Food as a substrate for micro organisms- Micro organisms important in food microbiology; Molds,

yeasts and bacteria - General Characteristics - Classification and importance. Principles of food
preservation - Asepsis - Removal of micro organisms, - High temperature - Low temperature - Drying -
Food additives. Nanoscience in food preservation; microencapsulation.
Contamination and spoilage of food products -Food borne infections (Bacillus cereus, Salmonellosis,
Shigellosis, Listeria monocytogenes and Campylobacter jejuni) and intoxications – (Staphylococcus
aureus, Clostridium botulinum, Clostridium perfringens and [Link] borne disease outbreaks -
newly emerging pathogens. Conventional and Novel technology in control of food borne pathogens and
preventive measures - Food sanitation - plant sanitation - Employees’ health standards. Regulatory
Agencies &criteria for food safety.
Microflora of raw milk - sources of contamination. Spoilage and preservation of milk and milk
products. - antimicrobial systems in raw milk. Importance of biofilms, their role in transmission of
pathogens in dairy products and preventive strategies.
Food fermentations: Indian Pickles Bread, vinegar, fermented vegetables (sauerkraut), fermented dairy
products (yoghurt, cheese, Acidophilus Milk, Kefir, Koumiss). Oriental fermented foods-Miso –
Tempeh Ontjom. Natto, Idli Spoilage and defects of fermented dairy products -. Functional fermented
foods and nutraceuticals, bioactive proteins and bioactive peptides, genetically modified foods.
Probiotic microorganisms, concept, definition safety of probiotic microorganisms, legal status of
probiotics Characteristics of Probiotics for selection: stability maintenance of probiotic microorganisms.
Role of probiotics in health and disease: Mechanism of probiotics. Application of bacteriocins in foods.
Biopreservation. Prebiotics: concept, definition, criteria, types and sources of prebiotics, prebiotics and
gut microflora - Prebiotics and health benefits: mineral absorption, immune response, cancer
prevention, elderly health and infant health, prebiotics in foods.

23UMICC62 - CORE – XIV: FOOD, DAIRY AND PROBIOTIC MICROBIOLOGY

UNIT-I

FOOD MICROBIOLOGY
Food microbiology deals with the study of interaction between the microbiology and
food, contamination and spoilage of food due to the microbes and preservation techniques to
reduce the load of microbes in the food.
Spoilage of food by micro organisms
Cause of spoilage:
Spoilage may be due to one or more of the following
1. Growth and activity of micro organisms
2. Insects
3. Action of the enzymes of the plant or animal food.

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 4. Purely chemical reactions i.e., those not catalysed by enzymes of the tissues or of
micro organisms
5. Physical changes such as those caused by freezing, burning, drying, pressure etc.
Classification of foods
1. Stable or non perishable foods
These foods which do not spoil unless handled carelessly include such products as sugar,
flour and dry beans.
2. Semi perishable foods
If these foods are properly handled and stored, they will remain unspoiled for a fairly
long period. Ex: Potatoes, apples etc.
3. Perishable foods
It includes most important daily foods that spoil readily Ex: Meats, fish, poultry, milk,
eggs, fruits and vegetables.

MICROORGANISMS AND ITS GROWTH IN FOOD

Microorganisms present in food use them as source of nutrients or their growth media
and grow. Growth of microorganisms in food can result in improving the quality of food and in
some cases can deteriorate their quality depending on the type of microorganism.
Growth of harmful microorganisms in food can result in spoilage and some times cause
several diseases on consumption of such food. Food spoilage by microorganisms is due to
increase in their numbers, utilizing nutrients, causing enzymatic changes resulting in bad flavors
due to breakdown of some food materials or synthesis of new compounds. Due to such microbial
activities, food become unfit for human consumption. Microorganisms bring about oxidation of
reduced carbon, nitrogen; sulfur compounds present in dead plants and animals in nature and
become important part in cycling of these elements.
Food acts as good medium for transmission of many diseases. If the food is contaminated
by pathogenic microorganisms, they can grow and increase their population and cause diseases
on consumption of such food. Some time microorganisms may not grow in food but they are
transported through food. Therefore, food act as good medium for spread of diseases. Several
food born diseases are the result of microorganism present in food or their growth in them.

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MICROORGANISMS INVOLVED IN FOOD MICROBIOLOGY
Food serves as growth medium for different kinds of microorganisms. Microorganisms
enter into food and grow as contaminants or intended additions. Growth of microorganisms in
food may spoil food quality or improve depending on the types of microorganisms and the
changes they bring about.
Food are assessed for their quality in terms of physical, chemical, sensory and
microbiological characteristics. Microbiological characteristics are assessed in terms of the
microorganisms present in food, their characters, ability to change the quality, their influence on
health of consumer. It is necessary for food microbiologists to become acquainted with the
microorganisms important in food at least to the extent that will enable them to identify the main
types with their characteristics.
Important microorganisms associated with food are molds, yeasts, bacteria, viruses and
some protozoa parasites. Knowledge of general characters and primary identification methods is
necessary for the people working with food microbiology.
MOLDS
Mold growth on foods, with its fuzzy or cottony appearance, sometimes colored, is
familiar to everyone, and usually food with a moldy or "mildewed" food is considered unfit to
eat. Special molds are useful in the manufacture of certain foods or ingredients of foods. Molds
have been grown as food or feed and are employed to produce products used in foods, such as
amylase for bread making or citric acid used in soft chinks. Some molds do produce various
toxic metabolites (mycotoxins).
General characteristics of molds
Morphological Characterstics
Hyphae and Mycelium
The mold thallus consists of a mass of branching, intertwined filaments called hyphae
(singular hypha), and the whole mass of these hyphae is known as the mycelium. The hyphae
may be submerged, or growing within the food, or aerial, or growing into the air above the food.
Molds are divided into two groups: septate, i.e., with cross walls dividing the hypha into
cells; and noncoenocytic, septate with the hyphae apparently consisting of cylinders without
cross walls. The non-septate hyphae have nuclei scattered throughout their length and are
considered multicellular.

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 Special, mycelial structures or parts aid in the identification of molds. Examples are the
rhizoids, or "holdfasts," of Rhizopus and Absidia, the foot cell in Aspergillus, and the
dichotomous, or Y-shaped, branching in Geotrichum.
Reproductive structures
Molds can grow from a transplanted piece of mycelium. Reproduction of molds is chiefly
by means of asexual spores. Some molds also form sexual spores.
Asexual Spores
The asexual spores of molds are produced in large numbers and are small, light, and
resistant to drying. They are readily spread through the air to alight and start new mold thallus
where conditions are favorable.
The three principal types of asexual spores are (1) conidia (singular conidium), (2)
arthrospores or oidia (singular oidium), and (3) sporangiospores.
Conidia are cut off, or bud, from special fertile hyphae called conidiophores and usually
are in the open, i.e., not enclosed in any container.
Sporangiospores are in sporangium (plural sporangia), or sac, at the tip of a fertile hypha,
the sporangiophore.
Arthrospores are formed by fragmentation of a hypha, so that the cells of the hypha
become arthrospores.
Chlamydospore or resting cell, can withstand unfavourable conditions better than
ordinary mold mycelium can and later, under favorable conditions, can grow into a new mold.

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24

Sexual Spores: The molds which can produce sexual spores are classified on the basis of the
manner of formation of these spores and the type produced. The non septate molds
(Phycomycetes) that produce.
1. Oospores are termed Oomycetes. These molds are mostly aquatic; however, included in this
group are several important plant pathogens. The oospores are formed by the union of a small
male gamete and a large female gamete.

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2. Zygospores: Zygomycetes form zygospores by the union of the tips of two hyphae which
often appear similar and which may come from the same mycelium or from different mycelia.
Both Oospores and zygospores are covered by a tough wall and can survive drying for long
periods.
3. Ascospores: The Ascomycetes (septate) form sexual spores known as ascopores, which are
formed after the union of two cells from the same mycelium or from two separate mycelia. The
ascospores, resulting from cell division after conjugation, are in an ascus, or sac, with usual eight
spores per ascus.
4. Basidiospores: The Basidiomycetes, which include most mushrooms, plant rusts, smuts, etc.,
form a fourth type of sexual spore, the basidiospore.
Cultural Characteristics
Some molds are loose and fluffy; others are compact. Some look velvety on the upper
surface, some dry and powdery, and others wet or gelatinous. Definite zones of growth in the
thallus distinguish some molds, e.g., Aspergillus niger.
Pigments in the mycelium-red, purple, yellow, brown, gray, black, etc - are characteristic,
as are the pigments of masses of asexual spores; green, blue-green, yellow, orange, pink,
lavender, brown, gray, black, etc.
The appearance of the reverse side of a mold on an agar plate may be striking, like the
opalescent blue-black or greenish-black color of the underside of Cladosporium.
Physiological characteristics
Moisture Requirements
An approximate limiting total moisture content of a given food for mold growth can be
estimated, and therefore it has been claimed that below 14 to 15 percent total moisture in flour or
some dried fruits will prevent or greatly delay mold growth.
Temperature Requirements
Most molds would be considered mesophilic i.e., able to grow well at ordinary
temperatures. The optimal temperature for most molds is around 25 to 30 0 C, but some grow well
at 35 to 370C or above, e.g., Aspergillus spp., and some at still higher temperatures.
A number of molds are psychrotrophic; i.e., they grow fairly well at temperatures of
refrigeration, and some can grow slowly at temoeratures below freezing. Growth has been

6
reported at as low as - 5 to - 10 0C. A few are thermophilic; i.e., they have a high optimal
temperature.
Oxygen and pH Requirements
Molds are aerobic; i.e., they require oxygen for growth; this is true at least for the molds
growing on foods. Most molds can grow over a wide range of hydrogen-ion concentration (pH 2
to 8.5), but the majority are favored by an acid pH.
Food Requirements
Molds in general can utilize many kinds of foods, ranging from simple to complex. Most
of the common molds possess a variety of hydrolytic enzymes, and some are grown for their
amylases, pectinases, proteinases, and lipases.
Inhibitors
Compounds inhibitory to other organisms are produced by some molds, such as penicillin
from Penicillium chrysogenum and clavacin from Aspergillus clavatus.
Classification and identification of molds
Molds are plants of the kingdom Myceteae. They have no roots, stems, or leaves and are
devoid of chlorophyll. They belong to the Eumycetes, or true fungi, and are subdivided further to
subdivisions, classes, orders, families, and genera.
The following criteria are used chiefly for differentiation and identification of molds:
1 Hyphae septate or non-septate
2 Mycelium clear or dark (smoky)
3 Mycelium colored of colorless
4 Whether sexual spores are produced and the type: oospores, zygospores, or ascospores
6 Characteristics of the spore head
a) Sporangia: size, color, shape, and location
b) Spore heads bearing conidia: single conidia, chains, budding conidia, or masses; shape and
arrangement of sterigmata or phialides; gumming together of conidia
7 Appearance of sporangiophores or conidiophores: simple or branched, and if branched the type
of branching; size and shape of columella at tip of sporangiophore; whether conidiophores are
single or in bundles
8 Microscopic appearances of the asexual spores, especially of conidia: shape, size, color;
smooth or rough; one-, two-, or many-celled

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9 Presence of special structures (or spores): stolons, rhizoids, foot cells, apo-physis,
chlamydospores, sclerotia, etc.
Molds of Industrial Importance
Mucor:
Mucor are involved in the spoilage of some foods and the manufacture of others. A
widely distributed species is M. racemosus; M. rouxii is used in the "Amylo" process for the
saccharification of starch, and mucors help ripen some cheese, (e.g., Gammelost) and are used in
making certain Oriental foods.

Zygorrhynchus These soil molds are similar to Mucor except that the zygo-spore suspensors are
markedly unequal in size.
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Rhizopus Rhizopus stolonifer, the so-called bread mold, is very common and is involved in the
spoilage of many foods: berries, fruits, vegetables, bread, etc.
Absidia: Similar to Rhizopus, except that sporangia are small and pear-shaped.
Thamnidium: Thamnidium elegans is found on meat in chilling storage, causing "whiskers" on
the meat.
Aspergillus: The aspergilli are very widespread. Many are involved in the spoilage of foods, and
some are useful in the preparation of certain foods. The molds grow well in high concentrations
of sugar and salt and hence in many foods of low moisture content. The A. niger group, with A.
niger as a leading species, is widespread and may be important in foods. The spore-bearing
heads are large, tightly packed, and globular and may be black, brownish-black, or purple-brown.
The A. flavus-oryzae group includes molds important in the making of some Oriental foods and
the production of enzymes. Conidia give various yellow to green shades to the spore heads, and
dark sclerotia may be formed.
Penicillium: This is another genus that is widespread in occurrence and important in foods. The
genus is divided into groups and subgroups, and there are numerous species. The genus is
divided into large groups on the basis of the branching of the spore-bearing heads, or penicillin
(little brushes). These heads, or verticillata, are a whorl or cluster of three or more elements:
sterigmata, metulae (subbranches), and branches. P. expansum, the blue-green-spored mold,
causes soft rots of fruits. Other important species are P. digitatum, with olive, or yellowish-green
conidia, causing a soft rot of citrus fruits; P. italicum, called the “blue contact mold” with blue
green conidia, also rotting citrus fruit; [Link], with grayish conidia, useful in the ripening
of Camembert cheese; and P. roqueforti, with bluish-green conidia, aiding in the ripening of blue
cheeses, e.g., Roquefort.
Trichothecium The common species, T. roseum (Figure 2-14), is a pink mold which grows on
wood, paper, fruits such as apples and peaches, and vegetables such as cucumbers and
cantaloupes.
Geotrichum (Oospora or Oidium) This genus is included with the yeast like fungi by some
writers and with the molds by others. Species may be white, yellowish, orange, or red, with the
growth appearing first as a firm, felt like mass that later becomes soft and creamy. Geotrichum
candidum (Oospora lactis), often called the "dairy mold,” gives white to cream-colored growth.

9
Neurospora (Monilia): This genus has been described under various names because of the
confusion concerning its classification. It is classed among the perfect molds (producing sexual
spores) and call the genus Neurospora. Neurospora (Monilia) sitophila, the most important
species in foods, sometimes is termed the "red bread mold" because its pink, loose-textured
growth often occurs on bread.

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11
Sporotrichum Among the saprophytic species is S. carnis, found growing on chilled meats,
where it causes “white spot."
Botrytis One species important in foods is B. cinerea . It causes a disease of grapes but may grow
saprophytically on many foods.
Cephalosporium: Cephalosporium acremonium is a common species.
Trichoderma T. viride is a common species. The mature mold plant is bright green because the
balls of green conidia are glued together, and tufts of white hyphae (sterile) stick up well above
the conidiophores.
Scopulariopsis S. brevicaulis is common species. Colonies are brownish and cottony.

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Pullularia: Ovate, hyaline conidia (blastospores or buds from preexisting cells) borne as lateral
buds on all parts of the mycelium. Colonies are pale and slimy and yeastlike when young,
becoming mycelial and dark and leathery in age. P. pullulans is a common species.
Cladosporium C. herbarum is a leading species. These dark molds cause "black spot" on a
number of foods, on cellar walls, etc. Colonies of C. herbarum are restricted in growth and are
thick, velvety, and olive - to gray-green; the reverse side of the plant is a striking opalescent
blue-black to greenish-black.
Helminthosporium Species of this genus are for the most part plant pathogens but ma grow
saprophytically on vegetable materials
Alternaria Molds of this genus are common causes of the spoilage of foods. A. citri (rotting
citrus fruits), A. tenuis, and [Link] common species.
Stemphylium: This, too, is a common genus, The conidia are dark and multicellular but have,
fewer cross - walls than those of Alternaria and are rounded at both ends.
Fusarium Molds of this genus often grow on foods. The species are very difficult to identify,
and the appearance of growth is variable.
Endomyces Yeast like fungi, forming mycelium and arthrospores. Some species rot fruits.
Monascus Colonies of M. purpureus are thin and spreading and reddish or purple in color.
Found on dairy products and on Chinese red rice (ang -khak).
Sclerotinia Some species cause rots of vegetables and fruits, where they are present in the
conidial stage. The lemon-shaped conidia are in chains, with a “plug” separating conidia.

YEASTS
Yeasts are generally not filamentous but unicellular and ovoid or spheroid and which
reproduce by budding or fission. Yeasts may be useful or harmful in foods. Yeast fermentations
are involved in the manufacture of foods such as bread, beer, wines, vinegar, and surface ripened
cheese, and yeasts are grown for enzymes and for food.
General characteristics of yeasts
Morphological Characteristics

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 The morphological characteristics of yeasts are determined by microscopic examination.
Form and Structure
The form of yeasts may be spherical to ovoid, lemon shaped, pearshaped, cylindrical,
triangular, or even elongated into a false or true mycelium. They also differ in size.
Visible parts of the structure are the cell wall, cytoplasm, water vacuoles, fat globules,
and granules, which may be metachromatic, albuminous, or Starchy. Special staining is
necessary to demonstrate the nucleus.
Reproduction
Most yeasts reproduce asexually by multilateral or polar budding, a process in which
some of the protoplasm bulges out the cell wall; the bulge grows in size and finally walls off as a
new yeast cell. The bud appears to grow from a tube like projection from the mother cell. A few
species of yeasts reproduce by fission, and one reproduces by a combination of fission and
budding.
Sexual reproduction of "true" yeasts (Ascomycotina) results in the production of
ascospores, the yeast cell serving as the ascus. The formation of ascospores follows conjugation
of two cells in most species of true yeasts, but some may produce ascospores without
conjugation, followed by conjugation of ascospores or small daughter cells. The usual number of
spores per ascus and the appearance of the ascospores are characteristic of the kind of yeast.
"False" yeasts, which produce no ascospores or other sexual spores, belong to the Fungi
Imperfecti. Cells of some yeasts become chlamydospores by formation of a thick wall about the
cell, for example, Candida, Rhodotorula, and Cryptococcus.

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Cultural Characteristics
Growth as a film on the surface of liquid media suggests an oxidative or film yeast, and
production of a carotenoid pigment indicates the genus Rhodotorula. The appearance of the
growth is important when it causes colored spots on foods.
The oxidative yeasts may grow as a film, pellicle, or scum on the surface of a liquid and
then are termed film yeasts. Fermentative yeasts usually grow throughout the liquid and produce
carbon dioxide.
Physiological Characteristics
Most yeast requires more moisture than molds, however on the basis of water activity or
aw, yeasts may be classified as ordinary if they do not grow in high concentrations of solutes,.
Lower limits of aw for ordinary yeasts range from 0.88 to 0.94. Osmophilic yeasts have been
found growing slowly in media with an aw as low as 0.62 to 0.65 in syrups, although some
osmophilic yeasts are stopped at about 0.78 in both salt brine and sugar syrup.

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 The range of temperature for growth of most yeasts is 25 to 30 0C and the maximum about
35 to 470 C. Some kinds can grow at 00C or less.
The growth of most yeasts is favored by an acid reaction in the vicinity of pH 4 to 4.5,
Classification and identification of yeasts
The true yeasts are in the subdivision Ascomycotina, and the false, or asporogennous,
yeasts are in the subdivision Fungi Imperfecti or Deuteromycotina. Certain yeasts are actually
represented in two different genera based on whether they reproduce sexually.
The principal bases for the identification and classification of genera of yeasts are as follows:
1 Whether ascospores are formed.
2 If they are spore-forming:
a) The method of production of ascospores:
(1) Produced without conjugation of yeast, cells (parthenogenetically). Spore formation
may be followed by
(a) Conjugation of ascospores. (b) Conjugation of small daughter cells.
(2) Produced after isogamic conjugation (conjugating cells appear similar).
(3) Produced by heterogamic conjugation (conjugating cells differ in appearance).
b) Appearance of ascospores: shape, size, and color. Most spores are spheroidal or ovoid, but
some have odd shapes, e.g., most species of Hansenula, which look like derby hats
c) The usual number of ascospores per ascus: one, two, four, or eight.
3 Appearance of vegetative cells: shape, size, color, inclusions.
4 Method of asexual reproduction:
a. Budding. b. Fission. c. Combined budding and fission. d. Arthrospores (oidia).
5. Production of a mycelium, pseudo mycelium, or no mycelium.
6. Growth as a film over surface of a liquid (film yeasts) or growth throughout medium.
7 Color of macroscopic growth.
8 Physiological characteristics (used primarily to differentiate species or strains within a
species): a. Nitrogen and carbon sources.
b. Vitamin requirements.
c. Oxidative or fermentative: film yeasts are oxidative; other yeasts may be
fermentative or fermentative and oxidative.
d. Lipolysis; urease activity, acid production, or formation of starch like compounds.

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Yeasts of industrial importance
Genus Schizosaccharomyces These yeasts, which reproduce asexually by fission and form four
or eight ascospores per ascus after isogamic conjugation, have been found in tropical fruits,
molasses, soil, honey, and elsewhere. A common species is S. pombe.
Genus Saccharomyces Cells of these yeasts may be round, ovate, or elongated and may form a
pseudo-mycelium. Reproduction is by multipolar budding or by ascospore formation. The
ascospores, one to four per ascus, are usually round or ovate. The leading species, S. cerevisiae,
is employed in many food industries, with special strains used for the leavening of bread, as top
yeasts for ale, for wines, and for the production of alcohol, glycerol, and invertase.
Top yeasts are very active fermenters and grow rapidly at 200C. The clumping of the cells and
the rapid evolution of CO2 sweep the cells to the surface, hence the term top yeast.
Bottom yeast do not clump, grow more slowly, and are best fermenters at lower temperatures
(10 to 150 C). The absence of clumping and the slower growth and evolution of CO 2 permit the
yeast to settle to the bottom, hence the term bottom yeast. These characteristics of brewers' yeast
are observations and do not explain why some yeast clump, or flocculate. S. cerevisiae var.
ellipsoideus is a high-alcohol-yielding variety used to produce industrial alcohol, wines, and
distilled liquors.
Genus Kluyveromyces. These yeasts reproduce by multilateral budding, and ascopores are
liberated upon maturity.
Genus Zygosaccharomyces. These yeasts are notable for their ability to grow in high
concentrations of sugar (hence they are termed osmophilic) and are involved in the spoilage of
honey, sirups, and molasses and in the fermentation of soy sauce and some wines.
Zygosaccharomyces nussbaumeri grows in honey.
Genus Pichia These oval to cylindrical yeasts may form pseudomycelia. Ascospores are round
or hat-shaped, and there are one to four per ascus. A pellicle is formed on liquids; e.g., P.
membranaefaciens grows a pellicle on beers or wines.
Genus Hansenula These yeasts resemble Pichia in appearance but are usuua1ly more
fermentative, although some species form pellicles. Ascospores are hat- or Saturn-shaped.
Genus Debaryomyces These round or oval yeasts form pellicles on meat brines. Ascospores
have a warty surface. D. kloeckeri grows on cheese and sausage.

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Genus Hanseniaspora These lemon-shaped (apiculate) yeasts grow in fruit juices. Nadsonia
yeasts are large and lemon-shaped.
False Yeasts (Fungi Imperfect)
Genus Torulopsis These round to oval fermentative yeasts with multilateral budding cause
trouble in breweries and spoil various foods. T. sphaerica ferments lactose and may spoil milk
products. Other species can spoil sweetened condensed milk, fruit-juice concentrates, and acid
foods.
Genus Candida These yeasts form pseudohyphae or true hyphae, with abundant budding cells or
blastospores, and may form chlamydospores. Many form films and can spoil foods high in acid
and salt. C. utilis is grown for food and feed. C. krusei has been grown with dairy starter cultures
to maintain the activity and increase the longevity of the lactic acid bacteria. Lipolytic
C. lipolytica can spoil butter and oleomargarine.
Genus Brettanomyces These ogive - or arch-shaped yeasts produce high amounts of acid and are
involved in the late fermentation of Belgian lambic beer and English beers. They also are found
in French wines. B. bruxellansis and B. lambicus are typical species.
Genus Kloeckera These are imperfect apiculate or lemon-shaped yeasts. K. apiculata is common
on fruits and flowers and in the soil.
Genus Trichosporon These yeasts bud and form arthrospores. They grow best at low
temperatures and are found in breweries and on chilled beef. T. pullulans is a common species.
Genus Rhodotorula These red, pink, or yellow yeasts may cause discolorations on foods, e.g.,
colored spots on meats or pink areas in sauerkraut.
BACTERIA
Morphological characteristics important in food bacteriology
Encapsulation
The presence of capsules or slime may account for sliminess or ropiness of a food. In
addition, capsules serve to increase the resistance of bacteria to adverse conditions, such as heat
or chemicals. To the organism they may serve as a source of reserved nutrients. Most capsules
are polysaccharides of dextrin, dextran, or levan.
Formation of Endospores
Bacteria of the genera Bacillus, Clostridium, Desulfotomaculum, Sporolactobacillus
(rods), and Sporosarcina (cocci) share the ability to form endospores. Bacillus - aerobic and

18
some facultative anaerobic and Clostridium - anaerobic. Endospores are formed at an
intracellular site, are very refractile, and are resistant to heat, ultraviolet light, and desiccation.
Formation of Cell Aggregates
It is characteristic of some bacteria to form long chains and of others to clump under
certain conditions. It is more difficult to kill all bacteria in intertwined chains or sizable clumps
than to destroy separate cells.
Cultural characteristics important in food bacteriology
Bacterial growth in and on foods often is extensive. Pigmented bacteria cause
discolorations on the surfaces of foods; films may cover the surfaces of liquids; growth may
make surfaces slimy; or growth throughout the liquids may result in undesirable cloudiness or
sediment.
Physiological characteristics important in food bacteriology
These changes include hydrolysis of complex carbohydrates to simple ones; hydrolysis of
proteins to polypeptides, amino acids, and ammonia or amines; and hydrolysis of fats to glycerol
and fatty acids. O-R reactions, which are utilized by the bacteria to obtain energy from foods
(carbohydrates, other carbon compounds, simple nitrogen-carbon compounds, etc.), yield such
products as organic acids, alcohols, aldehydes, ketones, and gases.
Genera of bacteria important in food bacteriology
Genus Acetobacter These bacteria oxidize ethyl alcohol to acetic acid. They are rod-shaped and
motile and are found on fruits, vegetables, souring fruits, and alcoholic beverages. They are a
definite spoilage problem in alcoholic beverages.
Genus Aeromonas These are gram-negative rods with an optimum temperature for growth of 22
to 28 C. They are facultative anaerobes and can be psychrophilic. They are frequently isolated
from aquatic environments. A. hydrophila can be a human pathogen; it is also pathogenic to fish,
frogs, and other mammals.
Genus Alcaligenes As the name suggests, an alkaline reaction usually is produced in the
medium of growth. A. viscolactis. causes ropiness in milk, and A. metalcaligenes gives a slimy
growth on cottage cheese. These organisms come from manure, feeds, soil, water, and dust. This
genus also contains organisms which were formerly classified in the genus Achromobacter.
Genus Alteromonas Several former species of Pseudomonas are now classified as Alteromonas.
They are marine organisms that are potentially important in sea foods.

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Genus Arthrobacter A predominant soil organism, it is inert in most foods. However, some
species can grow at 50 C and would be considered psychrotrophs.
Genus Bacillus The endospores of species of this aerobic to facultative genus usually do not
swell the rods in which they are formed. Different species may be mesophilic or thermophilic,
actively proteolytic, moderately proteolytic, or non proteolytic, gas-forming or not, and lipolytic
or not. In general the spores of the mesophiles, e.g., B. subtilis, are less heat-resistant than spores
of the thermophiles. Spores of the obligate thermophiles, e.g., B. stearothermophilus, are more
resistant than those of facultative thermophiles, e.g., B. coagulans. The actively proteolytic
species usually may also sweet-curdle milk; B. cereus is such a species.
Genus Brevibacterium B. linens is related to Arthrobacter globiformis and may be synonymous.
B. linens may be important in the surface smear of certain cheeses, e.g., brick or Limburger,
where the culture produces an orange red pigmentation and helps ripening.
Genus Brochotrix These are gram-positive rods which can form long filamentous like chains
that may fold into knotted masses. The optimum temperature for growth is 20 to 25 0C, but
growth can occur over a temperature range of 0 to 45 0C depending on the strain. They can spoil a
wide variety of meats and meat products when they are stored aerobically or vacuum packed and
held refrigerated.
Genus Campylobacter These bacteria were originally classified in the genus vibrio. Several
strains of C. fetus subsp. jejuni have been associated with gastroenteritis in humans.
Genus Clostridium The endospores of species of this genus of anaerobic to microaerophilic
bacteria usually swell the end or middle of the rods in which they are formed. Different species
may be mesophilic or thermophilic and proteolytic or non-proteolytic.
Genus Corynebacterium The diphtheria organism, C. diptheriae, may be transported by foods.
C. bovis, with the slender, barred, or clubbed rods characteristic of the genus, is commensal on
the cow's udder, can be found in aseptically drawn milk, and may be a cause of bovine mastitis.
Genus Dessulfotomaculum A gram, negative rod which swells when an endospore forms. They
are common inhabitants of the soil, fresh water, and the rumen. Sulfur compounds can serve as
the terminal electron acceptor in respiration and thereby be reduced to hydrogen sulfide.
Genus Enterobacter Some were formerly classified as Aerobacter. They are widely distributed
in nature; a member of the coliform group.

20
Genus Erwinia The species of this genus are plant pathogens that cause necrosis, galls, wilts, or
soft rots in plants and therefore damage the plants and vegetable and fruit products from them. E.
carotovora is associated with the market disease called "bacterial soft rot."
Genus Escherichia Found in feces, a predominant gram-negative rod isolated from the intestinal
tract of warm-blooded animals and widely distributed in nature. One of the "coliform group," the
genus is divided into many biotypes and serotypes, some of which can be pathogenic to humans.
Genus Flavobacterium The yellow to orange-pigmented species of this genus may cause
discolorations on the surface of meats and be involved in the spoilage of shellfish, poultry, eggs,
butter, and milk. Some of the organisms are psychrotrophic and have been found growing on
thawing vegetables.
Genus Gluconobacter (Formerly Acetomonas). Species can oxidize ethanol to acetic acid. G.
oxydans causes ropiness in beer following viscous growth in beer or wort.
Genus Halobacterium Bacteria of this genus, e.g., H. salinarium, are obligate halophiles and are
usually chromogenic. They may grow and cause discolorations on foods high in salt, such as
salted fish.
Genus Klebsiella Many are capsulated. Commonly associated with the respiratory and intestinal
tracts of humans. K. pneumoniae is the causative organism for a bacterial pneumonia in humans.
Family Lactobacillaceae
Genus Lactobacillus The lactobacilli are rods, usually long and slender, that form chains in most
species. They are microaerophilic, (some strict anaerobes are known), are catalase-negative and
gram-positive, and ferment sugars to yield lactic acid as the main product.
Homofermentation: They ferment sugar chiefly to lactic acid if they are homo-fermentative,
with small amounts of acetic acid, carbon dioxide, and trace products.
Heterofermentation: If they are hetero-fermentative, they produce appreciable amounts of
volatile products, including alcohol, in addition to lactic acid.
Genus Listeria These bacteria are gram-positive, non-spore-forming rods with tumbling
motility. L. monocytogenes can cause food-borne disease outbreaks.
Genus Microbacterium Bacteria of this genus are important because of their resistance to
adverse conditions and their use in production of vitamins. They are small, nonmotile,
grampositive, asporogenous, catalase-positive, aerobic, homo-fermentative, lactic acid-producing
rods, which sometimes produce palisade arrangements.

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Genus Micrococcus Most of the species prominent in foods are gram-positive, aerobic, and
catalase-positive.
Genus Mycobacterium The tubercle bacilli that cause tuberculosis, M. tuberculosis, has been
spread by foods, especially raw milk from infected cows.
Genus Pediococcus The cocci occur singly, in pairs or short chains, or in tetrads (division in two
planes) and are gram-positive, catalase-negative, and microaerophilic. They are
homofermentative.
Genus Photobacterium The genus includes coccobacilli and occasional rods which can be
luminescent. They are not widespread; however, P. phosphoreum has been known to cause
phosphorescence of meats and fish.
Genus Propionibacterium Members of this genus may be found in foods. In Swiss cheese
certain species (e.g., Propionibacterium freudenreichii) ferment the lactates to produce the gas
that helps form the holes or eyes, and also contribute to the flavor. Pigmented propionic bacteria
can cause color defects in cheese.
Genus Proteus Bacteria of this genus have been involved in the spoilage of meats, seafood, and
eggs. The presence of these bacteria in large numbers in non refrigerated foods has made them
suspect as a cause of food poisoning.
Genus Pseudomonas A number of species of Pseudomonas can cause food spoilage. These
bacteria are gram-negative, usually motile, asporogenous rods.
Genus Salmonella Species of these enteric pathogens may grow in foods and cause food
infections.
Genus Serratia Many species produce a pink or magenta pigment and may cause red
discolorations on the surface of foods. S. marcescens is the most common species.
Genus Shigella Species of Shigella, causing bacillary dysenteries, may be transported by foods .
Genus Sporolactobacillus Lactobacillus inulinus has been classified as S. inulinus because of its
ability to font endospores. It resemble Lactobacillus in many characteristics.
Genus Sporosarcina A gram-positive coccus that forms endospores. S. ureae and S. halophila
are the two species listed.
Genus Staphylococcus The gram-positive staphylococci grow singly, in pairs, in tetrads, or in
irregular, grapelike clusters. The most important species, S. aureus, usually gives yellow to

22
orange growth, although it may be white on occasion. Many of the beta-hemolytic, coagulase
positive strains are pathogenic, and some produce an enterotoxin which causes food poisoning
Genus Streptococcus The cocci in this genus are characteristically in pairs, in short chains, or in
long chains, depending on the species and the conditions of growth, and all is homofermentative.
The streptococci important in foods are divided in food four groups: the pyogenic, viridans,
lactic, and, enterococcus groups.
Genus Streptomyces Members of this genus can cause undesirable flavors and appearance when
growing on foods; musty or earthy odors and tastes from these organisms may be absorbed by
nearby foods when growth of the Streptomyces is near at hand.
Genus Vibrio Bacteria in this genus are widely distributed in fresh and salt water, in soil, and in
the alimentary canal of humans and animals. Some are moderately halophilic. Some species are
pathogenic to humans.
Genus Yersinia Can be found in the soil. Y. pestis is the causative-organism of plague in humans
and in rats and other rodents. Some strains of Y. enterocolitica are also pathogenic and causative
agents of food-borne disease outbreaks.
PRINCIPLES OF FOOD PRESERVATION
We get food from different sources like plant, animal and microbial. The nutritive quality
of food depends on quality and quantity of nutrients they possess. Food are prepared for
consumption by taking materials of plant, animal or microbial origin. Some food of plant origin
are directly consumed as fresh raw food such as vegetables, fruits etc and some are eaten after
cooking. Most of the animal based foods are consumed after cooking.
Microorganisms serve as food such as single cells proteins or makers of food such as
bread, cake and other fermented food. Due to activity of microorganisms, the nutritive value of
food becomes enriched in terms of more digestibility, increased mineral and vitamin content,
taste, flavour and keeping quality. Most of the plant based food such as grains are available in
dried form for long term preservation and consumption.
The preservation and keeping quality of food and food materials depend mainly on their
composition and water content. Long term preservation or storage of food or food materials
brings changes in their chemical composition and nutritive value. These changes could be due to
physical, chemical or microbiological activities occurring in food or food materials.

23
Microbiological changes occurring in food or food materials mainly depend on water content,
water activity (aw) and temperature of storage.

Food preservation is the process of physical, chemical or microbiological treatments of

food and beverages to prevent food spoilage and food borne diseases. Food have to be preserved
to increase their shelf life to avoid spoilage by microorganisms and to curtail pollution of land
and water with spoilt food. Food preservation should also take the type of food into consideration
before applying the preservation method. There could be perishable food, semi-perishable food
and non-perishable or stable food.

Methods of food preservation

Foods are preserved by different methods depending on the type of food, its composition
and period of preservation required. Some of the commonly practiced preservation methods
based on above explained principles are given below.

ASEPSIS
It is the process of keeping out microorganisms to avoid spoilage. Food, whether fruits or
vegetables, if in healthy condition, are free from microbes and are contaminated if damaged
during processing or if the protecting external covering is damaged.
The load of microorganisms in food is described as “bioburden”. The type and numbers
of organisms comprising the bioburden influence food spoilage and this also decides the quality
of food.
Asepsis in food industry is important to reduce bioburden of food to avoid spoilage.
Some methods of ‘asepsis’ are packaging of food, hygienic handling and processing of food,
adoption of sanitary methods in slaughtering, handling and processing of meat.
REMOVAL OF MICROORGANISMS
 It is not an effective method of food preservation but helpful during processing of food for
juice and soft drink production.
 The methods followed are filtration, centrifugation, sedimentation, washing and trimming.
 Filtration is used for fruit juices, beer, wine and water. Centrifugation is used for milk and
sedimentation for water.
 Washing and trimming are followed for fruits and vegetables.

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 Washing food may be dangerous if the water used is contaminated or has pathogenic
enterobacteriaceae members like E. coli, Salmonella, Shigella, etc.
MAINTENANCE OF ANAEROBIC CONDITIONS
 Food is preserved by sealing, packing, canning to create anaerobic condition. The seals,
packs or cans are completely filled, evacuated of air leading to creation of vacuum or they
are replaced with inert gases like CO2, N2, and H2.

 Anaerobic conditions created prevent growth of aerobes but obligate anaerobes could grow
like Cl. botulinum which is the most dangerous organism as it produces a potent neurotoxin.
 Precautions should be taken to avoid contamination by anaerobes when anaerobic
conditions are used for preservation.
USE OF HIGH TEMPERATURE
Exposure to heat denatures proteins leading to inactivation of enzymes required for
metabolism. The heat treatment necessary to kill microorganisms or their spores varies with the
kind of organism, its physiological state and the environment during heating.
The heat treatment selected depends on kind of organisms to be killed, other preservative
methods to be employed and the effect of heat on food. Certain factors affect the heat resistance
of cells or spores like
a) Initial concentration of cells / spores in food to be heated.
b) Previous history of cells/ spores, that is the conditions under which they have been grown
like culture medium, temperature, growth phage, desiccation etc.
Thermal death time
It is defined as the time taken at a given temperature to kill specified number of
microorganisms under defined conditions.
Thermal death point
It is the temperature necessary to kill all microorganisms present in a given substance in
defined time (minutes).
For efficient application of high temperature for preservation of food, the rate of heat
penetration into food should be efficient and this is influenced by material of the container,
consistency, size and shape of contents, initial temperature of food, prior treatments if any etc.
Material of container make like glass has slower rate of penetration than metal.

25
 Consistency of can contents and size and shape of food item, like larger one takes longer
time, smaller one takes shorter time. Food items could retain their identity, lose their form and
become viscous, or layer out.
Initial temperature of food, like lower the initial temperature, longer time of exposure and
higher the initial temperature, shorter the time of exposure required for treatment.
The various degrees of heating used on food can be classified as
(a) pasteurization

(b) heating at 1000 C and

(c) heating above 1000 C

Pasteurization
Pasteurization is a heat treatment that kills part but not all the microorganisms present

and usually involves the application of temperatures below 100 0 C. The heating may be by
means of steam, hot water, dry heat, or electric currents and the products are cooled promptly
after the heat treatment.
Pasteurization is used under these conditions like
 when more rigorous heat treatments might harm the quality of product, as with
market milk,
 when the aim is to kill pathogens, as with market milk,
 when the main spoilage organisms are not very heat resistant, such as the yeasts
in fruit juices, when any surviving spoilage organisms will be taken care of by
additional preservative methods to be employed, as in the chilling of market milk,
and
 when competing organisms are to be killed, allowing a desired fermentation,
usually by added starter organisms, as in cheese making.

Preservative methods used to supplement pasteurization include

(a) refrigeration, e.g., milk,
(b) keeping out microorganisms, usually by packaging the product in a sealed container
as in case of fruit juices,
(c) maintenance of anaerobic conditions, as in evacuated, sealed containers,

26
 (d) addition of high concentrations of sugars in sweetened condensed milk, and
(e) presence or addition of chemical preservatives, e.g., the organic acids on pickles.

Times and temperatures used in the pasteurizing process depend on the method employed
and the product treated. The high-temperature-short-time (HTST) method employs a
comparatively high temperature for a short time whereas the low-temperature- holding (LTH) for
long-time method uses a lower temperature for a longer time.

The minimal heat treatment of market milk is at 72 0C for 16 sec like in the HTST

method; and at 137.80C for at least 2 sec in the ultra pasteurized method. Ice cream mix may be

heated at 71.10C for 30 min or at 82.20 C for 16 to 20 sec.

Grape wines may be pasteurized for 1 min at 82 0 to 850 C in bulk. Dried fruits usually

are pasteurized in the package at 65.60 to 850 C in for 30 to 90 min. The average heat treatment

for carbonated juices would be 65.60 C for 30 min.

Heating at 1000C

 A temperature of approximately 100 0C is obtained by boiling a liquid food, by immersion

of the container of food in boiling water, or by exposure to flowing steam. Some very acid

food, e.g., sauerkraut, may be preheated to a temperature some what below 100 0C,
packaged hot, and not further heat-processed.
 Blanching fresh vegetables before freezing or drying involves heating briefly at about

1000C.
 During baking, the internal temperature of bread, cake, or other bakery products approaches

but never reaches 1000C as long as moisture is present, although the oven is much hotter.
The temperature of unsealed canned goods heated in the oven cannot exceed the boiling
temperature of the liquid present.

 Simmering is incipient or gentle boiling, with the temperature reaching about 1000C.

27
 In roasting meat the internal temperature reaches only about 60 0 C in rare beef, upto 800C

in well-done beef, and 850C in a pork roast.

 Frying gets the outside of the food very hot, but the center ordinarily does not reach

1000C .

Heating at above 1000C

Temperatures above 1000C usually are obtained by means of steam under pressure in
steam-pressure sterilizers (autoclaves) or retorts. With no pressure, the boiling temperature of

water at sea level is 100 0C; with 5 lb of pressure it is 109 0C, with 10 lb it is 115.5 0C, and with

15 lb it is 121.50C. Milk can be heated to temperatures up to 1500C by use of steam injection or

steam infusion followed by “flash evaporation” of the condensed steam and rapid cooling.
USE OF LOW TEMPERATURES
Low temperatures are used to retard chemical reactions and action of food enzymes and
to slow down or stop the growth and activity of microorganisms in food. The lower the
temperature, the slower will be chemical reactions, enzyme action, and microbial growth. A low
enough temperature will prevent the growth of any mesophilic or thermophilic microorganism.
As the temperature drops from its optimal temperature toward the minimal, the rate of
growth of the microorganism decreases and is slowest at the minimal temperature. Cooler
temperatures will prevent growth of an organism but slow metabolic activity may continue.
Therefore, the cooling of food from ordinary temperatures has a different effect on various
organisms present.
In general, freezing prevents the growth of most food-borne microorganisms and
refrigeration temperatures slow growth rates. Commercial refrigeration temperatures, i.e., lower

than 5 to 7.20C, effectively retard the growth of many food-borne pathogens.

One notable exception is Clostridium botulinum type E, which has a minimum

temperature for growth of about 3.30C, Yersinia enterocolitica can survive and grow at

temperatures as low as 0 to 30C. The molds, Cladosporium sp and Sporotrichum sp have been

28
found to be growing on food at -6.70C and Penicillium and Monilia at -40C. Growth by one

yeast has taken place at -340C, and two others grew at -180C.
Different methods commonly employed in use of low temperature for preservation are
given below:
Common, or cellar storage
The temperature in common, or celler, storage usually is not much below that of the

outside air and seldom is lower than 15 0C. Root crops, potatoes, cabbage, celery, apples, and
similar food can be stored at these temperatures for limited periods.
Chilling or cold storage
Chilling storage is at temperatures not far above freezing and usually involves, cooling by
ice or by mechanical refrigeration. Most perishable food, including eggs, dairy products, meats,
seafood, vegetables, and fruits, may be held in chilling storage (commonly used deep freezers)
for a limited time with little change from their original condition.
Factors to be considered in connection with chilling storage include the temperature of
chilling, the relative humidity, air velocity and composition of the atmosphere in the store room,
and the possible use of ultraviolet rays or other radiations.
Temperature
The lower the temperature of storage, the greater the cost, Therefore, although most food
will be kept best at a temperature just above their freezing point, they are not necessarily stored
at this low temperature. The chilling temperature is selected based on the kind of food, the time
and conditions of storage. Some varieties of apples undergo “ low-temperature breakdown” at

temperatures near freezing, and sweet potatoes keep best at 10 to 12.8 0C, and banana cannot be

refrigerated but stored at13.3 to16.70C.

PRESERVATION BY DRYING
Preservation of food by drying has been practiced for centuries. Drying usually is
accomplished by the removal of water, but any method that reduces the amount of available
water i.e., lowers aw in a food is also a form of drying. For example sugar may be added, as in

sweetened condensed milk will reduce the amount of available moisture.

29
 Moisture may be removed from food by different methods, ranging from the ancient
practice of drying by sun rays to the modern artificial ones like freeze-drying.
Drying can be accomplished by various methods.
In sun-dried food, moisture is removed by exposure of food to the sun rays without any
artificially produced heat and without controlled temperatures, relative humidifies, or air
velocities.
A dehydrated or desiccated food has been dried by artificially produced heat under
controlled conditions of temperature, relative humidity, and air flow. Condensed usually implies
that moisture has been removed from a liquid food, and evaporated may have a similar meaning
or may be used synonymously with the term dehydrated. Different methods of drying are solar
drying, mechanical drying, freeze-drying and smoking.
Solar drying
Solar drying is limited to mostly the tropical climates with a hot sun and a dry
atmosphere. It is applicable to certain fruits, such as raisins, prunes, figs, apricots, nectarines,
pears and peaches.
The fruits are spread out on trays and may be turned during drying. Fish, rice and other
grains may also be sun-dried. Many household preparations like papads are also sun-dried.
Drying by mechanical dryers
Most methods of artificial drying involve the passage of heated air with controlled
relative humidity over the food to be dried or the passage of the food through such air. A number
of devices are used for controlled air circulation. The simplest dryer is the evaporator or kiln.
Forced-draft drying systems employ currents of heated air that move across the food, usually in
tunnels.
Liquid food, such as milk, juices, and soups, may be evaporated by the use of
comparatively low temperatures and a vacuum in a vacuum pan or similar device. These food
could also be drum-dried by passage over a heated drum, with or without vacuum or spray-dried
by spraying the liquid into a current of dry, heated air.
Freeze drying
Freeze drying, or the sublimation of water from a frozen food by means of application of
both vacuum and heat is being used for a number of food, including meats, poultry, seafood,

30
fruits, and vegetables. Frozen thin layers of food of low sugar content may be dried without
vacuum by sublimation of moisture during passage of dry carrier gas.
Smoking
Food items like fish are subjected to smoking which removes moisture and also helps in
preservation as smoke like woods smoke has antimicrobial compounds.
Other Methods
Electronic heating has been suggested for the removal of still more moisture from a food
already fairly well dried. Foam-mat drying, in which liquid food is whipped to foam, dried with
warm air, and crushed to a powder, is significantly being used for most food.
Another method is pressure-gun puffing of partially dried food to give a porous structure

that facilitates and hastens drying. Tower drying is a process in which dehumidified air at 30 0C
or lower is used and has been successful with tomato concentrate, milk, and potatoes.
Factors that influence or control drying
Various factors influence the method of drying as given below:
a) The temperature employed varies with the food and the method of drying taking into
consideration the sensitivity of food to temperature.
b) The relative humidity of the air influences drying and is varied with the food, the method of
drying and also with the stage of drying. It usually is higher at the start of drying than it is at
later stage of drying.
c) The velocity of the air influences by removal of moisture at faster rate if velocity is more and
slower rate if velocity is less.
d) The time of drying determines the extent of dehydration. More the time, more is the drying,
but it again depends on other three factors.
Improper control of these factors may cause case-hardening, resulting from more rapid
evaporation of moisture from the surface than diffusion from the interior, with a resulting hard,
horny, impenetrable surface film that hinders further drying.
Pre-treatments of food before drying
Many of the food need pretreatments before drying for an effective effect of drying
method on the microbial population.
These pretreatments may include:
(a) selection and sorting for size, maturity, and soundness

31
 (b) washing, especially of fruits and vegetables
(c) peeling of fruits and vegetables by hand, machine, lye bath, or abrasion
(d) subdivision into halves, slices, shreds, or cubes
(e) alkali dipping, which is used primarily for fruits such as raisins, grapes, and prunes
(for sun drying) and employs hot 0.1 to 1.5 percent lye or sodium carbonate
(f) blanching or scalding of vegetables and fruits
(g) sulfuring of light-colored fruits and certain vegetables.

Fruits are sulfured by exposure to sulfur dioxide gas produced by the burning of sulfur so
that a level of 1,000 to 3,000 ppm, depending on the fruit, will be absorbed. Vegetables may be
sulfured after blanching in a similar manner or by dipping into or spraying with sulfite solution.
Sulfuring helps maintain an attractive light color, conserve vitamin C and perhaps vitamin A, and
repel insects; it also kills many of the microorganisms present.
Post-treatments of food after drying
The procedures after drying vary with the kind of dried food. They could be sweating,
packing, pasteurization.
Sweating
“Sweating” is a process of storage, usually in bins or boxes, for equalization of moisture
or re-addition of moisture to a desired level. It is used primarily with some dried fruits and some
nuts (almonds, English walnuts).
Packaging
Most food are packaged soon after drying for protection against moisture loss,
contamination with microorganisms, and infestation with insects, although some dried food (e.g.
fruits and nuts) may be held as long as a year before packaging.
Pasteurization
Most dried fruits are subjected to pasteurization to kill pathogens and spoilage organisms.

Fruits in package are treated generally at 65.6 to 85 0C for 30 to70 min at 70-100% relative
humidity.
Changes during the drying process
Heat applied during the drying process causes a reduction in total numbers of
microorganisms, but the effectiveness varies with the kinds and numbers of organisms originally

32
present and the drying process employed. Usually all yeasts and most bacteria are destroyed, but
spores of bacteria and molds commonly survive, as do vegetative cells of a few specie of heat –
resistant bacteria.
More microorganisms are killed by freezing than by dehydration during the freeze-drying
process. The dried food item remains un-spoilt after drying if storage conditions are adequate.
Microorganisms decrease in number but some resistant forms like spores of bacteria and molds
could survive and also increase, during storage

FACTORS AFFECTING THE GROWTH OF MICROORGANISMS IN FOOD

Growth of microorganisms in nature is dependent on various factors. The factors
influencing the growth of microorganisms are physical, chemical and biological in nature. The
important factors which contribute to the growth of microorganisms in food are intrinsic and
extrinsic factors.
Intrinsic Factors
 Nutrient content
 pH
 Redox potential
 Water activity
 Inhibitory substances

Extrinsic Factors
 Temperature, Growth rate
 Relative humidity , Gaseous atmosphere

Intrinsic Parameters
Nutrient content
Nutritional quality of food in terms of their chemical composition, nutritive value or
nutrients, their proportion and growth promoting ability are important for growth of
microorganisms.

33
 Food for energy have more carbohydrates and other carbon compounds like esters,
alcohols, peptides, amino acids , organic acids and their salts. Carbohydrates may include
polysaccharides like cellulose, starch and different sugars. Depending on the type of
carbohydrate and other energy compounds present, there will be variation in terms of
microorganisms growing in food.
Most of the sugary food have growth of yeasts and bacteria and starchy food promote
growth of molds which are able to hydrolyze starch by producing amylases. Cellulolytic and
pectinolytic molds grow on food containing these materials. Carbon for growth usually comes
from carbohydrates and other organic compounds in most of the heterotrophic microorganisms.
The total nitrogen content and different nitrogenous nutrients in food may vary.
Microorganisms differ in their ability to use various nitrogenous compounds as source of
nitrogen for growth. Many organisms are unable to hydrolyze proteins due to lack of proteolytic
enzymes.
Proteolytic organisms produce proteases and hydrolyze proteins to peptides and amino
acids which are used as source of nitrogen for growth by them and others. Proteolytic property is
different among microorganisms depending on the type of protease enzyme produced. Bacteria
are more proteolytic than fungi in general.
Protein rich food promote more growth of bacteria than molds and yeasts. Some of the
lactic acid bacteria grow best with polypeptides as nitrogen source, cannot attack casein, and do
not grow well with only a limited number and kinds of amino acids present.
Presence of fermentable carbohydrates in a food results in an acid fermentation and
suppression of proteolytic bacteria preventing production of obnoxious nitrogenous products.
Many proteolytic bacteria grow best at pH values near neutrality and are inhibited by acidity.
Some microorganisms are not able to prepare vitamins and other growth factors. They
need to be provided in growth medium. Food contain different vitamins, minerals and other
growth factors and their composition and content may vary.

Water Activity (aW)

Water is often the major constituent in foods. Even relatively ‘dry’ foods like bread and
cheese usually contain more than 35% water. The state of water in a food can be most usefully
described in terms of water activity.

34
 aW =Vapour pressure of a substance or solution

Vapour pressure of water at same temperature

Water activity of a food is the ratio between the vapour pressure of the food, when in a
completely undisturbed balance with the surrounding air, and the vapour pressure of pure water
under identical conditions.
Water activity, in practice, is measured as Equilibrium Relative
Humidity (ERH) and is given by the formula:
Water Activity (aW) = ERH / 100

Water activity is an important property that can be used to predict food safety, stability
and quality. The various applications of water activity includes; maintaining the chemical
stability of foods, minimizing non enzymatic browning reactions and spontaneous autocatalytic
lipid oxidation reactions, prolonging the desired activity of enzymes and vitamins in foods,
optimizing the physical properties of foods such as texture.
Water activity scale extends from 0 (bone dry) to 1.00 (pure water). But most foods have
a water activity in the range of 0.2 for very dry foods to 0.99 for moist fresh foods. Based on
regulations, if a food has a water activity value of 0.85 or below, it is generally considered as
non-hazardous. This is because below a water activity of 0.91, most bacteria including the
pathogens such as Clostridium botulinum cannot grow. But an exception is Staphylococcus
aureus which can be inhibited by water activity value of 0.91 under anaerobic conditions but
under aerobic conditions, it requires a minimum water activity value of 0.86. Most molds and
yeasts can grow at a minimum water activity value of 0.80. Thus a dry food like bread is
generally spoiled by molds and not bacteria.
In general, the water activity requirement of microorganisms decreases in the following
order: Bacteria > Yeast > Mold. Below 0.60, no microbiological growth is possible. Thus the
dried foods like milk powder, cookies, biscuits etc are more shelf stable and safe as compared to
moist or semi-moist foods.
Factors that affect water activity requirements of microorganisms include the following-
kind of solute added, nutritive value of culture medium, temperature, oxygen supply, pH,
inhibitors. Each microorganism has a minimal water activity for growth as shown in Table.
Minimum water activity values of spoilage microorganisms

35
 Microbial group Minimum aw Examples

Most bacteria 0.91 Salmonella spp.,

Clostridium botulinum
Most yeasts 0.88 Torulopsis spp.
Most molds 0.80 Aspergillus flavus
Halophilic bacteria 0.75 Wallemia sebi
Xerophilic molds 0.65 Aspergillus echinulatas
Osmophilic yeasts 0.60 Saccharomyces bisporus

Water activity of some foods and susceptibility to spoilage by microorganisms

Water activity Microorganisms grow at this Food examples

awand above

1.00 – 0.95 Pseudomonas, [Link], Proteus, Highly perishable fresh foods

Shigella, Klebsiella, Bacillus, & canned fruits, vegetables,
Clostridium perfringens & meat, fish, milk, eggs; foods
some yeasts containing up to 40% (w/w)
sucrose or 7% NaCl.
0.95 – 0.91 Salmonella, Vibrio Some cheeses (cheddar,
parahemolyticus, Clostridium Swiss), Cured meats; some
botulinum, Serratia, fruit juice concentrates; bread;
Lactobacillus, Pediococcus; high moisture prunes; foods
some molds and yeasts containing 55%(w/w) sucrose
or 12% NaCl
0.91 – 0.87 Many yeasts like Candida, Fermented sausages; sponge
Torulopsis, Hansenula; cakes; dry cheese; margarine;
Micrococcus foods containing 65%(w/w)
sucrose (saturated) or 15%
NaCl
0.87 – 0.80 Most molds, Staphylococcus Most fruit juice concentrates,
aureus, most Saccharomyces sweetened condensed milk,

36
 spp., Debaromyces flour, rice, pulses containing
15 –17% moisture, salami
0.80 – 0.75 Most halophilic bacteria, Jam, Marmalade, Glace fruits,
Mycotoxigenic Aspergilli Soy sauce
0.75 - 0.65 Xerophilic molds, Rolled oats containing 10%
Saccharomyces bisporus moisture; Fudge;
Marshmallows; Jelly; Some
dried fruits; Nuts, Peanut
Butter
0.65 - 0.60 Osmophilic yeasts, Few molds Dried fruits containing 15 –
20% moisture; Honey
0.50 No microbial proliferation Pasta containing 12%
moisture; Spices containing
10% moisture
0.40 No microbial proliferation Whole egg powder containing
5% moisture
0.30 No microbial proliferation Cookies, biscuits crackers,
bread crusts etc. containing 3
– 5% moisture

Water acts as an essential solvent that is needed for most biochemical reactions by the
microorganisms. Water activity of the foods can be reduced by several methods: by the addition
of solutes or hydrophilic colloids, cooking, drying and dehydration: (e.g., egg powder, pasta), or
by concentration (e.g. condensed milk) which restrict microbial growth so as to make the food
microbiologically stable and safe.
A wide variety of foods are preserved by restricting their water activity. These include:
Dried or Low Moisture Foods
These contain less than 25% moisture and have a final water activity between 0.0 and
0.60. e.g., Dried egg powder, milk powder, crackers, and cereals. These products are stored at

37
room temperature without any secondary method of preservation. These are shelf stable and do
not spoil as long as moisture content is kept low.
Intermediate Moisture Foods
These foods contain between 15% and 50% moisture content and have a water activity
between 0.60 and 0.85. These foods normally require added protection by secondary methods
such as pasteurization, pH control, refrigeration, preservatives, but they can also be stored at
room temperature. These include dried fruits, cakes, pastries, fruit cake, jams, syrups and some
fermented sausages. These products are usually spoiled by surface mold growth.
pH and Buffering capacity
The pH, or hydrogen ion concentration, [H+], of natural environments varies from about
0.5 in the most acidic soils to about 10.5 in the most alkaline lakes. Since the pH is measured on
a logarithmic scale, the [H+] of natural environments varies over a billion-fold and some
microorganisms are living at the extremes, as well as every point between the extremes.
The range of pH over which an organism grows is defined by three cardinal points: the
minimum pH, below which the organism cannot grow, the maximum pH, above which the
organism cannot grow, and the optimum pH, at which the organism grows the best.
Microorganisms which grow at an optimum pH well below neutrality (7.0) are called
acidophiles. Those which grow best at neutral pH are called neutrophiles and those that grow
best under alkaline conditions are called alkalophiles.
In general, bacteria grow faster in the pH range of 6.0- 8.0, yeasts 4.5-6.5 and
filamentous fungi 3.5-6.8, with the exception of lactobacilli and acetic acid bacteria with optima
between pH 5.0 and 6.0.
Approximate pH ranges of different microbial groups
Minimum Optimum Maximum

Most Bacteria 4.5 6.5 – 7.5 9.0

Yeasts 1.5 – 3.5 4.0 – 6.5 4.0 – 6.5

Molds 1.5 – 3.5 4.5 – 6.8 8.0 – 11.0

The approximate pH ranges of some common food commodities are shown in Table.
Approximate pH ranges of some common food commodities

38
 Product pH

Citrus fruits 2.0-5.0

Soft drinks 2.5-4.0

Apples 2.9-3.3

Bananas 4.5-4.7

Beer 3.5-4.5

Meat 5.6-6.2

Vegetables 4.0-6.5

Fish ( most spp) 6.6-6.8

Milk 6.5-6.8

Wheat flour 6.2-6.8

Egg white 8.5-9.5

Fermented shark 10.5-11.5

The acidity of a product can have important implications for its microbial ecology, and
the rate and character of its spoilage. For example, vegetables generally have a moderately acidic
pH and thus are spoiled by soft-rot producing bacteria such as Erwinia carotovora and
Pseudomonads, while in fruits, a lower pH prevents bacterial growth and spoilage is caused by
yeasts and molds.
Also, fish spoil more rapidly as compared to meat under chill conditions. The pH of meat
(5.6) is lower than that of fish (6.2-6.6) and this contributes to the longer storage life of meat.
The pH-sensitive genus Shewanella plays an important role in fish spoilage but has not been
reported in normal meat (pH<6.0).
The natural or inherent acidity of foods may be thought of as nature’s way of protecting
the respective plant or animal tissues from destruction by microorganisms. Thus, the ability of
low pH to restrict microbial growth has been deliberately employed since long in the
preservation of foods with acetic and lactic acids.

39
Redox Potential (Eh)
Redox Potential (Eh) is defined as the measure of whether microorganism has the
tendency to gain electrons (reduction) or lose electrons (oxidation).
Prevailing oxygen concentration of food in their environment, chemical composition and
type of microorganisms associated, contribute to the oxidation-reduction (O-R) potential of food
and affect growth of microorganisms in them.
The O-R potential of food is determined by characters such as
(a) oxygen tension of atmosphere above the food,
(b) access of atmosphere to the food,
(c) resistance of food to the changes occurring and
(d) O-R state of materials present in food.
Oxygen content in head space of the food contributes to the growth of microorganisms as
microorganisms are aerobic, micro-aerophilic, facultative and anaerobic for oxygen requirement
to grow. Aerobic and micro-aerophilic organisms require free oxygen in air but in different
concentrations, facultatives grow in presence or absence of oxygen and anaerobes grow only in
the absence of oxygen.
Most of the fungi, bacteria and yeasts grow in food under aerobic conditions and
anaerobic bacteria such as Clostridium botulinum require anaerobic conditions. Oxygen
concentration of head space on food depends on its access to air. Food stored under evacuation
or canned promote growth of anaerobic bacteria.
Redox potential of a system is expressed as Eh and expressed in millivolts (mV). Highly
oxidized substrates have positive Eh and reduced substrates have negative Eh. Therefore aerobic
organisms require positive Eh and anaerobic ones require negative Eh values for their growth.
Most of the fresh plant and animal food have low redox potential because of reducing
substances present in them. Fresh vegetables and fruits contain reducing substances such as
ascorbic acid, reducing sugars and animal tissues have sulfhydryl (-SH) and other reducing group
compounds considered as anti-oxidants.
Fresh vegetables, fruits and meat support growth of aerobic microorganisms at their
surfaces because of positive redox potential. However, the anaerobic microorganisms grow in
inner parts of vegetables, fruits and meat because of negative redox potential.

40
 Most of processed plant and animal food gain positive redox potential therefore promote
growth of aerobic organisms. Food subjected to heat and/or packed under negative pressure
promote growth of anaerobic organisms.
Microorganisms growing in food may create different redox potential states depending on
their oxygen requirement and redox changes they bring about in food contents. Quality of
processed food in terms of redox status depends on the changes brought about by processing
methods, type of microorganisms and chemical changes.
Redox potential of some foods
Food Eh(mV) pH

Raw meat -200 5.7

Minced meat +225 5.9

Whole wheat -350 6.0

Ground grain (Barley) +225 7.0

Fruit juices +300 to + 3.0-5.5

Grape +410 3.8

Lemon +380 2.2

Inhibitory substances
Some foods can resist the attack by microorganisms due to presence of certain naturally
occurring substances which possess antimicrobial activity such as essential oils in spices
( eugenol in cloves and cinnamon, allicin in garlic, cinnamic aldehyde in cinnamon, thymol in
sage); lactaferrin, lactoperoxidase and lysozyme in milk; and ovatransferrin, avidin, lysozyme
and ovoflavoprotein in hen’s egg albumin.
Similarly, casein as well as free fatty acids found in milk also exhibit antimicrobial
activity. The hydroxycinnamic acid derivatives (p-coumaric, feluric, caffeic and chlorogenic
acids) found in fruits, vegetables, tea and other plants possess antibacterial and antifungal
activity.

41
 Also natural covering of foods like shell of eggs and nuts, outer covering of fruits and
testa of seeds, hide of animals provide protection against entry and subsequent spoilage by
microorganisms.
QUESTION BANK

UNIT – I

2 MARKS

1. Define food microbiology?

2. Food as a substrate for microbes?
3. Molds in food?
4. Yeast in food?
5. Bacteria in food?
6. How affect temperature against microbes?
7. How moisture act against microorganisms?
8. What are the other factors affecting the microbial growth?

5 MARKS

1. Molds in industrial importance?

2. Classification and identification of yeast?
3. Genera of bacteria in food microbiology?
4. Details about water activity?
5. Discuss about the Redox potential?

10 MARKS

1. Microbes in food microbiology?

2. Factors affecting the growth of microbes?

UNIT - II

PRINCIPLES OF FOOD PRESERVATION

We get food from different sources like plant, animal and microbial. The nutritive quality
of food depends on quality and quantity of nutrients they possess. Food are prepared for
consumption by taking materials of plant, animal or microbial origin. Some food of plant origin
are directly consumed as fresh raw food such as vegetables, fruits etc and some are eaten after
cooking. Most of the animal based foods are consumed after cooking.

42
 Microorganisms serve as food such as single cells proteins or makers of food such as
bread, cake and other fermented food. Due to activity of microorganisms, the nutritive value of
food becomes enriched in terms of more digestibility, increased mineral and vitamin content,
taste, flavour and keeping quality. Most of the plant based food such as grains are available in
dried form for long term preservation and consumption.
The preservation and keeping quality of food and food materials depend mainly on their
composition and water content. Long term preservation or storage of food or food materials
brings changes in their chemical composition and nutritive value. These changes could be due to
physical, chemical or microbiological activities occurring in food or food materials.
Microbiological changes occurring in food or food materials mainly depend on water content,
water activity (aw) and temperature of storage.

Food preservation is the process of physical, chemical or microbiological treatments of

food and beverages to prevent food spoilage and food borne diseases. Food have to be preserved
to increase their shelf life to avoid spoilage by microorganisms and to curtail pollution of land
and water with spoilt food. Food preservation should also take the type of food into consideration
before applying the preservation method. There could be perishable food, semi-perishable food
and non-perishable or stable food.

Methods of food preservation

Foods are preserved by different methods depending on the type of food, its composition
and period of preservation required. Some of the commonly practiced preservation methods
based on above explained principles are given below.

ASEPSIS
It is the process of keeping out microorganisms to avoid spoilage. Food, whether fruits or
vegetables, if in healthy condition, are free from microbes and are contaminated if damaged
during processing or if the protecting external covering is damaged.
The load of microorganisms in food is described as “bioburden”. The type and numbers
of organisms comprising the bioburden influence food spoilage and this also decides the quality
of food.

43
 Asepsis in food industry is important to reduce bioburden of food to avoid spoilage.
Some methods of ‘asepsis’ are packaging of food, hygienic handling and processing of food,
adoption of sanitary methods in slaughtering, handling and processing of meat.
REMOVAL OF MICROORGANISMS
 It is not an effective method of food preservation but helpful during processing of food for
juice and soft drink production.
 The methods followed are filtration, centrifugation, sedimentation, washing and trimming.
 Filtration is used for fruit juices, beer, wine and water. Centrifugation is used for milk and
sedimentation for water.
 Washing and trimming are followed for fruits and vegetables.
 Washing food may be dangerous if the water used is contaminated or has pathogenic
enterobacteriaceae members like E. coli, Salmonella, Shigella, etc.
MAINTENANCE OF ANAEROBIC CONDITIONS
 Food is preserved by sealing, packing, canning to create anaerobic condition. The seals,
packs or cans are completely filled, evacuated of air leading to creation of vacuum or they
are replaced with inert gases like CO2, N2, and H2.

 Anaerobic conditions created prevent growth of aerobes but obligate anaerobes could grow
like Cl. botulinum which is the most dangerous organism as it produces a potent neurotoxin.
 Precautions should be taken to avoid contamination by anaerobes when anaerobic
conditions are used for preservation.
USE OF HIGH TEMPERATURE
Exposure to heat denatures proteins leading to inactivation of enzymes required for
metabolism. The heat treatment necessary to kill microorganisms or their spores varies with the
kind of organism, its physiological state and the environment during heating.
The heat treatment selected depends on kind of organisms to be killed, other preservative
methods to be employed and the effect of heat on food. Certain factors affect the heat resistance
of cells or spores like
a) Initial concentration of cells / spores in food to be heated.
b) Previous history of cells/ spores, that is the conditions under which they have been grown
like culture medium, temperature, growth phage, desiccation etc.
Thermal death time

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 It is defined as the time taken at a given temperature to kill specified number of
microorganisms under defined conditions.
Thermal death point
It is the temperature necessary to kill all microorganisms present in a given substance in
defined time (minutes).
For efficient application of high temperature for preservation of food, the rate of heat
penetration into food should be efficient and this is influenced by material of the container,
consistency, size and shape of contents, initial temperature of food, prior treatments if any etc.
Material of container make like glass has slower rate of penetration than metal.
Consistency of can contents and size and shape of food item, like larger one takes longer
time, smaller one takes shorter time. Food items could retain their identity, lose their form and
become viscous, or layer out.
Initial temperature of food, like lower the initial temperature, longer time of exposure and
higher the initial temperature, shorter the time of exposure required for treatment.
The various degrees of heating used on food can be classified as
(a) pasteurization

(b) heating at 1000 C and

(c) heating above 1000 C

Pasteurization
Pasteurization is a heat treatment that kills part but not all the microorganisms present

and usually involves the application of temperatures below 100 0 C. The heating may be by
means of steam, hot water, dry heat, or electric currents and the products are cooled promptly
after the heat treatment.
Pasteurization is used under these conditions like
 when more rigorous heat treatments might harm the quality of product, as with
market milk,
 when the aim is to kill pathogens, as with market milk,
 when the main spoilage organisms are not very heat resistant, such as the yeasts
in fruit juices, when any surviving spoilage organisms will be taken care of by

45
 additional preservative methods to be employed, as in the chilling of market milk,
and
 when competing organisms are to be killed, allowing a desired fermentation,
usually by added starter organisms, as in cheese making.

Preservative methods used to supplement pasteurization include

(a) refrigeration, e.g., milk,
(b) keeping out microorganisms, usually by packaging the product in a sealed container
as in case of fruit juices,
(c) maintenance of anaerobic conditions, as in evacuated, sealed containers,
(d) addition of high concentrations of sugars in sweetened condensed milk, and
(e) presence or addition of chemical preservatives, e.g., the organic acids on pickles.

Times and temperatures used in the pasteurizing process depend on the method employed
and the product treated. The high-temperature-short-time (HTST) method employs a
comparatively high temperature for a short time whereas the low-temperature- holding (LTH) for
long-time method uses a lower temperature for a longer time.

The minimal heat treatment of market milk is at 72 0C for 16 sec like in the HTST

method; and at 137.80C for at least 2 sec in the ultra pasteurized method. Ice cream mix may be

heated at 71.10C for 30 min or at 82.20 C for 16 to 20 sec.

Grape wines may be pasteurized for 1 min at 82 0 to 850 C in bulk. Dried fruits usually

are pasteurized in the package at 65.60 to 850 C in for 30 to 90 min. The average heat treatment

for carbonated juices would be 65.60 C for 30 min.

Heating at 1000C

 A temperature of approximately 100 0C is obtained by boiling a liquid food, by immersion

of the container of food in boiling water, or by exposure to flowing steam. Some very acid

food, e.g., sauerkraut, may be preheated to a temperature some what below 100 0C,
packaged hot, and not further heat-processed.

46
 Blanching fresh vegetables before freezing or drying involves heating briefly at about

1000C.
 During baking, the internal temperature of bread, cake, or other bakery products approaches

but never reaches 1000C as long as moisture is present, although the oven is much hotter.
The temperature of unsealed canned goods heated in the oven cannot exceed the boiling
temperature of the liquid present.

 Simmering is incipient or gentle boiling, with the temperature reaching about 1000C.

 In roasting meat the internal temperature reaches only about 60 0 C in rare beef, upto 800C

in well-done beef, and 850C in a pork roast.

 Frying gets the outside of the food very hot, but the center ordinarily does not reach

1000C .

Heating at above 1000C

Temperatures above 1000C usually are obtained by means of steam under pressure in
steam-pressure sterilizers (autoclaves) or retorts. With no pressure, the boiling temperature of

water at sea level is 100 0C; with 5 lb of pressure it is 109 0C, with 10 lb it is 115.5 0C, and with

15 lb it is 121.50C. Milk can be heated to temperatures up to 1500C by use of steam injection or

steam infusion followed by “flash evaporation” of the condensed steam and rapid cooling.
USE OF LOW TEMPERATURES
Low temperatures are used to retard chemical reactions and action of food enzymes and
to slow down or stop the growth and activity of microorganisms in food. The lower the
temperature, the slower will be chemical reactions, enzyme action, and microbial growth. A low
enough temperature will prevent the growth of any mesophilic or thermophilic microorganism.
As the temperature drops from its optimal temperature toward the minimal, the rate of
growth of the microorganism decreases and is slowest at the minimal temperature. Cooler
temperatures will prevent growth of an organism but slow metabolic activity may continue.
Therefore, the cooling of food from ordinary temperatures has a different effect on various
organisms present.

47
 In general, freezing prevents the growth of most food-borne microorganisms and
refrigeration temperatures slow growth rates. Commercial refrigeration temperatures, i.e., lower

than 5 to 7.20C, effectively retard the growth of many food-borne pathogens.

One notable exception is Clostridium botulinum type E, which has a minimum

temperature for growth of about 3.30C, Yersinia enterocolitica can survive and grow at

temperatures as low as 0 to 30C. The molds, Cladosporium sp and Sporotrichum sp have been

found to be growing on food at -6.70C and Penicillium and Monilia at -40C. Growth by one

yeast has taken place at -340C, and two others grew at -180C.
Different methods commonly employed in use of low temperature for preservation are
given below:
Common, or cellar storage
The temperature in common, or celler, storage usually is not much below that of the

outside air and seldom is lower than 15 0C. Root crops, potatoes, cabbage, celery, apples, and
similar food can be stored at these temperatures for limited periods.
Chilling or cold storage
Chilling storage is at temperatures not far above freezing and usually involves, cooling by
ice or by mechanical refrigeration. Most perishable food, including eggs, dairy products, meats,
seafood, vegetables, and fruits, may be held in chilling storage (commonly used deep freezers)
for a limited time with little change from their original condition.
Factors to be considered in connection with chilling storage include the temperature of
chilling, the relative humidity, air velocity and composition of the atmosphere in the store room,
and the possible use of ultraviolet rays or other radiations.
Temperature
The lower the temperature of storage, the greater the cost, Therefore, although most food
will be kept best at a temperature just above their freezing point, they are not necessarily stored
at this low temperature. The chilling temperature is selected based on the kind of food, the time
and conditions of storage. Some varieties of apples undergo “ low-temperature breakdown” at

48
temperatures near freezing, and sweet potatoes keep best at 10 to 12.8 0C, and banana cannot be

refrigerated but stored at13.3 to16.70C.

Relative Humidity
Too low a relative humidity results in loss of moisture and hence of weight. It also leads
to the wilting and softening of vegetables and the shrinkage of fruits. Too high a relative
humidity favors the growth of spoilage microorganisms. The highest humidity required for
surface growth of bacteria on food is above 92%, for yeasts it is 90-92% and for molds 85-90%.
Ventilation
Ventilation or control of air velocities of the storage room is important in maintaining a
uniform relative humidity throughout the room, removing odors, and preventing the development
of stale odors and flavors.
Composition of storage atmosphere
The amounts and proportions of gases in the storage atmosphere influence preservation
by chilling. Nowadays increased attention has been given to “gas storage” of food, where the
composition of the atmosphere has been controlled by the introduction of carbon dioxide, ozone
(experimentally) or other gas or the removal of carbon dioxide.
The presence of optimal concentrations of carbondioxide or ozone will influence food
preservation variably like
(i) a food will remain unspoiled for a longer period,
(ii) a higher relative humidity can be maintained without harm to the keeping quality
of certain food,
(iii) a higher storage temperature can be used without shortening the keeping time of
the food that is possible with ordinary chilling storage.
Irradiation
Use of ultraviolet irradiation along with chilling storage helps preserve some food at
higher temperature or by use of higher humidity. Ultraviolet lamps are normally installed in
rooms for storage of meat and cheese.
Freezing or frozen storage

49
 The storage of food in the frozen condition has been an important preservative method
for centuries where outdoor freezing temperatures were available. With the development of
mechanical refrigeration and the quick-freezing processes, the frozen food industry has expanded
rapidly. Even in the home, the freezing of food has become extensive, now that home deep
freezers are readily available.
The lower the storage temperature, the slower will be any chemical or enzymatic
reactions, but most of them will continue slowly at any temperature now used in storage. To
avoid this as a common practice the enzymes in vegetables are inactivated by scalding or
blanching.
The efficiency of freezing as a method of preservation depends on various factors like
selection and preparation of food, method of freezing, condition of food and its method of
harvest etc.

Selection and preparation of food for freezing

The quality of the food to be frozen is of prime importance. Fruits and vegetables are
selected on the basis of their suitability for freezing and their maturity and are washed,
trimmedcut, or otherwise pretreated as desired. Most vegetables are scaled or blanched, and
fruits may be packed in a syrup.
Meats and seafood are selected for quality and are handled so as to minimize enzymatic
and microbial changes. Most food are packaged before freezing. But strawberries, may be frozen
before packaging.

Freezing of food
The rate of freezing of food depends on a number of factors, such as the method
employed, the temperature, circulation of air or refrigerant, size and shape of package, and kind
of food. Sharp freezing usually refers to freezing in air with only natural air circulation or at best
with electric fans.

The temperature is usually -23.3 0C or lower but may vary from -15 to -29 0C, and
freezing may take from 3 to 72 hrs. also termed slow freezing. Quick freezing is variously
defined, but in general implies a freezing time 30 min or less and usually involves the freezing of
small packages or units of food.

50
 Quick freezing is accomplished by one of three general methods:
(i) direct immersion of the food or the packaged food in a refrigerant, as in the
freezing of fish in brine or of berries in special syrups,
(ii) indirect contact with the refrigerant, where the food or package is in contact

with the passage through which the refrigerant at -17.8 to –45.60C flows

(iii) air-blast freezing, where frigid air at -17.8 to -34.4 0C is blown across the
materials being frozen.

Certain fruits and vegetables, fish, shrimp, and mushrooms now are being frozen by

means of liquid nitrogen at -1960C. For dehydro-freezing, fruits and vegetables have about half
their moisture removed before freezing. The condition of the food at the time of freezing will
determine the potential quality of the frozen food. This is in turn dependent on condition of food
at harvesting or slaughter and methods of handling or processing etc.
Changes during freezing
The quick-freezing process rapidly slows chemical and enzymatic reactions in food and
stops microbial growth. The physical effects of freezing are of great importance. There is an
expansion in volume of the frozen food, and ice crystals may form and grow in size. These
crystals usually are larger with slow freezing, and more ice accumulates between tissue cells than
with quick freezing and may crush cells.
Water is drawn from the cells to form such ice, with a resultant increase in the
concentration of solutes in the unfrozen liquor. The increased concentration of solutes in the cells
hastens the salting out, dehydration, and denaturation of proteins and causes irreversible changes
in colloidal systems.
Changes during storage
Meat, poultry, and fish proteins may become irreversibly dehydrated, the red myoglobin
of meat may be oxidized, especially at surfaces, to brown metmyoglobin. Fats of meat and fish
may become oxidized and hydrolyzed. The unfrozen, concentrated solution of sugars, salts etc.,
may ooze from packages of fruits or concentrates during storage as a viscous material called the
metacryotic liquid.

51
 Fluctuation in the storage temperature results in growth in the size of ice crystals and in
physical damage to the food. Desiccation of the food is likely to take place at its surface during
storage. When ice crystals evaporate from an area at the surface, a defect called freezer burn is
produced on fruits, vegetables, meat, poultry, and fish.
Changes during thawing
Thawing is reverse of freezing when temperature is increased. Most of the changes that
seem to appear during thawing are the result of freezing and storage but do not become evident
earlier. When the ice crystals melt, the liquid either is absorbed back into the tissue cells or leaks
out from the food.
Slow, well-controlled thawing usually results in better return to the cells than does rapid
thawing and results in a food more like the original food that was frozen. The pink or reddish
liquid that comes from meat on thawing is called drip, or bleeding, and the liquid oozing from
fruits or vegetables on thawing is termed leakage. The wilting or flabbiness of vegetables and the
mushiness of fruits on thawing are chiefly the result of physical damage during freezing.
Lethal effects of freezing
One of the most widely used techniques for the preservation of cultures is by freezing and
frozen storage, usually in liquid nitrogen. Lethal effects are thought to be the result of
denaturation or flocculation of essential cell proteins or enzymes possibly as a result of the
increased concentration of solutes in the unfrozen water or perhaps in part because of physical
damage by ice crystals.

Repaid cooling of cells from an optimal temperature to 0 0C can also result in death,
referred to as cold shock and is thought to be related to alterations of lipids in the membrane with
damage to the permeability of the cell or to the release of repair enzyme inhibitors, e.g., a
ribonuclease inhibitor.
Response of microorganisms to freezing
Various factors influence microorganisms variably during low temperature preservation.
Some microorganisms die, some are injured, and some are not damaged. The factors that
influence are given below:
1. The kind of microorganism and its state - Resistance to freezing varies with the kind of
microorganism, its phase of growth, and whether it is a vegetative cell or a spore.

52
2. The freezing rate - Faster freezing rates are less destructive than slower rates since critical
range would be passed quickly.
3. The freezing temperature - High freezing temperatures are more lethal. More organisms are

inactivated at -4 to -100 C than at -15 to -300 C.

4. The time of frozen storage - The initial killing rate in freezing is rapid, but later there is
gradual reduction of microorganisms and this is called storage death. Storage of frozen food
in the critical range of temperatures results in more rapid reduction than at higher or lower
freezing temperatures.
5. The kind of food - Composition of the food influences rate of death of organisms during
freezing and storage. Sugar, salt, proteins, colloids, fat, and other substances may be
protective, whereas high moisture and low pH may hasten killing.
6. Alternate freezing and thawing - Alternate freezing and thawing is reported to hasten the
killing of microorganisms but apparently does not always do so.
As the temperature is lowered, more and more water freezes, and the remaining or
unfrozen free water in cells or food items at each temperature becomes more and more
concentrated with solutes (salts, proteins, nucleic acids, etc.) This can change the pH of cellular
matter, concentrate electrolytes, alter colloidal states, denature proteins, and increase viscosity.
Ice crystals can form outside the cell (“extra cellular ice”) and draw water out of the cell with a
resulting dehydration or concentration effect.
Intracellular crystals may form and grow or crystallize right through the cell, resulting in
altered permeability or “holes” in the membrane and cell wall. Intracellular ice is though to be
more harmful to cells than are extra cellular ice crystals.

PRESERVATION BY DRYING
Preservation of food by drying has been practiced for centuries. Drying usually is
accomplished by the removal of water, but any method that reduces the amount of available
water i.e., lowers aw in a food is also a form of drying. For example sugar may be added, as in

sweetened condensed milk will reduce the amount of available moisture.

Moisture may be removed from food by different methods, ranging from the ancient
practice of drying by sun rays to the modern artificial ones like freeze-drying.

53
Drying can be accomplished by various methods.
In sun-dried food, moisture is removed by exposure of food to the sun rays without any
artificially produced heat and without controlled temperatures, relative humidifies, or air
velocities.
A dehydrated or desiccated food has been dried by artificially produced heat under
controlled conditions of temperature, relative humidity, and air flow. Condensed usually implies
that moisture has been removed from a liquid food, and evaporated may have a similar meaning
or may be used synonymously with the term dehydrated. Different methods of drying are solar
drying, mechanical drying, freeze-drying and smoking.
Solar drying
Solar drying is limited to mostly the tropical climates with a hot sun and a dry
atmosphere. It is applicable to certain fruits, such as raisins, prunes, figs, apricots, nectarines,
pears and peaches.
The fruits are spread out on trays and may be turned during drying. Fish, rice and other
grains may also be sun-dried. Many household preparations like papads are also sun-dried.
Drying by mechanical dryers
Most methods of artificial drying involve the passage of heated air with controlled
relative humidity over the food to be dried or the passage of the food through such air. A number
of devices are used for controlled air circulation. The simplest dryer is the evaporator or kiln.
Forced-draft drying systems employ currents of heated air that move across the food, usually in
tunnels.
Liquid food, such as milk, juices, and soups, may be evaporated by the use of
comparatively low temperatures and a vacuum in a vacuum pan or similar device. These food
could also be drum-dried by passage over a heated drum, with or without vacuum or spray-dried
by spraying the liquid into a current of dry, heated air.
Freeze drying
Freeze drying, or the sublimation of water from a frozen food by means of application of
both vacuum and heat is being used for a number of food, including meats, poultry, seafood,
fruits, and vegetables. Frozen thin layers of food of low sugar content may be dried without
vacuum by sublimation of moisture during passage of dry carrier gas.
Smoking

54
 Food items like fish are subjected to smoking which removes moisture and also helps in
preservation as smoke like woods smoke has antimicrobial compounds.
Other Methods
Electronic heating has been suggested for the removal of still more moisture from a food
already fairly well dried. Foam-mat drying, in which liquid food is whipped to foam, dried with
warm air, and crushed to a powder, is significantly being used for most food.
Another method is pressure-gun puffing of partially dried food to give a porous structure

that facilitates and hastens drying. Tower drying is a process in which dehumidified air at 30 0C
or lower is used and has been successful with tomato concentrate, milk, and potatoes.
Factors that influence or control drying
Various factors influence the method of drying as given below:
a) The temperature employed varies with the food and the method of drying taking into
consideration the sensitivity of food to temperature.
b) The relative humidity of the air influences drying and is varied with the food, the method of
drying and also with the stage of drying. It usually is higher at the start of drying than it is at
later stage of drying.
c) The velocity of the air influences by removal of moisture at faster rate if velocity is more and
slower rate if velocity is less.
d) The time of drying determines the extent of dehydration. More the time, more is the drying,
but it again depends on other three factors.
Improper control of these factors may cause case-hardening, resulting from more rapid
evaporation of moisture from the surface than diffusion from the interior, with a resulting hard,
horny, impenetrable surface film that hinders further drying.
Pre-treatments of food before drying
Many of the food need pretreatments before drying for an effective effect of drying
method on the microbial population.
These pretreatments may include:
(a) selection and sorting for size, maturity, and soundness
(b) washing, especially of fruits and vegetables
(c) peeling of fruits and vegetables by hand, machine, lye bath, or abrasion
(d) subdivision into halves, slices, shreds, or cubes

55
 (e) alkali dipping, which is used primarily for fruits such as raisins, grapes, and prunes
(for sun drying) and employs hot 0.1 to 1.5 percent lye or sodium carbonate
(f) blanching or scalding of vegetables and fruits
(g) sulfuring of light-colored fruits and certain vegetables.

Fruits are sulfured by exposure to sulfur dioxide gas produced by the burning of sulfur so
that a level of 1,000 to 3,000 ppm, depending on the fruit, will be absorbed. Vegetables may be
sulfured after blanching in a similar manner or by dipping into or spraying with sulfite solution.
Sulfuring helps maintain an attractive light color, conserve vitamin C and perhaps vitamin A, and
repel insects; it also kills many of the microorganisms present.
Post-treatments of food after drying
The procedures after drying vary with the kind of dried food. They could be sweating,
packing, pasteurization.
Sweating
“Sweating” is a process of storage, usually in bins or boxes, for equalization of moisture
or re-addition of moisture to a desired level. It is used primarily with some dried fruits and some
nuts (almonds, English walnuts).
Packaging
Most food are packaged soon after drying for protection against moisture loss,
contamination with microorganisms, and infestation with insects, although some dried food (e.g.
fruits and nuts) may be held as long as a year before packaging.
Pasteurization
Most dried fruits are subjected to pasteurization to kill pathogens and spoilage organisms.

Fruits in package are treated generally at 65.6 to 85 0C for 30 to70 min at 70-100% relative
humidity.
Changes during the drying process
Heat applied during the drying process causes a reduction in total numbers of
microorganisms, but the effectiveness varies with the kinds and numbers of organisms originally
present and the drying process employed. Usually all yeasts and most bacteria are destroyed, but
spores of bacteria and molds commonly survive, as do vegetative cells of a few specie of heat –
resistant bacteria.

56
 More microorganisms are killed by freezing than by dehydration during the freeze-
drying process. The dried food item remains un-spoilt after drying if storage conditions are
adequate. Microorganisms decrease in number but some resistant forms like spores of bacteria
and molds could survive and also increase, during storage.
PRESERVATION BY RADIATION
Radiations are of various types and are divided into two categories one on each side of
visible light. Low–frequency, long-wavelength, low-quantum-energy radiation ranges from radio
waves to infrared. The effect of these radiations on microorganisms is related to their thermal
agitation of the food. Conversely, the high-frequency, shorter-wavelength radiation have high
quantum energies and actually excite or destroy organic compounds and microorganisms without
heating the product.
Microbial destruction without the generation of high temperatures is referred by the term
“cold sterilization”.
Different types of radiations used in food preservation are
 ultraviolet radiation,
 ionizing radiations like X-rays, gamma rays, beta (ß) rays and
 microwaves.
Ultraviolet irradiation
Ultraviolet irradiation has been the most widely used in the food industry. It is a short
wavelength, less penetrating radiation with wavelengths near 260 nm is absorbed strongly by
purines and pyrimidines and is therefore the most germicidal.
Ultraviolet radiation around 200nm is strongly absorbed by oxygen, this may result in the
production of ozone, and so could be ineffective against microorganisms. The usual source of
UV radiation in food industry is from quartz-mercury vapor lamps, that emit radiation at
[Link] lamps are available in various sizes, shapes and power.
UV radiation induces pyrimidine dimmer formation leading to replication errors followed
by mutations that could be lethal.

Various factors influence the effectiveness of UV radiation such as

Time

57
 The longer the time of exposure to a given concentration, the more effective the
treatment.
Intensity
The intensity of the rays reaching an object will depend on the power of the lamp, the
distance from the lamp to the object, and the kind and amount of interfering material in the path
of the rays.
Penetration
Nature of the object or material being irradiated has an important influence on the
effectiveness of the process. Penetration is reduced even by clear water, which also exerts a
protective effect on microorganisms. Dissolved mineral salts, especially of iron, and cloudiness
greatly reduce the effectiveness of the rays.
Distance
The intensity of destruction of microorganisms falls proportional to the square of the
distance from the lamp.
Action on Microorganisms
Each microorganism has a characteristic resistance to ultraviolet irradiation. This can
vary with the phase of growth and the physiological state of the cell. This is given in the
following table. UV radiation results in damage to the DNA of the cells and death.
Applications of UV radiation in the food industry
Examples of the successful use of these rays include treatment of water used for
beverages, aging of meats, treatment of knives for slicing bread, treatment of bread and cakes,
packaging of sliced bacon, sanitizing of eating utensils, prevention of growth of film yeast on
pickle, vinegar, and sauerkraut vats, killing of spores on sugar crystals and in syrups, storage and
packaging of cheese, prevention of mold growth on walls and shelves, and treatment of air used
for, or in, storage and processing rooms.
Ionizing radiations
Radiation classified as ionizing includes x-rays, gamma rays, cathode or beta rays,
protons, neutrons, and alpha particles. These are called ionizing radiations as they cause
ionization in the material exposed.

Terminology in usage of ionizing radiation

58
  Rad - It is a unit of radiation dosage, being equivalent to the absorption of 100 erg per
gram of irradiated material. A Megarad (Mrad) is 1 million rad, and a Kilorad
(Krad) is 1000 rads.
 Gray(Gy) - It equals 100 rads and is being used as a term to replace rads.
 Radappertization - It is a term used to define "radiation sterilization". The resulting
product is shelf-stable.
 Radurization - It refers to "radiation pasteurization", where low-dose treatments are used
to extend the shelf life of products.
 Radicidation - It is "radiation pasteurization", but used with intention of elimination of a
particular pathogen.
X-Rays
X-rays, gamma rays, and cathode rays are equally effective in sterilization for equal
quantities of energy absorbed. X-rays and gamma rays have good penetration, while cathode rays
have comparatively poor penetration. Though effective, X-rays are not commonly used for
treatment in food preservation because of the harmful effects while handling and cost.
Effects on microorganisms
The bactericidal efficacy of a given dose of irradiation depends on the following:
The kind and species of organism
The lethal doses of radiations are variable for various organisms like for example for
humans it is 0.0056-0.0075 KiloGrays, for yeast 4-9, molds 1-4, bacterial spores 12-20.
The numbers of organisms (or spores) originally present
The more organisms there are, the less effective a given dose will be.
The composition of the food
Some constituents, e.g., proteins, catalase, and reducing substances (nitrites, sulfites, and
sulfhydryl compounds), may be protective.
The presences or absence of oxygen
The effect of free oxygen varies with the organism, ranging from no effect to
sensitization of the organism.
The physical state of the food during irradiation
Both moisture content and temperature affect different organisms in different ways.
The condition of the organisms

59
 Age, temperature of growth and sporulation, and state-vegetative or spore – may affect
the sensitivity of the organisms.
Effect of ionizing radiation
It is supposed that irradiated microorganisms are destroyed by passage of an ionizing
particle or quantum of energy through them, or in close proximity to, a sensitive portion of the
cell, causing a direct “hit” on this target, ionization in this sensitive region, and subsequent death
of the organism (this is called the target theory).
It is assumed also that much of the germicidal effect results from ionization of the
surroundings, especially of water, to yield free radicals, some of which may be oxidizing or
reducing and therefore helpful in destruction of the organisms. Irradiation also may cause
mutations in the organisms present.
Radiation doses, if heavy, have been found to be producing undesirable side reactions in
many food like undesirable colours, odors, tastes etc. Some changes are such as:
(i) Meat - Rise in pH, destruction of glutathione, increase in carbonyl compounds, H 2S,

methyl mercaptan.
(ii) Fats - Destruction of natural antioxidants, oxidation, polymerization, increase in carbonyl
compounds.
(iii) Vitamins of food - Reduction in levels of thiamine, pyridoxin, vitamin B 12, C,D,E & K,

riboflavin and niacin are fairly stable.

Mechanical destruction of microorganisms
The microbes in food items are destroyed by subjecting them to mechanical stress. This
could destroy the cells or lead to their disintegration. The methods followed are grinding, high
pressures etc. They are not generally used at industrial level but followed at domestic levels.
Some examples of this method are grinding of batter for idli, wet milling of malt in beer
preparation, mincing of meat, lysis of cells by application of pressure etc.
Combined methods or Hurdle Technology
Combination of two or more methods could be used to preserve food items as only rarely
a single method is effective. When preservative methods are combined, the required intensity of
each is reduced to less than that when a single method is used like for example, when benzoate
or sorbate is added to fruit juices, less heat is required to sterilize them.

60
 If salt, sugar and vinegar are all added to pickles, sauces, each can be used at a lower
concentration than only if one were added. Food previously irradiated with gamma rays or
treated with antibiotic tylosin require less heat for sterilization than food not so treated.

CHEMICAL PRESERVATIVES AND FOOD ADDITIVES

A food additive is a substance or mixture of substances, other than the basic food stuff,
which is present in food as a result of any aspect of production processing, storage or packaging.
Those food additives which are specifically added to prevent the deterioration or decomposition
of food have been referred to as chemical preservatives.
Preservatives may inhibit microorganisms by interfering with their cell membranes, their
enzyme activity, or their genetic mechanisms. Other preservatives may be used as antioxidants to
hinder the oxidation of unsaturated fats, as neutralizers of acidity, as stabilizers to prevent
physical changes, as firming agents, and as coatings or wrappers to keep out microorganisms,
prevent loss of water, or hinder undesirable microbial enzymatic and chemical reactions.
An ideal anti-microbial preservative should have wide range of microbial activity, should
be non-toxic to consumers, should be economical, should not affect organoleptic properties of
food, should not be inactivated by food, should not promote the growth of resistant strains, and
should kill rather than inhibit microbes.
Factors that influence the effectiveness of chemical preservatives in killing microorganisms or
inhibiting their growth and activity are:
(i) concentration of the chemical,
(ii) kind, number, age, and previous history of the organism,
(iii) temperature,
(iv) time, and
(v) the chemical and physical characteristics of the substrate in which the organism is found
(moisture content, pH, kinds and amounts of solutes, surface tension, and colloids and
other protective substances).

A chemical agent may be bactericidal at a certain concentration, only inhibitory at a lower level,
and ineffective at still greater dilutions.
Antimicrobial preservatives added to food can be grouped as follows:-

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 1. Those added preservatives not defined as such by law - Natural organic acids (lactic,
malic, citric, etc.) and their salts, vinegars (acetic is a natural acid), sodium chloride,
sugars, spices and their oils, woods smoke, carbon dioxide, and nitrogen.
2. Substances generally recognized as safe (GRAS) for addition to food - Propionic acid and
sodium and calcium propionates, caprylic acid, sorbic acid and potassium, sodium, and
calcium sorbates, benzoic acid and benzoates and derivatives of benzoic acid such as
methylparaben and propylparaben, sodium diacetate, sulfur dioxide and sulfites,
potassium and sodium bisulfite and metabisulfite, and sodium nitrite.
3. Chemicals considered to be food additives - These include all not listed in the first two
categories. They can be used only when proved safe for humans or animals, and they then
fall into group 4.
4. Chemicals proved safe and approved by the Food and Drug Administration - Preservatives
added to inhibit or kill microorganisms may be classified on various other criteria, such
as their chemical composition, mode of action, specificity, effectiveness, and legality.
Organic acids and their salts
Lactic, acetic, propionic, and citric acids or their salts may be added to or developed in
food. Citric acid is used in syrups, drinks, jams, and jellies as a substitute for fruit flavors and for
preservation.
Propionates
Sodium or calcium propionate is used most extensively in the prevention of mold growth
and rope development in baked goods and for mold inhibition in many cheese food and spreads.
Propionic acid is a short-chain fatty acid (CH 3CH2COOH) and, like some other fatty acids,

perhaps affects the cell-membrane permeability, although its precise mode of fungistatic action is
not known.
Benzoates
The sodium salt of benzoic acid has been used extensively as an anti-microbial agent in
food. It has been incorporated into jams, jellies, margarine, carbonated beverage, fruit salads,
pickles, relishes, fruit juices, etc. The mechanism of action of the benzoates is not clear; it is
known, however, that the effectiveness of the benzoic acid esters increases with an increase in
the chain length of the ester group.
Sorbates

62
 Sorbic acid, as the calcium, sodium, or potassium salt, is used as a direct anti-microbial
additive in food and as a spray, dip, or coating on packaging materials. It is widely used in
cheeses, cheese products, baked goods, beverages, syrups, fruit juices, jellies, jams, fruit
cocktails, dried fruits, pickles, and margaine. Sorbic acid and its salts are known to inhibit yeast
and molds but are less effective against bacteria.
Acetates
Derivatives of acetic acid, e.g., monochloroacetic acid, peracetic acid, dehydroacetic
acid, and sodium diacetate, have been recommended as preservativesbut not all are approved by
FDA. Acetic acid in the form of vinegar is used in mayonnaise, pickles, catsup, pickled sausages,
and pigs’ feet. Acetic acid is more effective against yeasts and bacteria than against molds.
Nitrites and nitrates
Combinations of these various salts have been used in curing solutions and curing
mixtures for meats. Nitrites decompose to nitric acid, which forms nitrosomyoglobin when it
reacts with the pigments in meats and thereby forms a stable red color. Nitrites can react with
secondary and tertiary amines to form nitrosamines, which are known to be carcinogenic. Recent
work has emphasized the inhibitory property of nitrites towards Clostridium botulinum in meat
products, particularly in bacon and canned or processed hams. Nitrates have a limited effect on
limited number of organisms and would not be considered a good chemical preservative.
Sulfur dioxide and sulfites
Sulfur dioxide and sulfites are used in the wine industry to sanitize equipment and to
reduce the normal flora of the grape must. In aqueous solutions, sulfur dioxide and various
sulfites, including sodium sulfite, potassium sulfite, sodium bisulfite, potassium bisulfite, sodium
metabisulfite, and potassium metabisulfite, form sulfurous acid, the active antimicrobial
compound.
Ethylene and propylene oxide
Ethylene oxide kills all microorganisms, propylene oxide, although it kills many
microorganism, is not as effective as that of ethylene oxide. They are thought to act strong
alkylating agents attacking labile hydrogen. They have also been used successfully in dried
fruits, dries eggs, gelatin, cereals, dried yeast, and spices.
Sugar and salt

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 Sodium chloride is used in brines and curing solutions or is applied directly to the food.
Salt has been reported to have its action like
(a) Causes high osmotic pressure and hence plasmolysis of cells, the percentage of salt
necessary to inhibit growth or harm the cells is varying with the microorganism.
(b) It dehydrates food by drawing out and tying up moisture as it dehydrates microbial
cells
(c) It ionizes to yield the chlorine ion, which is harmful to organisms
(d) It reduces the solubility of oxygen in the moisture
(e) It sensitizes the cell against carbon dioxide and
(f) it interferes with the action of proteolytic enzymes.
Sugars, such as glucose or sucrose, owe their effectiveness as preservatives to their
ability to make water unavailable to organisms and to their osmotic effect. Examples of food
preserved by high sugar concentrations are sweetened condensed milk, fruits in syrups, jellies,
and candies etc.
Alcohol
Ethanol, a coagulant and denaturizer of cell proteins, is most germicidal in concentrations
between 70 and 95 percent. Flavoring extracts, e.g., vanilla and lemon extracts, are preserved by
their content of alcohol.
Formaldehyde
The addition of formaldehyde to food is not permitted, except as a minor constituent of
woods smoke, but this compound is effective against molds, bacteria, and viruses and can be
used where its poisonous nature and irritating properties are not objectionable.
Wood smoke
The smoking of food usually has main purposes like adding desired flavours and
preserving. Woods smoke contains a large number of volatile compounds that may have
bacteriostatic and bactericidal effect. Formaldehyde is considered the most effective of these
compounds, with phenols and cresols next in importance.
Other compounds in the smoke are aliphatic acids from formic through caproic; primary
and secondary alcohols, ketones, and acetaldehyde and other aldehydes; waxes; resins; guaiacol
and its methyl and propyl isomers; and catechol, methyl, catechol and pyrogallol and its methyl

64
ester. Wood smoke is more effective against vegetative cells than against bacterial spores, and
the temperature varies with the kind of wood employed.
Mostly hickory wood is used. Other than that apple, oak, maple, walnut are also used.
Spices and other condiment
Spices and other condiments do not have any marked bacteriostatic effect in the
concentrations customarily used but may help other agents in preventing the growth of
organisms in food. The inhibitory effect of spices differs with the kind of spice and the
microorganism being tested.
Mustard flour and the volatile oil of mustard, for example, are very effective against
Saccharomyces cerevisiae but are not as potent as cinnamon and cloves against most bacteria.
The essential oils of spices are more inhibitory than the corresponding ground spices. Cinnamon
and clove, containing cinnamic aldehyde and eugenol, respectively, usually are more
bacteriostatic than are other spices.
Other food additives
Halogens are added to water for washing food or equipment, for cooling, and for addition
to some products, i.e., washing butter; water for drinking may be chlorinated by the direct
addition of chlorine, or hypochlorites or chloramines may be used. Idoine-impregnated wrappers
have been employed to lengthen the keeping time of fruits.
Iodophors, which are combinations of iodine with non-ionic wetting agents and acid, are
being used in the sanitization of dairy utensils. Halogens kill organisms by oxidation, injury to
cell membranes, or direct combination with cell proteins.
Antibiotics
Most of the better-known antibiotics have been tested on raw food, chiefly proteinaceous
ones like meats, fish, and poultry, in an endeavor to lengthen the storage time at chilling
temperatures. Aureomycin (chlortetracycline) has been found superior to other antibiotics tested
because of its broad spectrum of activity. Terramycin (oxytetracycline) is almost as good for
lengthening the time of preservation of food.

Developed preservatives
Preservatives could be produced in food by microbes like lactic acid, alcohol, bacterixin,
etc. Their preservative effect is mostly supplemented by one or more additional preservative

65
methods like low temperature, high temperature, anaerobic conditions, sodium chloride, sugar
etc. Developed acidity plays a part in preservation of sauerkraut, pickles, green olives, fermented
milk and cheese.

UNIT – II

2 MARKS

1. What is food preservation?

2. Define – asepsis?
3. How remove the microorganisms in food?
4. Anaerobic conditions in food preservation?
5. How affect temperature factor?
6. What is drying?
7. Define radiation?
8. What are chemical preservatives?
9. What are the food additives?
10. Uses and examples for food additives?

5 MARKS

1. Principles of food preservation?

2. Uses of high temperature?
3. Details about the freezing of foods?
4. Preservation by drying?
5. Preservation by radiations?

10 MARKS

1. Discuss about the physical food preservation?

2. Discuss about the chemical food preservation?

66
 UNIT – III

FOOD SPOILAGE
Most foods serve as good growth medium for many different microorganisms.
Considering the variety of foods and the methods used for processing, it is apparent that
practically all kinds of microorganisms are potential contaminants. Given a chance to grow, the
microorganisms will cause changes in appearance, flavor, odour and other qualities of foods.
These degradation processes includes:
Putrefaction:
Protein foods + proteolytic microorganisms amino acids +amines+ ammonia+ H2S.

Fermentation:
Carbohydrate foods +saccharolytic microorganisms organic acids + alcohol + gases.
Rancidity:
Fatty foods + lipolytic microorganisms fatty acids + glycerol.

Microorganisms cause spoilage not only by degradation of foods, but also by synthesis of
various products like pigments and polysaccharides leading to discolorations and formation of
slimes.
CONTAMINATION, PRESERVATION AND SPOILAGE OF CEREAL AND CEREAL
PRODUCTS
Contamination
 Exteriors of harvested grains retain some of the micro-organisms while growing and also
adds contamination from soil, insects and other sources.
 Freshly harvested grains contain a few thousand to millions of bacteria per gram and
several hundred thousand mold spores. Bacteria are mostly in the families
Pseudomonadaceae, Micrococaceae, Lactobacillaceae and Bacillaceae.
 Souring and washing the grains remove some of the microorganisms, but most of the
microorganisms are removed with the outer portions of the grain during milling.
 There is a possibility of contamination during other procedures such as blending and
conditioning. Ex: Wheat flour contains spores of bacillus, coliform bacteria,

67
 Achromabacter, Flavobacterium, Sarcina, Micrococcus, Alcaligenes and Serratia. Fungi
include Aspergillus, Penicillium and Alternaria, Cladosporium.
 Cornmeal and flour contain 100 to several thousand bacteria and molds per gram.
Fusarium and Penicillium are the dominant molds.
 The surface of a freshly baked loaf of bread is practically free of viable micro organisms
but they subject to contamination by mold spores from the air during cooling and before
wrapping.
 During slicing, contamination may take place from micro organisms in the air, on the
knives or on the wrapper.
 The contamination of grains and cereal products with molds has become a significant
concern because of the presence of aflatoxins. (Produced by Aspergillus flavus and
Aspergillus parasiticus).
Preservation
Most cereals have low aW (water activity). Storage temperature of about 4.4 to 7.2°C is
recommended for the dry products.
Asepsis
Adequate cleaning and sanitization of equipment is essential for preservation. Improperly
sanitized equipment may be source of rope bacteria, acid forming bacteria that causes sourness
of doughs. Bread, cakes and other baked goods that may be subject to spoilage by molds should
be protected against contamination by mold should be protected against contamination by mold
spores.
Use of Heat
Bakery products which are unbaked, partially backed, fully baked. The complete baking
process ordinarily destroys all bacteria cells, yeasts, and mold spores, but not spores of the rope
forming or other bacteria. Mold spores in prober cloths in bakeries can build up enough heat
resistance to survive baking unbaked or partially baked products usually are kept on the retailers
shelf for only a short period or kept cool during longer storage. Ex: Boston brown bread and nut
bread have been successfully canned.
Use of Low Temperatures
Ordinary room temperature may be used by homemakers for short term storage of baked
goods. Bakery goods can be stored in freezers. Unbaked or partially baked products, waffles,

68
cheese cake, ice cream pie and fish, poultry and meat pies are usually frozen. Bread and rolls can
be stored successfully for months in the frozen condition.
Use of chemical preservatives
Ammonia, propionic acid are used for preservation of bakery products. 2 % Ammonia
and propionic acid (1%) reduce mold growth in high moisture corn. Sodium and calcium
propionate, sodium diacetate and sorbates are used extensively. Acidification of the dough with
acetic acid has been used to combat rope.
Use of Irradiation
U.V rays have been used to destroy or reduce numbers of mold spores in dough and proof
rooms, on the knives of slicing machines. Gamma and cathode rays have been applied
experimentally for the preservation of baked goods. Low level irradiation can also be used to
destroy insects in stored grains.
SPOILAGE
Cereal grains and meals
Cereal grains and meals are not processed to reduce their natural flora of microorganisms
greatly, they are likely to contain molds, yeasts and bacteria. These microorganisms readily grow
if moisture increases. Unavailable to many organisms, these grains contain some sugar and
available nitrogen compounds, minerals and accessory growth substances.
Molds will grow on the surface, where air is available. A wet mash of the grains or a
mash of the meals will undergo an acid fermentation, chiefly by the lactic acid and coliform
bacteria normally present on plant surfaces.
Ex: Aspergillus, Penicillium, Mucor, Rhizopus and Fusarium.
Flours
Dry cleaning and washing grains and the milling and sifting of flour reduce the content of
microorganisms. White wheat flour usually bleached by an oxidizing agent such as oxide of
nitrogen, chlorine, nitrosyl chloride, benzoyl peroxide. Moisture content of flour less than 13%
has been reported to prevent the growth of all micro organisms. 15% moisture permits growth of
mold growth. 17% moisture permits growth of bacteria & molds. Acid forming bacteria Ex:
Acetobacter sps permits acetic acid fermentation.
Bread

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 Acid fermentation by lactics and coliform bacteria that is normal in flour pastes or dough
may be too extensive if too much time is permitted resulting that the dough and bread made from
it may be too ‘sour’.

Mold / Moldiness
Molds are most common and most important cause of the spoilage of bread and most
bakery products. Temperatures attained in the baking procedure usually are high to kill mold
spores in and on the loaf, so that molds must reach the outer surface or penetrate after baking.
Contamination may come from air during cooling, handling, from wrappers and initiate
growth in the crease of load and between the slices. Chief molds involved in the spoilage of
bread are Bread mold – Rhizopus stolonifer or Rhizopus nigricans bread mold contains white
cottony mycelium and black dots of sporangia. Green spores are produced by Penicillium
expansum Penicillium stoloniferum, Aspergillus [Link] or purplish-brown to black
conidial heads and yellow pigment diffusing into the bread. Monilia sitophila – produces pink
conidia give a pink or reddish color to its growth. Species of Mucor or Geotrichum also
contaminate the bread and produces spores.
Mold spoilage is favored by
1. Heavy contamination after baking due to air heavily laden with mold spores, a long
cooling time, considerable air circulation or a contaminated slicing machine.
2. Slicing in that more air is introduced into the loaf.
3. Wrapping, especially if the bread is warm when wrapped.
4. Storage in a warm & humid place.
Rope / Ropiness
Ropiness of bread is fairly common in home baked bread, especially during weather.
Ropiness is uncommon in commercially baked bread, because of the preventive measures now
employed. Ropiness caused by mucoid variant of Bacillus subtilis, Bacillus licheniformis, (B.
mesentericus), B. panis spores of these bacteria can withstand the temperature of the bread
during baking, which does not exceed 100°C. Ropy condition due to capsulation of bacillus,
together with hydrolysis of the flour proteins (gluten) by proteinases of the organism and starch
by amylase.
i)Red bread

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 Red bread or bloody bread is due to the occurrence of red color from the growth of
pigmented bacteria Serratia marcescens. In ancient times, appearance of red color was
considered [Link] conditions favor growth. Molds such as Monilia sitophila may
impart red color to bread. Red color in the crumb of dark bread has been caused by Geotrichum
aurantiacum (syn, Oidium aurantiacum)
ii) Chalky bread
Chalky bread is due to white, chalk like spots. It is due to the yeast like fungi
Endomycopsis fibuligera and Trichosporon variable.
Cakes & Bakery products
Molds are the chief cause of spoilage of cakes and other bakery products. Topping or
fillings are more prone to microbial spoilage than actual baked portion. Frostings due to high
sugar content spoiled by molds or yeast upon storage. The deterioration of breads, cakes, pies is
referred to as staling and it is due to due to mostly by physical changes during holding and not by
micro organisms.
Pasta, Macroni and Tapioca
Pasta is egg based product containing flour, water and eggs, pasta is delivered and stored
dry. Macaroni usually contains only flour, water and other nutrients. Swelling of moist macaroni
has been reportedly caused by gas production by bacteria resembling Enterobacter cloacae.
Purple streaks are produced by Monilia. Tapioca prepared from the root starch of cassava will
spoiled by orange pigmented, starch hydrolyzing bacterium.
Breakfast cereals and other cereal snacks
Breakfast cereals are flaked, puffed or extruded. In the initial steps of manufacturing of
these products, there are high levels of moisture and therefore the possibility of microbial
growth. Finished products contain low number of micro organisms.
Prepared doughs
These products contains a yeast or a lactic acid bacteria inoculum. Microbial
contamination depends on quality of ingredients used and sanitary practices employed.

CONTAMINATION AND SPOILAGE OF FRUITS AND VEGETABLES

Contamination

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  Fruits and vegetables are gathered into boxes, lugs, baskets, trucks during harvesting and
they are subject to contamination with spoilage organisms. From each other and form the
containers unless these have been adequately sanitized.
 Mechanical damage during transportation to market may increase susceptibility to decay
and growth of microorganisms may take place. Precooling of the product and
refrigeration during transportation will slow such growth.
 Soaking, washing by agitation tend to distribute spoilage organisms from damaged to
whole foods. Recirculated or reused water is likely to add organisms and the washing
process may moisten surfaces enough to permit growth of organisms during a holding
period. Washing with detergent or germicidal solutions will reduce numbers on the foods.
 Sorting spoiled fruits or vegetables or trimming spoiled parts removes microorganisms,
but additional handling may result in mechanical damage. The spraying gives a fresh
appearance to the vegetables and delays decomposition but also adds organisms Ex:
Psychrotrophs
 In the processing plant, the fruits & vegetables are subjected to further contamination.
Adequate washing at the plant causes a reduction in numbers of micro organisms peeling
by steam, hot water or lye and blanching reduces micro organisms.
 Sweating of products during handling increases microbial numbers. Equipments or
instruments may add microorganisms during processing.
 Wooden surfaces are difficult to clean and sanitize and therefore are especially likely to
be sources of contamination. Hot water blanching reduces total numbers of organisms on
the food, but may cause the building of spores of thermophilic bacteria, causing the
spoilage of canned foods Ex: Flat sour spores in peas.
 Inclusion of decayed parts of fruits increases the numbers of micro organisms in fruit
juices. Added ingredients such as sugar and starch may add spoilage organisms,
especially spores of thermophilic bacteria.
Preservation of vegetables
Asepsis
Limited amount of contamination of vegetables takes place between harvesting and
processing or consumption i.e., gross contamination can be avoided. Boxes, lugs, baskets and

72
other containers should be practically free of the growth of micro organisms and some will need
cleaning and sanitation.
Vegetables if they contact with spoilage vegetables. Heat resistant spores of spoilage
bacteria may be present.
Ex: Flat sour bacteria, putrefactive anaerobes (Clostridium thermosaccharolyticum)
Removal of micro organisms
Washing of vegetables removes most contaminants on the surface but leaves natural
microbial surface flora. Wash water is of good bacteriological quality, it should not add
organisms. Chlorinated water is used for washing. Part of the mold growth on strawberries can
be removed by washing with a nonionic detergent solution.
Use of heat
Vegetables to be dried or frozen and some to be canned are scalded or blanched to
inactivate their enzymes. 1000 to 10,000 fold are decreased.
Use of Low temperatures
Root crops, potatoes, cabbage, celery can be preserved for a limited time by cellar
storage.
i) Chilling
Most vegetables are cooled and kept at chilling temperature before preservation. Chilling
is accomplished by use of cold water, vaccum cooling used for lettuce.
i) Hydrocooling
Cooling before normal cold storage is done immediately after harvesting by use of a cold
water spray, a practice referred to as hydrocooling. Freshening of leafy vegetables by a water
spray will cool the products and help in preservation. Potatoes turn sweet at temperatures below
2.2 – 4.4°C. Sweet potatoes and onions are subjected to special curing treatments before storage.
iii) Freezing
Freezing process reduces the number of organisms. Micrococcus are predominant on
thawing vegetables such as sweet corn and peas. Achromobacter and Enterobacter spp are also
commonly present. Lactobacillus are also common on peas under such conditions.
Drying
Dried vegetables and vegetable products are used in dried soups. Dried spices and
condiments are used as flavoring materials.

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Explosive puffing
Many vegetables can be dried by a process called explosive puffing. Usually small pieces
of the diced, partially dehydrated vegetables are placed in a closed rotating chamber. Heat is
applied and the chamber is pressurized to a pre determined level, then the pressure is released
instantaneously. It results in additional loss of water, but more important, a porous network of
capillaries is formed in the product. The increased porosity simplifies further drying and imports
good reconstituting ability.
Use of preservatives
Rutabagas and turnips sometimes are paraffined to lengthen their keeping time. Zinc
carbonate has been reported to eliminate most mold growth on lettuce, beets, and spinach.
Biphenyl vapors will control Fusarium on potatoes. Controlled atmosphere of CO2 or ozone
about chilled vegetables have little practical use.
Added preservative
Sodium chloride is the only added chemical preservative in common use. The amount
added to vegetables may vary from the 2.25 – 2.5% (Ex: Sauerkraut). Lower concentrations of
salt permit an acid fermentation by bacteria to take place, as the percentage of salt is increased,
the rate of acid production becomes slower until a level of salt is reached that will permit no
growth or production of acid.
Preservation by irradiation
Treatment with gamma rays to inactivate micro organisms causing decay, followed by
storage has resulted in discoloration, softening or other deterioration of most vegetables.
Irradiation used successfully to delay spouting of potatoes, onions and garlic and to kill insect
reproduction on some vegetables.
Preservation of fruits and fruit products
Asepsis
Care should be taken to avoid contamination during harvesting, processing and from
containers. Before harvest, fruits are usually exposed to insecticides and fungicides may have
their flora altered by such treatments.
Removal of Micro organisms

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 Washing – Removes dirt, micro organisms and poisonous sprays. Washing may be with
water, detergent solutions, bactericidal solutions such as chlorinated water. Trimming also
removes micro organisms. Clear fruit juices may be sterilized by filtration.
Use of Heat:
Fruits seldom are blanched before other processing because blancing causes excessive
physical damage. On the basis of their pH fruits are divided into
Acid foods High acid foods
Tomatoes, pears and pine apples Berries Steam pressure sterilizer is not required for most
fruits, since heating at about 100°C is sufficient and can be accomplished by flowing steam or
boiling water . More acid the fruit, less heat required for its preservation.
Use of low temperatures
Apples can be preserved for a limited time in common or cellar storage.
Chilling
Each fruit has its own optimal temperature and relative humidity for chilling storage.
Fruits are treated with hypochlorites, sodium bicarbonate, borax, propionates, biphenyl, Ophenyl
phenols, sulfur dioxide, thiourea, thiabendazole, dibromo tetrachloro ethane before or during
storage.
Fruits also have been enclosed in wrappers treated with chemicals Ex: Sulfite paper on
grapes, iodine paper on grapes and tomatoes, borax paper on oranges. Waxed wraps, paraffin oil,
paraffin, waxes and mineral oil have been applied for mechanical protection.
CA Storage
It implies the altering of various gases from normal atmospheric concentrations; usually
this is done by increasing the CO2 concentration and decreasing the O2 concentration. MA
storage (modified atmosphere storage) used to describe CA conditions which are not accurately
maintained or conditions where the air is initially replaced with gas but no further measures are
taken to keep the gas atmosphere constant.
Gas storage means CA or MA storage.
Under certain circumstances only one gas is used. Ex: packaging a product in 100% N2.
Optimal concentrations of CO2 and O2 vary with kind and variety of fruits. CO2 storage
employed with apples, pears, bananas, citrus fruits, plums, peaches, grapes and other fruits.

75
 Ozone in concentrations of 2 to 3 ppm in the atmospheres has been reported to double the
storage time of loosely packed small fresh fruits, such as straw berries, raspberries, currants and
grapes and delicate varieties of apples.
Ethylene in the atmosphere is used to hasten ripening or produce a desired color change
and it is not considered preservative, although a combination of this gas and activated
hydrocarbons has been suggested for the preservation of fruits.

Freezing
During preparation of fruits for freezing, undesirable changes may take place such as
darkening, deterioration in flavor and spoilage by micro organisms especially molds. Washing
the fruit removes most of the soil micro organisms some fruits are frozen in large drums.
Drying
Number of microorganisms in dried fruits are comparatively low and that spores of
bacteria and molds are likely to be the most numerous. Alkali treatment, sulfuring, blanchig and
pasteurization reduce numbers of micro organisms.
Use of preservatives
Chemicals applied to the fruits chiefly as dips or spray or impregnated in wrappers for the
fruits among substances that have been applied to the outer surfaces of fruits are waxes,
hypochlorites, biphenyl and alkaline sodium O- phenylphenate. Wrappers for fruits have
impregnated with a variety of chemicals include iodine, sulfite, biphenyl, O-phenylphenol plus
hexamine and others.
SPOILAGE
Deterioration of raw vegetables and fruits due to
1. Physical factors
2. Action of their own enzymes
3. Microbial action
4. Combinations of above
5. Mechanical damage due to
6. Action of animals, birds, insects.
7. Wounding
8. Bursting

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 9. Freezing
10. Dessication
11. Other mishandling
Plant pathogen infection may takes place and unfit for consumption. Improper
environmental conditions during harvesting, transit, storage and marketing may favor spoilage.
Plant enzymes continue their activity in raw plant foods. If O2 is available, the plant cells will
respire as long as they are alive and hydrolytic enzymes can continue their action after death of
the cell.
General types of microbial spoilage:
The most commonly occurring types of spoilage are as follows:
1. Bacterial soft rot: Caused by Erwinia caratovora. These are fermenters of pectins.
Pseudomonas marginalis and Clostridium, Bacillus sps have also been isolated from these rots.
It results in a water soaked appearance, a soft mushy consistency and often a bad odor.
2. Gray mold rot: Botrytis cinerea. Botrytis name derived from the gray mycelium of the mold.
It is favored by high humidity and a warm temperature.
3. Rhizopus soft rot: It is caused by sps of Rhizopus Ex: Rhizopus stolonifer. This rot is often
soft and mushy. The cottony growth of the mold with small, black dots of sporangia often covers
masses of the foods.
4. Anthrachose: Usually caused by Collectotrichum lindemuthianum, Collectotrichum
coccodes, defect is spotting of leaves and fruits or seed pods.
5. Alternaria Rot: Caused by Alternaria tenuis. Areas become greenish brown early in the
growth of the mold and later turn to brown or black spots.
6. Blue mold rot: Caused by Penicillium digitatum. Bluish green color spores are produced.
7. Downey mildew: Caused by Phytopthora bremia.
8. Watery soft rot: Caused chiefly by Sclerotinia sclerotiorum. Found in mostly vegetables.
9. Stem end rots: Caused by Diplodia, Alternaria, Phomopsis, Fusarium.
10. Black mold rot: Caused by Aspergillus niger. Dark brown to black masses of spores of the
mold termed “Smut” called by layperson.
11. Black rot: Caused by Alternaria, Ceratostomella, Physalospora.
12. Pink mold rot: Caused by pink spored Trichothecium roseum.
13. Fusarium rots: Caused by Fusarium sps.

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14. Green mold rot: Caused by Cladosporium, Trichoderma.
15. Brown rot: Caused chiefly by Sclerotinia (Monilinia fructicola) sps.
16. Sliminess or souring: Caused by Saprophytic bacteria in piled, wet, heating vegetables.

CONTAMINATION, PRESERVATION, SPOILAGE OF EGGS

CONTAMINATION OF EGGS
 Most freshly laid eggs are sterile, at least inside, but the shells soon become contaminated
by fecal matter from the hen, by the cage or nest, by wash water if the eggs are washed, by
handling, and perhaps by the material in which the eggs are packed.
 The total number of microorganisms per shell of a hen's egg has been reported to range
from 102 to 107 with a mean of about 105.
 The types of microorganisms recovered from the shell are diverse. Salmonella sps may be
on the shell or in the egg as laid, build up during processing, and appear in significant
numbers in frozen or dried eggs.
Preservation
The egg has various ways of protecting itself from microbial invasion. The shell and the
thin surface layer of proteinaceous material known as the cuticle or bloom are the first line of
defense and serve to retard entry. However, the shell is porous for gas exchange during
embryonic development. The membranes inside the shell also tend to serve as a mechanical
barrier. Albumen is an inadequate growth medium for many microorganisms. The characteristics
of albumen that hinder microbial growth include a pH of 9 to 10 that may be reached during
storage. Low levels of simple nitrogenous compounds like apoprotein ties up riboflavin, avidin
ties up biotin, antiproteolytic factor prevent bacterial proteinases from releasing nitrogenous
compounds necessary for growth, conalbumin (ovotransferrin) chelates iron and lysozyme is an
enzyme which degrades the cell walls of gram-positive bacteria.

Asepsis
Care should be taken to reduce the contamination of the outside of the shell by hen feces
and dirt from the nests. When the eggs are broken for drying or freezing care is taken to discard
those in which microbial growth has taken place. Reduce contamination from equipment by
cleaning and sanitizing it.

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Removal of microorganisms
Dirty eggs command a lower price than clean ones, various methods have been tried for
the removal of soil. Dry cleaning, as by sandblasting, removes dirt and also the bloom (mucin).
Washing with warm, plain water removes dirt, the bloom, and part of the microorganisms
but encourages the penetration of bacteria into the egg through the pores in the shell.
Cleaning disinfecting the washing machines with 1 % hypochlorite solution reduces
contamination with rot bacteria. The use of 1 to 3% acetic acid was effective in removing the
flora but resulted in a reduction of shell thickness and egg quality. Lye, acids, formalin,
hypochlorites, quaternary ammonium compounds, various detergents, and detergent-sanitizer
combinations have been tried in washing solutions.
Use of heat
The heat coagulability of the egg white determines to a great extent the maximal heat
treatment that can be given shell eggs. Rotting was well controlled by heating at 60 0C for 320
seconds. Heating in water was superior to heating in oil. A thermostabilization method of
dipping eggs into hot water reduces evaporation of moisture from the egg by a slight coagulation
of the outermost part of the egg albumen. Pasteurization is required for most egg products.
Pasteurization is not done for salt yolk for use in salad dressings.
Use of low temperatures
Chilling
Most shell eggs are preserved by chilling. They are selected for storage on the basis of
their general appearance and the result of candling.

Candling
To candle an egg, it is held and rotated in front of a light to examine it for defects such as
cracks, rots, molds, blood, developing embryo, crusted or sided yolk, weak white, or large air
cell. The eggs should be cooled after production and held at a temperature and relative humidity
that depend on the anticipated time of storage.
Freezing
The egg meats whole or separated, are filtered to remove pieces of shell and stringy
material (chalazae), mixed or churned, standardized as to solids content, and frozen in 30- or 50-
lb tin cans or other containers, usually by a sharp-freezing process. Yolks frozen separately and

79
stored frozen form a jelly which does not become fluid on thawing. The frozen eggs are stored at
-17.8 to -20.50C. Frozen eggs may contain high numbers of bacteria, up to millions per gram
from contamination of egg pulps, contamination from pieces of shell and from equipment in the
breaking room, and growth before freezing

Preservation by drying
Most of the drying of eggs done by spray dryer, where the liquid is sprayed into a current
of dry, heated air. Another method is the roller or drum process, in which the liquid egg is passed
over a heated drum, with or without vacuum. Air drying is accomplished by means of open pans,
as used by the Chinese, or by the belt system, where the egg liquid is on a belt that passes
through a heated tunnel (60 to 71.10 C). Spray drying or pan drying, combined with tunnel
drying, is used for egg white. Formerly, eggs were dried to a moisture of about 5 percent, but the
keeping quality of dried white or whole egg is improved as the moisture content is decreased
toward 1 percent, and the trend is in that direction.
Use of preservatives
Preservatives may be used on the shells of eggs, in the atmosphere around them, or on
wraps or containers for eggs. An enormous number of different substances have been applied to
the surface of the shells of eggs or used as packing material about eggs to aid in their
preservation.
Materials used for the dry packing of eggs in the home include salt, lime, sand, sawdust,
and ashes. Immersion in water glass containing solution of sodium silicate. It is a successful
home method of preservation. The solution is inhibitory because of its alkalinity. Other
inhibitory chemicals that have been tried are borates, permanganates, benzoates, salicylates,
formates.
Use of Irradiation
Salmonella in liquid, frozen, and dried eggs can be inactivated by means of ionizing
radiation.
Spoilage of Eggs
Defects in the fresh egg

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 Fresh eggs may exhibit cracks, leaks, loss of bloom or gloss, or stained or dirty spots on
the exterior as well as "meat spots" (blood clots), general bloodiness, or translucent spots in the
yolk when candled.
Changes during storage
The changes that take place in eggs while they are being held or stored may be divided
into those due to non microbial causes and those resulting from the growth of microorganisms.
Changes Not Caused by Microorganisms
1. Untreated eggs lose moisture during storage and hence lose weight. The amount of
shrinkage is shown to the candler by the size of the air space or air cell at the blunt end of
the egg, a large cell indicating much shrinkage.
2. The change in the physical state of the contents of the egg, as shown by candling or by
breaking out the egg.
3. The white of the egg becomes thinner and more watery as the egg ages, and the yolk
membrane becomes weaker.
4. When an old egg is broken onto a flat dish, the thinness of the white is more evident
and the weakness of the yolk membrane permits the yolk to flatten out or even break. By
contrast, a broken fresh egg shows a thick white and a yolk that stands up strongly in the
form of a pemisphere.
5. During storage, the alkalinity of the white of the egg increases from a normal pH of
about 7.6 to about 9.5. Any marked growth of the chick embryos in fertilized eggs also
serves to condemn the eggs.
Changes Caused by Microorganisms
To cause spoilage of an undamaged shell egg, the causal organisms must do the following
(1) contaminate the shell,
(2) penetrate the pores of the shell to the shell
(3) grow through the shell membranes to reach the white
(4) grow in the egg white, despite the previously mentioned unfavorable conditions there,
to reach the yolk, where they can grow readily and complete spoilage of the egg.
In general, more spoilage of eggs is caused by bacteria than by molds. The types of
bacterial spoilage, or "rots," of eggs go by different names.

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1. Green rots, caused chiefly by Pseudomonas fluorescens, a bacterium that grows at 00C; the
rot is so named because of the bright-green color of the white during early stages of
development. This stage is noted with difficulty in candling but shows up clearly when the egg is
broken. Odor is lacking or is fruity or "sweetish." The contents of eggs so rotted fluoresce
strongly under ultraviolet light.
2. Colorless rots, which may be caused by Pseudomonas, Acinetobacter, Alcaligenes, certain
coliform bacteria, or other types of bacteria. These rots are detected readily by candling, for the
yolk usually is involved, except in very early stages, and disintegrates or at least shows a white
incrustation. The odor varies from a scarcely detectable one to fruity to "highly offfensive."
3. Black rots, where the eggs are almost opaque to the candling lamp because the yolks become
blackened and then break down to give the whole-egg contents a muddy-brown color. The odor
is putrid, with hydrogen sulfide evident, and gas pressure may develop in the egg. Species of
Proteus most commonly cause these rots, although some species of Pseudomonas and
Aeromonas can cause black rots. Proteus melanovogenes causes an especially black coloration in
the yolk and a dark color in the white. The development of black rot and of red rot usually means
that the egg has at some time been held at temperatures higher than those ordinarily used for
storage.
4. Pink rots: Pink rots occur less often, and red rots are still more infrequent. Pink rots are
caused by strains of Pseudomonas. They resemble the colorless rots, except for a pinkish
precipitate on the yolk and a pink color in the white.
5. Red rots: Red rots caused by species of Serratia, are mild in odor and are not offensive.
Spoilage of Eggs by Fungi
The spoilage of eggs by fungi goes through stages of mold growth that give the defects
their names.
1. Pin spot molding
Very early mold growth is termed pin-spot molding because of the small, compact
colonies of molds appearing on the shell and usually just inside it. The color of these pin spots
varies with the kind of mold. Penicillium species cause yellow or blue or green spots inside the
shell, Cladosporium species give dark-green or black spots, and species of Sporotrichum produce
pink spots.
1. Superficial fungal spoilage

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 In storage atmospheres of high humidity a variety of molds may cause superficial fungal
spoilage, first in the form of a fuzz or "whiskers" covering the shell and later as more luxuriant
growth. Molds causing spoilage of eggs include species of Penicillium, Cladosporium,
Sporotrichum, Mucor, Thamniidium, Botrytis, Alternaria, and other genera.
2. Fungal rotting
The final stage of spoilage by molds is fungal rotting, after the mycelium of the mold has
grown through the pores or cracks in the egg. Jellying of the white may result, and colored rots
may be produced, e.g., fungal red rot by Sporotrichum and a black color by Cladosporium, the
cause of black spot of eggs as well as of other foods.
3. Off-flavors:
Off-flavours sometimes are developed in eggs. Mustiness may be caused by any of a
number of bacteria, such as Achromobacter perolens, Pseudomonas graveolens, and P.
mucidolens. The growth of Streptomyces on straw or elsewhere near the egg may produce musty
or earthy flavors that are absorbed by the egg. Molds growing in the shell also give musty odors
and tastes. A hay odor is caused by Enterobacter cloacae. Fishy flavors are produced by certain
strains of Escherichia coli.

DAIRY AND FERMENTED FOODS

YOGURT

Yogurt is fermented milk product characterized by viscous consistency, acidulous taste

(pH 4.6) and distinct aroma – acetaldehyde

Spoilage – bloom cartons / frothy consistency and yeasty off flavor, odour

Prevention

Sterilization of filling equipment, careful in storage and packaging, use of LAF & UV in filling
areas & periodic fumigation, control of spillages, use of heat and preservatives in final products

Production

Whole milk / skim milk + water

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 Pasteurization (85°C / 30 minutes batch process). (90 - 95°C / 10 minutes continuous process)

Homogenize (60-65°C) for smooth texture

Addition of emulsifiers (agar, gums, alginate to increase the viscosity)

Add sweetners (5% sugar inhibit lactic acid production)

Heat (90-95°C) and cool

Inoculate starter culture (Streptococcus, Lactobacillus)

Incubate (4-16 hours at 30- 45°C) and cool (10 – 15 °C)

Add fruits & flavoring agents

Packaging (maintain at 4-5°C for 2 weeks)

Nutritive value of yoghurt

Fermented milk, minerals remain unchanged while proteins, carbohydrates, vitamins and fats
are subjected to change, substance formed are lactic acid, alcohol, CO2, antibiotics and vitamins.

i) Proteolysis- breakdown of proteins into peptides, peptones and amino acids, Essential
amino acids such as leucine, isoleucine, methionine, phenyl alanine, tyrosine,
tryptophan and valine
ii) Hydrolysis of lactose- lactose hydrolysed into lactic acid it inhibits the growth of
putrifactants
iii) Lipolysis- reduces the size of globules (ie) fats into free fatty acids
iv) Changes in vitamins- 2 folds increase in vitamin B such as thiamine, riboflavin and
nicotinamide
v) Antibacterial activities- materials responsible for antibiotic properties are lactic acid,
hydrogen peroxide and lactobacilline

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 vi) Therapeutic properties- improves appetite, high digestive capacity relief to chronic
diarrhea and ulcerative colitis, potent antitumour activity, remove fat from liver and
enhance bile secretion.

BUTTER

Butter is a dairy product that consists of butterfat, milk proteins, and water. It is made by
churning fresh or fermented cream or milk. It is used as a spread and a condiment—and in
cooking, such as baking, sauce making, and pan frying.

Commonly made from cows' milk, butter can also be manufactured from the milk of
other mammals, including sheep, goats, buffalo, and yaks. Producers sometimes add salt,
flavorings, or preservatives. Rendering butter produces clarified butter or ghee, which is almost
entirely butterfat.

Butter is a water-in-oil emulsion that results from an inversion of the cream, an oil-in-
water emulsion. The milk proteins are the emulsifiers. Butter remains a solid when refrigerated,
but softens to a spreadable consistency at room temperature, and melts to a thin liquid
consistency at 32–35 °C (90–95 °F). The density of butter is 911 g/L. It generally has a pale
yellow color, but varies from deep yellow to nearly white.

Nutritional content

As butter is essentially just the milk fat, it contains only traces of lactose, so moderate
consumption of butter is not a problem for the lactose intolerant.

It is a good source of Vitamin A equivalent.

Production

Cream separation (unhomogenized milk) was pasteurized at 88 - 93°C cooled to 16-21°C

Starter culture ( Streptococcus lactis, Streptococcus cremoris – acid producers, Streptococcus

diacetylactis, Leuconostoc – flavor producers)

85
 Churning (adding colour)braining buttermilk,

Adding salt (40 – 60 %), washing, packed and Stored at 5°C

Unhomogenized milk and cream contain butterfat in microscopic globules. These

globules are surrounded by membranes made of phospholipids and proteins, which prevent the
fat in milk from pooling together into a single mass. Butter is produced by agitating cream,
which damages these membranes and allows the milk fats to conjoin, separating from the other
parts of the cream.

Butter contains fat in three separate forms: free butterfat, butterfat crystals, and
undamaged fat globules. In the finished product, different proportions of these forms result in
different consistencies within the butter; butters with many crystals are harder than butters
dominated by free fats.

Almost all commercially-made butter begins with pasteurized cream, which is commonly
heated to a relatively high temperature above 80 °C (180 °F). Before it is churned, the cream is
cooled to about 5 °C (40 °F) and allowed to remain at that temperature for at least eight hours;
under these conditions about half the butterfat in the cream crystallizes.

Churning produces small butter grains floating in the water-based portion of the cream.
This watery liquid is called buttermilk—although the buttermilk most common today is instead a
directly fermented skimmed milk. The buttermilk is drained off; sometimes more buttermilk is
removed by rinsing the grains with water. Then the grains are "worked": pressed and kneaded
together. When prepared manually, this is done using wooden boards called scotch hands. This
consolidates the butter into a solid mass and breaks up embedded pockets of buttermilk or water
into tiny droplets.

Types

Cultured butter

Butter made from a fermented cream is known as cultured butter. During fermentation,
the cream naturally sours as bacteria convert milk sugars into lactic acid. The fermentation

86
process produces additional aroma compounds, including diacetyl, which makes for a fuller-
flavored and more "buttery" tasting product.

Cultured butter is usually made from pasteurized cream whose fermentation is produced
by the introduction of Lactococcus and Leuconostoc bacteria.

Butter is produced from fresh cream and then incorporate bacterial cultures and lactic
acid. Using this method, the cultured butter flavor grows as the butter is aged in cold storage. For
manufacturers, this method is more efficient, since aging the cream used to make butter takes
significantly more space than simply storing the finished butter product. A method to make an
artificial simulation of cultured butter is to add lactic acid and flavor compounds directly to the
fresh-cream butter; while this more efficient process is claimed to simulate the taste of cultured
butter, the product produced is not cultured but is instead flavored.

Sweet cream butter

Dairy products are often pasteurized during production to kill pathogenic bacteria and
other microbes. Butter made from pasteurized fresh cream is called sweet cream butter.
Production of sweet cream butter first became common in the 19th century, with the
development of refrigeration and the mechanical cream separator.

Raw cream butter.

Butter made from fresh or cultured unpasteurized cream is called raw cream butter. While
butter made from pasteurized cream may keep for several months, raw cream butter has a shelf
life of roughly ten days.

Clarified butter

Clarified butter is butter with almost all of its water and milk solids removed, leaving
almost-pure butterfat. Clarified butter is made by heating butter to its melting point and then
allowing it to cool; after settling, the remaining components separate by density. At the top,
whey proteins form a skin, which is removed. The resulting butterfat is poured off from the
mixture of water and casein proteins that settle to the bottom.

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 Ghee is clarified butter that is brought to higher temperatures of around 120 °C (250 °F)
after the water evaporates, so the milk solids brown. This flavors the ghee, and also produces
antioxidants that help protect it longer from rancidity. Because of this, ghee can keep for six to
eight months under normal conditions.

Cream can be skimmed from whey instead of milk, as a byproduct of cheese-making.

Whey butter can be made from whey cream. Whey cream and butter have a lower fat content and
taste more salty, tangy and "cheesy." They are also cheaper than "sweet" cream and butter.

Spoilage
Butter readily absorbs volatile materials from the air. Microorganisms in the butter
can cause the following defects:
1. Surface taint, also called "rabbito" and "putridity," which is blamed on Pseudomonas
putrefaciens, introduced usually by the wash water, churns, or equipment. It is worse in unsalted
or low-salt butter. The "sweaty-feet" odor is due chiefly to isovaleric acid.
2. Fishiness, caused by Aeromonas hydrophila.
3. Esterlike flavors, resulting from the action of P. fragi.
4. Skunk like flavors, caused by P. mephitica.
5. Roquefort like flavors, produced by molds.
Chemically produced flavors include
(1) Rancidity produced by lipase in the cream,
(2) Tallowiness from oxidations of unsaturated fats catalyzed by copper and bacterial
enzymes and favored by a low pH, low-temperature pasteurization, salt, air, and ozone,
and
(3) Fishiness, where trimethylamine is produced from lecithin. This defect is favored by
high acidity, salt, overworking of the butter, and the presence of copper.

CHEESE

Cheese is a food derived from milk that is produced in a wide range of flavors, textures,
and forms by coagulation of the milk protein casein. It comprises proteins and fat from milk,
usually the milk of cows, buffalo, goats, or sheep. During production, the milk is usually

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acidified, and adding the enzyme rennet causes coagulation. The solids are separated and pressed
into final form. Some cheeses have molds on the rind or throughout. Most cheeses melt at
cooking temperature.

For a few cheeses, the milk is curdled by adding acids such as vinegar or lemon juice.
Most cheeses are acidified to a lesser degree by bacteria, which turn milk sugars into lactic acid,
then the addition of rennet completes the curdling. Vegetarian alternatives to rennet are
available; most are produced by fermentation of the fungus Mucor miehei.

Cheese is valued for its portability, long life, and high content of fat, protein, calcium,
and phosphorus. Cheese is more compact and has a longer shelf life than milk, although how
long a cheese will keep may depend on the type of cheese; labels on packets of cheese often
claim that a cheese should be consumed within three to five days of opening. Generally, hard
cheeses last longer than soft cheeses, such as Brie or goat's milk cheese.

Cheese makers near a dairy region may benefit from fresher, lower-priced milk, and
lower shipping costs. The long storage life of some cheese, especially if it is encased in a
protective rind, allows selling when markets are favorable.

Production

Raw milk

Pasteurize at 71 - 75° C for 15 seconds

Cool and incubate in cheese vat (add starter culture Lactobacillus lactis)

Milk with 0.19 – 0.21%lactic acid (rennet added)

Curd formation

Cutting of curd

Whey release

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 Scalding 38 - 40° C and stirring (acid production)

Cheddaring – squeezing and stretching the curd (lactic acid 0.6 – 0.8% - primary
mrtabolism)

Milling and salting (add flavor and further release of whey)

Moulding and Pressing

Ripening for 2 – 24 months (secondary metabolism – proteinase enzyme released from

starter culture produce amino acid, indole, sulphur and phenol, hydrogen peroxide and
bacteriocin)

A required step in cheesemaking is separating the milk into solid curds and liquid whey.
Usually this is done by acidifying (souring) the milk and adding rennet. The acidification can be
accomplished directly by the addition of an acid, such as vinegar, in a few cases, but usually
starter bacteria are employed instead. These starter bacteria convert milk sugars into lactic acid.
The same bacteria also play a large role in the eventual flavor of aged cheeses. Most cheeses are
made with starter bacteria from the Lactococci, Lactobacilli, or Streptococci families.

Curd processing

The cheese has set into a very moist gel. Some soft cheeses are now essentially complete:
they are drained, salted, and packaged. For most of the rest, the curd is cut into small cubes. This
allows water to drain from the individual pieces of curd.

Some hard cheeses are then heated to temperatures in the range of 35–55 °C (95–131 °F).
This forces more whey from the cut curd. It also changes the taste of the finished cheese,
affecting both the bacterial culture and the milk chemistry. Cheeses that are heated to the higher
temperatures are usually made with thermophilic starter bacteria that survive this step—either
Lactobacilli or Streptococci.

Salt has roles in cheese besides adding a salty flavor. It preserves cheese from spoiling,
draws moisture from the curd, and firms cheese’s texture in an interaction with its proteins.

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Some cheeses are salted from the outside with dry salt or brine washes. Most cheeses have the
salt mixed directly into the curds.

Other techniques influence a cheese's texture and flavor. Some examples are :

 Stretching: The curd is stretched and kneaded in hot water, developing a stringy, fibrous
body.
 Cheddaring: The cut curd is repeatedly piled up, pushing more moisture away. The curd
is also mixed (or milled) for a long time, taking the sharp edges off the cut curd pieces
and influencing the final product's texture.
 Washing: The curd is washed in warm water, lowering its acidity and making for a
milder-tasting cheese.

Most cheeses achieve their final shape when the curds are pressed into a mold or form.
The harder the cheese, the more pressure is applied. The pressure drives out moisture the molds
are designed to allow water to escape and unifies the curds into a single solid body.

Ripening

A newborn cheese is usually salty yet bland in flavor and, for harder varieties, rubbery in
texture. These qualities are sometimes enjoyed—cheese curds are eaten on their own—but
normally cheeses are left to rest under controlled conditions. This aging lasts from a few days to
several years. As a cheese ages, microbes and enzymes transform texture and intensify flavor.
This transformation is largely a result of the breakdown of casein proteins and milkfat into a
complex mix of amino acids, amines, and fatty acids.

Processed cheeses

Processed cheese is made from traditional cheese and emulsifying salts, often with the
addition of milk, more salt, preservatives, and food coloring. It is inexpensive, consistent, and
melts smoothly. It is sold packaged and either pre-sliced or unsliced, in a number of varieties. It
is also available in aerosol cans in some countries.

Eating and cooking

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 At refrigerator temperatures, the fat in a piece of cheese is as hard as unsoftened butter,
and its protein structure is stiff as well. Flavor and odor compounds are less easily liberated
when cold. For improvements in flavor and texture, it is widely advised that cheeses be allowed
to warm up to room temperature before eating.

Above room temperatures, most hard cheeses melt. Rennet-curdled cheeses have a gel-
like protein matrix that is broken down by heat. When enough protein bonds are broken, the
cheese itself turns from a solid to a viscous liquid. Soft, high-moisture cheeses will melt at
around 55 °C (131 °F), while hard, low-moisture cheeses such as Parmesan remain solid until
they reach about 82 °C (180 °F). Acid-set cheeses, including halloumi, paneer, some whey
cheeses and many varieties of fresh goat cheese, have a protein structure that remains intact at
high temperatures. When cooked, these cheeses just get firmer as water evaporates.

As its temperature continues to rise, cheese will brown and eventually burn. Browned,
partially burned cheese has a particular distinct flavor of its own and is frequently used in
cooking.

SPOILAGE

During manufacturing

 Off flavor – Clostridium, Coliforms, yeasts

 Gassiness - Clostridium,Bacillus polymyxa (produce gas and defects in ripening)
 Bitter flavor – Coliforms, Micrococci, yeasts (acid proteolytic bacteria)
Leuconostoc – produce holes / open in cheddar cheese
 Sliminess – Pseudomonas fragi, Alcaligenes

During ripening

 Physical changes- hole formation, change nature of texture

 Chemical changes – undesirable end product, metal discoloration
 Gas holes / eyes / cracks / splitting

SPOILAGE FLAVOURED MILK:

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 Milk is an excellent culture medium for many kinds of microorganisms, being high in
moisture, nearly neutral in pH, and rich in microbial foods. A plentiful supply of food energy is
present in the form of milk sugar (lactose), butterfat, citrate, and nitrogenous compounds in
many forms (proteins, amino acids, ammonia, urea, and other compounds), and the accessory
foods and minerals required by microorganisms are available. Some inhibitory substances
(lactoperoxidase and agglutinins) are present in freshly drawn milk but soon become
comparatively ineffective.
Gas Production
Gas production by bacteria usually is accompanied by acid formation and it is
undesirable in milk and milk products. The chief gas formers are the coliform bacteria,
Clostridium spp., gas forming Bacillus species that yield both hydrogen and carbon dioxide.
Yeasts, propionics, and hetero fermentative lactics produce only carbon dioxide.
Proteolysis
The hydrolysis of milk proteins by microorganisms usually is accompanied by the
production of a bitter flavor caused by some of the peptides released.
Proteolysis is favored by
1. Storage at a low temperature.
2. by the destruction of lactic and other acid formers by heat.
3. By the destruction of formed acid in the milk by molds and film yeasts.
4. The neutralization of acids by products of other organisms.
The types of change produced by proteolytic microorganisms include
(1) Acid proteolysis, in which acid production and proteolysis occur together.
(2) Proteolysis with little acidity or even with alkalinity.
(3) Sweet curdling, caused by rennin like enzymes of the bacteria at an early stage of
proteolysis.
(4) Slow proteolysis by intracellular enzymes of bacteria after their autolysis.
(5) Residual proteolytic activity of heat-stable proteinase.
Ropiness
Ropiness and sliminess can occur in milk, cream, or whey but are important mostly in
market milk and cream.
Nonbacterial ropiness or sliminess may be due to

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 (1) Stringiness caused by mastitis and in particular by fibrin and leukocytes from the
cow's blood (in contrast to ropiness produced by bacteria, it is present when the milk is
drawn, not developed during holding of the milk).
(2) Sliminess resulting from the thickness of cream, e.g., at the top of a bottle.
(3) Stringiness due to thin mms of casein or lactalbumin during cooling, as sometimes is
observed on surface coolers. This effect is only temporary.

Bacterial ropiness is caused by slimy capsular material from the cells, usually gums or
mucins, and develops best at low storage temperatures. The ropiness decreases as the acidity of
the milk or cream increases.
Surface ropiness is caused most often by Alcaligenes viscolactis, an organism chiefly
from water or soil that can grow fairly well in the vicinity of 100C. Some of the thermoduric
microococci, e.g., Micrococcus freudenreichii, can cause surface ropiness.
Alkali Production
The group of alkali formers includes bacteria which cause an alkaline reaction in milk
without any evidence of proteolysis. The alkaline reaction may result from the formation of
ammonia, as from urea, or of carbonates, as from organic acids such as citric acid. Examples of
alkali formers are Pseudomonas fluorescens and A. viscolactis.
Flavor Changes
As drawn from the cow, milk may be abnormal in flavor because of the individual cow, mastitis,
the stage of lactation of the cow, or feed. Some of the off-flavors caused by microorganisms are
described as follows.
1. Sour or Acid Flavor: The acidity may be described as "clean," as produced by Streptococcus
lactis and other lactic. "aromatic;" when lactic streptococci and aroma-forming Leuconostoc
species are growing together. "sharp," when appreciable amounts of volatile fatty acids (formic,
acetic, or butyric) are produced by coliform bacteria, Clostridium spp., and other organisms.
2. Bitter Flavors: Bitterness usually results from proteolysis but may follow lipolysis or even
fermentation of lactose. Milk from cows late in their lactation period sometimes is slightly bitter.
Other organisms causing bitterness are certain strains of coliform bacteria and of asporogenous
yeasts. Some cocci cause very bitter milk. Actinomycetes sometimes give bitter musty flavors.

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3. Burnt or Caramel Flavor: Certain strains of Streptococcus lactis var. maltigenes produce
this flavor, which resembles the cooked flavor of overheated milk.
4. Miscellaneous Other Flavors
 A barny flavor by Enteroobacter oxytocum
 Soapiness by ammonia formers like Pseudomonas sapolactica
 A turnip like flavor by Escherichia coli and [Link]
 A malty flavor by yellow micrococci from the udder
 Fruity flavor by P. fragi
 A potato like flavor by P. mucidolens,
 Fishiness by Aeromonas hydrophila or various cocci that produce trimethylamine from
lecithin
 Earthy or musty flavors by actinomycetes
 Fruity, ester like, and alcoholic flavors by yeasts
 An amyl alcohol flavor by white and orange micrococci
 Putrefaction by species of Clostridium, P. putrefaciens, and other putrefactive bacteria.
Other flavors have been termed unclean, stale, astringent, oily, weedy, carroty, etc.
Inadequately cooled raw milk that has been held in a tightly covered can so that volatile products
of bacterial metabolism have collected above the milk has an undesirable odor that varies in
nature. Milk in this condition is termed smothered.
Color Changes
Color changes caused by microorganisms may occur along with other changes. The color
may be due to the surface growth of pigmented bacteria or molds in the form of a scum or ring or
may be present throughout the milk.
1. Blue Milk: Pseudomonas syncyanea produces a bluish-gray to brownish color in milk in pure
culture but when growing with an acid former like Streptococcus lactis causes a deep-blue color.
2. Yellow Milk: Pseudomonas synxantha may cause a yellow color in the cream layer of milk,
coincident to lipolysis and proteolysis. Species of Flavobacterium can also give yellowness.
3. Red Milk: Red milk usually is caused by species of Serratia, e.g., S. marcescens, but is rare
because other bacteria ordinarily outgrow the red-pigmented species. Brevibacterium
erythrogenes produces a red layer at the top of the milk, followed by proteolysis. Micrococcus
roseus may grow and produce red sediment, and yeast may produce pink or red colonies on the

95
surface of sour milk or cream. Blood in milk will give it a red color. The red blood cells settle
out or can be centrifuged out.
4. Brown Milk: A brown color may result from Pseudomonas putrefaciens or by the enzymic
oxidation of tyrosine by P. fluorescens.

MICROBIAL SPOILAGE OF CANNED FOODS

Introduction
 Canning is one of important method of packaging food for long term storage.
Normally food is stored in metallic containers along with heat treatment. The
heat treatment varies depending upon type of food. There is always a chance
that microorganisms may survive if the heat treatment is not proper thereby
leading to spoilage of food. Usually the incidences of food spoilage in cans are
low. The spoilage of can could be due to biological or chemical reasons or
combination of both. The biological spoilage is primarily due to microbial
growth while chemical spoilage is due to hydrogen produced due to reaction of
acid in food and iron on can. The degree of swelling can also be increased by
high summer temperature and high altitudes. Certain other factors such as
overfilling, buckling, denting or closing the can while cool may also be
responsible for spoilage of foods in cans.
 Chemical spoilage
 The chemical spoilage in most cases is due to production of hydrogen gas
produced in can because of action of acid of food on iron of can. This spoilage
is termed as Hydrogen swell. It occurs due to following factors:
 a) Increased storage temperature.
 b) Increased acidity of food
 c) Improper exhaust
 d) Presence of soluble sulfur and phosphorous compounds
 e) Improper timing and lacquering of can at internal surfaces
 Biological spoilage
 The cause of biological spoilage is microbial activity. In heat treated cans, the
growth of microorganisms occur due to:
Leakage of can
 It occurs because of manufacturing defects, punctures or rough handling.
Bacteria are introduced into can by either in holes or improper seams. In this
type, the microorganisms are not usually heat resistant and wide array of
organisms had been found to cause spoilage as it is post processing
contamination. Microbes may also get entry into can due to cold water, used to
cool cans after heat treatment. Leakage may also be responsible for release of
vacuum, which can favor the growth of microorganisms. Presence of low heat
resistance organisms usually indicates leakage of can.

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 Under processing
 It includes sub-optimal heat treatment, faulty retort operations, excessive
microbial load and contamination in product, change in consistency of the
product.
 Stages in Appearance of Can
 A can undergo different transformations from being a normal can to completely
spoilt can as it depend upon various factors. All these stages are described
based on appearance of can from outside.

 Microbial Spoilage of Canned Foods

 The microbial spoilage of canned food is classified as caused by thermophilic
bacteria and mesophilic organisms. Most common spoilages of microbial origin
are known as flat sour spoilage, Thermophilic anaerobic (TA) spoilage and
putrefaction. These different types are briefly described here.
 Flat sour spoilage
 This is caused by souring bacteria. One characteristic of this spoilage is that
ends of can remain flat during souring. Because of this condition, the detection
of spoilage from outside is not possible thereby culturing of contents become
necessary to detect the type of organisms. Main organisms involved
are Bacillus, while it occurs more frequently in low acid foods. Bacillus spp.
has ability to produce acid without gas formation.
 TA spoilage
 This type of spoilage is caused by thermophilic anaerobe not producing
hydrogen sulfide. Clostridium thermosaccharolyticum is the main organism
involved. It produces acid and gas in foods. Spoiled food produces sour or
cheesy smell.
 Sulfur stinker spoilage
 This type of spoilage occurs in low acid foods and primarily Desulfotomaculum
nigricans is involved. The spores of these organisms are destroyed at optimal
heat treatment, thus presence of this organism usually indicates under
processing in terms of heat treatment. It produces hydrogen sulfide which
produce typical odour.
 Spoilage By Mesophillic Spore formers
 Bacillus and Clostridium are involved in this type of spoilage which is usually
indicative of under spoilage.
 Spoilage by Non-Spore Formers
 Presence of non spore formers in cans indicate post processing contamination.
The organisms whose vegetative cells are heat resistant are more readily found.
Following organisms are more prominent:

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  Presence of these organisms indicates leakage of container. Cooling water is
one of the important source of contamination, thus coilforms also gain entry
into the can through leakage.
 Spoilage by Fungi
 Yeasts
 Yeasts and their spores are not thermo tolerant, thus they are not found in
suitably heat treated cans. Their presence indicates under processing or post
pasteurization contamination through leakage. Fermentative yeasts are more
prominent and they produce carbon dioxide, thus causing swelling of cans.
Film yeasts too can grow on the surface of the food products.
 Molds
 Among molds, Aspergilus and Penicillium are most spoiling organisms. These
can grow at high sugar concentration. Acidification is considered method of
preventing growth of molds. Some of the molds are resistant to heat. Molds are
more common in home canned foods where heating as well as sealing is not
under total aseptic conditions.
CONTAMINATION, PRESERVATION, SPOILAGE OF FISH AND OTHER SEA
FOODS
Sea foods include fresh, frozen, dried, picked and salted fish as well as various shell fish
and fresh water.
Contamination
 Microbial flora of living fish depends on the water in which they live. Slime on the outer
surface of fish contain Pseudomonas, Acinetobacter, Moraxella, Alcaligenes,
Micrococcus, Flavobacterium, Corynebacterium, Sarcina, Serratia, Vibrio and Bacillus.
 Bacteria on fish from northern waters are mostly psychrophilies, where as fish from
tropical water plus species of Aeromonas, Lactobacillus, Brevibacterium, Alcaligenes and
Streptococcus.
 Intestines of fish from both sources are found bacteria of the genera Alcaligenes,
Pseudomonas, Flavobacterium, Vibrio and Bacillus, Clostridium and Escherichia. Boats,
boxes, bins, fish houses and fishers soon become heavily contaminated with these
bacteria.
 Oyster and other shell fish contain Alcaligenes, Flavobacterium, Moraxella,
Acinetobacter. These fish thrush water bodies and pick up soil and micro organisms.
 Number of micro organisms on the skin of fish can be influenced by the method of
catching. Trawling fish nets along the bottom for long periods results in exposure of the
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 fish to high bacterial cants in the sediment. Fish cakes and fish sticks may have
contamination from potatoes, spices and flavorings etc.
Preservation
Of all the flesh foods, fish is the most susceptible to autolysis, oxidation and hydrolysis
of fats and microbial spoilage.
Use of Heat
Live crabs are cooked in retorts at temperatures up to 121°C to facilitate removing the
meat from the shell. Hand picking and hand packing of the meat is common. Processing times
and temperatures for canned crabmeat range from 85.6 to 87.2°C for 92 – 150 min. These
processes are considered pasteurization and cans are preserved by refrigeration.
Use of Low Temperature
Chilling
Fish flesh autolyzes and the fats become oxidized at temperatures above freezing rapidly
at summer temperatures and more slowly as the temperature is dropped toward freezing.
Preservation by chilling is temporary. Necessity of chilling is required from the catching area to
processing plant. Small fish are more perishable than whole dressed fish autolyze more slowly
than whole fish but are spoiled more readily by bacteria.
Freezing
In earlier days ice with added salt was employed. Now mechanical refrigeration includes
sharp freezing and the fish were “glazed” i.e., a layer of ice was frozen around the outside.
Whole fish – usually sharp frozen in air or in salt brine. Quick freezing is applied to wrapped
fillets or steaks. Quick frozen fish may thaw to like their original condition than fish frozen more
slowly. During storage the fats of frozen fish are subject to hydrolysis and oxidation. Fatty fish
deteriorate more rapidly than lean ones.
Use of Irradiation
Gamma or cathode irradiation applied to some kinds of fish. U.V rays has been tried but
not in practice.
Preservation by Drying
Dry salting of fish or immersion in brine constitutes a method of drying. In this, moisture
is removed or tied up. Oxidation of fish oils is not retarded and may cause deterioration. Salt cod

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is prepared by a combination of salting and air drying. Flesh is removed from bones and skin.
Smoking of fish results in drying of the fish.
Use of preservatives
Salting or marination of fish by dry salt or in brine is effective not only because of the
drying effect mentioned but also because of the effect of sodium chloride. Fish may be drysalted
so as to contain 4-5% salt. Salts contributes halophiles which may discolor the fish. Ex: Red
colour due to Serratia salinaria. Micrococcus sps may grow on the fish and decrease in
flavobacterium, Alcaligenes, pseudomonas and others.
Benzoic acid and benzoated are moderately successful.
Sodium and potassium nitrites and nitrates lengthens the keeping time. These chemicals
are permitted in some countries only.
Sorbic acid delays spoilage of smoked or salted fish.
Boric acid has been used in Europe with some improvement in the keeping quality, but
use of boric acid is illegal in united stated.
Other chemicals used as preservatives but very limited are formaldehyde, hypochlorited,
hydrogen peroxide, sulfer oxide, undecylenic acid, capric acid, p – oxybenzoic acid and
chloroform.
Antibiotics also have been used as dip or in ice. Chlortetracycline and oxytetracycline are
best and their use is permitted.
Preservatives incorporated in ice:
Germicidal ices are prepared by adding a chemical preservative to water before freezing.
Eutectic ice Non eutectic ice
When the added chemical is uniformly distributed throughout. Ex: Nacl
When the added chemical not uniformly distributed throughout. Ex: Sodium benzoate
Antioxidants:
Fats and oils of fatly fishes (Ex: Herring, mackerel, mullet and salmon) are comosed of
unsaturated fatty acids and hence are subject to oxidative changes producing oxidative rancidity
and sometimes undesirable alterations in colour. To counteract these undesirable changes,
antioxidants may be applied as dips, coatings, glazes or gases. Good results have been reported
with nordihydroguaiaretic acid, ethyl gallate, ascorbic acid and other compounds and with
storage in carbon dioxide.

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SPOILAGE
Fish spoiled by autolysis, oxidation or bacterial activity or combination of these. Fish
flesh is more perishable than meat because autolysis by the fish enzymes and because of the less
acid reaction of fish flesh that favors microbial growth.
Factors influencing kind and rate of spoilage
1. The kind of fish
Flat fish spoil more readily than round fish because they pass through rigor mortis more
rapidly. But flat fish like halibut keeps longer because of the low PH (5.5) of its flesh. Certain
fatty fish deteriorate rapidly because of the oxidation of the unsaturated of their oils.
2. Condition of the fish when cought
Fish that are exhausted as the result of struggling, lack of oxygen and excessive handling
spoil more rapidly. “Feedy fish” i.e., those full of food when caught, are more perishable than
those with an empty intestinal tract.
3. The kind and extent of contamination of the fish flesh with bacteria
Contamination occurs from mud, water, handlers, exterior slime and intestinal content of
the fish. Contamination takes place in the net, fishing boat on the docks etc.

4. Temperature
Chilling the fish is the most commonly used method for preventing or delaying bacteria
growth and hence spoilage until the fish is used or otherwise processed. Cooling should be rapid
as possible 0 to -1°C and this low temperature should be maintained.

Evidences of spoilage
The change is gradual from a fresh condition to staleness and then to inedibility it is
difficult to determine or agree on the first appearance of spoilage. Estimate of volatile acids or
volatile bases or a test for pH, H2S, ammonia etc. are used. Bacteriological tests are too slow to
be useful..
Bacteria causing spoilage
Bacteria involved in the spoilage of fish are natural flora of the external slime of fishes
and their intestinal contents. Predominant kinds of bacteria causing spoilage vary with the
temperatures at which the fish are held, but at the chilling temperatures usually employed, sps of

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pseudomonas are most likely to predominate, with Acinetobacter, Moraxella and
Flavobacterium sps. Microccus and Bacillus are less often appears.
A musty or muddy odor and taste of fish has been attributed to the growth of
Streptomyces sps in the mud at the bottom of the body of water and absorption of the flavor by
the fish.
Discolorations of the fish flesh may occur by Pseudomonas flourescens, yellow
micrococci and others.
`Red or pink color produce from growth of Sarcina, Micrococcus, Bacillus or by molds
or yeasts.
Chocolate brown color occurs by asporogenous yeast. Pathogens parasitizing the fish
may produce discolorations or lesions.
Spoilage of special kinds of fish and sea foods:
 Salt fish are spoiled by salt tolerant or halophilic bacteria (Serratia, Micrococcus, Bacillus,
Alcaligenes, Pseudomonas and others). These bacteria cause discolorations, a red color
being common. Molds are the chief spoilage organisms on smoked fish. In chilled shrimps,
Acinetobacter, Moraxella and Vibrio are chief responsible for spoilage. Crabmeat
deteriorated by Pseudomonas, Acinetobacter, Proteus at higher temperatures.
 Raw lobsters are spoiled by pseudomonas, Alcaligenes and Flavobacterium.
 Crabs and oysters may contain sps of Vibrio including Vibrio parahaemolyticus.
 Oysters remain in good conditions as long as they are kept alive in the shell at chilling
temperature, but they decompose rapidly when they are dead, as in shucked oysters.

UNIT – III

2 MARKS

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 1. What is food spoilage?
2. What is Rancidity?
3. What is ropiness?
4. What is explosive puffing?
5. What are black rots?
6. What is curing?
7. Define – Putrefaction?
8. Define – Taint?
9. What is Ham?
10. Antioxidants in spoilage of fish?

5 MARKS

1. Examples for food spoilage causal organisms?

2. Contamination and spoilage of fruits?
3. Preservation and spoilage of flavored milk?
4. Contamination of youghurt?
5. Spoilage of Cheese?

10 MARKS

1. Discuss in details about the cereal product?

2. Write in detail notes about the sea foods?
3. Discuss in detail about the canned foods?

UNIT – IV
THE FOOD SAFETY AND QUALITY ASSURANCE

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Food Processing program is a two-semester, full-time graduate certificate program designed for
students having a previous degree or diploma in science, food or engineering, and who want to
be prepared for a career in the food processing industry in the area of food safety.
The program offers a suite of courses designed to enhance skills and knowledge gained in
previous education stages for direct applicability in the food processing environment with respect
to food safety and quality assurance.
The Food Safety and Quality Assurance program is unique in Ontario because students apply
their learning in Conestoga's pilot plant facility, enabling learners to incorporate theoretical
knowledge in quality assurance and food safety to the practical environment of food
manufacturing.

Quality assurance (QA) is a set of activities for ensuring quality in the processes by which
products are developed. It’s a proactive process and aims to prevent defects by concentrating on
the process used to make the product. The goal of QA is to improve development and test
processes so that defects don’t arise when the product’s being developed.

QA can be achieved by establishing a good quality management system and assessing its
adequacy. What’s more, everyone on the team involved in developing a product is responsible
for quality assurance.

Rigorous quality assurance and quality control processes are implemented to supply safe food
and beverage products to the consumer. Produce must adhere and be compliant with the national
regulatory guidelines and limits, for example the United States has the Food and Drug
Administration (FDA), Europe has the Food Safety Authority (EFSA) and China has the Food
and Drug Administration (CFDA). Each regulatory body require zero tolerance in the strict
levels of consented contaminants allowed in a food product.
MICROBIAL TESTING OF FOODS
The examination of foods for the presence of microorganisms is one of the important
areas of food microbiology. Number of bacteria in foods is an indicator of their handling,
processing and ultimately quality. The various types of methods used can be grouped broadly
into two classes: Conventional methods and Rapid methods. The conventional methods include
cultural and microscopic methods, physical methods, chemical methods and the immunological
methods, while the rapid methods includes the use of DNA based methods.
DYE REDUCTION METHOD (MBRT)

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 Dye reduction test involve the use of redox dyes like methylene blue to determine the
quality of milk. Methylene blue is reduced and loses its color in the presence of actively growing
bacteria. The time taken for the reduction of methylene blue is inversely proportional to the
number of viable bacteria. The shorter the methylene blue reduction time poorer is the quality of
the milk. The advantages of the method are as follows:
1. It is a simple and rapid method for grading bulk supplies of raw milk;
2. It is an inexpensive method;
3. No skilled personnel are needed to perform the test.
LITMUS MILKTEST
Litmus milk is a complex medium that can produce a variety of results. Because of this,
litmus milk can give quite unreliable results.
Introduction
Litmus milk is a complex medium that can potentially distinguish among many species of
bacteria. Litmus milk has several components that can be metabolized:lactose (milk sugar);
casein (milk protein); and litmus (a pH indicator that is purple to blue at neutral to alkaline pH
and pink under acidic conditions).If lactose is fermented, the solution should turn [Link] gas is
produced during fermentation, you may be able to see bubblesor cracks in the milky medium
(but this is often difficult to observe).If lactose is not fermented and proteins are metabolized
instead, the solution will become alkaline and turn blue in color. If casein is digested, the milk
will coagulate to form a curd (a solid).Casein may be metabolized all the way down to individual
amino acids. This process, called peptonization, results in a clear (not milky) liquid that is
usually brown in color. Finally, the litmus may be reduced and become colorless. The culture
will then look milk white.
Methods
1. Obtain two Litmus Milk broths from the back shelf.
2. Inoculate one broth with cells from your stock unknown culture.
Leave the other broth uninoculated (this will act as a negative control).
3. Incubate both broths at an appropriate temperature (which ever is a better growth temperature
for your organism).
4. Observe your cultures over the next several days. Changes should be visible in five to seven
days.

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Limitations

1. Litmus milk is a complex medium that can produce a diversity of results. Because of this,
litmus milk can give quite unreliable results (i.e., the results often give negative results
when something positive should happen). Thus, you would be advised to use litmus milk
as a confirmatory test but not a definitive test (except as a last resort).
2. A clot formation is simply recorded as “clot” and cannot clearly differentiate between a
clot and curd formation in this medium.

ALKALINE PHOSPHATASE

Alkaline phosphatase is a hydrolase enzyme responsible for removing phosphate groups

from many types of molecules, including nucleotides, proteins, and alkaloids. The process of
removing the phosphate group is called dephosphorylation. As the name suggests, alkaline
phosphatases are most effective in an alkaline environment. It is sometimes used synonymously
as basic phosphatase.

Bacterial

In Gram-negative bacteria, alkaline phosphatase is located in the periplasmic space,

external to the cell membrane. Since this space is much more subject to environmental variation
than the actual interior of the cell, bacterial alkaline phosphatase is comparatively resistant to

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inactivation, denaturation, and degradation, and also has a higher rate of activity. In the
laboratory, however, mutant Escherichia coli lacking alkaline phosphatase survive quite well, as
do mutants unable to shut off alkaline phosphatase production.

The optimal pH for the activity of the E. coli enzyme is 8.0 while the bovine enzyme
optimum pH is slightly higher at 8.5.

Dairy Industry

Alkaline phostphatase is commonly used in the dairy industry as an indicator of

successful pasteurization. This is because the most heat stable bacterium found in milk,
Mycobacterium paratuberculosis, is destroyed by temperatures lower than those required to
denature ALP. Therefore ALP presence is ideal for indicating successful pasteurization.

Pasteurization verification is typically performed by measuring the fluorescence of a

solution which becomes fluorescent when exposed to active ALP. Fluorimetry assays are
required by milk producers in the UK to prove alkaline phosphatase has been denatured, as p-
Nitrophenylphosphate tests are not considered accurate enough to meet health standards.

Alternatively the colour change of a para-Nitrophenylphosphate substrate in a buffered

solution (Aschaffenburg Mullen Test) can be used. Raw milk would typically produce a yellow
colouration within a couple of minutes, whereas properly pasteurised milk should show no
change. There are exceptions to this, as in the case of heat-stable alkaline phophatases produced
by some bacteria but these bacteria should not be present in milk.

FOOD SANITATION

Food sanitation refers to the measures undertaken to prevent food contamination to

ensure that it is safe to consume. Food sanitation is recommended at every step of a supply chain
within the food industry. These rules and procedures are normally put in place by public health
agencies.

Food sanitation is defined as aseptic practices in preparation, processing & packaging

unit, sanitation of plants, fermenters, utensils, health of employees.

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HAZARD ANALYSIS AND CRITICAL CONTROL POINTS

HAZARD

Hazard is defined as anything associated with food to cause harm to consumer.

 Unacceptable contamination, growth, survival of food poisoning organism.

 Presence of microbial toxin
 Presence of microbial enzyme that can cause spoilage

It occurs during raw material production, manufacturing, distribution and may also be
microorganisms, chemical, pesticide residues or physical substance.

CRITICAL CONTROL POINTS

CCP is defined as the procedures that identify during raw material production,
manufacturing, retailing, distribution or consumer use in which hazards can be controlled. It
focuses attention on specific prevention & control measures.

 CCP1 – Will assure the control of hazard

 CCP2 – Will minimize a hazard but cannot assure its control

Risk - Risk is defined as likelihood of hazard occurring. It is ranked as low, medium or high.

Severity – severity is level of danger presented by a hazard, a risk can be very low and the
severity high. Eg: bovine tuberculosis in dairy industry is extremely low risk but the severity of
the disease is high

Monitering – Checking whether CCP is under control. It involves visual inspection, smell,
physical measurement, chemical measurement, microbiological testing. Eg: Canned food, UHT
milk and frozen foods.

Verification – Application of tests that are additional to those used for monitoring. It determines
whether HACCP system is operating correctly. Eg: Chlorine in can water can be monitored
ensure that the level of contamination are kept to an acceptable level.

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Stages of HACCP

Raw materials

Stages of manufacturing process

Retailing

Education of the workforce

Distribution system

Target consumer

Way in which consumer handle the product in relation to hazard or risk

Monitoring

Verification

Application

 Milk industry
 Fish and fishery products
 Fresh-cut produce
 Drinks and nectary products
 Food outlets
 Meat and poultry products
 School food and services

HACCP in milk industry

CCP

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  Pre-process contamination with pathogens or spoilage organism can occur.
 Post-processes recontamination with pathogens or spoilage organism can occur.
 Unacceptable growth of spoilage organism can occur.
 Pasteurization is used to kill pathogens

Monitoring & verification

Milk is potential hazard with number of important pathogens. Eg: Mycobacterium bovis,
Salmonella, Campylobacter, Listeria monocytogenes and Yersinia

 Pasteurization – 71.7°C for 15 seconds (HTST)

 Monitoring – temperature in pasteurizer holding time observation of correct plant
functioning by trained operators.
 Verification – Examination of thermographic records, phosphatase test – phosphatase
enzyme in milk, destroyed if pasteurization has been carried out correctly.

Food and Drug Administration

Introduction
The Food and Drug Administration (FDA or USFDA) is a federal agency of the United
States Department of Health and Human Services, one of the United States federal executive
departments. The FDA is responsible for protecting and promoting public health through the
control and supervision of food safety, tobacco products, dietary supplements, prescription and
over-the-counter pharmaceutical drugs (medications), vaccines, biopharmaceuticals, blood
transfusions, medical devices, electromagnetic radiation emitting devices (ERED), cosmetics,
animal foods & fee and veterinary products. As of 2017, 3/4th of the FDA budget
(approximately $700million) is paid by people who consume pharmaceutical products, due to the
Prescription Drug User Fee Act.

Guidance & Regulation (Food and Dietary Supplements)

FDA guidance and regulatory information with links to Federal Register documents. You can
also access information about food safety programs, manufacturing processes, industry systems,
and import/export activities.
Guidance Documents & Regulatory Information:
Guidance Documents: Guidance documents represent FDA's current thinking on a topic. They
do not create or confer any rights for or on any person and do not operate to bind FDA or the
public. You can use an alternative approach if the approach satisfies the requirements of the
applicable statutes and regulations.

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 Regulatory Information: FDA issues regulations to implement its statutory authority. The
regulations can create binding obligations and have the force of law. Links to Federal Register
documents (advance notices of proposed rulemaking, proposed rules, interim final rules, and
final rules) are posted in this section.

Food Facility Registration

Information on the requirement that owners, operators, or agents in charge of domestic or
foreign facilities that manufacture, process, pack, or hold food for consumption in the United
States must register with FDA.

Retail Food Protection

More than 3000 state, local, and tribal agencies have primary responsibility to regulate the
retail food and foodservice industries in the United States. FDA assists regulatory agencies and
the industries they regulate by providing a model Food Code, guidance, training, program
evaluation, and technical assistance.

Imports & Exports

Information on:
Importing food products into the United States, including Prior Notice of Imported Food
Exporting food products from the United States, including export certificates

Federal/State Food Programs

Information about milk and seafood safety cooperative programs with FDA and state and local
health and regulatory agencies.

FDA Act:

Food Protection Plan 2007

FDA developed the Food Protection Plan to address the changes in food sources,
production, and consumption. The plan presents a robust strategy to protect the nation's food
supply from both unintentional contamination and deliberate attack.

The Biologics Control Act was enacted in 1902

The food and drug law did not come into place until 1906. Although it was fairly
comprehensive, it was more of an enforcement statute. It did not require premarket authorization
at that time.

FDA Food Safety Modernization Act (FSMA)

FSMA is the most sweeping reform of FDA's food safety authority in more than 70 years.

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 This act gives FDA new and enhanced mandates and authorities to protect consumers and
promote public health.

Federal Food, Drug, and Cosmetic Act in 1938 .

And this, again, came about as a result of a tragedy involving a drug, specifically elixir
sulfanilamide, which contained an unlabeled solvent that resulted in a number of people dying.
Congress realized that the existing law did not make illegal the failure to disclose that
information. So Congress enacted a new law that extended control over products, beyond drugs,
such as cosmetics and therapeutic devices, although the term device was not used. The statute
required evidence of safety before a product could be marketed.

World Health Organization(WHO)

Introduction
Every year, World Health Day is celebrated on 7 April to mark the anniversary of the
founding of the World Health Organization (WHO) in 1948. World Health Day is a global
campaign, inviting everyone from global leaders to the public in all countries to focus on a
single health challenge with global impact. Focusing on new and emerging health issues, World
Health Day provides an opportunity to start collective action to protect people's health and well-
being. It is an opportunity to engage in finding solutions that benefit us all. Constitution of the
World Health Organization. Geneva: WHO 1946

WHO guideline:
A WHO guideline is any document developed by WHO containing recommendations for
clinical practice or public health policy. A recommendation tells the intended end-user of the
guideline what he or she can or should do in specific situations to achieve the best health
outcomes possible, individually or collectively. It offers a choice among different interventions
or measures having an anticipated positive impact on health and implications for the use of
resources.
Recommendations help the user of the guideline to make informed decisions on whether to
undertake specific interventions, clinical tests or public health measures, and on where and when
to do so. Recommendations also help the user to select and prioritize across a range of potential
interventions.
Principles for WHO guidelines:
WHO's strategic vision, set out in The Twelfth General Programme of Work of WHO (2014),
specifies that in its normative and standard-setting work, WHO is and will remain a science- and
evidence-based organization with a focus on public health. Guidelines are the fundamental means
through which the Organization fulfils its technical leadership in health, as identified in this
strategic document. It states that “WHO’s legitimacy and technical authority lie in its rigorous
adherence to the systematic use of evidence as the basis for all policies”.

To do this, WHO helps Member States build capacity to prevent, detect and manage
foodborne risks by:
providing independent scientific assessments on microbiological and chemical hazards
that form the basis for international food standards, guidelines and recommendations, known as
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the Codex Alimentarius, to ensure food is safe wherever it originates;
assessing the safety of new technologies used in food production, such as genetic modification
and nanotechnology;helping improve national food systems and legal frameworks, and implement
adequate infrastructure to manage food safety risks.
The International Food Safety Authorities Network (INFOSAN) was developed by WHO
and the UN Food and Agriculture Organization (FAO) to rapidly share information during food
safety emergencies;
promoting safe food handling through systematic disease prevention and awareness
programmes, through the WHO Five Keys to Safer Food message and training materials; and
advocating for food safety as an important component of health security and for integrating food
safety into national policies and programmes in line with the International Health Regulations
(IHR - 2005).
Works closely with FAO, the World Organization for Animal Health (OIE) and other
international organizations to ensure food safety along the entire food chain from production to
consumption.
Food Safety and Nutrition
General considerations
General principles for the safe handling and preparation of food apply to all contexts, including
the household, mass-feeding centers for disaster-affected people, and targeted feeding centers,
such as therapeutic-feeding centers.
Golden rules for safe food preparation:(WHO 1991d)
1. Cook raw foods thoroughly. Under normal circumstances raw foodstuffs and water may
become contaminated with pathogens, but in times of disaster the risk of contamination is even
greater. Thorough cooking will kill the pathogens, which means the temperature of all parts of
the food must reach at least 70°C. Uncooked fruits or vegetables should not be eaten, unless they
can be peeled. If milk has not been pasteurized, it should be boiled before use. Cooking will not
necessarily destroy biotoxins.
2. Eat cooked food immediately. When cooked foods cool to room temperature, bacteria begin
to grow. The longer the wait, the greater the risk. To be on the safe side, eat cooked foods as
soon as they come off the heat.
3. Prepare food for only one meal. Foods should be prepared freshly and for one meal only, as
far as possible. If foods have to be prepared in advance, or if there are leftovers, they should be
stored cold, i.e. below 5°C (in a refrigerator or in a cold box), or hot, i.e. above 60°C. This rule is
vitally important when it is planned to store food for more than 4–5 hours. Cooked foods that
have been stored must be thoroughly reheated before eating, i.e. all parts reheated to at least
70°C. Thorough reheating of foods is essential if refrigerators have ceased to operate for some
hours due to power cuts.
4. Avoid contact between raw foods and cooked foods. Safely cooked food can become
contaminated through even the slightest contact with raw food. This cross-contamination can be
direct, e.g. when raw fish comes into contact with cooked foods. It can also be indirect. For

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example, preparing raw fish and then using the same unwashed cutting surface and knife to slice
cooked food should be avoided, or all the potential risks of illness that were present before
cooking may be reintroduced. Cross-contamination may also occur in a freezer when the power
has been off for some time and this should be checked for. The juice of raw meat and poultry
may drip onto other foods.
5. Choose foods processed for safety. Many foods, such as fruits and vegetables, are best in
their natural state. However, in disasters and emergencies, they may not be safe and should be
peeled before consumption if eaten raw. Foods that have been processed (e.g. canned food and
packed dried food) and that have not been affected by the disaster may be safer. Dry rations may
be easier to keep safe, as they do not need cold-storage, but they do need to be kept dry.
6. Wash hands repeatedly. Hands should be washed thoroughly before preparing, serving or
eating food and after every interruption, especially after use of the toilet or latrine, changing a
baby or touching animals. After preparing raw foods, especially those of animal origin, hands
should be washed again before handling cooked or ready-to-eat foods.

7. Keep all food preparation premises meticulously clean. Since foods are so easily
contaminated, any surface used for food preparation must be kept absolutely clean. Scraps of
food and crumbs are potential reservoirs of germs and can attract insects and animals. The
immediate surrounding of the temporary shelter, especially the kitchen and food storage areas,
should be cleaned and sullage and solid kitchen waste should be disposed of properly. Food
should be stored in closed containers to protect it from insects, rodents and other animals. Fly
and rat traps should be used if necessary

Food Safety and Standard Authority of India (FSSAI)

Introduction

Food Safety and Standards Authority of India (FSSAI) is an autonomous body

established under the Ministry of Health & Family Welfare, Government of India. The FSSAI
has been established under the Food Safety and Standards Act, 2006 which is a consolidating
statute related to food safety and regulation in India. FSSAI is responsible for protecting and
promoting public health through the regulation and supervision of food safety. The FSSAI is
headed by a non-executive Chairperson, appointed by the Central Government, either holding or
has held the position of not below the rank of Secretary to the Government of India. Rita Teotia
is the current Chairperson for FSSAI and Pawan Kumar Agarwal is the current Chief Executive
Officer for FSSAI.

The FSSAI has its headquarters at New Delhi. The authority also has 6 regional offices
located in Delhi, Guwahati, Mumbai, Kolkata, Cochin, and Chennai. 14 referral laboratories
notified by FSSAI, 72 State/UT laboratories located throughout India and 112 laboratories are
NABL accredited private laboratories notified by FSSAI.

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 The FSSAI is headed by a non-executive Chairperson, appointed by the Central Government,
either holding or has held the position of not below the rank of Secretary to the Government of
India. Rita Teotia is the current Chairperson for FSSAI and Pawan Kumar Agarwal is the current
Chief Executive Officer for FSSAI.

History

FSSAI was established by Former Union Health Minister Dr Anbumani Ramadoss,

Government of India on 5 August 2011 under Food Safety and Standards Act, 2006 which was
operationalized in year 2006. The FSSAI consists of a chairperson & 22 members. The FSSAI is
responsible for setting standards for food so that there is one body to deal with and no confusion
in the minds of consumers, traders, manufacturers, and [Link] of Health & Family
Welfare, Government of India is the Administrative Ministry of Food Safety and Standards
Authority of [Link] following are the statutory powers that the FSS Act, 2006 gives to the
Food Safety and Standards Authority of India (FSSAI).

1. Framing of regulations to lay down food safety standards

2. Laying down guidelines for accreditation of laboratories for food testing
3. Providing scientific advice and technical support to the Central Government
4. Contributing to the development of international technical standards in food
5. Collecting and collating data regarding food consumption, contamination, emerging risks
etc.
6. Disseminating information and promoting awareness about food safety and nutrition in
India

The FSSAI is headed by a non-executive Chairperson, appointed by the Central

Government, either holding or has held the position of not below the rank of Secretary to the
Government of India.[4] Rita Teotia is the current Chairperson for FSSAI and Pawan Kumar
Agarwal is the current Chief Executive Officer for FSSAI.

Regulatory framework

The Food Safety and Standards Authority of India is a statutory body under Food Safety and
Standards Act, 2006. The Food Safety and Standards Act (FSS), 2006 is the primary law for
regulation of food products. This act also sets up the formulation and enforcement of food safety
standards in India. The FSSAI appoints food safety authorities on the state level.

The FSSAI functions under the administrative control of the Ministry of Health and Family
Welfare. The main aim of FSSAI is to

1. Lay down science-based standards for articles of food

2. To regulate manufacture, storage, distribution, sale and import of food
3. To facilitate food safety

Food Laws and Regulations

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 The requirement of food regulations may be based on several factors such whether a
country adopts international norms developed by the Codex Alimentarius Commission of the
Food and Agriculture Organization of the United Nations and the World Health Organization of
a country may also has its own suite of food regulations. Each country regulates food differently
and has its own food regulatory framework.

Food laws in our country

The Indian Parliament has recently passed the food Safety and Standards Act,2006 that
overrides all other food related laws. Such as.,

1. Prevention of Food Adulteration Act, 1954

2. Fruit products Order, 1955
3. Meat food products Order,1973
4. Vegetable oil Products (Control) order,1947
5. Edible Oils Packaging (Regulation)Order 1988
6. Solvent extracted Oil, De-oiled Meal and Edible Flour (Control)Order,1967
7. Milk and Milk products order,1992 etc are repeated after commencement of FSS
Act,2006

Indian standard Institution (ISI)

Introduction

Indian standard Institution came into being on January 6, 1947. Symbolic of the role, the
Indian Standards Institution (I.S.I.) was to play the first standard drawn for independent India.
In the Bureau of Indian Standards (BIS), the National Standards Body of India formulates
standards through technical committees which have representatives from the manufacturers,
technical experts and users. The standards are documents of consensus which are finalized after
taking the views of all those who may have an interest in it. BIS plays the key role of holding
the secretariat for the Technical Committees and collating and analyzing the data and other
information which may be required for formulating the standards. BIS has formulated nearly
18,000 standards which may be categorized as basic standards, product specifications and
methods of test and codes of practices.

ISI Standard Mark

With the objective of satisfying the consumer in terms of product quality, the BIS has
undertaken various quality certification activities. The domestic consumer is familiar with the ISI
mark on a product which is an assurance that the product conforms to the requirements as laid
down in the specification. Conformity to the standard is ensured through regular surveillance of
the manufacturing process, surprise inspections and testing of samples drawn from the factory as
well as from the market. Fraudulent and unauthorized use of the ISI mark is a violation of the

116
law punishable under the BIS Act.

Quality Management System

There is a world-wide movement for installing Quality Management Systems in accordance

with the IS/ISO 9000 series. In India also this has become a prime requisite for manufacturers
and service sector units which wish to make an impact in the domestic and global markets. The
concept of Quality Management Systems aims at quality control mechanism at every stage of a
manufacturing or a service system and not just the quality of the end product as is the case with
product certification.
With the growing concern for environment-friendly industrial activity, the ISO 14,000 series
of standards were developed. After adopting these standards as national standards the BIS has
also launched the Environment Management System Certification under which units may
demonstrate their compliance with the ISO 14000 standards. The Hazard Analysis Critical
Control Points (HACCP) Certification which again has acquired international recognition for
certifying the safety of food manufacturing process has also been undertaken by the BIS. The
Bureau of Indian Standards has also formulated IS 15000: 1998 which is equivalent to the
internationally accepted Codex standards.
Wrong use of ISI mark
It is very common in India to find products with fake ISI marks, that is, affixing ISI marks on the
product without actually getting certified. Fake ISI marks usually do not carry (i) the mandatory
7-digit license number(written as CM/L-xxxxxxx) required by BIS; and (ii) IS number on top of
the ISI mark which signifies the number of the Indian Standard for the particular [Link] is
a punishable offense by the law, but the practice is uncommon. Some of the products use fake
ISI marks. For example if a mixer grinder box has a small ISI mark in it with the ISI code of the
wire means that the wire has been ISI certified but the mixer grinder is not an ISI certified
product.
The AGMARK Standard

The AGMARK standard was set up by the Directorate of Marketing and Inspection of the
Government of India by introducing an Agricultural produce Act in 1937. The word
‘AGMARK’ seal ensures quality and purity. A sample AGMARK seal is as below

AGMARK BESAN

[Link]. B-162002

GRADE-STANDARD

PLACE OF PACKAGING…….

DATE OF PACKAGING……

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NET WEIGHT……..

The quality of a product is determined with reference to the size, variety, weight, colour,
moisture, fat content and other factors are taken into account. The grades incorporated are
grades 1,2,3 and 4 or special, good, fair and ordinary.

Environmental Protection Agency (EPA)

The Environmental Protection Agency (EPA) was established in December 1970 by an executive
order of United States President Richard Nixon. The EPA is an agency of the United States
federal government whose mission is to protect human and environmental health. Headquartered
in Washington, D.C., the EPA is responsible for creating standards and laws that promoting the
health of individuals and the environment.

The EPA was established in response to widespread public environmental concerns that gained
momentum in the 1950s and 1960s. The EPA seeks to protect and conserve the natural
environment and improve the health of humans by researching the effects of and mandating
limits on the use of pollutants. The EPA regulates the manufacturing, processing, distribution,
and use of chemicals and other pollutants. In addition, the EPA is charged with determining safe
tolerance levels for chemicals and other pollutants in food, animal feed, and water.

Governing of Environmental laws and policies:

US Environmental Protection Agency (EPA):

The governing body for major statues or laws

National Environmental Policy Act (NEPA-1969):

Establishes policies, set goals and facilitate the implementation of such policies

NEPA requires federal agencies to assess environmental impact of implementing their

major programs (work with NASA and army)

Concerns about regulations

1. Structured so cost effective compared to benefits

2. Command and control
3. US free market choice
4. Uneven application of regulations
5. Lack properly structured incentives
6. Generate inefficiencies
7. Paper work, costs
8. Duplicate(Federal, state,etc)

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Step 1: EPA Proposes a Regulation
The Agency researches the issues and, if necessary, proposes a regulation, also known as a
Notice of Proposed Rulemaking (NPRM). The proposal is listed in the Federal Register (FR) so
that members of the public can consider it and send their comments to us. The proposed rule and
supporting documents are also filed in EPA's official docket on [Link]

Step 2: EPA Considers Your Comments and Issues a Final Rule

Generally, once we consider the comments received when the proposed regulation was issued,
we revise the regulation accordingly and issue a final rule. This final rule is also published in the
FR and in EPA's official docket on [Link].

Step 3: The Regulation is Codified in the Code of Federal Regulations

Once a regulation is completed and has been printed in the FR as a final rule, it is codified when
it is added to the Code of Federal Regulations (CFR). The CFR is the official record of all
regulations created by the federal government. It is divided into 50 volumes, called titles, each of
which focuses on a particular area. Almost all environmental regulations appear in Title 40. The
CFR is revised yearly, with one fourth of the volumes updated every three months. Title 40 is
revised every July 1.

UNIT – IV

2 MARKS

1. Define HTST?
2. Define HACCP?
3. Define WHO?
4. Define FSSAI?
5. What is EPA?
6. What is food standards?
7. What is Quality control?
8. Define – FDA?
9. Define – MBRT?
10. Define alkaline phosphatase?

5 MARKS

1. MBRT test?
2. Quality assurance of food products?
3. Details about FDA?

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 4. Write the notes about the Litmus milk test?
5. Discuss about the alkaline phosphates test?

10 MARKS

1. Write the brief notes about the HACCP?

2. Write in details about the FSSAI?

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 UNIT – V

FOOD BORNE DISEASE

Foodborne disease is any illness resulting from the consumption of food contaminated
with one or more disease-producing agents. These include bacteria, parasites, viruses, fungi and
their products as well as toxic substances not of microbial origin. More than 250 different food
borne diseases have been described. These different diseases have many different symptoms, so
there is no one "syndrome" that can be indicated as specific to food borne illness. However, the
microbe or toxin enters the body through the gastrointestinal tract, and often causes the first
symptoms there, so nausea, vomiting, abdominal cramps and diarrhea is common symptoms in
many food borne diseases.

• Viable pathogenic micro organisms (Bacteria, viruses, fungi) or their preformed toxins

• Pathogenic algae, parasites, protozoa and their preformed toxins

• Toxins naturally present or formed in some foods e.g, toxic mushrooms, some sea foods, red kidney
bean poisoning, biological amines in cheese and fermented meats etc.

• Toxic chemicals in contaminated food and water, such as heavy metal and some pesticides

• Nutritional disorders such as rickets due to calcium deficiency

• Allergy to or inability to utilize some normal components of food

• Indigestion from over eating or other reasons

Types of Microbial Food Borne Diseases

On the basis of mode of illnesses, food borne disease (FBD) can be arbitrarily divided
into three groups.

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Intoxication
Illness in this case occurs as a consequence of ingestion of a pre formed bacterial or a
mold toxin due to its growth in a food. A toxin has to be present in the contaminated food. Once
the micro organism have grown and produced toxin in a food, there is no need of viable cells
during the consumption of the food for illness to occur .e.g, Staphylococcal food poisoning.
Infection
Illness occurs as a result of the consumption of food and water contaminated with
enteropathogenic bacteria. It is necessary for the cells of enteropathogenic bacteria to remain
alive in the food or water during consumption. The viable cells even if present in small numbers
have the potential too establish and multiply in the digestive tract to cause the illness. e.g,
Salmonellosis.
Toxicoinfection
Illness occurs from the ingestion of a large number of viable cells of some pathogenic
bacteria through contaminated food and water. Generally the bacterial cells either sporulate or
die and release toxin(s) to produce the symptoms. e.g. Bacillus cereus Gastroenteritis.
Bacteria
 [Link]
 Salmonellosis
 Bacillus cereus
 C. perfringens
 Listeriosis
 Shigellosis
 Yersiniosis
Enteric viruses
Protozoan
Helminthic
The main factors responsible for the food borne illness includes:
a) Improper holding temperature during processing.
b) Inadequate cooling during storage.
c) Contaminated equipments and utensils.
d) Food from unsafe source.

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 e) Poor personal hygiene.
f) Adding contaminated ingredients to cooked foods.

FOOD POISONING

People infected with food borne organisms may be symptom-free or may have symptoms
ranging from mild intestinal discomfort to severe dehydration and bloody diarrhea. Depending
on the type of infection, people can even die as a result of food poisoning.

More than 250 different diseases can cause food poisoning. Some of the most common
diseases are infections caused by bacteria, such as Campylobacter, Salmonella, Shigella, E. coli,
Listeria, and botulism.

Bacteria responsible for food borne illness

Type of disease Causative bacteria Kind and nature of Major symptom(s)

the bacteria type
Intoxication Staphylococcus Gram-positive cocci, GIT
Staph poisoning aureus strains present in pairs, short Non-gastric
Botulism Clostridium chains or bunched
botulinumstrains grape like clusters.
Anaerobic, Gram-
positive spore
forming rod.
Infection Over 2000 Rod shaped, motile, GIT
Salmonellosis Salmonella non spore forming GIT
Campylobacter spp. (except S. typhi Gram-negative GIT
enteritis and bacteria GIT and non-gastric
Yersiniosis S. paratyphi) Gram-negative, GIT
Enterohemorrhagic E. Campylobacter jejuni slender, curved, GIT and non-gastric
coli colitis & Cam. coli strains motile, GIT
Non-hemorrhagic E. Yersinia microaerophillic rod GIT
coli strains enterocolitica Small rod-shaped,

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Listeriosis E. coli 0157:H7 Gram-negative GIT and non-gastric
Shigellosis Shiga-like toxin bacterium GIT and non-gastric
Vibrio producing Gram-negative,
parahemolyticus E. coli strains like E. motile, non-
gastroenteritis coli 026: H11 sporulating, rod
Vibrio vulnificus Listeria shaped facultative
infection monocytogenes anaerobic bacterium.
Brucellosis Four shigella spps. Gram-negative,
e.g. motile, non-
Sh. dysenteriae sporulating, rod
Vibrio shaped facultative
parahemolyticus anaerobic bacterium.
Vibrio vulnificus Motile, Gram-positive
Brucella abortus bacteria
Gram-negative, non-
motile, non-spore
forming rods.
Gram-negative,
curved bacteria
Lactose-fermenting,
halophilic, gram-
negative,
opportunistic
pathogen.
Gram-negative
bacteria
Toxicoinfection Clostridium anaerobic, Gram- GIT
Clostridium perfringens positive, spore GIT
perfringens Bacillus cereus forming rod GIT
gastroenteritis Enteropathogenioc Gram-positive, GIT
Bacillus cereus and enterotoxigenic facultative aerobic GIT

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gastroenteritis E. coli spore former rods GIT
E. coli gastroenteritis Vibrio cholerae Gram-negative,
Cholera Aeromonas motile,
Gastroenteritis by hydrophila non-sporulating, rod
opportunististic Plesiomonas shaped, facultative
pathogens shigelloides anaerobic bacterium
Aeromonas Gram-negative,
hydrophila curved bacteria
gastroenteritis Gram-negative
Plesiomonas bacteria
shigelloides Gram-negative, rod-
gastroenteritis shaped bacterium
A detailed account of major bacterial pathogens is as follows:
1) Staphylococcal Food Poisoning
Caused by bacterium Staphylococcus aureus, Staphylococcal Food Poisoning is one of
the most common types of food poisoning.
Biology of Staphylococcus aureus
Staphylococcus aureus is a spherical bacterium (coccus) which on microscopic
examination appears singly, in pairs, or bunched, grape-like clusters. They are Gram-positive,
facultative anaerobes, but grow rapidly under aerobic conditions. They are mesophiles with a

growth temperature range of 7 to 480C and have the ability to grow at low a W (0.86), low pH

(4.8), and high salt and sugar concentrations of 15% and in the presence of NO2.

S. aureus are naturally present in the nose, throat, skin, and hair of healthy humans,
animals and birds. They can be present in infections such as cuts, wounds, abscesses and facial
acne etc. The contamination generally occurs through these sources because of improper
handling by infected persons.
Toxins, Disease and Symptoms
Only certain strains of S. aureus can cause food poisoning, namely, those that produce an
enterotoxin. Enterotoxigenic strains of S. aureus produce six different enterotoxins: A, B, C, D,

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E, F etc which are serologically distinct heat stable proteins of molecular weight of 26 to 30 kDa.
Staph toxins are enteric toxins and cause gastroenteritis.
The primary symptoms are salivation, nausea and vomiting, abdominal cramps &
diarrhea. Secondary symptoms are sweating, chills, headache and dehydration.
Enterotoxin ingested along with food affects cells

Enterotoxin affects vomit receptors water & sodium pumps out of the cell

Vomiting center in the brain stimulated Diarrhoea, fluid & electrolyte loss

Vomiting Dehydration
Identification methods
Enumeration technique in one or more selective differential media like Baird-Parker agar
and G.C. Gioletti Cantoni broth to determine the load of viable cells followed by several
biochemical tests, such as hemolysis, coagulase, thermonuclease reactions are performed to link
the potential cause of food poisoning outbreaks. Sensitive immunological tests viz., ELISA etc
has been developed for this purpose.
2) Botulism
Although the disease is very rare, the disease has a high fatality rate. Botulism is caused
by the bacterium Clostridium botulinum, which produces an exotoxin that is most potent of all
known poisons. 1 gram is sufficient to kill approximately 30 billion mice.
Biology of Clostridium botulinum
Clostridium botulinum is an anaerobic, Gram-positive, spore-forming rod that produces a
potent neurotoxin (Fig. 4). Spores of C. botulinum are widely distributed in soil, sewage, mud,
sediments of marshes, lakes and coastal waters, plants- fruits and vegetables and intestinal
contents of animals and fishes. Cells are sensitive to low pH (<4.6), lower Aw (0.93), and

moderately high salt (5.5%). The spores are heat-resistant (killed at 115 0C) and can survive in
foods that are incorrectly or minimally processed. Foods most commonly associated are low acid
vegetables (green beans, corn, spinach, asparagus, pepper and mushrooms) and fruits (figs and
peaches) and also fermented, improperly cooked and smoked fish, meat, poultry and fish eggs in
hermatically sealed containers.

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Toxins, Disease and Symptoms
Seven types (A, B, C, D, E, F and G) of botulism are recognized, based on the antigenic
specificity of the neurotoxin produced by each strain. As they are highly potent toxins only a
very small amount (1ng/kg body weight) is necessary for severe symptoms and even death. The

toxins are heat labile and can be destroyed by high and uniform heat- 95 0C for 15 minutes or
boiling for 5 minutes.
At the initial stage (generally 12h to 36h, but can be 2h), some gastrointestinal symptoms
(nausea, vomiting, diarrhea and constipation) are evident. Neurological symptoms appear in a
short time particularly when the amount of toxin consumed is more.
Neurotoxin ingested with food

Passes through gut mucosa into blood stream

Toxin spread throughout the body

Binds to nerves at the unction

Result in paralysis

Identification methods
The presence of C. botulinum can be determined by enumeration techniques using
selective agar media and followed by anaerobic incubation. The presence of toxins is more often
tested by immunological methods like ELISA etc.
3) Salmonella Gastroenteritis
Salmonella typhi and S. paratyphi were considered the major causes of worldwide
foodborne and waterborne diseases caused by Salmonella. With the introduction of
pasteurization of milk and chlorination of water supplies, the spread of typhoid and paratyphoid
fever was greatly reduced. However, the incidence of foodborne cases of Salmonellosis is still on
the rise, which is quite puzzling.
Biology of Salmonella

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 Salmonella is a rod-shaped, Gram negative, non-sporulating, facultative anaerobic motile

bacterium. They are mesophiles with a growth temperature range of 5 to 46 0C and optimum

growth temperature of 35 to 37 0C. They are sensitive to low a W (<0.94) and low pH (4.5 or

below). There are over 2000 serovars of Salmonella based on flagellar, somatic and capsular
antigens, which are capable of causing Salmonellosis in humans. Salmonella are natural
inhabitants of gastrointestinal tracts of animals, birds, pets, frogs, turtle and insects. They are
also present in soil, water and sewage contaminated with fecal matter from where they can
contaminate foods directly or indirectly and can cause Salmonellosis. Foods generally associated
with outbreaks of Salmonella includes raw meats, poultry, eggs, milk and dairy products, fish,
shrimp, frog legs, yeast, coconut, sauces and salad dressing, cake mixes, cream-filled desserts
and toppings, dried gelatin, peanut butter, cocoa, and chocolate. Various Salmonella species have
long been isolated from the outside of egg shells.
Toxins, Disease and Symptoms

Generally 105 to 106 cells need to be consumed for food borne salmonellosis but it can
be as few as 15- 20 cell in some cases. The disease starts with the penetration and passage of
Salmonella organisms from gut lumen into epithelium of small intestine where inflammation
occurs; there is evidence that an enterotoxin may be produced, perhaps within the enterocyte. S.
typhi and the paratyphoid bacteria normally cause septicemia and produce typhoid or typhoid-
like fever in humans. Other forms of salmonellosis generally produce milder symptoms which
includes nausea, vomiting, abdominal cramps, diarrhea, chills, fever, headache and prostration.

Bacteria ingested along with food

Organism grows in host gut cells invade the tissues & release endotoxin

Affects gut leads to loss of water & sodium ions

Gastroenteritis Fever, Vomiting, Diarrhoea

Identification methods

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 The identification & enumeration of Salmonella can be done using selective differential
agar media. Although conventional culture methods require 5 days for presumptive results,
several rapid methods are available which require only 2 days. Immunological methods which
are used to detect Salmonella include ELISA, enrichment serology and Fluorescent Antibody
technique.
4) Shigellosis (Bacillary Dysentery)
The genus Shigella contains four species: Shigella dysenteriae, Sh. flexneri. Sh. boydii,
and Sh. sonnei and each species have several serovars. Children below 5 years of age are most
affected.
Biology of Shigella
Shigella is Gram-negative, non-motile, non-spore forming rod-shaped bacteria. The

strains grow between 7 and 460C, with an optimum at 370C. They can survive for days under
refrigeration, freezing, and 5% NaCl and pH4.5, but are sensitive to pasteurization. The organism
is frequently found in water polluted with human feces. The foods implicated in outbreaks
include salads, raw vegetables, milk and dairy products, and poultry. Contamination of these
foods is usually through the fecal-oral route.
Toxins, Disease and Symptoms
The strains are believed to produce a plasmid encoded invasive factor-Shiga toxin, which
is an exotoxin having enterotoxigenic property. The infective dose is very low as few as 10 cells
depending on age and condition of host.
The symptoms include abdominal pain, cramps, diarrhea, fever, vomiting, accompanied
by blood, pus, or mucus in the stools. Infections are associated with mucosal ulceration, rectal
bleeding, drastic dehydration; fatality may be as high as 10-15% with some strains.
Identification methods
Organisms are difficult to demonstrate in foods because methods are not developed or are
insensitive. A genetic probe to the virulence plasmid has been developed by FDA and is
currently under field test. However, the isolation procedures are still poor.
5) Escherichia coli enteritis
Since its discovery in 1805, E. coli was considered a harmless, Gram negative, motile,
non-sporulating; rod shaped, facultative anaerobic bacterium, a normal inhabitant of Gastro-
intestinal Tract (GIT) of humans and warm blooded animals and birds.

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i) Enterotoxigenic Escherichia coli (ETEC)
Disease
Gastroenteritis/ travelers' diarrhea are the common name of the illness caused by ETEC.
Infective dose
A relatively large dose (100 million to 10 billion bacteria) of ETEC is probably necessary
to establish colonization of the small intestine, where these organisms proliferate and produce
toxins which induce fluid secretion.
Symptoms
The most frequent clinical syndrome of infection includes watery diarrhea, abdominal
cramps, low-grade fever, nausea and malaise.
Associated food
Contamination of water with human sewage may lead to contamination of foods. Infected
food handlers may also contaminate foods. These organisms are infrequently isolated from dairy
products such as semi-soft cheeses.
Identification methods
With the availability of a gene probe method, foods can be analyzed directly for the
presence of ETEC, and the analysis can be completed in about 3 days. Alternative methods
which involve enrichment and plating of samples for isolation of E. coli and their subsequent
confirmation as toxigenic strains by conventional toxin assays may take at least 7 days.
ii) Enteropathogenic Escherichia coli (EPEC)
Disease
Infantile diarrhea is the name of the disease usually associated with EPEC.
Infective dose
EPEC are highly infectious for infants and the dose is presumably very low. In the few

documented cases of adult diseases, the dose is presumably similar to other colonizers (> 10 6
total dose).
Symptoms
Occasionally, diarrhea in infants is prolonged, leading to dehydration, electrolyte
imbalance and death (50% mortality rates have been reported in third world countries).
Associated food

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 Common foods implicated in EPEC outbreaks are raw beef and chicken, although any
food exposed to fecal contamination is strongly suspect.
Identification methods
The isolation and identification of E. coli in foods follows standard enrichment and
biochemical / immunological procedures.
iii) Enterohemorrhagic E. coli (EHEC)
Disease
Hemorrhagic colitis is the name of the acute disease caused by E. coli O157:H7.
Infective dose
Unknown, but may be similar to that of Shigella spp. (as few as 10 organisms).
Symptoms
The illness is characterized by severe cramping (abdominal pain) and diarrhea which is
initially watery but becomes grossly bloody. Occasionally vomiting occurs. Fever is either low-
grade or absent.
Associated food
Undercooked or raw hamburger (ground beef) has been implicated in many of the
documented outbreaks, however E. coli O157:H7 outbreaks have implicated unpasteurized fruit
juices, dry-cured salami, lettuce, game meat, and cheese curds.
Identification methods
Several microbiological methods can be used to isolate E. coli O157:H7 from foods.
Sorbitol-MacConkey agar has been used extensively to isolate this organism from clinical
specimens.
iv) Enteroinvasive Escherichia coli (EIEC)
Disease
Enteroinvasive E. coli (EIEC) may produce an illness known as bacillary dysentery.
Following the ingestion of EIEC, the organisms invade the epithelial cells of the intestine,
resulting in a mild form of dysentery, often mistaken for dysentery caused by Shigella species.
Infective dose
The infectious dose of EIEC is thought to be as few as 10 organisms (same as Shigella).
Symptoms

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 The illness is characterized by the appearance of blood and mucus in the stools of
infected individuals, abdominal cramps, diarrhea, vomiting, fever, chills, and a generalized
malaise.
Associated food
It is currently unknown what foods may harbor EIEC, but any food contaminated with
human feces from an ill individual, either directly or via contaminated water, could cause disease
in others.
Identification methods
Foods are examined as are stool cultures. Detection of this organism in foods is
extremely difficult because undetectable levels may cause illness.
6) Bacillus cereus Gastroenteritis
The causal organism is a bacterium Bacillus cereus, which is a Gram-positive, facultative
aerobic spore former whose cells are large rods and whose spores do not swell the sporangium
situated centrally or sub-terminally. A wide variety of foods including meats, milk, vegetables,
and fish have been associated with the diarrheal type food poisoning.
Toxins, Disease and Symptoms
The diarrheal type of illness is caused by a large molecular weight protein, while the
vomiting (emetic) type of illness is believed to be caused by a low molecular weight, heat-stable
peptide.
The symptoms of B. cereus diarrheal type food poisoning are the onset of watery
diarrhea, abdominal cramps, and pain after 6-15 hours of consumption of contaminated food.
Nausea may accompany diarrhea, but vomiting (emesis) rarely occurs
Identification methods
B. cereus can be enumerated by surface plating on a mannitol, egg yolk agar medium
with polymyxin B as the selective medium. Colonies surrounded by precipitation resulting from
lecithinase are indicative of B. cereus.
7) Campylobacter Enteritidis/ Campylobacteriosis
Campylobacteriosis caused by C. jejuni was first recognized as the cause of an outbreak
in 1979. It is also often known as campylobacter enteritis or gastroenteritis. It is caused by
several species of Campylobacter, however Campylobacter jejuni and Campylobacter coli are

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more common. Campylobacter jejuni is a Gram-negative slender, curved, and motile rod. It is a
microaerophilic organism, which means it has a requirement for reduced levels of oxygen.
Toxins, Disease and Symptoms
C. jejuni produces thermolabile enterotoxins that cross reacts with cholera toxin. The
infective dose of C. jejuni is about 400-500 bacteria. C. jejuni infection causes diarrhea, which
may be watery or sticky and can contain blood (usually occult) and fecal leukocytes (white
cells). Other symptoms often present are fever, abdominal pain, nausea, headache and muscular
pain.
Identification methods
The identification methods require an enrichment broth containing antibiotics, special
antibiotic-containing plates and a microaerophilic atmosphere generally a microaerophilic
atmosphere with 5% oxygen and an elevated concentration of carbon dioxide (10%). Isolation
can take several days to a week.
8) Vibrio species
Two species of Vibrio which are of prime concern and are emerging as food borne
pathogens are:
A. Vibrio parahemolyticus Gastroenteritis
Gastroenteritis caused by Vibrio parahemolyticus is quite common in Japan and accounts
for 40 to 70% for the total foodborne poisonings. The cells are Gram-negative, non-sporulating

motile curved rods with an optimum temperature of 30 to 37 0C, but can grow over a temperature

range of 5 and 420C. The cells can grow quickly in a salt concentration of 3 to 5% NaCl but are
sensitive to 10% salt. Infections with this organism have been associated with the consumption
of raw, improperly cooked, or cooked, recontaminated fish and shellfish.
Toxins, Disease and Symptoms
V. parahemolyticus associated gastroenteritis is the name of the infection caused by this
organism. The pathogenic strains are capable of producing a heat-stable hemolysin factor and are
designated as Kangawa positive. Disease is caused when the organism attaches itself to an
individuals' small intestine and excretes an heat stable hemolysin factor.

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 Diarrhea, abdominal cramps, nausea, vomiting, headache, fever, and chills may be
associated with infections caused by this organism. The illness is usually mild or moderate,
although some cases may require hospitalization.
B. Vibrio vulnificus Septicemia
Vibrio vulnificus, a lactose-fermenting, halophilic, gram-negative, opportunistic
pathogen, is found in estuarine environments and associated with various marine species such as
plankton, shellfish (oysters, clams, and crabs), and finfish. Consumption of these products raw or
recontaminated may result in illness.
Toxins, Disease and Symptoms
This organism causes wound infections, gastroenteritis, or a syndrome known as
"primary septicemia". The infective dose for gastrointestinal symptoms in healthy individuals is
unknown but for predisposed persons, septicemia can presumably occur with doses of less than
100 total organisms.
The ingestion of V. vulnificus by healthy individuals can result in gastroenteritis. The
"primary septicemia" form of the disease follows consumption of raw seafood containing the
organism by individuals with underlying chronic disease, particularly liver disease. It may also
cause distinctive bulbous skin lesions.
General Control Measures for Prevention of Food Borne Diseases
a) By employing proper sanitary measures.
b) By the use of chemical agents in preventing the growth of microorganisms.
c) Removing leftover food promptly from work surfaces and utensils in food preparation area.
d) By cooking or applying proper and desired processing treatment to the food before
consumption.
e) By avoiding chances of re-contamination of food, hence storing it in proper package.
f) Maintaining good personal hygiene, like food handlers should not have any open cuts and
wounds.
g) Maintaining proper sanitation and preventing contamination of water to be used.
h) Use of proper storage condition restricting recontamination and growth.

Mycotoxin

Mycotoxicosis and Mycetismus

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 Many fungi produce toxic to poisonous substance called mycotoxins that cause acute or
chronic intoxication and damage and sometimes death.

The mycotoxins are secondary metabolites and their effect is not dependent on fungal infection.

A variety of mycotoxins are produced by poisonous mushroom called Amanita species.

The ingestion of toxins present in food material is called mycotoxicosis and the ingestion of
toxins along with the fungi is called mycetismus.

Mycotoxin can be detected in bile, urine, faces, milk, feeds and food stuffs by various methods

Mycotoxicosis

Aflatoxicosis

Aflatoxins are the toxic metabolites produced by Aspergillus flavus.

This is one of the most potent toxin related to contaminated food such as peanuts, corn, grains
[Link] were discovered as the cause of strange manifestations of Turkey –X-Disease during
early 1960’s in England.

There is a series of compound with similar activity they are designated as aflatoxin B1, B2,
G1&G2 because of the Blue(B) and green(G) fluorescence exhibited when irradiated with
ultraviolet light and their metabolites on thin layer chromatography .Aflatoxins B1 and B2,G2
are dihydroderivatives of B1 and G1.

[Link] produces G1 and G2 toxins. These toxins are aflatoxins M1 and M2 respectively.
These are found in the milk (M stands for milks) of the lactating mammals fed on aflatoxin
contaminated food stuffs.

Aflatoxins B1 is important because of its toxicity. It produces hepatomas (liver cancer) and
herpatic necrosis in duckling, trout and other animals.

Signs and symptoms are lethargy, anorexia and muscle weakness of the patients. Other features
are fever, jaundice, hepatomegaly and portal hypertension.

Corn,wheat and rice contain more aflatoxins than sorghum, peanut and soyabean

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Acute aflatoxicosis Reye’sSyndrome in which the patients presents with symptoms of
encephalopathy with fatty degeneration of the viscera, is an epidemic disease of children in the
developing countries.

Indian children cirrhosis is caused by aflatoxicosis.

Ochratoxicosis

The natural occurrence of ochratoxins in grains and other products has been widely
reported .Ochratoxin A is naturally occurring mycotoxin produced by certain species of
Aspergillus and Penicillium

This toxin causes a nephrotoxin immunosuppressant,teratogen. Mutagen and carcinogen.

Human exposure to ochratoxins can occur directly by consumptionof contaminated plant food or
indirectly by consumption of animal tissues exposed to contaminated materials.

It produces Balkan endemic neuropathy (BEN) is Bulgaria and urinary tract,tumours are
associated with this disease.

Fumonisins toxicosis

Contamination of cereals with fungus.

Fusarium moniliforme is a common contaminant of corn throught out the world.

It is also produces Fusarium moniliforme.

It is responsible for causing the leukoencephalo malaria in horses, hepatotoxic and carcinogenic
affects in rats and pulmonary edema in pigs.

Trichothecenes

Trichothecenes are mainly produced by Fusarium species but other fungal genera like
Myrothecium,[Link] and cephalosporium can also produce these
compounds.

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[Link] produce primarily toxin and [Link] produce nivalenol and
deoxynivalenol(an emetic vomit inducing)

Alimentary toxic aleukia is caused by cereals infected with [Link] in which there are
necrotic lesions of the oral cavity,oesophagus , stomach and marked leucopenia.

Mycetismus

It is also known as mushroom poisoning.

This is due to the consumption of fleshy [Link] produce gastro intestinal disturbances to
hallucinations delirium, cramps and death.

Ergotism

Ergot is a fungus found on rye,a kind of cereal ergot poisoning is the oldest known mycetismuc
and is caused by the species of genus [Link] toxic facts in human beings results from the
consumption of the bread made of rye flour,contaminated with Claviceps purpurea.

There is inflammation of the infected tissues followed by necrosis and gangrene.

They produce αadrenergic blockage that inhibits respone to epinephrine and 5-hydroxy
tryptamine.

If not treated, it produce vasoconstrictum which results in necrosis and gangrene .

i. There is an epileptic fits ,vomiting and etc.,

There are 2 groups
Ergometrine
It is a water soluble either lysergic acid or its isomer linked to an amino acid.
Ergometrine is clinically used for the prevention of bleeding during surgery.
Ergotamine
It is a water insoluble group. This alkaloid is a polypeptide in which isolysergic
acid is linked to the amino acid. Used for the treatment of migranine.
Asprine poisoning

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 Coprinus atrementarius, when prepared as butter sauce, if the alcohol is used after
disulfuram. Like reaction occur. Disulfuram is a sulphur component that is the presence
of alcohol it causes nausea and vomiting
Symptoms
Hot flushed sensation due to dilation of blood vessels, chest pain, headache, vertigo,
nausea, weakness and confusion.
Muscarine poisoning
Incocybe species, particularly [Link] large quantities of muscarine.
It causes perspiration, lacrimation.
It is type of Mushroom poisoning.
There may be diarrhea, headache, blurred vision followed by contraction of pupils
hypotension etc.,
Muscionol,Ibotenic acid and Musacazone
The edible mushrooms are the most innocent of foods,aflavours , the slightest of textures
and nothing, more .By contrast,some closely related fungi may produce dramatic
symptoms of [Link] patherina, Amanita muscaria are two poisonous fungi
which are eaten by mistakes.
Symptoms
Sweating, twitching, mycosis, muscular in co-ordination,convulsions and coma.
Cyclopeptides,Amatoxins in Phallotoxins
1. Amatoxins are present in different toadstool,causing fatal human poisonings.
2. They produce cellular necrosis through the inhibition of mRNA synthesis.
3. The phallotoxins are cyclic heptapeptides Amanita phallides is cytotoxic. The
phallotoxins are not absorbed through gastrointestinal tract but injected in
experimental animals it remains active.
4. Amanita produces several toxins like phalloidin, phalloin,α,β,γ amanitin.
FOOD POISOINING OUTBREAKS
Foodborne disease outbreaks are investigated to prevent both ongoing transmission of
disease and similar outbreaks in the future.
Specific objectives include:
- control of ongoing outbreaks;

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- detection and removal of implicated foods;
- identification of specific risk factors related to the host, the agent and the
environment;
- identification of factors that contributed to the contamination, growth, survival and
dissemination of the suspected agent;
- prevention of future outbreaks and strengthening of food safety policies;
- acquisition of epidemiological data for risk assessment offoodborne pathogens;
- stimulation of research that will help in the prevention of similar outbreaks.
The scale of an outbreak may range from a local outbreak of a small number of linked
cases with mild disease to a nationwide or international outbreak of severe disease
involving the mobilization of public health resources from all levels.
Irrespective of the scale, a full investigation of a food borne disease outbreak will
normally include:
- epidemiological investigations;
- environmental and food investigations;
- laboratory investigations.
Investigation of a potential outbreak starts with the assessment of all available
information; this should confirm or refute the existence of an outbreak and allow a
working case definition to be established.
This assessment must be initiated quickly and completed promptly in order to prevent
further illnesses, and should include: - checking the validity of the information;
- obtaining reports of applicable laboratory tests that have been performed;
- identifying cases and obtaining information about them;
- ensuring the collection of appropriate clinical specimens and food samples. Once
the validity of the reporting source has been verified, a group of the initial cases
– perhaps 5 to 10 persons – should be identified and interviewed as soon as possible.
This critical step helps to provide a clearer picture of the clinical and epidemiological
features of the affected group.
Delays in conducting these interviews can lead to recall bias or to people’s inability to
remember what they ate or what they did. The interviews should be open and
comprehensive and include questions about: - demographic details, including

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occupation; - clinical details, including date of onset, duration and severity of
symptoms; - visits to health care providers or hospitals; - laboratory test results; -
contact with other ill persons; - food consumption history; the respondent’s thoughts
on what caused their illness; - whether the respondent knows others with the same or
a similar illness; - potential common exposures among those who have the same or a
similar illness; - date of exposure to suspected foods.
ESTABLISHING CASE DEFINITION
A case definition is a set of criteria for determining whether a person should be
classified as being affected by the disease under investigation.
As such, it is an epidemiological tool for counting cases – it is not used to guide
clinical practice. A case definition should be simple and practical and should include
the following four components: - clinical and laboratory criteria to assess whether a
person has the illness under investigation; the clinical features should be significant or
hallmark signs of the illness; - a defined period of time during which cases of illness
are considered to be associated with the outbreak; - restriction by “place” – for
example, limiting the group to patrons of a particular restaurant, employees of a
particular factory or residents of a particular town; - restriction by “person”
characteristics – limiting the group to, for example, persons over one year of age,
persons with no recent diarrhoeal disease, etc.
Ideally, a case definition will include all cases (high sensitivity) but exclude any
person who does not have the illness (high specificity). A sensitive case definition
will detect many cases but may also count as cases individuals who do not have the
disease. A more specific case definition is more likely to include only persons who
truly have the disease under investigation but also more likely to miss some cases.
Because a single case definition that suits all needs is rare, it is quite common for case
definitions to change during an investigation or for different case definitions to be
used for different purposes. Many investigators use the following (or similar) case
definitions in parallel: ƒ Confirmed cases – have a positive laboratory result (isolation
of the causative agent or positive serological test). This case definition has high
specificity. ƒ Probable cases – have the typical clinical features of the illness but

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 without laboratory [Link] cases – have fewer or atypical clinical
features. This case definition has high sensitivity.

UNIT – V

2 MARKS

1. What is the food borne diseases?

2. Define – Intoxication?
3. Define – Infection?
4. Infective dose of ETEC?
5. What is food poisoning?
6. What is food sanitation?
7. Control measures for food borne diseases?
8. What is severity?
9. What is CCP?
10. Define Mycotoxins?

5 MARKS

1. Details about the food borne diseases?

2. What is Gastroenteritis?
3. Details about the Botulism?
4. Discuss about the food poisoning?
5. Details about the Mycetisums?

10 MARKS

1. Write the brief notes about the Escherichia coli enteritis?

2. Discuss in details about the food poisoning?
3. Investigation of food poisoning outbreaks?

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