Introduction to Food

UNIT 1 INTRODUCTION TO FOOD Microbiology

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

1.7 Important Micro-organisms in Food

1.7.1 Important Mold Genera
1.7.2 Important Yeast Genera
1.7.3 Important Viruses
1.7.4 Important Bacterial Genera

1.8 Normal Microflora of some Common Foods

1.9 Let Us Sum Up
1.10 Key Words
1.11 Answers to Check Your Progress Exercises
1.12 Suggested Reading

1.0 OBJECTIVES
After reading the Unit, we shall be able to:
z explain the applied aspects of microbiology;
z comprehend the need and scope of food microbiology;
z enumerate the important genera of micro-organisms associated with food;
and
z specify normal microflora of some common foods.

1.1 INTRODUCTION
Micro-organisms are living entities of microscopic size and include bacteria,
viruses, yeasts and molds (together designated as fungi), algae, and protozoa.
While bacteria are classified as prokaryotes (cells without definite nuclei), the
fungi, algae, and protozoa are eukaryotes (cells with nuclei); viruses do not
have regular cell structures and are classified separately. Micro-organisms are 7
Fundamentals of Food present everywhere on earth, which includes humans, animals, plants and
Microbiology
other living creatures, soil, water, and atmosphere, and they can multiply
everywhere except in the atmosphere. Together, their numbers far exceed all
other living cells on this planet. They were the first living cells to inhabit the
earth over 3 billion years ago; and since then they have played important
roles, many of which are beneficial to the other living systems.
Among the micro-organisms, some molds, yeasts, bacteria, and viruses have
both desirable and undesirable roles in our food. In this unit, the scope of
food microbiology, importance of microbes in food and predominant micro-
organisms associated with food have been discussed.

1.2 THE SCIENCE OF MICROBIOLOGY

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

1.3 FOOD MICROBIOLOGY - ITS ORIGIN AND

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

Food Laboratory
fermentations management

Water Food
quality hygiene

Topics of major interest

to the food microbiologist

Food Food-borne
preservation disease

Food Quality
spoilage control

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

Fundamentals of Food Not all groups of micro-organisms are of equal interest to the food
Microbiology
microbiologist. Bacteria come very much on top of the list with molds and
yeasts also of considerable importance and viruses less so. The associations
that these organisms have with the manufacture and consumption of foods are
summarized in Fig. 1.2.

Food-borne disease Food-borne disease

Food spoilage
Food spoilage

Bacteria Moulds Production of single cell protein

Food fermentations
Food fermentations

Production of food additives and enzymes Production of enzymes

Food spoilage Food-borne disease

Yeasts Food fermentations Viruses Identification of food

poisoning bacteria

Production of food additives and enzymes

Failure of dairy fermentations

Fig. 1.2: Various groups of micro-organisms and their associations with food

Protozoa and algae have minimum direct impact on the production, processing
and consumption of food. Food-borne disease can be caused by some protozoa
and others belonging to this group are important in the treatment of wastes.
Algae are used to produce alginates; some have the potential for use in the
production of single-cell protein and some marine species produce toxins that
might enter our food along with sea foods.

1.4 IMPORTANCE OF MICRO-ORGANISMS IN

FOODS

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

food has increased greatly. Their role in food can be either desirable (food
bioprocessing) or undesirable (food borne diseases and food spoilage), which
is briefly discussed here.

1.4.1 Food-borne Diseases

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

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

1.4.2 Food Spoilage

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

1.4.3 Food Bioprocessing

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

1.4.4 Food Additives

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

1.4.5 Food Biopreservation

Antimicrobial metabolites (e.g. bacteriocins and organic acids like acetic,
propionic and lactic acids) of desirable Micro-organisms are being developed
and used in foods in place of preservatives of non-food (chemical) origin to
control pathogenic and spoilage micro-organisms in food. Economic production
of these antimicrobial compounds and their effectiveness in food systems have
generated wide interest.

1.4.6 Probiotics
Consumption of foods containing live cells of bacteria and that have apparent
health benefits has generated interest among consumers. The role of these
bacteria for health and bacterial efficacy benefits is being critically investigated.
11
Fundamentals of Food
Microbiology Check Your Progress Exercise 1
Note: a) Use the space below for your answer.
b) Compare your answers with those given at the end of the unit.
1) What are Micro-organisms?
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2) What is Food Microbiology?
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3) What is a probiotic?
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1.5 CLASSIFICATION AND NOMENCLATURE

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

1.6.1 Bacteria
Bacteria are unicellular Micro-organisms that are approximately one micro
meter (10-3mm) in diameter with variations in morphology from short and

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

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

Check Your Progress Exercise 2

Note: a) Use the space below for your answer.
b) Compare your answers with those given at the end of the unit.
1) What is Systematics?
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2) What are the two most common Micro-organisms found in food?
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Fundamentals of Food 3) Classify bacteria on the basis of their morphology.
Microbiology
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4) What is a bacteriophage?
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1.7 IMPORTANT MICRO-ORGANISMS IN FOOD

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

SPOILAGE
Structure
2.0 Objectives

2.1 Introduction

2.2 Food Contamination

2.2.1 Contamination of Living Plants and Animals
2.2.2 Sources of Contamination

2.3 Food Spoilage

2.3.1 Types of Spoilage
2.3.2 Classification of Foods on the Basis of Stability

2.4 Role of Micro-organisms

2.4.1 Micro-organisms Involved in Spoilage
2.4.2 Growth of Micro-organisms

2.5 Factors Affecting Spoilage

2.5.1 Extrinsic Factors
2.5.2 Intrinsic Factors
2.5.3 Interaction Among Growth Factors

2.6 Deteriorative Effect of Micro-organisms

2.6.1 Physical Changes
2.6.2 Chemical Changes

2.7 Different Types of Spoilage

2.8 Common Methods of Food Preservation
2.9 Let Us Sum Up
2.10 Key Words
2.11 Answers to Check Your Progress Exercises
2.12 Suggested Reading

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

2.2 FOOD CONTAMINATION

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

Aerosols
Air
Spoiled Water
foods

Processing Soil
Equipment Dust
Raw material
with natural
micro flora
Diseased plants Sewage
and animals

Packing Humans
Materials

Pests
Faeces
30 Fig. 2.1: Sources of contamination of food
iv) Food storage: Food that is stored incorrectly, for instance an uncooked Food Contamination
and Spoilage
chicken thigh resting next to a bunch of grapes, can be a source of
transferring bacteria and other contaminants from one food to another.
v) Food preparation: A great deal of food contamination occurs during the
preparation stage. A sick person can pass on germs, ranging from flu to
gastroenteritis. A chopping board used for meat that is not washed and
then used for vegetables is another source of possible contamination.
Unwashed hands, dirty kitchen spaces, insects and rodents in the kitchen
etc. are all possible sources of food contamination.

2.3 FOOD SPOILAGE

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

The criteria for assurance in foods suitable for consumption are:

• The desired stage of development or maturity of the food.

• Freedom from pollution at any stage in the production and subsequent

handling of the food.

• Freedom from objectionable, chemical and physical changes resulting

from action of food enzymes; activity of microbes, insects, rodents, invasion
of parasites; and damage from pressure, freezing, heating, drying, and the
like.

• Freedom from micro-organisms and parasites causing food borne illnesses.

Enzymatic and microbial activities are undesirable when they are unwanted or
uncontrolled. An example is the souring of milk; if unwanted, it is spoilage, yet
the same process is purposely used in the production of certain cheeses and
other fermented products made from milk.

2.3.1 Types of Spoilage

The food may become unacceptable due to the following factors:

1) Growth and activities of micro-organisms principally bacteria, yeasts and

moulds (This is by far the most important and common cause of
food spoilage).

2) Activities of food enzymes (enzymatic browning is a common example).

3) Infestation by insects, parasites and rodents.

4) Chemical changes in a food (i.e. not catalyzed by enzymes of the tissues 31

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

Classified as semi perishables are usually the dry goods, such as flour, dry
legumes, baked goods, hard cheeses, dried fruits and vegetables, and even
waxed vegetables. Frozen foods, though basically perishable, may be classified
as semi perishables provided they are freezer-stored properly.

The majority of our food materials must be classified as perishables. This

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

2.4 ROLE OF MICRO-ORGANISMS

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

2.4.1 Micro-organisms involved in Spoilage

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

Yeasts are unicellular organisms of small sizes which multiply by budding. In

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

2.4.2 Growth of Micro-organisms

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

Resting Phase
Logof Numbers of Bacteria

Growth
(Log)

Lag Death Phase

Phase

Time

Fig. 2.1: A typical bacterial growth curve

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

Check Your Progress Exercise 1

Note: a) Use the space below for your answer.
b) Compare your answers with those given at the end of the unit.
1) What are spoilage bacteria?
.............................................................................................................
.............................................................................................................
.............................................................................................................
2) Classify foods on the basis of their stability.
.............................................................................................................
.............................................................................................................
.............................................................................................................
3) What are the types of food spoilage?
.............................................................................................................
.............................................................................................................
.............................................................................................................
4) What causes food spoilage?
.............................................................................................................
.............................................................................................................
.............................................................................................................

2.5 FACTORS AFFECTING SPOILAGE

Food is a chemically complex matrix, and predicting whether, or how fast,

micro-organisms will grow in any given food is difficult. Most foods contain
sufficient nutrients to support microbial growth. Several factors encourage,
prevent or limit the growth of micro-organisms in foods; the most important
are water activity aw, pH and temperature.
Factors affecting microbial growth are divided into two groups -intrinsic and
extrinsic parameters (Table 2.1). These factors affect the growth of micro-
organisms on foods. When spoilage of a food occurs under a given set of
circumstances, not all of the different types of organisms contaminating a food
are associated with the spoilage process. In fact, the spoilage flora always is

34
dominated by just a few and sometimes only one organism. Components of Food Contamination
and Spoilage
the micro flora compete with one another for the available nutrients and the
organism(s) with fastest growth under a particular set of circumstances will
become dominant and give rise to the spoilage symptoms.

Table 2.1: Intrinsic and Extrinsic Parameters Affecting Microbial Growth

Intrinsic parameters Extrinsic parameters

Water activity, humectant identity Temperature

Oxygen availability Relative humidity

pH, acidity, acidulant identity Atmospheric composition

Buffering capacity Packaging

Available nutrients

Natural antimicrobial substances

Presence and identity of natural microbial flora

Colloidal form

The component of the micro flora which becomes dominant is determined by

a complex interaction between the components of the contaminating micro
flora (implicit factors), the storage environment (extrinsic factors) and the
physico-chemical properties of the food (intrinsic factors).

All of the factors that influence the growth of micro-organisms have been
dealt with as under. Time is included because, under any given set of
circumstances, spoilage takes a finite period to occur and equates with the
storage life of a product.

A knowledge of the intrinsic and extrinsic parameters should enable you to

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

2.5.1 Extrinsic Factors

Extrinsic factors relate to the environmental factors that affect the growth rate
of micro-organisms. They are as follows:

A. Temperature

Microbes have an optimum temperature as well as minimum and maximum

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

The optimal temperature for the proliferation of most micro-organisms is from

150 to 400C. However, many genera of microbes are capable of growth from
00 to 150C and other even micro-organisms will grow at subzero temperatures.
Still other genera will grow at temperatures up to and exceeding 1000C.

Microbes classified according to temperature of optimal growth include:

• Thermophiles (high-temperature-loving micro-organisms), with growth

optima at temperatures above 450C (e.g., Bacillus stearothermophilus,
Bacillus coagulans, and Lactobacillus thermophilus).

• Mesophiles (medium-temperature-loving micro-organisms), with growth

optima between 200 and 450C (e.g., most Bactobacilli and Staphylococci).

• Psychrotrophs (cold-temperature-tolerant micro-organisms), which

tolerate and thrive at temperatures below 200C (e.g., Pseudomonas and
Acinetobacter).

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

C. Relative Humidity

This extrinsic factor affects microbial growth and can be affected by

temperature. All micro-organisms have high requirements for water to support
their growth and activity. A high relative humidity can cause moisture
condensation on food, equipment, walls, and ceilings. Condensation causes
moist surfaces, which are conducive to microbial growth and spoilage. Also,
microbial growth is inhibited by a low relative humidity. Micro-organisms
bacteria require the highest relative humidity of the various. Optimal relative
humidity for bacteria is 92% or higher, whereas yeasts need 90% or higher
and for molds, the value of relative humidity is 85-90%.
36
2.5.2 Intrinsic Factors Food Contamination
and Spoilage

Intrinsic factors that affect the rate of proliferation relate more to the
characteristics of the substrates (foodstuff or debris) that support or affect
growth of micro-organisms. These major intrinsic factors are:
A. Water Activity (a w )
Water is required by micro-organisms, and a reduction of water availability
constitutes a method of food preservation through reduction of microbial
proliferation. It is important to recognize that it is not the total amount of
moisture present that determines the limit of microbial growth, but the amount
of moisture which is readily available for metabolic activity of microbes.
The unit of measurement for water requirement of microorganism is
usually expressed as water activity (a w ). Water activity is defined as the
vapor pressure of the subject solution divided by the vapor pressure of the
pure solvent: aw = p/p0, where p is the vapor pressure of the solution and po is
the vapor pressure of pure water. The approximate optimal aw for the growth
of many micro-organisms is 0.99, and most microbes require an aw higher
than 0.91 for growth. The relationship between relative humidity (RH) and aw
is RH = aw × 100. Therefore an aw of 0.95 is equivalent to an RH of 95%.
Generally, bacteria have the highest water activity requirements of the micro-
organisms. Molds normally have the lowest aw requirements, with yeasts being
intermediate. Most spoilage bacteria do not grow at an aw below 0.91, but
molds and yeasts can grow at an aw of 0.80 or lower. Molds and yeasts are
more likely to grow in partially dehydrated surfaces (including food), whereas
bacterial growth is retarded.

Approximate Minimum (aw) Values for Growth

ORGANISMS GROUPS WATER ACTIVITY

MOST SPOILAGE BACTERIA 0.90

MOST SPOILAGE YEASTS 0.88

MOST SPOILAGE MOLDS 0.80

B. pH

The pH for optimal growth of most micro-organisms is near neutrality (7.0).

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

37
Fundamentals of Food
Microbiology

Fig. 2.2: pH and growth of micro-organisms

C. Oxidation – Reduction Potential

The oxidation – reduction (redox) potential is an indication of the oxidizing

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

D. Nutrient Requirements

In addition to water and oxygen (except for anaerobes), micro-organisms

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

E. Inhibitory Substances

Microbial proliferation can be affected by the presence or absence of inhibitory

substances. Substances or agents that inhibit microbial activity are called
38
Fundamentals of Food
Microbiology UNIT 3 FOOD-BORNE DISEASES
Structure
3.0 Objectives
3.1 Introduction
3.2 What is a Disease?
3.3 How Do Micro-organisms Cause Disease?
3.4 Food-borne Diseases and the Agents
3.4.1 Diseases by Bacteria
3.4.2 Diseases by Molds
3.4.3. Diseases by Viruses
3.4.4. Diseases by Parasites
3.4.5 Diseases by Natural toxins
3.4.6 Diseases by Prions

3.5 Types of Food-borne Diseases

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

3.9 Let Us Sum Up

3.10 Key Words
3.11 Answers to Check Your Progress Exercises

3.12 Suggested Reading

3.0 OBJECTIVES
After reading this Unit, we shall be able to:
z specify food-borne diseases: their types and causative agents;
z enumerate common food-borne diseases and their symptoms; and
z state emerging food-borne pathogens and reasons for their emergence.

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

3.2 WHAT IS DISEASE?

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

3.3 HOW DO MICRO-ORGANISMS CAUSE

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

3.4 FOOD-BORNE DISEASES AND THE AGENTS

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

Agent Example

Bacteria Salmonella, Staphylococcus aureus, Clostridium

perfringens, C.botulinum, Bacillus cereus

Viruses Hepatitis A virus, Parvovirus, Norwalk virus

Molds Claviceps purpurea, Aspergillus flavus (foodborne

aflatoxins)

Marine protozoa Gonyaulax tamareusis (paralytic crustacean

poisoning). Gambierdiscus toxicus (poisoning with
ciguatera). Prorocentrum lima and some species
of Dinophys (diarrheic shellfish poisoning)

Parasites Trichinella spiralis, Taenia solium, Giardia

(protozoa, amoebas) lamblia, Cryptosporidium, Entamoeba histolytica

Chemical substances Heavy metals (lead, zinc, copper, cadmium, mercury,

arsenic, antimony), pesticides, herbicides, fungicides,
substances used for cleaning and disinfections.

Toxic plants Germinated potatoes, apricot core, red bean

(Phaseolus vulgaris)

Toxic animals Scombrotoxin from fish insufficiently cooked –

herring – (histamine)

3.4.1 Diseases by Bacteria

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

3.4.3 Diseases by Viruses

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

3.4.5 Diseases by Natural Toxins

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

6) Grayanotoxin (Honey intoxication)

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

• Scombroid Poisoning (also called Histamine Poisoning)

Scombroid poisoning is caused by the ingestion of foods that contain high levels
of histamine and possibly other vasoactive amines and compounds. Histamine and
other amines are formed by the growth of certain bacteria (Proteins morganii,
Klebsiella pneumoniac) and the subsequent action of their decarboxylase enzymes
on histidine and other amino acids in food, either during the production of a
product such as Swiss cheese or by spoilage of foods such as fishery products,
particularly tuna or mahi mahi. However, any food that contains the appropriate
amino acids and is subjected to certain bacterial contamination and growth may
56 lead to scombroid poisoning when ingested.
3.4.6 Diseases by Prions Food Borne Diseases

Prions are normal proteins of animal tissues that can misfold and become infectious:
they are not cellular organisms or viruses. Prions are associated with a group of
diseases called Transmissible Spongiform Encephalopathies (TSEs). In humans,
the illness suspected of being Food-borne is variant Creutzfeldt-Jakob disease
(vCJD). The human disease vCJD and the cattle disease, bovine spongiform
encephalopathy (BSE), also known as “mad cow” disease, appear to be caused
by the same agent.

Check Your Progress Exercise 1

Note: a) Use the space below for your answer.
b) Compare your answers with those given at the end of the unit.
1) What are pathogens?
............................................................................................................
............................................................................................................
............................................................................................................
2) Define FBD.
............................................................................................................
............................................................................................................
............................................................................................................
............................................................................................................
3) Mycotoxins are ..................... metabolites.
4) What is an enterotoxin?
............................................................................................................
............................................................................................................
............................................................................................................

3.5 TYPES OF FOOD-BORNE DISEASES (FBDs)

Types of food poisoning include the following :
• Intoxications
• Infections.
• Toxi-infections
Table 3.2 enlists the food-borne diseases and causative pathogens

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

Type of disease Causative microorganism Microbial group Major symptom(s)

type

Intoxication

Staph. poisoning Staphylococcus aureus Bacteria, Gm +* Gastric

strains

Botulism Clostridium botulinum Bacteria, Gm + Non gastric

strains

Mycotoxin Mycotoxins producing mold Molds Non gastric

poisoning strains, e.g., Aspergillus
flavus

Infection

Salmonellosis Over 2000 Salmonella Bacteria, Gm - * Gastric

serovars (except S. typhi
and S. paratyphi)

Campylobacter Campylobacter jejuni and C. Bacteria, Gm - Gastric

enteritis coli strains

Yersiniosis Pathogenic strains of Bacteria, Gm - Gastric

Yersinia enterocolitica
58
 Food Borne Diseases
Type of disease Causative microorganism Microbial group Major symptom
(s) type

Intoxication Intoxication Intoxication

Enterohemorrhagic Escherichia coli Bacteria, Gm - Gastric and

E. coli colitis O157:H7 nongastric

Nonhaemorrhegic Shiga-like toxin (verotoxin) Bacteria, Gm - Gastric

E. coli colitis producing
E. coli strains like
E. coli 026:H11

Listeriosis Listeria monocytogenes Bacteria, Gm + Gastric and

(Pathogenic strains) nongastric

Shigellosis Four Shigella species e.g., Bacteria, Gm - Gastric

S. dysenterie

Vibrio Pathogenic strains of Bacteria, Gm - Gastric

parahaemolyticus V. parahaemolyticus
gastroenteritis

Vibrio vulnificus Vibrio vulnificus strains Bacteria, Gm - Gastric and

infection nongastric

Brucellosis Brucella abortus Bacteria, Gm - Gastric and

nongastric

Viral infections Pathogenic enteric viruses, Viruses Gastric and

e.g., Hepatitis A. virus nongastric

Toxi infection

Clostridium Clostridium perfringens Bacteria, Gm+ Gastric

perfringens strains
gastroenteritis

Bacillus cereus Bacillus cereus strains Bacteria, Gm + Gastric

gastroenteritis

E. coli Enteropathogenic and Bacteria, Gm - Gastric

gastroenteritis enterotoxigenic Escherichia
coli strains

Cholera Pathogenic strains of Vibrio Bacteria, Gm - Gastric

cholerae

Gastroenteritis by Opportunist Pathogens

Aeromonas Aeromonas hydrophila Bacteria, Gm - Gastric

hydrophila strains
gastroenteritis

Plesiomonas Plesiomonas shigelloidies Bacteria, Gm - Gastric

shigelloides strains
gastroenteritis

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

59
 Beneficial Role of
UNIT 4 BENEFICIAL ROLE OF Micro-organisms

MICRO-ORGANISMS
Structure
4.0 Objectives
4.1 Introduction
4.2 Fermentation
4.3 Fermented Foods and their Importance
4.4 Food Fermentation-Science and Technology
4.5 Types of Food Fermentations
4.5.1 Acid Food Fermentation
4.5.2 Yeast food fermentation
4.5.3 Solid State Fermentation

4.6 Common Examples of Food Fermentation

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

4.7 Fermented foods as Functional Foods

4.7.1 Probiotics
4.7.2 Prebiotics
4.7.3 Synbiotics
4.7.4 Use of Probiotics
4.7.5 Health Benefits of Probiotics

4.8 Let Us Sum Up

4.9 Key Words
4.10 Answers to Check Your Progress Exercises
4.11 Suggested Reading

4.0 OBJECTIVES

After reading this Unit, we shall be able to:

z describe beneficial role of microorganisms;
z explain fermentation – its science and technology;
z comprehend the needs and types of food fermentations;
z state common examples of different types of food fermentations; and
z attribute fermented foods as functional foods.

71
Fundamentals of Food
Microbiology 4.1 INTRODUCTION
Food microbiology can be divided into three focus areas; beneficial microorganisms,
spoilage microorganisms, and disease causing microorganisms. Beneficial
microorganisms are those used in food fermentation to produce products such as
cheese, fermented meat (pepperoni), fermented vegetables (pickles), fermented
dairy products (yoghurt), and ethnic fermented products such as sauerkraut, idli
and kimchi. In fermented products (produced by natural or controlled fermentation),
microorganisms metabolize complex substrates to produce enzymes, flavor
compounds, acids, and antimicrobial agents to improve product shelf-life and to
prevent growth of pathogens and to provide product attributes. Micro organisms
with their enzymes, also breakdown indigestible compounds to make the product
more palatable and easy to digest. In addition, the beneficial microorganisms also
serve as probiotics to impart direct health benefit by modulating the immune
system to provide protection against chronic metabolic diseases, bacterial infection,
atherosclerosis, and allergic responses. Examples of beneficial microorganisms
are Lactobacillus acidophilus, Lactobacillus arabinosus, Lactobacillus lactis,
and Pediococcus cerevisiae. Food spoilage microorganisms are those which
upon growth in a food, produce undesirable flavour (odour), texture and
appearance, and make food unsuitable for human consumption. Sometimes
uncontrolled growth of many of the beneficial microorganisms can also cause
spoilage. Food spoilage is a serious issue in developing countries because of
inadequate processing and refrigeration facilities. Examples of food spoilage
microorganisms are Brochotrix, Lactobacillus, Bacillus, Pseudomonas spp.
and some molds. The micro-environment created in a spoilt food generally
discourages the growth of the pathogenic microorganisms, which are considered
poor competitors.

4.2 FERMENTATION
The uniqueness of several microorganisms and their often unpredictable nature
and biosynthetic capabilities, in a specific set of environmental conditions, have
made them ideal candidates, in attempts to solve difficult problems in life sciences
and other fields. Microorganisms have been used in various ways over the past
many decades, to advance medical technology, human and animal health, food
processing, food safety and quality, genetic engineering, environmental protection
and agricultural biotechnology. The use of beneficial microorganisms in the food
sector has been a long tradition, namely lactic acid bacteria and yeasts in
fermentation processes; the former are widely used in the manufacture of fermented
food and are among the best studied microorganisms. The fermentations may be
by yeasts, bacteria, molds or combination of these organisms. Detailed knowledge
of a number of physiological traits has opened novel potential applications for
these organisms in the food industry, while other traits might be specifically, beneficial
for human health.

The food and beverage industry exploits non-pathogenic microorganisms for the
production of fermented foods. These foods are prepared from raw materials and
acquire their characteristic properties by a process that involves microorganisms.
In certain cases the endogenous enzymes of the wild micro flora nature to the raw
material may play a decisive role. It is believed that fermented foods originated
72
from the Orient and date back to the prehistoric times. Originally, these were Beneficial Role of
Micro-organisms
fermented “spontaneously” by autochthonous strains found in the raw materials
or the environment; This was the start of traditional biotechnology. The most
important were cheese, yoghurt, wine, vinegar, beer, bread and the traditional
fungal fermentations used in Asia and Africa, for the production of food.

In the biochemical sense, the term fermentation refers to a metabolic process in

which organic compounds (particularly carbohydrates) are broken down to release
energy without the involvement of a terminal electron acceptor such as oxygen.
Partial oxidation of the substrate occurs so that only a relatively small amount of
ATP energy is released compared to the energy generated if a terminal electron
acceptor is involved. Partial oxidation of a carbohydrate can give rise to a variety
of organic compounds such as alcohols organic acids and acetone. The compounds
produced by micro-organisms vary from organism to organism and are produced
through different metabolic pathways.

The term fermentation can also be applied to any industrial process that produces
a material that is useful to humans and if the process depends on the activity of
one or more micro-organisms (consortia). These processes, known as industrial
fermentations, are usually carried out on a large scale and in vessels in which the
organisms are normally grown in liquid media. Some industrial fermentations are
fermentation in the biochemical sense but the majority of microorganisms involved
are aerobic and use oxygen (as terminal electron acceptor) and thereby metabolize
carbohydrates completely.

A vast range of materials are produced by industrial fermentations. These include

1) Organic chemicals used as fuels, food additives, antibiotics and enzymes for
use in the food and other industries. Vinegar is an example of a food additive
produced by an industrial fermentation.

2) Organisms are produced on a large scale for the extraction of protein (single
cell protein) that can be used as a part of the human diet. Quorn is an
example of a single cell protein, It is produced from the fungus Fusarium
graminearum. This mycoprotein, purified from the fungus, is currently available
for use as a food and is incorporated into a range of dishes that appear on
supermarket shelves. Meatless dishes with a high protein content made from
Quorn are particularly appealing to vegetarians.

3) Yeast cells produced for use in industries such as the baking industry, which
relies on the mass production of large amounts of baker’s yeast.

4) Foods produced on a large scale as a result of the activities of micro-

organism, e.g. cheese, yogurt and bread.

5) Production of alcoholic beverages, e.g. beer and wines.

6) Cellular extracts used as food additives, e.g., yeast extracts from yeast cells
produced as a by product of the brewing industry.

7) Industrial fermentations are now often considered under the heading of

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

Food Safety
- antimicrobials
- bacteriocin
- cyclic
dipeptides

Organoleptic
Nutritional
- texture
- vitamins Starter - aroma
- biogenic amine Cultures - flavour
production

Health Technological
- nutraceuticals - phase resistance
- bioactive peptides - lactose-negative
- accelerated ripening

Fig. 4.1: Functional properties of starter cultures for food fermentation

7) Detoxification of raw materials : The fermentation process may remove

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

Pretreatment
Medium nutrients
Sterilization
Biocatalyst Substrate and nutrients
(immobilized or free)
Bioreactor
Purification
Products Waste Utilization
Processing treatment
(Thermal, Chemical,
Freezing, Drying etc.)

Packaging By-products
Useful products
Labelling

Marketing
Fig 4.2: A generalized scheme of various operations employed in food
76
fermentation technology
 Beneficial Role of
Check Your Progress Exercise 1 Micro-organisms

Note: a) Use the space below for your answer.

b) Compare your answers with those given at the end of the unit.
1) What is fermentation?
............................................................................................................
............................................................................................................
............................................................................................................
............................................................................................................
2) What is the purpose of fermentation?
............................................................................................................
............................................................................................................
............................................................................................................
............................................................................................................
3) Define zymology.
............................................................................................................
............................................................................................................
............................................................................................................
............................................................................................................

4.5 TYPES OF FOOD FERMENTATIONS

A number of different types of food fermentation can be recognized.

4.5.1 Acid Food Fermentation

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

4.6 COMMON EXAMPLES OF FOOD

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

1. KANIMA Nepal, Sikkim, Soyabeans — Solid snack food

& Darjeeling

2. KHAMAN India Bengal gram — Solid, cake like

breakfast food

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

Japan substitute

4. PAPADAM India Black gram Saccharomyces Solid, crisp,

sp. condiment

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

6. TEMPEH Indonesia Soyabeans Rhizopus sp. Fried in oil, roasted,

and nearby as a meat substitute
regions

7. WARIES India Black gram Candida sp. Spongy, spicy

flour Saccharomyces condiment
sp.

8. JALEBIES India, Nepal Wheat flour Saccharomyces Syrup filled

& Pakistan bayanus confectionary

9. KANJI India Rice & Hansenula Sour liquid added to

Carrots anomala vegetables

10. MISO Japan, China Rice & Aspergillus Paste, soup base
soyabeans oryzae
Torulopsis
etchellsii
Lactobacillus sp.
Saccharomyces
rouxii

11. NAN India Unbleached —- Solid snack food/

Pakistan wheat flour bread
Afghanistan
Iran

12. DHOKLA India Bengal gram —- Spongy condiment

& wheat

13. DOSAI India Black gram & Yeasts Spongy breakfast

rice food

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

rice T. candida breakfast food
Trichosporon
pullulans

15. TORANI India Rice H. anomala Seasoning for

C. guilliermondii vegetables
C. tropicalis
G. candidum

4.6.2 Fermented Vegetable Foods

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

4.6.4 Fermented Dairy Products

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

Dahi India Milk (Cow/ L. lactis subsp. lactis, S. salivarius

(Dadhi) Persia Buffalo) subsp. thermophilus, L. delbrueckii
subsp. bulgaricus, plantarum, lactose
fermenting yeast, Mixed culture

Srikhand India Milk (Cow/ S. salivarious subsp.thermophilus, L.

(chakka) Buffalo) delbrueckii subsp. bulgaricus.

Lassi India Milk (Cow/ S. salivarius subsp. thermophilus, L.

Buffalo) delbrueckii subsp.bulgaricus.

Cultured butter Scandinevian Milk (Cow/ L. lactis subsp. lactis, L. lactic subsp.
milk and European Buffalo) diacetylactis, Leuconostoc
countries dextranicum subsp. citrovorum

Acidophilus milk Australia Cow’s milk L. acidophilus

Yoghurt Middle Asia, Cow’s milk S, salivarius subsp.thermophilus, L.

(bio-ghurt) Balkans Goat’s or delbrueckii subsp. bulgaricus,
mixed milk Micrococcus and other lactic acid,
80 bacteria, cocci, yeasts, molds