculture products and associated harmful health effects in humans and
requires control measures to ensure consumer protection.

(iii) MICROBIAL CONTAMINATION

If food is consumed that has been contaminated by certain, harmful bacteria or their
toxins (poisons produced by some of these bacteria), food poisoning may result. Bacteria
are responsible for most food poisoning cases. Symptoms of food poisoning may include
vomiting, diarrhea, fever and abdominal pain. The symptoms may take some time to occur
depending on the type of bacteria. In general, the bacteria must grow in the food to
produce sufficient numbers to infect the body, multiply with in the intestine and cause
illness. Alternatively, toxins may be produced in the foodstuff or within the intestine, to
produce symptoms very soon after ingestion. It is important to remember that food
contaminated with pathogenic bacteria will look, taste and smell perfectly normal. Steps
Agriculture & GK Page 86
Mob: 9473826765 [Link]
must therefore be taken to prevent pathogenic bacteria getting onto food and multiplying
to levels that will cause food poisoning.

Food poisoning due to microbes is very common, some food borne disease are given
below:

S. No. Organism Disease

1. Staphylococcus aureus Gastroenteritis
2. Clostridium botulinum Botulism
3. Bacillus cereus Diarrhea
4. Salmonella typhi Enteric fever typhoid
5. Escherichia coli Gastroenteritis,
diarrhea

Prevention and Control of Microbial Contamination:

• Areas must be regularly cleaned and, where necessary, disinfected.
• Interior surfaces, e. g. floors, walls, ceilings should be smooth, free from cracks and
should permit easy and effective cleaning.
• A high level of cleansing and hygiene should be practiced in every aspect of the
manufacture of drug product.
• Suitable grade of cleaning agents should be used to minimize health risks.
• Contact time, application, temperature, mechanical action and the chemistry of the
cleaning agents should all be considered during the design of the cleaning process.
• Materials used for cleaning should not come in direct contact with the product.
• Validation of cleaning practices must be carried out to provide evidence that the
process is effective in controlling microbial contamination.
• Steam used for cleaning and sanitization of production tools and equipment, supply
for autoclaves and humidification must be clean and free of additives.

Chemicals including pesticides, bleach and other cleaning materials can contaminate
food if not used carefully. For example, store cleaning fluids separate to foods to prevent
tainting and contamination if there is a spillage.

ANTIMICROBIAL RESISTANCE (AMR)

Antimicrobial resistance (AMR) has emerged as a major threat to public health
estimated to cause 10 million deaths annually by 2050. India carries one of the largest
burdens of drug-resistant pathogens worldwide. AMR develops when microbes develop
mechanisms to evade the action of antimicrobials. The driving factors that contribute to
AMR include:

• Misuse /overuse of antimicrobial in Human and Animal sectors

• Environmental contamination (including sewage and heavy metals)
• Use of antibiotics for non-therapeutic purpose o Few regulations against non-
therapeutic use of antibiotics.

Agriculture & GK Page 87

Mob: 9473826765 [Link]
• No stringent implementation protocols even when there are regulations.
• Poor sanitation plays a major role in the spread of antibiotic-resistant bacteria and
acquiring of Antibiotic Resistant Genes (ARG).
• No proper surveillance system in place.

FOOD POISONING & FOOD BORNE DISEASE

The term “food poisoning” is however restricted only to acute gastroenteritis due to
bacterial pollution of food or drink. The term “food-borne” disease is defined as,

“A disease, usually either infectious or toxic in nature, caused by agents that enter
the body through the ingestion of food”

Food poisoning syndrome results from ingestion of water and wide variety of food
contaminated with pathogenic microorganisms (bacteria, viruses, protozoa, fungi), their
toxins and chemicals. Food poisoning must be suspected when an acute illness with
gastrointestinal or neurological manifestation affects two or more persons, who have
shared a meal during the previous 72 hours. Some microbiologists consider microbial food
poisoning to be different from food-borne infections. In microbial food poisoning, the
microbes multiply readily in the food prior to consumption, whereas in food-borne
infection, food is merely the vector for microbes that do not grow on their transient
substrate. Others consider food poisoning as intoxication of food by chemicals or toxins
from bacteria or fungi. Consumption of poisonous mushroom leads to mycetism, while
consumption of food contaminated with toxin producing fungi leads to mycotoxicosis.

Some microorganisms can use our food as a source of nutrients for their own growth.
By growing in the food, metabolizing them and producing by-products, they not only render
the food inedible but also pose health problems upon consumption. Many of our foods will
support the growth of pathogenic microorganisms or at least serve as a vector for their
transmission. Food can get contaminated from plant surfaces, animals, water, sewage, air,
soil, or from food handlers during handling and processing.

Intoxication; Illness caused by the consumption of bacterial toxin formed in the food
e.g. Clostridium botulinum, Staphylococcus aureus.

Infection: Illness caused by the entrance of the bacteria into the body through ingestion of
contaminated food and reaction of the body to their metabolites.

CLASSIFICATION OF FOOD POISONING

Food Poisoning may be of two types:

Bacterial: Caused by the ingestion of foods, contaminated by living bacteria or their toxins.
Thus, they are two types, namely:

1. Food-Borne Intoxications

Agriculture & GK Page 88

Mob: 9473826765 [Link]
2. Food-Borne Infections

1. Food-Borne Intoxications:
Food-borne intoxications are caused

1. Due to naturally occurring toxins in some foods, including:

a) Lathyrism (beta-oxalyl amino-alanine)
b) Endemic Ascites (Pyrrolizidine alkaloids)

2. Due to toxins produced by certain bacteria, including:

a) Botulism
b) Staphylococcus toxins

3. Due to toxins produced by some fungi, including:

a. Aflatoxin
b) Ergot
c) Fusarium toxins

4. Due to toxins produced by some algae, like:

a. Planktonic dinoflagellates
b) Diatoms
c) Cyanobacteria

5. Food-Borne Infections:
Food-borne infections include

1. Bacterial infections such as:

• Salmonellosis
• Shigellosis
• E. coli diarrhea
• Cholera
• Streptococcal infection
• Brucellosis ▪ Listeriosis

2. Viral infections such as:

• Viral gastroenteritis
• Hepatitis A

3. Parasitic infections such as:

• Taeniasis
• Trichinosis

Agriculture & GK Page 89

Mob: 9473826765 [Link]
BACTERIAL FOOD POISONING
Food-Borne Diseases and Food Poisoning:
Food-borne disease is a disease caused by ingestion of food contaminated by any
agent, chemical or biological. Food poisoning is an acute enteritis caused by the ingestion of
food, characterized by diarrhea, vomiting, with or without fever and abdominal pains. Food
poisoning is normally associated with the small and large intestine. Certain types of food
poisoning are described as intoxications and others as infections.

Bacterial Etiology of Food Poisoning:

Food infections by bacteria can be divided into two types:

(i) Those in which the food does not ordinarily support the growth of pathogens but
merely carries them. E. g. Salmonella, Shigella, Vibrio etc.
(ii) Those in which the food can serve as a culture medium for growth of pathogens to
numbers that can infect the person.

Food borne infections by bacteria can also be classified as toxicosis and food-infections. In
toxicosis, the toxins are released by bacteria such as Clostridia, Bacillus and Staphylococcus.
In foodinfections, the bacteria are ingested, which later initiate the infection.

FOOD POISONING BACTERIA

Staphylococcus aureus:
S. aureus is gram positive cocci that occurs in singles, pairs, short chains, tetrads and
irregular grape like clusters. It is present ubiquitously in the environment. Only those strains
that produce enterotoxin can cause food poisoning. Food is usually contaminated from
infected food handler. The food handler with an active lesion or carriage can contaminate
food.

➢ Incriminated food: Custard and cream filled bakery food, ham, chicken, meat, milk,
fish, salads, puddings, pie etc.
➢ Pathogenesis: If the food is stored for some time at room temperature the
organism may multiply in the food and produce toxin. The bacteria produce
enterotoxin while multiplying in food. S. aureus is known to produce six serologically
different types of enterotoxins (A, B, C, C2, D and E) that differ in toxicity. Most food
poisoning is caused by enterotoxin A. Isolates commonly belong to phage type III.
These enterotoxins tend to be heat stable, with type B being most heat resistant. Low
temperature heat inactivated enterotoxin can undergo reactivation in some food.
Ingestion of as little as 23 μg of enterotoxin can induce vomiting and diarrhea.

Staphylococcal enterotoxins act as superantigens, binding to MHC II molecules and

stimulating T cells to divide and produce lymphokines such as IL-2 and TNF-alpha, which

Agriculture & GK Page 90

Mob: 9473826765 [Link]
induces diarrhea. The toxin acts on the receptors in the gut and sensory stimulus is carried
to the vomiting center in the brain by vagus and sympathetic nerves.

➢ Incubation period: Since the ingested food contains preformed toxin, the
incubation period is usually 1-6 hours.
➢ Clinical features: The onset is sudden and is characterized by vomiting and diarrhea
but no fever. The illness lasts less than 12 hours. There are no complications and
treatment is usually not necessary.
➢ Laboratory diagnosis: The presence of a large number of S. aureus organisms in a
food may indicate poor handling or sanitation; however, it is not sufficient evidence to
incriminate a food as the cause of food poisoning. Staphylococcal food poisoning can
be diagnosed if they are isolated in large numbers from the food and their toxins
demonstrated in the food or the isolated S. aureus must be shown to produce
enterotoxins. Dilutions of food may be plated on Baird Parker agar or Mannitol Salt
agar. Enterotoxin may be detected and identified by gel diffusion.

Bacillus cereus:
[Link] is a gram positive aerobic spore bearing bacilli. It is found abundantly in
environment and vegetation.

➢ Incriminated food: Commonly associated with rice and vegetables.

➢ Pathogenesis: During the slow cooling, spores germinate and vegetative bacteria
multiply, then they sporulate again. Sporulation is also associated with toxin
production. The toxin is heat-stable, and can easily withstand the brief high
temperatures used to cook fried rice. The short-incubation form is most often
associated with fried rice that has been cooked and then held at warm temperatures
for several hours. Long-incubation food poisoning is frequently associated with meat
or vegetable-containing foods after cooking. The short-incubation form is caused by a
preformed heat-stable enterotoxin of molecular weight less than 5,000 Dalton. The
long incubation form of illness is mediated by a heat-labile enterotoxin (molecular
weight of approximately 50,000 Daltons), which activates intestinal adenylate cyclase
and causes intestinal fluid secretion.
➢ Incubation period: 1-6 hours in short-incubation form and 8-16 hours in long
incubation form.
➢ Clinical features: B. cereus causes two types of food-borne intoxications. The
‘emetic-type’ or the short incubation type has an incubation period of 1 to 6 hours. It
is characterized by nausea, vomiting and abdominal cramps and resembles S. aureus
food poisoning in its symptoms and incubation period. Within 16 hours of eating
contaminated fried rice, patients suffer a bout of vomiting that generally lasts for less
than a day. The second type is manifested primarily by abdominal cramps and
diarrhea with an incubation period of 8 to 16 hours. Diarrhea may be a small volume
or profuse and watery. This type is referred to as the "long-incubation" or diarrheal

Agriculture & GK Page 91

Mob: 9473826765 [Link]
form of the disease, and it resembles food poisoning caused by Clostridium
perfringens. In either type, the illness usually lasts less than 24 hours after onset.
➢ Laboratory diagnosis: The short-incubation or emetic form of the disease is
diagnosed by the isolation of B. cereus from the incriminated food. The long-
incubation or diarrheal form is diagnosed by isolation of the organism from stool and
food. Isolation from stools alone is not sufficient because 14% of healthy adults have
been reported to have transient gastrointestinal colonization with B. cereus.

Clostridium perfringens:
It is a gram positive anaerobic spore bearing bacilli that is present abundantly in the
environment, vegetation, sewage and animal feces.

➢ Incriminated food: food-borne outbreaks of C. perfringens involve meat products

that are eaten 1- 2 days after preparation. Meats that have been cooked, allowed to
cool slowly, and then held for some time before eating are commonly incriminated.
Fish pastes and cold chicken too have been incriminated.
➢ Pathogenesis: Spores in food may survive cooking and then germinate when they
are improperly stored. When these vegetative cells form endospores in the intestine,
they release enterotoxins. The bacterium is known to produce at least 12 different
toxins. Food poisoning is mainly caused by Type A strains, which produces alpha and
theta toxins. The toxins result in excessive fluid accumulation in the intestinal lumen.
➢ Incubation period: 8-24 hours
➢ Clinical features: Illness is characterized by acute abdominal pain, diarrhea, and
vomiting. Illness is self-limiting and patient recovers in 18-24 hours.
➢ Laboratory diagnosis: Since the bacterium is present normally in the intestine,
their isolation from feces may not be sufficient to implicate it. Similarly, isolation from
food except in large numbers (>105/gram) may not be significant. The homogenized
food is diluted and plated on selective medium as well as Robertson cooked meat
medium and incubated anaerobically. The isolated bacteria must be shown to produce
enterotoxin.

Clostridium botulinum:
It is a gram-positive anaerobic spore bearing bacilli that is widely distributed in soil,
sediments of lakes and ponds, and decaying vegetation.

➢ Incriminated food: Most cases of botulism are associated with home canned or
bottled meat, vegetables and fish. In general, the low and medium acid canned foods
are often incriminated. The anaerobic environment produced by the canning process
may further encourage the outgrowth of spores.
➢ Pathogenesis: Not all strains of C. botulinum produce the botulinum toxin. Seven
toxigenic types of the organism exist, each producing an immunologically distinct form

Agriculture & GK Page 92

Mob: 9473826765 [Link]
of botulinum toxin. The toxins are designated A, B, C1, D, E, F, and G. Lysogenic phages
encode toxin C and D serotypes. Food-borne botulism is not an infection but an
intoxication since it results from the ingestion of foods that contain the preformed
clostridia toxin. If contaminated food has been insufficiently sterilized or canned
improperly, the spores may germinate and produce botulinum toxin. The toxin is
released only after the death and lysis of cells. The toxin resists digestion and is
absorbed by the upper part of the GI tract and then into the blood. It then reaches the
peripheral neuromuscular synapses where the toxin binds to the presynaptic
stimulatory terminals and blocks the release of the neurotransmitter acetylcholine.
This results in flaccid paralysis. Even 1- 2 μ g of toxin can be lethal to humans.
➢ Incubation period: 12-36 hours
➢ Clinical features: Common features include vomiting, thirst, dryness of mouth,
constipation, ocular paresis (blurred-vision), difficulty in speaking, breathing and
swallowing. Coma or delirium may occur in some cases. Death may occur due to
respiratory paralysis within 7 days.
➢ Laboratory diagnosis: Spoilage of food or swelling of cans or presence of bubbles
inside the can indicate clostridia growth. Food is homogenized in broth and inoculated
in Robertson cooked meat medium and blood agar or egg-yolk agar, which is
incubated anaerobically for 3-5 days at 37oC. The toxin can be demonstrated by
injecting intraperitoneally the extract of food or culture into mice or guinea pig.

Enterotoxigenic E. coli (ETEC):

E. coli are gram negative enteric bacilli that are carried normally in the intestine of
humans and animals. Some specific serotypes harbor plasmids that code for toxin
production. The enterotoxin production is limited to following O serotypes: O6, O8, O15,
O25, O63, O78, O148 and O159.

➢ Pathogenesis: The bacteria colonize the GI tract by means of fimbriae to specific

receptors on enterocytes of the proximal small intestine. Enterotoxins produced by
ETEC include the LT (heatlabile) toxin and or the ST (heat-stable) toxin. LTs are similar
to cholera toxin in structure and mode of action. LTs are holotoxin consisting of A
subunit and B subunit. The B subunit of LTs binds to specific ganglioside receptors
(GM1) on the epithelial cells of small intestine and facilitates the entry of A subunit
where it activates adenylate cyclase. Stimulation of adenylate cyclase causes an
increased production of cAMP, which leads to hypersecretion of water and
electrolytes into the lumen and inhibition of sodium reabsorption.
➢ Incubation period: 16-72 hours
➢ Clinical features: Sudden onset of watery diarrhea associated with nausea,
vomiting, abdominal cramping and bloating is commonly observed. This bacterium is
responsible for majority of traveler’s diarrhea. The disease is self-limiting and resolves
in few days.

Agriculture & GK Page 93

Mob: 9473826765 [Link]
➢ Laboratory diagnosis: The sample of feces is cultured on MacConkey’s agar. The ETEC
stains are indistinguishable from the resident E. coli by biochemical tests. These
strains are differentiated from nontoxigenic E. coli present in the bowel by a variety of
in vitro immunochemical, tissue culture, or DNA hybridization tests designed to detect
either the toxins or genes that encode for these toxins. With the availability of a gene
probe method, foods can be analyzed directly for the presence of enterotoxigenic E.
coli in about 3 days. LTs can be detected by Ligated rabbit ileal loop test,
morphological changes in Chinese hamster ovary cells and Y1 adrenal cells, ELISA,
immune diffusion, coagglutination etc.

Enterohemorrhagic E. coli (EHEC):

E. coli are gram negative enteric bacilli that are carried normally in the intestine of
humans and animals. EHEC strains have been associated with many serogroups including
O4, O26, O45, O91, O111, O145 and O157. The most serotype is O157:H7.

➢ Incriminated food: Cattle appear to be the main source of infection; most cases
being associated with the consumption of undercooked beef burgers and similar
foods. This disease is often associated with ingestion of inadequately cooked
hamburger meat, raw milk, cream and cheeses made from raw milk.
➢ Pathogenesis: EHEC strains may produce one or more types of cytotoxins, which are
collectively referred a Shiga-like toxins (SLT) since they are antigenically and
functionally similar to Shiga toxin produced by Shigella dysenteriae. SLTs were
previously known as verotoxin. The toxins provoke cell secretion and kill colonic
epithelial cells.
➢ Incubation period: 72-120 hours
➢ Clinical features: Initial symptoms may be diarrhea with abdominal cramps, which
may turn into grossly blood diarrhea in a few days. There is however, no fever.
➢ Laboratory diagnosis: Laboratory diagnoses involve culturing the faeces on
MacConkey's agar or on sorbitol MacConkey's agar, where they don't ferment
sorbitol. Strains can then be identified by serotyping using specific antisera. SLTs can
be detected by ELISA and genes coding for them can be detected by DNA hybridization
techniques.

Vibrio parahemolyticus:
They are straight or curved gram-negative halophilic bacilli. In morphology and
staining it resembles V. cholerae and is actively motile in liquid cultures. It is commonly
found in coastal seas, where it has been isolated from marine fauna such as crabs, shrimps,
fishes and mollusca.

➢ Incriminated food: Infections are associated with consumption of uncooked or

undercooked crabs, prawns, shrimps and other seafood.

Agriculture & GK Page 94

Mob: 9473826765 [Link]
➢ Pathogenesis: No enterotoxin has been demonstrated in the bacterium. The
infection is thought to result from invasion of intestinal epithelium.
➢ Incubation period: 7-48 hours
➢ Clinical features: The clinical infection is characterized by a sudden onset of acute
gastroenteritis. Infection may also result in diarrhea, abdominal pain, vomiting and
fever.
➢ Laboratory diagnosis: Homogenized food may be inoculated into TCBS agar or into
double strength alkaline peptone water and incubated overnight at 37oC. This
bacterium is positive for Kanagawa phenomenon where isolates from human feces
show hemolysis on blood agar.

Salmonella enteritidis:
These are gram-negative rod shaped bacteria that are classified under family
enterobacteriaceae. This species does not occur normally in humans but several animals act
as reservoirs.

➢ Incriminated food: Most important sources are chicken and poultry. Chicken, duck,
turkey and goose may be infected with Salmonella, which then find its way into its
feces, eggs or flesh of dressed fowl. Milk and milk products including ice creams have
been incriminated.
➢ Pathogenesis: Organism penetrates and passes through the epithelial cells lining
the terminal portion of the small intestine. Multiplication of bacteria in the lamina
propria produces inflammatory mediators, recruits neutrophils and triggers
inflammation. Release of LPS causes fever. Inflammation causes release of
prostaglandins from epithelial cells. Prostaglandins cause electrolytes to flow into
lumen of the intestine. Water flows into lumen in response to osmotic imbalance
resulting in diarrhea.
➢ Incubation period: 12-36 hours
➢ Clinical features: Sudden onset of abdominal pain, nausea, vomiting and diarrhea,
which may be watery, greenish and foul smelling. This may be preceded by headache
and chills. Other findings include prostration, muscular weakness and moderate fever.
In most cases the symptoms resolve in 2-3 days without any complications.
➢ Laboratory diagnosis: Homogenized food is cultured in selenite F broth and then
sub-cultured on deoxycholate citrate agar. Plates are incubated at 37oC overnight and
growth identified by biochemical tests and slide agglutination test.

Yersinia enterocolitica:
It is a gram-negative psychrophilic rod shaped bacterium that is motile only at
temperature below 30oC. Yersini enterocolitica is widely distributed in environment and
have been isolated frequently from soil, water and animals. The major animal reservoir for
Y. enterocolitica strains that cause human illness is pigs, but may also found in many other

Agriculture & GK Page 95

Mob: 9473826765 [Link]
animals including rodents, rabbits, sheep, cattle, horses, dogs, and cats. Serogroups that
predominate in human illness are O:3, O:8, O:9, and O:5.

➢ Incriminated food: Infection is most often acquired by eating contaminated food,

especially raw or undercooked pork products. Drinking contaminated unpasteurized
milk or untreated water can also transmit the infection.
➢ Pathogenesis: This organism may survive and grow during refrigerated storage.
Strains that cause human yersiniosis carry a plasmid that is associated with a number
of virulence traits. Ingested bacteria adhere and invade M cells or epithelial cells. They
exhibit resistance to complement and phagocytosis. They produce ST only at
temperatures below 30ºC. The role of ST in the disease process remains uncertain.
➢ Incubation period: 4-7 days
➢ Clinical features: Disease produced by Y. enterocolitica is a typical gastroenteritis
characterized by fever, abdominal pain, and diarrhea, which is often bloody. Illness
generally lasts from 1 to 2 weeks but chronic cases may persist for up to a year. Apart
from gastroenteritis it may also cause pseudoappendicitis, mesenteric lymphadenitis,
and terminal ileitis.
➢ Laboratory diagnosis: Suspected food is homogenized in phosphate-buffered
saline and inoculated into selenite F broth and held at 4oC for six weeks. The broth is
sub-cultured at weekly intervals on DCA or Yersinia selective agar plates. This is
termed as cold enrichment technique.

Campylobacter jejuni:
These are small, curved-spiral gram-negative bacilli with polar flagella. Campylobacter jejuni
appear in comma, S-shaped or “gull-wings/sea-gull” form. Campylobacter are harbored in
reproductive and alimentary tracts of some animals.

➢ Incriminated food: Transmission to humans occurs via a fecal-oral route,

originating from farm animals, birds, dogs, and processed poultry, with chicken
preparation comprising 50-70% of all campylobacter infections. The organism is
transmitted to man in milk, meat products and contaminated water. Undercooked
poultry and unpasteurized dairy are most often implicated as a source of C. jejuni.
➢ Pathogenesis: As few as 500 organisms can cause enteritis. The organism is invasive
but generally less so than Shigella. Campylobacter produces adenylate cyclase-
activating toxins same as of [Link] LT and cholera.
➢ Incubation period: Ranges from 2 to11 days.
➢ Clinical features: Patients present with abdominal pain and cramps, diarrhea,
malaise, headache, and usually fever. Typically, the diarrhea is watery, but in severe
cases bloody diarrhea may occur. Diarrhea may last 2-7 days and the organism may be
shed in the patients stool for up to 2 months. Bacteremia is observed in a small
minority of cases. The disease is usually self-limiting.

Agriculture & GK Page 96

Mob: 9473826765 [Link]
➢ Laboratory diagnosis: The feces may be inoculated in enrichment medium or on
selective media such as Campy BAP or Skirrow's medium. The plates are incubated in
microaerophilic conditions at 42° for 2-5 days.

FOOD POISIONING BY VIRUS

Viruses are not living organisms but bits of reproductive material that attack human
cells and hijack them. The most important viruses that cause foodborne disease are
Hepatitis A, Norwalk virus, Norovirus and some of the Calici viruses. Viruses don’t grow in
food, and one particle is enough to make you sick. Symptoms can be severe gastroenteritis
or similar to the ‘flu’. Generally, the illness only lasts one or two days. The exception is
Hepatitis A which can be a severe illness and last for many weeks.

➢ Viral Gastroenteritis: Viral gastroenteritis is usually regarded as a mild self-limiting

disease lasting 24-48 hours.
However, people can feel debilitated for 2 to 3 weeks, which has considerable
economic implications in terms of working days lost and impaired performance
Symptoms include, malaise, abdominal pain, pyrexia, diarrhea and/or vomiting. A
range of symptoms occurring in an outbreak should alert investigations to the
possibility of a viral cause. The viruses are usually transmitted by the faecal-oral route,
but they are also present in vomitus. Virus will be disseminated over a wide area in
aerosol droplets, which is a particular hazard where food is being prepared and laid
out. Although most transmission is directly from person to person, contaminated food
and water can give rise to common-source outbreaks. For instance, it has been
estimated that NLVs have an infective dose of between 10 and 100 virus particles.

➢ Norwalk-like Viruses: This group of viruses infects all age groups. There is a
variable incubation period of 12-60 hours, which is thought to be dose-dependent.
These viruses are responsible for both sporadic cases of gastroenteritis in the
community and for outbreaks in schools, hospitals, old age homes, hotels and cruise
ships.

➢ Rotavirus: Rotaviruses mainly infect young children. It is estimated that they cause
one million deaths a year in children under 5 years of age, mostly in developing
countries. In developed countries deaths are relatively rare, but rotavirus
gastrotenteritis is the most frequent reason for admission of young children to
hospital. Rotaviruses consistently account for around 80% of all gastroenteritis viruses.

➢ Astro virus: The Astro virus form a morphologically distinct group of viruses, been
associated with illness in children, often under 1 year of age, although outbreaks have
been reported in the elderly. The use of more sensitive molecular detection methods
is required to assess the incidence and epidemiology of these viruses. Astroviruses

Agriculture & GK Page 97

Mob: 9473826765 [Link]
have been seen in some adults following the consumption of shellfish or contaminated
water, but these incidents appear to be comparatively rare.

HEPATITIS

There are two forms of enterically transmitted hepatitis, hepatitis A and hepatitis E.

Hepatitis A
The most characteristic symptom of hepatitis A is jaundice, but milder symptoms of
nausea and general malaise without jaundice are common. Patients may feel unwell for
several weeks, but recovery is complete. Deaths are rare. Some infections, particularly in
children, may be asymptomatic. Like viral gastroenteritis, transmission is by the faecal-oral
route, but the primary site of viral replication is the liver. Virus excretion may commence up
to a week before symptoms are apparent, making control difficult.

Hepatitis E
Hepatitis E has been associated with large water-borne outbreaks in some developing
countries, notably in Asia. Africa and Central America. Food-borne transmission has been
suggested, but not proved conclusively. Illness appears more severe than hepatitis A,
particularly in pregnant women where a death rate of 17-33% has been observed. The
primary source of infection appears to be contaminated water rather than person-to-person
spread. Secondary person-toper son transmission is estimated at only 0.7-8%.

Some Ways of Preventing Food Poisoning:

➢ Good personal hygiene; such as thoroughly washing and drying hands when handling
food.
➢ Avoid cross-contamination; such as keeping raw foods and ready-to-eat foods
separate and using separate and clean utensils, containers and equipment.
➢ Cook foods thoroughly; make sure foods such as meats and poultry are cooked until
core temperature reaches 75°C.
➢ Avoid the Temperature Danger Zone; keep chilled foods cold at 5°C or colder, and hot
food hot at 60°C or hotter.
➢ Avoid spoiled foods; foods past their use-by dates or food in damaged containers or
packaging.
➢ If working in a food business: Follow the business Food Safety Program.
➢ Follow the advice given by the Food Safety Supervisor.
➢ Be trained in safe food handling.

Agriculture & GK Page 98

Mob: 9473826765 [Link]

FOOD SAFETY MANAGEMENT SYSTEM

FSMS (Food Safety Management System) can be defined as a system that uses GMP,
GHP and HACCP and other requirements as defined under Schedule IV of FSS (Licensing &
Registration of Food Businesses) Regulations, 2011. Since FSMS ensures food safety hence it
is the requirement for FBOs to submit a FSMS Plan specific for each FBO as per FSS
Regulations.

The FSMS programme helps FBOs to ensure that food is safe for human consumption.
FSSAI has made the FSMS programme in the regulations and FBOs are required to follow it.
It is not possible to get a license unless the FBO provides details of the FSMS programme to
FSSAI.

In India Food safety management system (FSMS) is a set of systems that are
interrelated and which when used in combination ensure that food is safe for human
consumption. FSMS includes procedures and controls laid down by FSSAI. It incorporates
Good Manufacturing Practices (GMP), Good Hygienic Practices (GHP), Hazard Analysis and
Critical Control Point (HACCP) and some other practices as may be specified in the FSSAI
regulations. For the FSMS system to be effective food businesses have to adopt all these
regulatory procedures to ensure food safety.

There are several voluntary Food Safety Certifications that also help to meet food
safety and to strengthen it, food safety system like; HACCP, ISO 22000, FSSC 22000, GMP,
GHP etc.

Schedule IV requirements and its significance:

To provide assurance of food safety, FBOs should strive to implement an effective
Food Safety Management System (FSMS) based on Hazard Analysis and Critical Control
Point (HACCP) and suitable pre- requisite programmes by actively controlling hazards
throughout the food chain starting from food production till final consumption. Every
licensed FBO must have a documented Food Safety Management System (FSMS) plan and is
required to comply with Schedule 4 of FSS (Licensing and Registration of Food Business)
Regulation, 2011. Schedule 4 introduces the concept of FSMS based on implementation of
Good Manufacturing Practices (GMP) and Good Hygiene Practices (GHP) by food businesses
and is divided into five parts. (We have already discussed about Schedule IV in detail in Food
Safety and Standards (Licensing & Registration) Regulation,2011.)

ADVANTAGES OF FSMS

• Continuous prevention of foodborne illness and related public relations disasters

• Food safety compliance during routine inspections
• Improved inventory control
• Reduction in product loss

Agriculture & GK Page 99

Mob: 9473826765 [Link]
• More consistency in product preparation and improved product quality
• Increased employee understanding and involvement in food safety
• More effective communication and collaboration with industry regulators

FSMS principles:
Food safety is related to the presence of food safety hazards at the time of
consumption (intake by the consumer). Food safety hazards can occur at any stage of the
food chain. Therefore, adequate control throughout the food chain is essential. Food safety
is ensured through the combined efforts of all the parties in the food chain. This document
specifies the requirements for a FSMS that combines the following generally recognized key
elements:

• interactive communication;
• system management;
• prerequisite programmes;
• hazard analysis and critical control point (HACCP) principles.

In addition, this document is based on the principles that are common to ISO
management system standards. The management principles are:

• customer focus;
• leadership;
• engagement of people;
• process approach;
• improvement;
• evidence-based decision making;
• relationship management.

FSMS GUIDANCE DOCUMENTS

A series of sector specific Food Safety Management System (FSMS) Guidance

Documents have been developed with the help of domain experts with the intent to provide
implementation guidance to food businesses (especially the small and medium businesses)
involved in manufacturing, packing, storage and transportation to ensure that critical food
safety related aspects are addressed throughout the supply chain.

These documents are primarily based on Schedule 4 of Food Safety & Standards
(Licensing & Registration of Food Businesses) Regulation, 2011 and lay down general
requirements on good hygienic practices to be followed by Food Business Operators &
indicate practical approaches which a business should adopt to ensure food safety. The
documents are recommendatory in nature and provide the basic knowledge and criteria for
implementation of Hazard Analysis and Critical Control Point (HACCP) system by the food
businesses. Sample HACCP Plans have been included from some established practicing

Agriculture & GK Page 100

Mob: 9473826765 [Link]
industries. These plans could be used as reference by the industry and modified or altered
based on their operations. Inspection checklists for FBOs to audit their facility & operations
are also included in these documents. The FBOs can evaluate themselves based on the
indicative scoring. Also, these documents provide important templates and forms to
facilitate the FBOs to maintain the records. These include mandatory forms as prescribed by
FSSAI & few templates for maintaining records of processes critical for food safety. FSSAI
has developed FSMS documents as guidance to the food businesses of following sectors:

• Spices
• Fish and Fish Products.
• Meat and Meat Products (Poultry).
• Health Supplements and Nutraceuticals.
• Vegetable edible oils and fats sectors
• Bakery sector
• Food grain warehouses
• Flour milling sector
• Catering sector.

Pre-requisite programmes such as GMP and GHP must be working effectively within a
commodity system before HACCP is applied. If these pre-requisite programmes are not
functioning effectively then the introduction of HACCP will be complicated, resulting in a
cumbersome, over-documented system.

GENERAL PRINCIPLES OF FOOD HYGIENE

➢ Food safety and suitability should be controlled using a science-based preventive
approach, for example a food hygiene system. GHPs should ensure that food is
produced and handled in an environment that minimizes the presence of
contaminants.
➢ Properly applied prerequisite programmes, which include GHPs, should provide the
foundation for an effective HACCP system.
➢ Each FBO should be aware of the hazards associated with the raw materials and other
ingredients, the production or preparation process, and the environment in which the
food is produced and/or handled, as appropriate to the food business.
➢ Depending on the nature of the food, food process, and the potential for adverse
health effects, to control hazards it may be sufficient to apply GHPs, including, as
appropriate, some that require more attention than others, as they have a greater
impact on food safety. When the application of GHPs alone is not sufficient, a
combination of GHPs and additional control measures at CCPs should be applied.
➢ Control measures that are essential to achieve an acceptable level of food safety,
should be scientifically validated.
➢ The application of control measures should be subject to monitoring, corrective
actions, verification, and documentation, as appropriate to the nature of the food
product and the size of the food business.

Agriculture & GK Page 101

Mob: 9473826765 [Link]
➢ Food hygiene systems should be reviewed to determine if modifications are needed.
This should be done periodically and whenever there is a significant change that could
impact the potential hazards and/or the control measures (e.g. new process, new
ingredient, new product, new equipment, new scientific knowledge) associated with
the food business.
➢ Appropriate communication about the food and food process should be maintained
among all relevant parties to ensure food safety and suitability across the entire food
chain.

Food safety and quality can be ensured through:

• Good Manufacturing Practices (GMP)
• Good Hygienic Practices (GHP)
• Hazard Analysis Critical Control Points (HACCP)
• ISO 22000

GMP - GOOD MANUFACTURING PRACTICES

Food manufacturing and producing industries should

follow and adopt Good Manufacturing Practices (GMPs) to
make sure that all products are manufactured under a safe
and healthy environment, which ensures the safety and
quality of their products to fulfil requirement of standards
regulations. GMPs must follow for various practices of product
testing, manufacturing, storage, handling, and distribution.
GMPs should fulfil the standards of Safety, Integrity, Purity,
Quality, and Composition (SISPQC).The practices of Hazard
Analysis and Critical Control Points (HACCP) and GMP programs give confidence and faith to
consumers that proper testing consistency and safety and quality checks have been
maintained throughout manufacturing, packaging, and distribution of products. GMP has
science and technology based rules, regulations and standards. It also has integrated
systems approach for quality, facilities and equipment, materials, production, packaging,
labelling and laboratory control. It keeps proper records for proposed amendments
regarding validation and cross-contamination.

Who should follow GMP?

All food processer, medicinal drug manufacturer, food product manufacturers,
packagers, labellers, and distributors, warehouse/storage facilities keepers should strictly
follow GMP and given different regulations of food safety and standards.

Purpose of GMP:

Agriculture & GK Page 102

Mob: 9473826765 [Link]
Drinking and other food preparation purposes used water, facilities for personal
hygiene, air quality and ventilation lighting storage operation controls time and
temperature control cross contamination raw materials packaging product information
traceability pest control personal hygiene transportation training food marketing food
services, verification. The brief outline of the GMP structure for a company is given below.

a. Quality assurance – Every industry should have its quality management department.
b. GMP for food, medicine and pharmaceutical products.
c. Sanitation and hygiene- This very important and crucial part of GMP.
d. Qualification and validation.
e. Complaints.
f. Product recalls.
g. Contract production and analysis- it can be subcategorized as general, contract giver,
contract accepter and the contract.
h. Self-inspection (SI) and quality audits:
i. Items for SI
ii. SI team
iii. Repetition of SI
iv. SI report
v. Follow-up action
vi. Audit for quality assurance and
vii. Suppliers’ audits and approval.
i. Personnel – In any industry there are two types of personal
i. General Worker and Operator and
ii. Key personnel.
j. Training.
k. Personal hygiene.
l. Premises -It may include general area, ancillary areas, storage areas, weighing areas,
production areas and quality control area.
m. Equipment.
n. Materials can be general materials, starting materials, packaging materials,
intermediate and bulk products, finished products, rejected, recovered, reprocessed
and reworked materials, recalled products, returned goods, reagents and culture
media, reference standards, waste materials and miscellaneous.
o. Documentation can include general records, documents on labels, testing procedures
records, specifications for starting and packaging materials, for intermediate and bulk
products and for finished products records, files on master formulae and Batch
Processing Records, Packaging instructions and Batch Packaging Records, Standard
Operating procedures (SOP's) and records and finally Logbooks.
p. GMP in production are general neat and clean practices, GMP in prevention of cross
contamination and bacterial contamination during production, GMP in processing
operations and packaging operations.

Agriculture & GK Page 103

Mob: 9473826765 [Link]
q. GMP in quality control are control of starting materials and intermediate, bulk and
finished products, test requirements, batch record review and stability studies.

FUNDAMENTALS OF GMP

• Quality Control - Product meets specifications.

• Quality Assurance -Systems ensure control and consistency; validation.
• Documentation-If it is not documented, it did not happen or it is just rumor.
• Verification and self-inspection.

PRINCIPLES OF GMP FOR FOOD

GMP consists of following 10 principles that introduce employees to critical behaviors
established by food regulations and industry leaders to maintain GMP in plants.

1. Writing procedures.
2. Following written procedures.
3. Documenting for traceability.
4. Designing facilities and equipment.
5. Maintaining facilities and equipment.
6. Validating work.
7. Job competence.
8. Cleanliness.
9. Component control.
10. Auditing for compliance.

FIVE P’s OF GMP

The place, Primary Materials, people, process and product are five important factors
of production and processing of food product that affect the quality and safety while
following GMPs. Places and premises must be clean and well organized. All surfaces and
roofs, side walls should be well cleaned and designed to prevent any kind of contamination.

First ‘P’: Place / Premises –Building and facilities:

Premises must be clean and equipment should be orderly arranged; surfaces should
allow for effective cleaning and should be designed to prevent contamination of food.
Equipment and containers should be easy to carry and convey for adequately cleaned,
disinfected and maintained to avoid the contamination of food.

Second ‘P’: Primary Materials:

All incoming materials should be checked, tested and recorded. Impure, adulterated,
broken, toxic etc. raw materials should reject and sent back.

Agriculture & GK Page 104

Mob: 9473826765 [Link]
Third ‘P’:
People Working people must be in adequate in number, qualified by education,
training, or experience to perform their respective [Link] Assurance is
responsible to release each product for sale, release raw material and packaging
components for use, approve all operational procedures, master formulas and
specifications, review all returns prior to resale, and investigate each complaint.

People known, or suspected, to be suffering from, or to be a carrier of a disease or

illness likely to be transmitted through food, it leads to contaminate the food and it can
transmit illness to consumers; So that all food handlers shall be medically examined by a
registered medical practitioner and vaccinated according to current legislative requirements
for avoiding the food contamination during the process of food [Link] outer
garments, light colored and suitable to the operation in a manner that protects against the
contamination of food, food-contact surfaces, or food packaging materials. Remove all
jewellery and other objects that might fall into food, equipment, or [Link] an
effective manner, hair nets, headbands, hand gloves, caps, beard covers, or other effective
hair restraints.

Wash hands properly in an adequate handwashing facility before starting work, after
each absence from the work station, immediately after using toilet and at any other time
when the hands may have become contaminated. Store clothing or other personal
belongings in areas other than where food is exposed or where equipment or utensils are
washed. Take any other necessary precautions to protect against contamination of food.
They should be in good health and capable of handling the duties assigned to them.

Fourth ‘P’:
Process Sanitation programs include documented procedures for effective cleaning of
the premises, equipment, handling of substances, and the health and hygienic behavior of
personnel. Raw materials and primary packaging materials are stored and handled in a
manner which prevents their mix-up, contamination with microorganisms or other
chemicals.

Raw materials, in-process samples and finished products are tested or examined to
verify their identity. Lot or batch identification is essential in traceability and product recall
and also helps effective stock [Link] foods should be labeled with clear
instructions to enable the next person in the food chain to handle, display, store and use the
product safely.

Fifth ‘P’:
Products Specifications are available for every product and include purity, quantity,
and identity of medicinal ingredients, potency, and test methods. Every lot must be tested
for conformance with its finished product specifications. Importers may follow a reduced
testing program in which - The first lot of each product is fully tested; and for each
Agriculture & GK Page 105
Mob: 9473826765 [Link]
subsequent lot, A Certificate of Analysis with actual test results is [Link] lot is
positively identified upon receipt. Transportation and storage conditions do not adversely
impact the product. Full confirmatory testing is conducted on at least one lot per year
dosage form, per supplier. Stability results demonstrate that the product will meet
specifications under the recommended storage conditions. Samples are retained under the
recommended storage conditions for one year past the expiry date of product. There are
sufficient samples available to perform complete testing of the product. Records must be
retained for one year past the expiration date of the product to which they refer. Recall
procedure is documented and effective by performing a Mock Recall. Sterile NHPs must be
manufactured and packaged in a separate enclosed area under the supervision of a trained
person using scientifically proven methods to ensure sterility.

BENEFITS OF GMP/HACCP

• Operating cost drops as rework and penalties due to non-compliance reduces and
efficiencies increases.
• Increases the customer satisfaction, employees, stockholders, regulators.
• Respect for an organization that demonstrates commitment to NHP safety.
• GMP/HACCP covers all safety and written procedures (SOP), which makes employees
more efficient and reduces errors during the manufacturing process.

GOOD HYGIENIC PRACTICES (GHP)

All consumers have the right to expect safe, hygienically
prepared and good quality food. This is the reason that the
handling of food requires care to prevent the hazards. Good
Hygiene Practices are the set of requirements to prevent
contamination of food in order to provide safe food to the
consumers. Food borne illnesses can result from contamination due
to improper practices like when there is:

• lack of environmental hygiene and poor sanitation

• mixed and inappropriate transportation
• poor storage • poor personal hygiene,
• unsafe source of food

CONTAMINANTS ARE IDENTIFIED AS

➢ Biological: bacteria, viruses or parasites that are present in air, food, water, soil,
animals or humans
➢ Physical: Foreign bodies in food are usually due to accidental contamination and / or
poor handling practices, these are visible particles like; pebbles, stones, metal, glass,
wood, insects, soil, dirt, jewellery, hair, fingernails etc.

Agriculture & GK Page 106

Mob: 9473826765 [Link]
➢ Chemical: chemicals used for cleaning and sanitizing food contact surfaces, pest
control chemicals, paints and water treatment chemicals, pesticides, fertilizers,
fungicides and also a group of some naturally occurring harmful chemicals like
Mycotoxins.

BASIC REQUIREMENTS OF GHP

There are basic 8 requirements of GHP:

• Primary Production
• Establishment Design and Facilities
• Control of operations
• Maintenance and sanitation
• Personal Hygiene
• Transportation
• Product information and consumer awareness
• Training

GOOD AGRICULTURAL PRACTICES (GAP)

Food and Agricultural Organization of the United Nations (FAO) defines Good
Agriculture Practice as a collection of principles to apply for on-farm production and post-
production processes, resulting in safe and healthy food and non-food agricultural products,
while taking into account economic, social and environmental sustainability. The pillars of
GAPs are economic viability, environmental sustainability, social acceptability and food
safety and quality.

Good Agricultural Practices (GAP) and Good Handling Practices (GHP) are voluntary
audits that verify that fruits and vegetables are produced, packed, handled, and stored to
minimize risks of microbial food safety hazards.

GAP & GHP audits verify adherence to the recommendations made in the U.S. Food
and Drug Administration’s Guide to Minimize Microbial Food Safety Hazards for Fresh Fruits
and Vegetables (pdf) and industry recognized food safety practices.

GAPs apply scientific recommendations and available knowledge to address

environmental, economic and social sustainability for on-farm production and post-
production processes resulting in safe and healthy food and non-food agricultural products.

FAO, 2007 published technical manual on guidelines of "Good Agricultural Practices

for Family Agriculture". According to this GAPs and GMPs (Good Manufacturing Practices)
are a set of principles, regulations and technical recommendations applicable to production,
processing and food transport, addressing human health care, environment protection and
improvement of worker conditions and their families. Initially the GAPs are more focused on
post-harvest handling, processing, packaging and quality standards. Now the concept of
GAPs covers all practices (starting from production level to ultimate consumer) which are

Agriculture & GK Page 107

Mob: 9473826765 [Link]
environmentally sustainable, economical feasible and socially acceptable. The
implementation of GAPs requires active role of farmers, food regulatory authorities,
retailers, and consumers in advocating food safety and sustainable food production.

GAPs may be applied to a wide of farming systems and at different scales. They are
applied through sustainable agriculture methods.

OBJECTIVES OF GAPS

The major objectives are:

• To improve sustainable natural resources use, workers' health and working conditions.
• To ensures quality and safety of produce in' the food chain.
• Creating new market opportunities for farmers and exporters
• Capturing new market advantages by modifying supply chain mechanism.

KEY ELEMENTS OF GAPS

The key elements of GAPs have been enumerated as:

a. Prevention of problems before they occur
b. Risk assessments
c. Commitment to food safety at all levels
d. Communication throughout the production chain
e. Mandatory employee education program at the operational level
f. Field and equipment sanitation
g. Integrated pest management
h. Oversight and enforcement
i. Verification through independent, third-party audit

BENEFITS OF GAPS

The benefits of GAPs are:

➢ Quality and safety of food and other agricultural products are improved with
appropriate adoption and monitoring of GAPs.
➢ It helps to reduce the risk of non-compliance with national and international
regulations, standards and guidelines (in particular of the Codex Alimentarius
Commission, World Organization for Animal Health and the International Plant
Protection Convention (IPPC) regarding permitted pesticides, maximum levels of
contaminants including pesticides, veterinary drugs, radionuclide and mycotoxins in
food and non-food agricultural products, as well as other chemical, microbiological
and physical contamination hazards.
➢ GAPs help to promote sustainable agriculture and contributes to meeting national and
international environment and social development objectives.

Agriculture & GK Page 108

Mob: 9473826765 [Link]
CHALLENGES OF GAPS

Many challenges are also associated with GAPs:

➢ GAPs implementation and record keeping and certification processes in particular will
increases the production costs.
➢ Lack of coordination between existing GAPs-related schemes and availability of
affordable certification systems has often led to increased confusion and certification
costs for farmers and exporters.
➢ Standards of GAPs can be used to serve competing interests of specific stakeholders in
agri-food supply chains by modifying supplier-buyer relations.
➢ Small scale farmers may face high risk of non-availability of export market
opportunities unless they are adequately informed, technically prepared, organized
and facilitate by governments and public agencies to meet this new challenge.
➢ Follow of all GAPs standards does not always promote all the environmental and social
benefits, which are claimed.

HAZARD ANALYSIS CRITICAL CONTROL POINT (HACCP)

HACCP is a means of providing assurance about safety of food. HACCP is an approach
to food manufacture and storage in which raw materials and each individual step in a
specific process are considered in detail and evaluated for its potential to contribute to the
development of pathogenic micro-organisms or other food hazards. It involves identification
of hazards, assessment of chances of occurrence of hazards during each step /stage in the
food chain – raw material procurement, manufacturing, distribution, usage of food products
and defining measures for hazard(s) control.

Why implementation of HACCP is necessary?

It is a preventive approach to ensure food safety.
End product inspection and testing, although important, is time consuming, expensive
and detects the problems only after they occur. In contrast, HACCP enables us to detect
hazards at any stage of processing or manufacture in order to ensure a good quality end
product, by taking appropriate action at the stage where the problem occurs.
It enables producers, processors, distributors and exporters to utilize resources efficiently
and in a cost-effective manner for assuring food safety.
FSSA, 2006 places primary responsibility for safe food with producers and suppliers through
HACCP, GMP, GHP. This is important for consumer protection and international food trade.
It assures consistently good quality products.

HACCP -HAZARD ANALYSIS CRITICAL CONTROL POINT SYSTEM

HACCP is a system that should lead to the production of microbiologically safe foods
by analyzing for the hazards of raw materials-those that may appear throughout processing
and those that may occur from consumer abuse.

Agriculture & GK Page 109

Mob: 9473826765 [Link]

It is a proactive, systematic approach to controlling food borne hazards. Although

some classic approaches to food safety rely heavily on end product testing, the HACCP
system places emphasis on the quality of all ingredients and all process steps on the premise
that safe products will result if these are properly controlled. The system is thus designed to
control organisms at the point of production and preparation.

➢ Control point:
Any point in a specific food system where loss of control does not lead to an
unacceptable health risk.

➢ Critical control point (CCP):

Any point or procedure in a food system where control can be exercised and a hazard
can be minimized or prevented.

➢ Critical limit:
One or more prescribed tolerances that must be met to ensure that a, CCP effectively
controls a microbiological health hazard.

➢ CCP decision tree:

A sequence of questions to assist in determining whether a control point is a CCP.

➢ Corrective action:
Procedures followed when a deviation occurs.

➢ Deviation:
Failure to meet a required critical limit for a CCP.

➢ HACCP plan:
The written document that delineates the formal procedures to be followed in
accordance with these general principles.

➢ Hazard:
Any biological, chemical, or physical property that may cause an unacceptable
consumer health risk (unacceptable contamination, toxin levels. growth, and/or
survival of undesirable organisms).

➢ Monitoring:
A planned sequence of observations or measurements of critical limits designed to
produce an accurate record and intended to ensure that the critical limit maintains
product safety.

Agriculture & GK Page 110

Mob: 9473826765 [Link]
➢ Validation:
That element of verification focused on collecting and evaluating scientific and
technical information to determine whether the HACCP plan, when properly
implemented, will effectively control the hazards.

➢ Verification:
Methods, procedures, and tests used to determine whether the HACCP system is in
compliance with the HACCP plan.

7 PRINCIPLES OF HACCP

PRINCIPLES OF HACCP

1. Conduct a hazard analysis:

Plan to determine the food safety hazards and identify the preventive measures the
plan can apply to control these hazards. A food safety hazard is any biological,
chemical, or physical property that may cause a food to be unsafe for human
consumption.
2. Identify critical control points:
A critical control point (CCP) is a point, step, or procedure in a food manufacturing
process at which control can be applied and, as a result, a food safety hazard can be
prevented, eliminated, or reduced to an acceptable level.

3. Establish critical limits for each critical control point:

Agriculture & GK Page 111

Mob: 9473826765 [Link]
A critical limit is the maximum or minimum value to which a physical, biological, or
chemical hazard must be controlled at a critical control point to prevent, eliminate, or
reduce that hazard to an acceptable level.

4. Establish critical control point monitoring requirements:

Monitoring activities are necessary to ensure that the process is under control at each
critical control point. In the United States, the FSIS requires that each monitoring
procedure and its frequency be listed in the HACCP plan.

5. Establish corrective actions:

These are actions to be taken when monitoring indicates a deviation from an
established critical limit. The final rule requires a plant's HACCP plan to identify the
corrective actions to be taken if a critical limit is not met. Corrective actions are
intended to ensure that no product is injurious to health or otherwise adulterated as a
result if the deviation enters commerce.

6. Establish verification procedure:

Verification procedures may include such activities as review of HACCP plans, CCP
records, critical limits and microbial sampling and analysis. FSIS is requiring that the
HACCP plan include verification tasks to be performed by plant personnel. Verification
tasks would also be performed by FSIS inspectors. Both FSIS and industry will
undertake microbial testing as one of several verification activities. Verification also
includes 'validation' – the process of finding evidence for the accuracy of the HACCP
system (e. g. scientific evidence for critical limitations).

7. Establish record keeping procedures:

The HACCP regulation requires that all plants maintain certain documents, including
its hazard analysis and written HACCP plan, and records documenting the monitoring
of critical control points, critical limits, verification activities, and the handling of
processing deviations. Implementation involves monitoring, verifying, and validating
of the daily work that is compliant with regulatory requirements in all stages all the
time. The differences among those three types of work are given by Saskatchewan
Agriculture and Food.

STEP INVOLVED IN HACCP APPLICATION

Logic Sequence for Application of HACCP:

Step 1 Assemble HACCP TEAM

Step 2 Describe Product
Step 3 Identify Intended Use
Step 4 Construct Flow Diagram

Agriculture & GK Page 112

Mob: 9473826765 [Link]
Step 5 On-site Confirmation of Flow Diagram
Step 6 List all Potential Hazards Conduct a Hazard Analysis to identify the significant
hazard(s) Consider Control Measures
Step 7 Determine CCPs
Step 8 Establish validated Critical Limits for each CCP
Step 9 Establish a Monitoring System for each CCP
Step 10 Establish Corrective Actions
Step 11 Validate the HACCP plan and establish Verification Procedures
Step 12 Establish Documentation and Record Keeping

1. Assemble HACCP Team and Identify Scope (Step 1):

The FBO should ensure that the appropriate knowledge and expertise are available for
the development of an effective HACCP system. This may be achieved by assembling a
multidisciplinary team responsible for different activities within the operation, e.g.
production, maintenance, quality control, cleaning and disinfection. The HACCP team is
responsible for developing the HACCP plan. The HACCP team should identify the scope of
the HACCP system and applicable prerequisite programmes. The scope should describe
which food products and processes are covered.

2. Describe product (Step 2):

A full description of the product should be developed, including relevant safety
information such as composition (i.e. ingredients), physical/chemical characteristics (e.g.
aw, pH, preservatives, allergens), processing methods/technologies (heat-treatment,
freezing, drying, brining, smoking, etc.), packaging, durability/shelf life, storage conditions
and method of distribution. Within businesses with multiple products, it may be effective to
group products with similar characteristics and processing steps for the purpose of
development of the HACCP plan. Any limits relevant to the food product already established
for hazards should be considered and accounted for in the HACCP plan, e.g. limits for food
additives, regulatory microbiological criteria, maximum allowed veterinary medicines
residues, and times and temperatures for heat treatments prescribed by competent
authorities.

3. Identify intended use and users (Step 3):

Describe the use intended by the FBO and the expected uses of the product by the
next FBO in the food chain or the consumer; the description may be influenced by external
information, e.g. from the competent authority or other sources on ways in which
consumers are known to use the product other than those intended by the FBO. In specific
cases (e.g. hospitals), vulnerable groups of the population may have to be considered.
Where foods are being produced specifically for a vulnerable population, it may be
necessary to enhance process controls, monitor control measures more frequently, verify
controls are effective by testing products, or conduct other activities to provide a high level
of assurance that the food is safe for the vulnerable population.

Agriculture & GK Page 113

Mob: 9473826765 [Link]
4. Construct flow diagram (Step 4):
A flow diagram that covers all steps in the production of a specific product, including
any applicable rework, should be constructed. The same flow diagram may be used for a
number of products that are manufactured using similar processing steps. The flow diagram
should indicate all inputs, including those of ingredients and food contact materials, water
and air if relevant. Complex manufacturing operations can be broken down into smaller,
more manageable modules and multiple flow diagrams that link together can be developed.
The flow diagrams should be used when conducting the hazard analysis as a basis for
evaluating the possible occurrence, increase, decrease or introduction of hazards. Flow
diagrams should be clear, accurate and sufficiently detailed to the extent needed to conduct
the hazard analysis. Flow diagrams should, as appropriate, include but not be limited to the
following, the sequence and interaction of the steps in the operation;

➢ where raw materials, ingredients, processing aids, packaging materials, utilities and
intermediate products enter the flow;
➢ any outsourced processes;
➢ where applicable reworking and recycling take place;
➢ where end products, intermediate products, waste and by-products are released or
removed.

5. On-site confirmation of flow diagram (Step 5):

Steps should be taken to confirm the processing activities against the flow diagram
during all stages and hours of operation and amend the flow diagram where appropriate.
The confirmation of the flow diagram should be performed by a person or persons with
sufficient knowledge of the processing operation.

6. List all potential hazards that are likely to occur and associated with
each step, conduct a hazard analysis to identify the significant hazards,
and consider any measures to control identified hazards (Step 6/
Principle 1):
Hazard analysis consists of identifying potential hazards and evaluating these hazards
to determine which of them are significant for the specific food business operation. An
example of a hazard analysis worksheet.

The HACCP team should list all potential hazards. The HACCP team should then
identify where these hazards are reasonably likely to occur at each step (including all inputs
into that step) according to the scope of the food business operation. Hazards should be
specific, e.g. metal fragments, and the source or reason for presence should be described,
e.g. metal from broken blades after chopping. The hazard analysis can be simplified by
breaking down complex manufacturing operations and analysing steps in the multiple flow
diagrams described in step 4.

Agriculture & GK Page 114

Mob: 9473826765 [Link]
7. Determine the Critical Control Points (Step 7/ Principle 2):
The FBO should consider which among the available control measures listed during
step 6, Principle 1 should be applied at a CCP. Critical Control points are to be determined
only for hazards identified as significant as of the result of a hazard analysis. CCPs are
established at steps where control is essential and where a deviation could result in the
production of a potentially unsafe food. The control measures at CCPs should result in an
acceptable level of the hazard being controlled. There may be more than one CCP in a
process at which control is applied to address the same hazard (e.g. the cook step may be
the CCP for killing the vegetative cells of a pathogenic spore-former, but the cooling step
may be a CCP to prevent germination and growth of the spores). Similarly, a CCP may
control more than one hazard (e.g. cooking can be a CCP that addresses several microbial
pathogens). Determining whether or not the step at which a control measure is applied is a
CCP in the HACCP system can be helped by using a decision tree. A decision tree should be
flexible, given whether it is for use in production, slaughter, processing, storage, distribution
or other processes. Other approaches such as expert consultation may be used.

8. Establish validated critical limits for each CCP (Step 8/ Principle 3):
Critical limits establish whether a CCP is in control, and in doing so they can be used to
separate acceptable products from unacceptable ones. These critical limits should be
measurable or observable. In some cases, more than one parameter could have a critical
limit designated at a particular step (e.g. heat treatments commonly include critical limits
for both time and temperature). Criteria often used include minimum and/or maximum
values for critical parameters associated with the control measure such as measurements of
temperature, time, moisture level, pH, aw, available chlorine, contact time, conveyor belt
speed, viscosity, conductance, flow rate, or, where appropriate, parameters that can be
observed, such as a pump setting. A deviation from the critical limit indicates that it is likely
that unsafe food has been produced.

9. Establish a Monitoring System for Each CCP (Step 9/ Principle 4):

Monitoring of CCPs is the scheduled measurement or observation at a CCP relative to
its critical limits. The monitoring procedures should be able to detect a deviation at the CCP.
Further, the monitoring method and frequency should be capable of timely detection of any
failure to remain within critical limits, to allow timely isolation and evaluation of the
product. Where possible, process adjustments should be made when monitoring results
indicate a trend towards a deviation at a CCP. The adjustments should be taken before a
deviation occurs. All records and documents associated with monitoring CCPs should be
signed or initialled by the person performing the monitoring and should also report the
results and timing of the performed activity.

10. Establish corrective actions (Step 10/ Principle 5):

Specific written corrective actions should be developed for each CCP in the HACCP
system in order to effectively respond to deviations when they occur. When critical limits at

Agriculture & GK Page 115

Mob: 9473826765 [Link]
CCPs are monitored continuously and a deviation occurs, any product being produced at the
time the deviation occurs is potentially unsafe. When a deviation in meeting a critical limit
occurs and monitoring was not continuous, then the FBO should determine what product
may have been impacted by the deviation. The corrective actions taken when a deviation
occurs should ensure that the CCP has been brought under control and food that is
potentially unsafe is handled appropriately and does not reach consumers. Actions taken
should include segregating the affected product and analysing its safety to ensure proper
disposition.

11. Validation of the HACCP Plan and Verification Procedures (Step 11/
Principle 6):
Before the HACCP plan can be implemented, its validation is needed; this consists of
making sure that the following elements together are capable of ensuring control of the
significant hazards relevant to the food business: identifying the hazards, critical control
points, critical limits, control measures, frequency and type of monitoring of CCPs,
corrective actions, frequency and type of verification and the type of information to be
recorded. Validation of control measures and their critical limits is performed during the
development of the HACCP plan. Validation could include a review of scientific literature,
using mathematical models, conducting validation studies, and/or using guidance developed
by authoritative sources.

Where HACCP guidance developed by external experts, instead of the HACCP team,
has been used to establish the critical limits, care should be taken to ensure that these limits
fully apply to the specific operation, product or groups of products under consideration.
During the initial implementation of the HACCP system and after verification procedures
have been established, evidence should be obtained in operation to demonstrate that
control can be achieved consistently under production conditions. Any changes having a
potential impact on food safety should require a review of the HACCP system, and when
necessary a revalidation of the HACCP plan.

After the HACCP system has been implemented, procedures should be established to
confirm that the HACCP system is working effectively. These include procedures to verify
that the HACCP plan is being followed and controlling hazards on an ongoing basis, as well
as procedures that show the control measures are effectively controlling the hazards as
intended. Verification also includes reviewing the adequacy of the HACCP system
periodically and, as appropriate, when changes occur.

12. Establish Documentation and Record Keeping (Step 12/ Principle 7):
Efficient and accurate record keeping is essential to the application of a HACCP
system. HACCP procedures should be documented. Documentation and record keeping
should be appropriate to the nature and size of the operation and sufficient to assist the
business to verify that the HACCP controls are in place and being maintained. Expertly
developed HACCP guidance materials (e.g. sector-specific HACCP guides) may be utilized as
Agriculture & GK Page 116
Mob: 9473826765 [Link]
part of the documentation, provided that those materials reflect the specific food
operations of the business.

➢ HACCP requires the education of nonprofessional food handlers, especially in the food
service industry and in homes; whether this will be achieved remains to be seen. The
failure of these individuals to get a proper understanding of HACCP could lead to is
failure.
➢ To be effective, this concept must be accepted not only by food processors but also by
food inspectors and the public. Its ineffective application at any level can be
detrimental to is overall success for a product.
➢ It is anticipated that experts will differ as to whether a given step is a CCP and how
best to monitor such steps. This has the potential of eroding the confidence of others
in HACCP.
➢ The adoption of HACCP by industry has the potential of giving false assurance to
consumers product is safe, and, therefore, there is no need to exercise the usual
precautions between the purchase and consumption of a product. Consumers need to
be informed that most outbreaks of food borne illness are caused by errors in food-
handling in homes and food service establishments and that no matter what steps a
processor takes, principles must be observed after foods are purchased for
consumption.

Typical monitoring systems include:

• Preventive maintenance inspections.

• Cleaning efficiency inspections.
• Hygiene audits and inspections.
• Ensuring compliance with food safety specifications to suppliers.
• Inspections to detect the presence of infestation and to monitor the effectiveness of
current pest control activities.
• Temperature checks of food commodities.
• Food supplier audits and inspections.

Useful information on checking performance against control standards can also be

obtained reactively from the following activities:

a. investigation of food safety incidents

b. food sampling activities;
c. observation of food handling practices; and
d. health surveillance of food handlers.

Reviewing the risk assessment:

The frequency of review depends upon the level of risk in the operation. Further, if a
serious food safety incident occurs in the organization, or elsewhere, but is possible in the

Agriculture & GK Page 117

Mob: 9473826765 [Link]
organization, and where a check on the risk assessment shows no assessment or a gap in
assessment procedures, then a review is necessary.

Risk communication:
Risk communication is defined for the purposes of the Codex Alimentarius
Commission as "The interactive exchange of information and opinions throughout the risk
analysis process concerning hazards and risks, risk-related factors and risk perceptions,
among risk assessors, risk managers, consumers, industry, the academic community and
other interested parties, including the explanation of risk assessment findings and the basis
of risk management decisions."

PREVENTING OR CONTROLLING THE RISKS

Once the food safety hazards have been identified and risks assessed, an operator of a
food business must either prevent the risk arising or, alternatively, control same. Much will
depend upon the magnitude of the risk in terms of the controls applied. in certain cases, the
level of competence of food handlers and others may need to be assessed prior to their
undertaking certain work. e. g. preparation of high risk foods. A range of controls must be
considered, including:

➢ Well-managed cleaning procedures.

➢ Temperature monitoring of freezers, display units, cold stores and refrigerators and or
frozen foods at time of delivery.
➢ Planned preventive maintenance systems covering, for instance, structural
equipment, hot water supplies and mechanical ventilation systems.
➢ Effective structural proofing against various forms of infestation, together with
monitoring the performance of pest control contractors.
➢ Food labelling, dating and stock rotation.
➢ Measures to protect food from risk of contamination and cross contamination.
➢ The provision of information, instruction and training for all staff.
➢ Health surveillance of food handlers in particular, including the use of health
questionnaires for newly-appointed food handlers.
➢ Hygienic storage and disposal of waste, together with strict control over rejected food.
➢ Checking the quality and fitness of fresh high risks food supplied from external sources
e.g. wholesaler, butcher.
➢ The operation of a formally documented product recall system.

Some other International Standards and Regulations:

Quality of the food is major concern worldwide now-a-days. So, each country has
formulated. its own standards and created agencies for strict quality control measures of
the food products. Some of them are internationally accepted standards. International
standards may apply to a certain region of the world or any trade between parties of
different countries.
Agriculture & GK Page 118
 Mob: 9473826765 [Link]

• Setting up of international standard for the purpose of food safety depends upon the
following agreements.
• Agreement on Agriculture (AoA)
• Agreement on the Application of Sanitary and Phytosanitary, Measures (SPS
Agreement)
• Agreement on Technical Barriers to Trade (TBT Agreement)
• International Health Regulation (2005)

ISO 22000 STANDARD

ISO 22000 is the most popular voluntary food safety standard in the food industry. The
ISO 22000 family are international voluntary consensus standards which define the
requirements for a Food Safety Management System (FSMS) and incorporates the following
elements which as defined as FSMS principles:

• Interactive communication
• System management
• Prerequisite programs
• HACCP principles.

CHARACTERISTICS OF ISO 22000

ISO 22000 specifies the characteristics of a management system design to:

• Carry out the Hazard Analysis
• Design the HACCP plan.
• Identify the prerequisite programmes
• Select the operational prerequisite programmes

ISO 22000 sets out the requirements for a food safety management system and can be
certified to it. It maps out what an organization needs to do to demonstrate its ability to
control food safety hazards in order to ensure that food is safe. It can be used by any
organization regardless of its size or position in the food chain.

The 10 Clauses of ISO 22000:2018

i. Scope
ii. Normative References
iii. Terms And Definitions
iv. Context of the Organization
v. Leadership 6. Planning
vi. Support
vii. Operation
viii. Performance Evaluation

Agriculture & GK Page 119

 Mob: 9473826765 [Link]
ix. Improvement

RELATION BETWEEN HACCP & ISO 22000

ISO 22000 integrates the principles of the Hazard Analysis and Critical Control Point
(HACCP) system and application steps developed by the Codex Alimentarius Commission. By
means of auditable requirements, it combines the HACCP plan with prerequisite
programmes. Hazard analysis is the key to an effective food safety management system,
since conducting a hazard analysis assists in organizing the knowledge required to establish
an effective combination of control measures. ISO 22000 requires that all hazards that may
be reasonably expected to occur in the food chain, including hazards that may be associated
with the type of process and facilities used, are identified and assessed. Thus, it provides the
means to determine and document why certain identified hazards need to be controlled by
a particular organization and why others need not.

ISO 9001 vs ISO 22000

In comparison with ISO 9001, the standard is a more procedural orientated guidance
than a principle based one. Apart from that, ISO 22000 is an industrial-specific risk
management system for any type of food processing and marketing, which is designed using
the ISO high level structure (HLS), also referred to as Annex SL, to be integrated with the
quality management system of ISO 9001. The detailed similarities and differences of the two
standards can be found elsewhere. Both ISO 9001 and ISO 22000 follow the process
approach. The process approach involves the systematic definition and management of
processes, and their interactions, so as to achieve the intended results in accordance with
the food safety policy and strategic direction of the organization. Management of the
processes and the system as a whole can be achieved using the PDCA cycle, with an overall
focus on risk-based thinking aimed at taking advantage of opportunities and preventing
undesirable results.

Both ISO 9001 and ISO 22000 have common management system principles used to
provide a unifying basis for values and beliefs which shape the organization’s purpose,
mission and culture.

• Customer focus
• Leadership
• Engagement of people
• Process approach
• Improvement
• Evidence-based decision making
• Relationship management

Agriculture & GK Page 120

Mob: 9473826765 [Link]
STEPS INVOLVED IN IMPLEMENTATION OF ISO 22000

ISO-22000
IMPLEMENTATION
STEPS

1. Nomination of the food

safety team

2. Setting up prerequisite
programmes

3. Development of the HACCP

Plan

[Link]

[Link], awareness

6. Internal FSMS audit

Agriculture & GK Page 121

Mob: 9473826765 [Link]

7. Management review

8. Certification

Agriculture & GK Page 122