UNIT-I FOOD MICROBIOLOGY

DEFINATION OF FOOD
Food is defined as any substance or materials that are consumed to provide nutritional support
for the body or for pleasure. It is usually of plant or animal origin, and contains essential nutrients,
such as carbohydrates, fats, proteins, vitamins, or minerals. It is ingested and assimilated by an
organism to produce energy, stimulate growth, and maintain life.

Almost all foods are of plant or animal origin. Cereal grain is a staple food that provides more
food energy worldwide than any other type of crop. Maize, wheat, and rice together account for
87% of all grain production worldwide.

Animals are used as food either directly or indirectly by the products they produce. Meat is an
example of a direct product taken from an animal, which comes from either muscle systems or from
organs. Food products produced by animals include milk produced by mammary glands, which in
many cultures is drunk or processed into dairy products such as cheese or butter. In addition, birds
and other animals lay eggs, which are often eaten, and bees produce honey, a reduced nectar from
flowers, which is a popular sweetener in many cultures.

Other foods not from animal or plant sources include various edible fungi, especially
mushrooms. Fungi and ambient bacteria are used in the preparation of fermented and pickled foods
such as leavened bread, alcoholic drinks, cheese, pickles, kombucha, and yogurt. Another example is
blue-green algae such as Spirulina. Inorganic substances such as baking soda and cream of tartar
are also used to chemically alter an ingredient.

COMPOSITION OF FOOD
Food composition is a term that is used to describe an analysis of the vitamins, minerals, and
other nutritive substances in a given food. In addition to vitamins and minerals, many food
composition reports include information on and analysis of phytonutrients and macronutrients
within a food. Food composition information can be used for a number of purposes. It can be used
to make sure that a certain person, family, or population is getting enough of one kind of nutrient,
vitamin, or mineral. It can also be used to define certain foods that may or may not have allergens.
An abbreviated version of a food composition report appears on the labels of most kinds of
packaged food and is intended for the same sorts of uses described above.

There are a number of organizations including the United States Department of Agriculture
(USDA) that compile and disseminate information on food composition. The food composition
information that is available from an organization like the USDA is usually much more detailed than
the information that is printed on packaged foods. The full-length version of a food composition
report for creamed corn baby food will list many — in some cases, dozens of — vitamins, minerals,
lipids, and amino acids. This information can be used to find out, for example, how much lysine is
present in one cup of this particular kind of baby food.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 1
 SIGNIFICANCE OF MICROORGANISMS IN FOOD

Three types of microorganisms occur in foods. They may be beneficial, pathogenic, or

cause spoilage. Beneficial microorganisms include those that may produce new foods or food
ingredients through fermentation(s) (e.g., yeasts and lactic acid bacteria) and probiotics. Spoilage
microorganisms, through their growth and ultimately enzymatic action, alter the taste of foods
through flavor, texture, or color degradation. Pathogenic microorganisms can cause human illness.
Two types of pathogenic microorganisms that grow in or are carried by foods are those that cause:
(1) intoxication and (2) infection. Intoxication results from microorganisms growing and
producing toxin (which causes the illness) in a food. An infection is an illness that results from
ingestion of a disease-causing microorganism. Infectious microorganisms may cause illness by the
production of enterotoxins in the gastrointestinal tract or adhesion to and/or invasion of the
tissues.
A major challenge for the sanitarian is to protect the production area and other involved
locations against microbes that can reduce the wholesomeness of food. Microorganisms can
contaminate and affect food, with dangerous consequences to consumers. The microorganisms
most common to food are bacteria and fungi. The fungi, which are less common than bacteria,
consist of two major microorganisms: molds (which are multicellular) and yeasts (which are
usually unicellular). Bacteria, which usually grow at the expense of fungi, are unicellular. Viruses,
although transmitted more from person to person than via food, should also be mentioned because
they may contaminate food as a consequence of poor worker hygiene.
Molds
Molds are multicellular microorganisms (eukaryotic cells) with mycelial (filamentous)
morphology. They consist of tubular cells, ranging from 30 to 100 μm in diameter, called hyphae,
which form a macroscopic mass called a mycelium. Molds generally withstand greater variations in
pH than do bacteria and yeasts and can frequently tolerate greater temperature variations.
Although molds thrive best at or near a pH of 7.0, a range from 2.0 to 8.0 can be tolerated, though an
acid-to-neutral pH is preferred.Molds have been considered beneficial and troublesome, ubiquitous
microorganisms. They often work in combination with yeasts and bacteria to produce numerous
indigenous fermented foods and are involved in industrial processes to produce organic acids and
enzymes. Molds are a major contributor to food product recalls. Most do not cause health hazards,
but some produce mycotoxins that are toxic, carcinogenic, mutagenic, or teratogenic (Able to
disturb the growth and development of an embryo or fetus) to humans and animals. Because molds
are difficult to control, food processors have encountered spoilage problems cauesd by these
microorganisms.
Yeasts
Yeasts are generally unicellular. They differ from bacteria in their larger cell sizes and
morphology, and because they produce buds during the process of reproduction by fission. The
generation time of yeasts is 2 to 3 hours in foods, leading from an original contamination of one
yeast/g of food to spoilage in approximately 40 to 60 hours. Like molds, yeasts can be spread
through the air or by other means and can alight on the surface of foodstuffs. Yeast colonies are
generally moist or slimy in appearance and creamy white. These microorganisms grow best in the
intermediate acid range, a pH of from 4.0 to 4.5. Yeasts are more likely to grow on foods with lower
pH and on those that are vacuum packaged. Food that is highly contaminated with yeasts will
frequently have a slightly fruity odor.
Bacteria
Bacteria produce pigments ranging from variations of yellow to dark shades, such as brown or
black. Certain bacteria have pigmentation of intermediate colors—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. Some species of
bacteria produce spores, which may be resistant to heat, chemicals, and other environmental
conditions. Some of these spore-forming bacteria are thermophilic microorganisms that produce a

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 2
 toxin that can cause foodborne illness.
Viruses
Viruses cannot reproduce outside of another organism and are obligate parasites of all living
organisms, such as bacteria, fungi, algae, protozoa, higher plants, and invertebrate and vertebrate
animals. Foodborne viruses cause diseases through viral gastroenteritis or viral hepatitis. A virus
that has caused a major increase in outbreaks in restaurants during the past 10 years is hepatitis A.
Intravenous drug use is one factor that accounts for some of this rise. Infectious hepatitis A can be
transmitted through food that has not been handled in a sanitary manner. The onset is 1 to 7 weeks
with an average length of 30 days. Symptoms include nausea, cramps, vomiting, diarrhea, and,
sometimes, jaundice, which can last from a week to several months. A major source of hepatitis is
raw meat from polluted waters.

SOURCES OF CONTAMINATION

Most foods are highly perishable because they contain nutrients required for microbial growth.
To reduce food spoilage and to eliminate foodborne illness, microbial proliferation must be
controlled. Food deterioration should be minimized to prolong the time during which an acceptable
level of flavor and wholesomeness can be maintained. If proper sanitation practices are not
followed during food processing, preparation, and serving, the rate and extent of the deteriorative
changes that lead to spoilage will increase.
Equipment
Contamination of equipment occurs during production, as well as when the equipment is idle.
Even with hygienic design features, equipment can collect microorganisms and other debris from
the air, as well as from employees and materials. Product contamination of equipment is reduced
through improved hygienic design and more effective cleaning.
Employees
Of all the viable means of exposing microorganisms to food, employees are the largest
contamination source. Employees who do not follow sanitary practices contaminate food that they
touch, with spoilage and pathogenic microorganisms that they come in contact with through work
and other parts of the environment. The hands, hair, nose, and mouth harbor microorganisms that
can be transferred to the food during processing, packaging, preparation, and service by touching,
breathing, coughing, or sneezing. Because the human body is warm, microorganisms proliferate
rapidly, especially in the absence of hygienic practices. After the chain of infection is broken the
spreading of bacteria from one location to another can be prevented. Generally, the mishandling of
food by people perpetuates the chain of infection until someone becomes ill or dies before
corrective actions are taken to prevent additional outbreaks. If every person that handles food
could achieve appropriate personal hygiene, food contamination could be minimized. Every
employee involved with food manufacturing can play a very important role in preventing food
contamination.
Air, Water and Soil
Water serves as a cleaning medium during the cleaning operation and is an ingredient added in
the formulation of various processed foods. It can also serve as a source of contamination. If
excessive contamination exists, another water source should be obtained, or the existing source
should be treated with chemicals (such as ultraviolet units) or other methods.
Airborne microorganisms cause contamination in food processing, packaging, storage, and
preparation areas. This contamination can result from unclean air surrounding the food plant or
from contamination through improper sanitary practices. The most effective methods of reducing
air contamination are through sanitary practices, filtering of air entering the food processing and
preparation areas, and protection from air by appropriate packaging techniques and materials.
Soil Normal flora gets contaminated by animal and human feces, sewage water and rain water
which contain high population of pathogenic bacteria like coliforms, Clostridium, Salmonella and
Vibrio. Soil also contains opportunistic plant pathogenic bacteria like Pseudomonas, Agrobacterium

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 3
and Xanthomonas. Pathogens like clostridia persist in soil for long periods as sporulated bacteria.
Soil contaminant bacteria are desiccation resistant and actively compete for soil nutrients with
native bacterial population. Soil pathogenic bacteria are causative agents of water and food borne
diseases of humans.
Sewage
Raw, untreated sewage can contain pathogens that have been eliminated from the human body,
as well as other materials of the environment. Examples are microorganisms causing typhoid and
paratyphoid fevers, dysentery, and infectious hepatitis. Sewage may contaminate food and
equipment through faulty plumbing. If raw sewage drains or flows into potable water lines, wells,
rivers, lakes, and ocean bays, the water and living organisms such as seafood are contaminated. To
prevent this contamination, privies and septic tanks should be sufficiently separated from wells,
streams, and other bodies of water. Raw sewage should not be applied to fields where fruits and
vegetables are grown.
Insects and Rodents
Flies and cockroaches are associated with living quarters, eating establishments, and food
processing facilities, as well as with toilets, garbage, and other filth. These pests transfer filth from
contaminated areas to food through their waste products; mouth, feet, and other body parts; while
the regurgitation of filth onto clean food during consumption. To stop contamination from these
pests, eradication is necessary, and food processing, preparation, and serving areas should be
protected against their entry. Rats and mice transmit filth and disease through their feet, fur, and
intestinal tract. Like flies and cockroaches, they transfer filth from garbage dumps and sewers to
food or food processing and foodservice areas.
Animal Hides
In the case of milk cows, the types of organisms found in raw milk can be a reflection of the
biota of the udder when proper procedures are not followed in milking and of the general
environment of such animals. From both the udder and the hide, organisms can contaminate the
general environment, milk containers, and the hands of handlers.

MICROBIAL EXAMINATIONS OF FOOD

The examination of foods for the presence, types, and numbers of microorganisms and/or their
products is basic to food microbiology. In spite of the importance of this, none of the methods in
common use permits the determination of exact numbers of microorganisms in a food product.
Although some methods of analysis are better than others, every method has certain inherent
limitations associated with its use. The four basic methods employed for "total" numbers are as
follows:
1. Standard plate counts (SPC) for viable cells
2. The most probable numbers (MPN) method as a statistical determination of viable cells
3. Dye reduction techniques to estimate viable cells that possess reducing capacities
4. Direct microscopic counts (DMC) for both viable and nonviable cells.

MEMBRANE FILTERS
Membranes with a pore size that will retain bacteria (generally 0.45 um) but allow water or
diluent to pass are used. Following the collection of bacteria upon filtering a given volume, the
membrane is placed on an agar plate or an absorbent pad saturated with the culture medium of
choice and incubated appropriately. Following growth, the colonies are enumerated. Alternatively,
a DMC can be made. In this case, the organisms collected on the membrane are viewed and counted
microscopically following appropriate staining, washing, and treatment of the membrane to render
it transparent. These methods are especially suited for samples that contain low numbers of
bacteria. Although relatively large volumes of water can be passed through a membrane without
clogging it, only small samples of dilute homogenates from certain foods can be used for a single
membrane.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 4
 DRY FILM METHOD
A rehydratable dry film method consisting of two plastic films attached together on one side
and coated with culture media ingredients and a cold-water-soluble jelling agent was developed by
Private Company and designated Petrifilm. The method can be used with nonselective ingredients
to make aerobic plate counts (APCs), and, with selective ingredients, certain specific groups can be
detected. Use of this method to date indicates that it is an acceptable alternative to SPC methods
that employ Petri dishes, and it has been approved by AOAC.
MOST PROBABLE NUMBERS
In this method, dilutions of food samples are prepared as for the SPC. Three serial aliquots or
dilutions are then planted into 9 or 15 tubes of appropriate medium for the three- or five-tube
method, respectively. Numbers of organisms in the original sample are determined by use of
standard MPN tables. The method is statistical in nature, and MPN results are generally higher than
SPC results. This method was introduced by McCrady in 1915. It is not a precise method of analysis;
the 95% confidence intervals for a three-tube test range from 21 to 395. When the three-tube test is
used, 20 of the 62 possible test combinations account for 99% of all results. Among the advantages
it offers are the following:
• It is relatively simple.
• Results from one laboratory are more likely than SPC results to agree with those from another
laboratory.
• Specific groups of organisms can be determined by use of appropriate selective and
differential media.
• It is the method of choice for determining fecal coliform densities. Among the drawbacks to its
use are the large volume of glassware required (especially for the five-tube method), the lack of
opportunity to observe the colonial morphology of the organisms, and its lack of precision.
DYE REDUCTION
Two dyes are commonly employed in this procedure to estimate the number of viable
organisms in suitable products: methylene blue and resazurin.To conduct a dye-reduction test;
properly prepared supernatants of foods are added to standard solutions of either dye for
reduction from blue to white for methylene blue and from slate blue to pink or white for resazurin.
The time for dye reduction to occur is inversely proportional to the number of organisms in the
sample.
DIRECT MICROSCOPIC COUNT In its simplest form, the DMC consists of making smears of food
specimens or cultures onto a microscope slide, staining with an appropriate dye, and viewing and
counting cells with the aid of a microscope (oil immersion objective). DMCs are most widely used in
the dairy industry for assessing the microbial quality of raw milk and other dairy products, and the
specific method employed is that originally developed by R.S. Breed (Breed count). Briefly, the
method consists of adding 0.01 mL of a sample to a 1-cm2 area on a microscope slide, and following
fixing, defatting of sample, and staining, the organisms or clumps of organisms are enumerated. The
latter involves the use of a calibrated microscope. The method lends itself to the rapid
microbiological examination of other food products, such as dried and frozen foods. Among the
advantages of DMC are that it is rapid and simple, cell morphology can be assessed, and it lends
itself to fluorescent probes for improved efficiency. Among its disadvantages are that it is a
microscopic method and therefore fatiguing to the analyst, both viable and nonviable cells are
enumerated, food particles are not always distinguishable from microorganisms, microbial cells are
not uniformly distributed relative to single cells and clumps, some cells do not take the stain well
and may not be counted, and DMC counts are invariably higher than counts by SPC. In spite of its
drawbacks, it remains the fastest way to make an assessment of microbial cells in a food product.
VIABLE BUT NONCULTURABLE ORGANISMS (VBNC)
Under certain conditions and in some environments, standard plate count results suggest either
an absence of colony-forming units or numbers that may be considerably lower than the actual
viable population. These counts are due to Viable But Not Cultivable microorganisms. It is believed
that 99.9% of microorganisms existing are not cultivable by man made laboratory techniques.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 5
 MICROBIAL SPOILAGE OF FOOD
Fruits, vegetables, meats, poultry, sea foods, milk and dairy products and various other food
products differ in their biochemical composition and therefore are subject to spoilage by different
microbial populations. Such changes depend upon the nature of the microbes involved in the
spoilage. Thus degradation of apple juice by yeast gives an alcoholic taste to the juice. Yeasts
convert the carbohydrate into ethanol.

Bacteria which attack food proteins, convert these into amino acids which are broken down again
into foul smelling end products. Digestion of cystein, for example, yields hydrogen sulphide, giving a
rotten egg smell to food. Digestion of tryptophan yields indole and skatole which give food a fecal
odour.

Two other products of the microbial metabolism of carbohydrates are (a) acid that causes foods to
become sour, and (b) gas which causes sealed cans to swell. Digestion of fats, as in spoiled butter,
yields fatty acids giving a rancid odour or taste to food. Food may become slimy due to production
of capsules in bacteria. There may be pigment development giving some colour to foods.

The mold forms a toxin aflatoxin. Another grain spoilage occurs by Claviceps purpurea, in rye, wheat
and barley grains causing ergot disease. The mold toxin may induce convulsions and hallucinations.
The drug LSD is derived from this toxin.

The chemical properties of a food product influence the type of microorganisms that can grow, and
hence determine the changes in appearance, flavour, odour, and other qualities of food.

Composition: Proteins are degraded by proteolytic organisms. Many bacterial species, especially
spore formers, gram negative rods such as Pseudomonas and Proteus, and a few cocci can attack
proteins. Mould spoilage is also common.

Carbohydrate foods are spoiled by carbohydrate fermenting. microorganisms, particularly by yeast

and moulds. Bacterial species of the genera Streptococcus, Leuconostoc and Micrococcus are
saccharolytic and can also attack carbohydrates.

Fats are digested by relatively few microorganisms, mainly moulds and few gram negative bacteria.
Fats undergo hydrolytic decomposition and become rancid, all malodorous fatty acids are set free.

Acidity: The reaction of nearly all foods is below pH 7.0. Foods are classified as acid or nonacid.
The reaction of acid food is below pH 4.5, and that of nonacid food above pH 4.5. Most fruits are acid
foods, while nearly all vegetables, fish, meats, and milk products are nonacid.

Acid foods have sufficiently low pH and, therefore, prevent the growth almost bacterial species.
They are spoiled mainly by yeast and moulds. Nonacid foods are particularly subject to bacterial
spoilage, but will also support growth of moulds under proper conditions.

Moisture and osmotic pressure: Growth of microorganisms require at least 13 per cent free
water in foods, Moulds require the least free water and bacteria require the most. Foods of high
sugar and salt concentration do not, support the growth of most microorganism). Bacteria are
generally inhibited (by 5 to 11 percent salt, whereas many moulds and some yeasts can tolerate salt
concentrations greater than 15 percent.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 6
Effects of Storage Conditions on Spoilage - Oxygen and temperature are the two most important
factors that influence the type of microbial growth and spoilage of food during storage.

Oxygen: The presence or absence of oxygen determines the types of organisms tbat can multiply
and the kind of spoilage produced. Moulds and aerobic bacteria: grow only where there is plenty of
air and chiefly cause surface spoilage. Yeast and facultative bacteria can grow in closed containers
as well as when exposed to the air. Clostridia and other organisms bring about spoilage of food
under strict anaerobic condition.

Temperature: Low temperature retards spoilage, but even subfreezing temperatures do not
prevent multiplication of all microorganisms until about -7°C is reached. Refrigerated foods are
therefore subject to spoilage by moulds and by some yeasts and bacteria. Foods stored at -18°C
remain free from microbial growth and a slow decrease in population may even occur. Foods and
food products stored at room temperature or in warm locations are easily spoiled by mesophilic
arid thermophilic organisms.

Types of Food Spoilage with Causative Organisms

Food Types of Causative microorganisms

spoilage
Fresh meat Putrefaction Clostridium, Pseudomonas, Porteus, Alcaligenes,
Souring Chromobacterium, Lactobacillus,Pseudomonas.
Cured meat Mouldy Penicillium, Aspergillus,Rhizopus.
Souring Pseudomonas, Micrococcus, Bacillus.
Greening Lactobacilli Streptococci,Pediococci.
Slimy Leuconostoc
Fish Discolouration Pseudomonas
Putrefaction Chromobacterium, Halobacterium, Micrococcus
Poultry Odour, Slime Pseudomonas, Alcaigenes, Xanthomonas.
Eggs Green rot Pseudomonas Fluorescens
Colourless rot Pseudomonas, Alcaigenes, Chromobacterium,
Black rot Coliform.
Fungal rot Proteus, Penicillium, Mucor
Fresh fruits and vegetables Bacterial soft rot Erwinia carotovera, Pseudomonas spp.
Gray mould rot Botryitis cinerea
Rhizopus soft rot Rhizopus nigrican
Blue mould rot Penicillium italicum
Black mould rot Aspergillus niger, Alternaria
Sliminess or Saprophytic bacteria
Souring
Pickles,Sauer, kraut Black pickles Bacillus nigricans
Soft pickles Bacillus spp.
Slimy kraut Lactobacillus Plantarum, L. cucumeris
Pink kraut Rhodotorula(asporogenous yeasts)
Sugar products, Honey, Ropy syrup Aerobacter aerogenes
Syrups Yeasty Saccharomyces, Torula,Zygosaccharomyces
Pink syrup Micrococcus roseus
Green syrup Pseudomonas fluorescens
Mouldy Aspergillus, Penicillium
Bread Mouldy Rhizopus, Aspergillus
Ropy Penicillium
Red bread Bacillus spp. Serratia marcesens

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 7
Bacteria Involved in Food Spoilage

Group Speices of
Lactics Lactobacillus, Leuconostoc, Pediococcus,Streptococcus
Acetics Acetobacter and Gluconobacter
Butyrics Clostridium
Proteolytics Bacillus, Pseudomonas, Clostridium, Proteus etc.
Lipolytics Pseudomonas, Alcaligenes, Serratia, Micrococcus Bacillus, Clostridium etc.
Sacccharolytic Bacillus, Clostridium etc.
Pectolytic Erwinia, Bacillus, Clostridium
Thermophiles Bacillus, Clostridium, Lactobacillus thermophilus.
Psychrophiles Pseudomonas, Flavobacterium, Alcaligenes, Micrococcus
Halophiles Halobacterium sarcina, Micrococcus, Pseudomonas
Osmophiles Leuconostoc
Pigment formers Flavobacterium, Serratia, Micrococcus
Slime or rope formers Alcaligenes, Enterobactor, Streptococcus, Lactobacillus etc
Gas formers Leuconostoc, Lactobacillus, proteus, Clostridium, Enterobactor etc
Coliforms E.coli, Enterobactor aerogenes.

FOOD POISONING
To control and prevent the development of food poisoning and to prevent the spread of disease by
foods is an important aspect of food microbiology. All food borne diseases are classified as food
infections or food intoxications. These classifications are somewhat random.

Food infections are those in which microorganisms present in the food at the time of eating grow in
the host and cause disease. Food intoxications are those diseases in which microorganism grow in
the food, producing a substance therein which is toxic to man and animals. Food poisoning is the
toxicity introduced into food by microorganisms and their products. This does not include the
toxications which follow the consumption of noxious plants (mushrooms),poisonous fish (mussels),
or decomposed foods containing certain chemicals (arsenic, lead, fluorides, antimony, mercury
salts).

At one time it was believed that the products of putrefaction were the cause of food poisoning.
These were called ptomaine, and were produced in putrefied meat and other proteinaceous foods.
These are amines and result chiefly from the decarboxylation of amino acids. It could be demons-
trated that ptomaines injected into animals were toxic, but when taken experimentally by mouth,
they were not especially toxic. The food poisoning, therefore, could not be due to ptomaines.

People generally do not eat foods that show visible spoilage, but sometimes if the appearance,
flavour and taste are normal, they do eat them. On the other hand, certain foods may show no signs
of spoilage and yet be responsible for food poisoning.

The fact that organisms are present in food does not necessarily mean that they are harmful. Most
of the organisms that cause spoilage of foods are harmless saprophytes. However, there are some
forms of illness caused by organisms growing in foods. Staphylococcal food poisoning and botulism
are defined as food intoxications, since ingestion of the toxin causes the disease symptoms.

The term food infection is used to describe diseases like Salmonellosis and Enteritis caused by
Salmonella spp and Cl. perfringens, since these diseases are caused by the ingestion of organisms.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 8
Staphylococcal Poisoning - It is the most common type of food poisoning. Certain strains of
Staphylococcus aureus produce a potent enterotoxin. Ingestion of food containing this toxin causes a
sudden onset of illness within 3 to 4 hours. Symptoms include nausea, vomitting and diarrhoea.
Recovery is rapid, usually within 24 to 48 hours. Death rarely occurs.

The organisms are widely distributed upon the skin and mucous membranes of the human body.
People who handle foods contaminate them ignorantly of carelessly. Foods most commonly
involved include those which are eaten cold, e.g. salads, bakery products, hams, pressed meat, and
dairy products.

A food having millions of Staphylococci may taste, smell, and appear to be little different from that
in which none of these organisms have grown. Secondly, staphylococcal enterotoxin is heat
resistant and can withstand boiling for 30 minutes. So recooking the food does not help. Control is
by preventing the entry of the bacteria to food, by destruction of the bacteria by heat, and by
restriction of their growth by refrigeration.

Botulism - Botulism is caused by the exotoxin of Clostridium botulinum, an anaerobic spore-

forming rod. It is one of the most potent exotoxins known to man. Botulism is a neuroparalylic
disease affecting humans and animals. Sixty to seventy percent of, cases are fatal.

The foods frequently implicated are those which have been smoked, pickled, or canned, allowed to
stand for a time and then eaten without cooking or with insufficient cooking. This means that the
preservative treatment is inadequate and has failed to destroy the spores of Clostridium botulinum.
Most cases are associated with home canned vegetables, infected sausages, ham, preserved meats
fowl, or fish.

These foods have the common property of being high in protein, near neutral in pH, and have little
residual available oxygen, all of which are factors contributing to toxin production. Fortunately, the
toxin is heat labile and is destroyed very quickly by boiling. Home canned food should always be
heated at the boiling point for several minutes before use.

Salmonellosis - Salmonellosis is caused through the ingestion of living bacteria of the Salmonella
group. The general symptoms are the same as in Staphylococcus poisoning, but with a longer
incubation period. The disorder is the result of the growth of Salmonella in the intestine. An
inoculum of about 600,000 cells is required for organisms to become established and cause illness.

Different species of Salmonella may be ingested with improperly cooked meat, eggs, and puddings
that have been contaminated by rodents or human carriers. Almost any food may, at times, be
contaminated with Salmonella, and under conditions of mishandling may become involved in the
transmission of salmonellosis.

The major reason for the widespread occurrence of Salmonella in meats is cross infect ion at the
killing plant. The animals on the farm that are infected with Salmonella may be only 0.5%, but this
may increase to 35% after 2 to 5 days in slaughterhouses.

Similarly, in poultry only a few per cent of the birds entering the plant may harbour Salmonella in
their intestinal tract, but after processing 30 to 35% may be infected. Salmonella is found on the
shell, and in the contents of' clean fresh eggs. After processing to egg powder, a high percentage of
the samples show the presence of Salmonella.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 9
 FOOD PRESERVATION

Principal Method of Food Preservation

1. Prevention or delay of microbial decomposition.
(a) By keeping microbes out (asepsis).
(b) By removal of microbes (e.g. filtration).
(c) By reducing the rate of microbial growth
(e.g. by low temperature, drying, anaerobic conditions and chemical inhibitors),
(d) By killing microbes (e.g. by heat or radiation).
2. Prevention or delay of self-decomposition of food
(a) By inactivation of food enzymes (e.g. blanching).
(b) By prevention of chemical reactions (e.g. by using antitoxidants).

Food Preservation Methods

Food preservation aims at preventing the microbial spoilage of food products and the growth of
the food borne pathogens. Thus, the two principal goals of food preservation methods are,
(i) Increasing the shelf life of the food and
(ii) Ensuring the safety for human consumption.

There are a variety of food preservation methods:

1. Heat: Heat kills microorganisms by changing the physical and chemical properties of their
proteins. The most common use of heat is in the process of canning. The food product is washed,
sorted, and graded and then subjected to steam heat for three to five minutes. This last process
called Blanching, destroys many enzymes in the food product and prevents further cellular
metabolism. The food is then peeled and cored, and diseased portions are removed. For canning,
containers are evacuated and placed in a pressurised steam steriliser, similar to an autoclave at
121°C. This removes especially Bacillus and Clostridiurn spores.

If canning is defective, foods may become contaminated by anaerobic; bacteria which produce gas.
These are species of Clostridium, and coliform bacteria (a group of Gram-negative nonspore-
forming rods which ferment lactose to acid and gas at 32°C in 48 hours).

2. Low temperature: Exposure of microorganisms to low temperatures reduces their rates of

growth and reproduction. This principle is used in refrigeration and freezing. Microbes are not
killed. In refrigerators at 5°C, foods remain unspoiled. In a freezer at -5°C the cellular crystals
formed tear and shred microorganisms. It may kill many of the microbes.

However, some are able to survive. Salmonella spp. and Streptococci survive freezing. For these
types rapid thawing and cooking is necessary. Deep freezing at -60°C forms smaller crystals. It
reduces biochemical activities of microbes.

Blanching of fruits and vegetables, by Scalding with hot, water or steam prior to deep freezing,
inactivate plant enzymes that may produce toughness, change in colour etc. A brief scalding prior to
freezeing also reduces the number of microorganisms on the food surface by up to 99 per cent,
enhances the colour of green vegetables.

3. Drying or Desiccation: Water from foods is removed in different ways. It may be done by spray
dryer which expels a fine mist of liquid such as coffee into a barrel cylinder containing hot air.
There may be used a heated drum onto which liquids like soup may be poured. Another machine is
a belt heater that exposes liquids as milk to a steam of hot air that evaporates water and produces
dried milk solids.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 10
A common process of freeze drying or Lyophilization is used these days. The food is deep frozen,
after which the water is drawn off by a vacuum pump in a machine. They dry product is then sealed
in foil and is reconstituted with water. This method is very useful for storing, transporting and
preserving bacterial cultures.

4. Osmotic pressur: The principle of osmosis is applied. Foods are preserved by adding salts and
sugars to them. These chemicals remove the water out of microbial cells causing them to shrink.
Thus stopping their metabolism. Jams, jellies, fruit syrups, honey etc. are preserved by high sugar
concentration. Fish, meat beef and vegetable products are preserved with salt.

6. Radiation: UV is used in meat storage facilities which reduce surface contamination, on meat
products. Gamma rays are also used for some meat products.

7. Anaerobiosis: Packaging of food products under anaerobic conditions - anaerobiosis is effective

in preventing aerobic spoilage process. Vacuum packing in an airtight container is used to eliminate
air.

8. Controlled atmospheres: Such atmospheres containing 10% CO2 are used to preserve stored
food products as apples and pears. This checks fungal growth. Ozone can also be added.

9. Other methods: These are asepsis i.e. washing utensils that come in contact with food; and
filtration and centrifugation, used, to remove microbes. Filtration is used for fruit juices, other
drinks etc. Bacteriological filters are used in industries.

5. Chemical preservatives: The most commonly used are the acids, such as sorbic acid, benzoic
acid and propionic acid. These check mainly the growth of yeasts and molds. Sorbic acid is used for
preservation of syrups, salads jellies and some cakes. Benzoic acid is used for beverages, margarine,
apple cider etc. Propionic acid is an ingredient of bread and bakery products. Sulphur dioxide, as
gas or liquid is also used for dried fruits, molasses and juice concentrates. Ethylene oxide is used for
spices, nuts and dried fruits.

Chemical Max.permitted
Preservatives concentration
Benzoic acid 0.1%
Methyl paraben 0.1%
Propyl paraben 0.1%
Sodium nitrite 500 ppm
Sodium nitrite 200 ppm
Sorbates 0.1%
Acetic acid 0.12-0.14 mg/ml in wines
Propylene oxide 300ppm
Ethylene oxide residue not to exceed 50ppm
Sulfites Variable with food stuff
Propionates 0.1%
Potassium 0.05%
metabisulphite

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 11
 Agent Source Symptoms & usual duration
Salmonella Raw meat, poultry, eggs, Diarrhoea, vomiting,
unpasteurised milk, pets, fever,
terrapins, infected food abdominal pain 1-5 days
handlers.
Campylobacter Raw meat, poultry, raw/bird Abdominal pain,
pecked milk, untreated diarrhoea 2-5 days
water, pets.
Listeria Found in environment, cattle, Fever, affects central
sheep, silage, unpasteurised nervous system
Monocytogenes milk products including soft Variable
cheeses, pates.

Staphylococcus Human nose, mouth, cuts and Vomiting, abdominal pain,

wounds. diarrhoea,
aureus fainting 6-24 hours

Clostridium Faeces of animal and man, Diarrhoea,

soil (on vegetable), dust, abdominal pain
Perfringens sewage. 12-48 hours

Bacillus cereus Cereal products, especially Vomiting, abdominal pain,

rice, spices, dust, soil. some diarrhoea 1-2 days
(toxin in food)

Bacillus cereus Cereal products, especially Diarrhoea, abdominal pain,

rice, spices, dust, soil. some vomiting 1-2 days
(toxin in gut)

Vibrio Sea water, shellfish. Diarrhoea, abdominal pain,

some vomiting 2-5 days
Paraheamolyticus

Escherichia coli Aminal origins – cattle, Diarrhoea, abdominal pain,

(infective) sheep, humans, sewage, meat fever 2-3 days
& raw milk.
Clostridium Soil, meat, fish and Central nervous system
botulinum vegetables (difficulty breathing,
double vision, nerve paralysis),
diarrhoea, vomiting
Variable can be fatal
Chemicals Vomiting, abdominal
(e.g. metallic pain, possibly effects on
poisons, central nervous system
pesticides, etc)
Poisonous Vomiting, abdominal pain,
plants/animals possibly effects on central
nervous system
Norwalk Virus Vomiting, diarrhoea,

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 12
 Bacteria Description Habitat Types of Cause
Responsible Foods
Staphylococcus Produces a heat- Nose and throat of 30 Meat and Poor personal hygiene
aureus stable toxin to 50 percent of seafood salads, and subsequent
healthy population; sandwich temperature abuse.
also skin and spreads and
superficial wounds. high salt foods.
Salmonella Produces an Intestinal tracts of High protein Contamination of
intestinal animals and man foods - meat; ready-to-eat foods,
infection poultry, fish and insufficient cooking
eggs. and recontamination
of cooked foods.
Clostridium Produces a spore dust, soil and Meat and Improper temperature
perfringens and prefers low gastrointestinal poultry dishes, control of hot foods,
oxygen tracts of animals and sauces and and recontamination.
atmosphere. Live man. gravies.
cells must be
ingested.
Clostridium Produces a spore Soils, plants, marine Home-canned Improper methods of
botulinum and requires a sediments and fish. foods. home-processing
low oxygen foods.
atmosphere.
Produces a heat-
sensitive toxin.
Vibrio Requires salt for Fish and shellfish Raw and Recontamination of
parahaemolyticus growth. cooked seafood. cooked foods or eating
raw seafood.
Bacillus cereus Produces a spore soil, dust and spices. Starchy food. Improper holding and
and grows in stroage temperatures
normal oxygen after cooking.
atmosphere.
Listeria Survives adverse Soil, vegetation and Milk, soft Contaminated raw
monocytogenes conditions for water. Can survive cheeses, products.
long time periods. for long periods in vegetables
soil and plant fertilized with
materials. manure.
Campylobacter Oxygen sensitive, Animal reservoirs Meat, poulty, Improper
jejuni does not grow and foods of animal milk, and pasteuriztion or
below 86o F. origin. mushrooms. cooking. cross-
contamination.
Versinia Not frequent Poultry, beef, swine. Milk, tofu, and Improper cooking.
enterocolitica cause of human Isolated only in pork. Cross-contamination.
infection. human pathogen.
Enteropathogenic Can produce Feces of infected Meat and Inadequate cooking.
E. coli toxins that are humans. cheeses. Recontamination of
heat stable and cooked product.
others that are
heat-sensitive.

Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI (Maharashtra) INDIA Page 13