Chapter 15

Chapter 15
Food Microbiology
Muhammad Saeed and Tahir Zahoor*
Abstract
Microbiology deals with the study of the microorganisms which are the ancestors
of life on earth. Food microbiology is the division of microbiology which deals
with those actions and reactions involved in food preservation, fermentation, value
additions (probiotics) and spoilage. Microorganisms are the main requirement for
fermentation and culture preparation process in different food items. Food
microbiology is defiantly a practical science and food microbiologist guarantees
hygienic and safe food to the consumers. It encompasses the complete knowledge
of food contamination, its sources and control. Food preservation depends upon the
presence and availability of favorable conditions for the growth of the
microorganisms. Microbially contaminated food can cause serious food borne
illnesses. Food spoilage can be prevented by using different techniques like use of
temperature, irradiation and preservative. All the features of food microbiology are
briefly discussed in this chapter.
Keywords: Preservation, Fermentation, Contamination, Spoilage
15.1. Introduction to Food Microbiology

Conventionally, microbiology can be defined as study of living organisms that
cannot be seen without microscope like viruses, bacteria, protozoa, etc. Food
microbiology deals with the food spoilage, food poisoning, microbial metabolism
and physiology. Microbiology plays role to establish recent procedures for quick
and operative finding of harmful bacteria. Recombinant DNA technology can be
*
Muhammad Saeed˧ and Tahir Zahoor
National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan.
˧
Corresponding author’s e-mail: drmseed@uaf.edu.com.pk
Managing editors: Iqrar Ahmad Khan and Muhammad Farooq

Editors: Tahir Zahoor and Masood Sadiq Butt
University of Agriculture, Faisalabad, Pakistan.
390 M. Saeed and T. Zahoor
used to develop desirable microbial strain, to produce safe and wholesome food for
the consumers (Frazier and Westhoff 2008). The study of microorganisms is
important to determine foodborne pathogens in order to control food spoilage that
can initiate food poisoning and infection. On the other side, knowledge of
beneficial microbes helps in the product development with respect to their isolation,
identification and utilization in the fermentation processes of various foods.
15.2. Scope of Food Microbiology

The presence and survival of microorganisms in food depends on physical and
chemical characteristics and their population depends on physical characteristics of
the food, storage conditions, type of the organisms and effects of processing. The
microorganisms can enter in food from natural sources that includes water, air, soil,
animals and sewage water either into raw material or during processing and
handling. (Adams and Moss 2008).
15.2.1. Food Spoilage

Microorganisms (yeast, bacteria, mold) when contaminate foods causes food
spoilage as a result of enzymatic hydrolysis and oxidation reduction reactions. The
factors of temperature, water activity, pH and other chemical reactions also add to
the food spoilage if not appropriately controlled. Bacteria break down the food into
various waste materials that can spoil the food and lead to unsafe food for
consumption. Yeast is responsible for the spoilage of food containing high sugar
content, beverages, bread, yogurt, apple cider, vinegar, beer, wine, etc. (Jay et al.
2005).
Appearance of spoiled food items is different from those of fresh foods i.e., in color
change, unpleasant flavor, soft texture and unacceptable taste. In case of mold
contamination, spoiled meat produces toxic compounds and consumption may
result in disease or death. Toxic effects and health hazards produced by the
consumption of spoiled foods are known as food poisoning or foodborne illness
comprising of food intoxication and food infection respectively.
15.2.2. Food Preservation

Food can be prevented from spoilage with extended shelf life by adopting various
preservation techniques like refrigeration, freezing, irradiations, use of high
temperature, chemical preservatives and irradiations etc.. Another way of
preservation is canning of food that preserves food for a long time spans. Canned
food is vacuum packed to keep oxygen out of the can that is surely needed to hinder
the growth of bacteria. Fermentation is another way of preservation in which
organic acids produced during the process prevents spoilage of food (Adams and
Moss 2008).
Food safety controls handling, processing and storage of food in a way to avoid
food poisoning. Food safety covers food handling, food processing, food additives,
food labeling, and food hygiene. It may also include policies, guidelines, rules and
15. Food Microbiology 391
regulations for the management of import, export and certification of foods. Food
can be a potential source of disease spread from person to person. It also serves as a
medium for the growth of microorganisms that can cause food spoilage and food
poisoning. In developed countries, there are standard methods for the food
processing, whereas in many developing countries, there is even a shortage of safe
drinking water. According to WHO, the most widespread health issues are now
related to foodborne diseases and as a result population gets affected throughout the
world (WHO 1992).
According to WHO, the five key principles of food hygiene are, prevention of food
from contamination through people, pets, and pests, separate raw and cooked foods,
cooking of foods at proper time and temperature to kill pathogens, suitable storage
temperature and water and cooking materials safety. Combination of two or more
methods can be used for food preservation (Frazier and Westhoff 2008).
15.2.3. Fermentation
Conversion of complex molecules into simpler compound by the action of
microorganisms or their enzymes is called fermentation. By fermentation process,
energy is produced in the form of ATP. In fermentation process microbial activity
is desirable. Fermentation enhances food digestion and ultimately increases
nutrients bioavailability (Adams and Moss 2008). Fermentation increases the shelf
life of perishable foods. Different commodities which undergo fermentation
process are cereals, milk, fish, meat, honey, fruits and vegetables. Glycolysis,
lipolysis and proteolysis are three important processes involved in the production of
flavors in fermented foods (Caplice and Fitzgerald 1999). Fermentation mainly
depends upon the raw material, starter culture and conditions under which
fermentation is carried out. Some examples of fermented products are beer, wine,
bread, vinegar, soy sauce, kimchi, sauerkraut, pickled vegetables, sausages,
buttermilk, yoghurt and cheese (Hansen 2002). Fermentation is of three types,
i. lactic acid fermentation such as kimchi, sauerkraut, pickled
vegetables, sausages, buttermilk, yoghurt and cheese,
ii. acetic acid fermentation like apple cider vinegar and
iii. alcoholic fermentation such as wine, beer and alcoholic beverages.
15.3. Origin of Microorganisms

Following the development of eukaryotic cell, the complexity has reached to its
peak. Imaginary concept of microbes must be cleared in the mind of cell and
molecular biologists to command the cellular background of microbes.
According to evolution, present form of life is much different from its initial form.
The exact design of alterations is significantly uncertain, and molecular biology
suggests that procedure of bacterial evolution should be investigated from its
background. Evolution suggests that prokaryotes modified themselves to compete
with ever changing surroundings to maintain their life on earth. As radiation and
oxygen concentrations in atmosphere changes, the microorganisms change
themselves accordingly to survive in nature. Multicellular organisms grown in the

old oceans start to consume other organisms as a food. Eventually, when oxygen
and other parameters of environment are suitable, the energy production processes;
photosynthesis is possible. Photosynthesis allowed fungi and plants evolution on
earth. The sexual reproduction method offered another benefit that was exploited
by certain organisms. The invention and improvements in microscopy brought the
discovery and revolution of microorganisms. In 1658, Athanasius Kircher observed
the presence of very small viable worms in spoiled milk and meat under
microscope. In 1664, Robert Hooke studied the molds under microscope and gave
useful information about their structure. Antony Wan Leeuwenhoek was the first
scientist who observed and described these small creatures as microorganisms. He
examined the samples of rain water, vinegar and saliva under the microscope and
reported the presence of bacteria under microscope in these samples. At that time
powerful microscopes were not invented so other microbiologists had to study the
microbes according to the ideas of Leeuwenhoek. New progress had not come into
being until 19th century started. In the nineteenth century, more progressive studies
about microorganisms were made with the powerful microscopes.
15.4. Importance of Food Microorganisms

Microorganisms are necessary in food applications and their importance can be
judged by three categories: a) Production of cultures for food application, b)
Application in food fermentation and c) microorganisms as a source of food. These
categories are discussed in detail as:
15.4.1. Production of Cultures for Food Application

General principles of microbial culture production include selection, preparation,
maintenance and purity of cultures. The microbial cultures which are used for food
applications include bacterial, yeast and mold cultures.
i) Bacterial Cultures
Most of the bacterial cultures are applied as starters for dairy products, sausages
and bread etc. These are pure cultures of lactic acid bacteria, except the propionic
acid bacteria added to Swiss cheese as well as acetic acid bacteria which are being
used for vinegar production.
ii) Lactic Acid Cultures
Some dairy industries maintain their own cultures and use them successfully for
years, on the other hand most operators take new cultures periodically or use
frozen, concentrated cultures prepared by commercial culture laboratories. The
majority of commercially available cultures is now in form of liquid nitrogen
frozen culture concentrates. These may be prepared by growing the cultures for
harvesting purpose, concentrate by centrifugation, standardized for activity and
then packaged in cans filled with liquid nitrogen. This process has replaced the
older practice of preparing bulk starter culture inocula over a succession of days
because number of cells is so high that 16-oz can be used for direct inoculation of a
cheese vat containing up to 5000 lbs. of milk. The most common dairy starters
consist of a mixture of Streptococcus lactis subsp. Lactis and S. mesenteroindes
subsp. Cremoris for lactic acid production while Leuconostoc cremoris or
Streptococcus lactis subsp. Diacetlactis for flavor and aroma development.
Lactobacilli and pediococcus acidilactici or P. pentosaceus cultures are used as
starter for fermented sausages.
iii) Propionic Cultures
Lyophilized or spray dried cultures of Propionibacterium freudenreichii are mixed
in milk and used in production of Swiss cheese to assist eye formation and flavor
perfection.
iv) Acetic Acid Cultures
In vinegar production, impure mixed cultures are allowed to develop naturally,
instead of pure cultures of Gluconobacter or Acetobacter which are not much
efficient in acetic acid production, are added in previous run of vinegar as raw
vinegar or transferred in vinegar generator.
v) Yeast Cultures
Industrially important yeasts include Baker’s yeast, yeast for malt beverage, wine
yeast and distiller yeast. These types can be used as guard against possible
undesirable changes.
vi) Baker’s Yeast
It can be obtained from the grain mashes, wood hydrolyse, waste sulfite liquor from
paper mills and other materials. Strains of S. cerevisiae should have characteristics
such as remain stable and viable in the cake and produce carbon dioxide in the
dough when used for leavening. About 3 to 5 lbs. of yeast is added per 100 gallon
of medium. Active dry yeast has less than 8% moisture level that is made by drying
the yeast cells.
vii) Yeast for Malt Beverages
It may be pure culture obtained from the specialized laboratories when needed.
Special bottom yeast for beer which includes the strains of S. uvarum
(carlsbergensis), top yeast for ale which include the strains of S. cerevisiae or
sometimes a mixture of bottom and top yeast is used for stout and porter.
viii) Wine Yeast
For wine making strain of S.cerevisiae versus ellipsoideus are adopted to make the
specific type of wine such as Burgundy, Tokay and Compiegne. The starter culture
is grown and builds up in juice of grapes or other fruits used in the fermentation.
ix) Distiller’s Yeast
It includes the use of high alcohol yielding strains of S. cerevisiae versus
ellipsoideus. The medium would be malted grain usually corn or rye for whiskey,
molasses for rum or fruit juices for brandy.
x) Mold Cultures
Spores of Penicillium roqueforti for blue cheeses, stilton, gorgonzola, roquefort,
etc. are usually grown on the cubes of sterilized, moistened cheeses. The cheese is
white, tangy, crumbly and slightly moist, with distinctive veins of blue mold. Mold
starters are prepared in the form of pallets or commonalities of mycelium that are
produced during the submerged growth. These cultures are used for industrial sub
merged fermentation. The soy sauce koji is generally a combined pure culture of
Aspergillus oryzae with yeast and Lactobacillus dellbrueckii. In this case, mold
culture is allowed to grow on cooked and sterile rice.
Different ways of preparing the mycelia or spores cultures include
1) thin layered growth on the media surface,
2) growth on acidified hydrated enriched wheat bran,
3) growth by the submerged technique in oxygen containing liquid medium,
usually resulting in pallets with or without spores,
4) growth in a flask containing medium or similar container.
The prepared mold growth can be recovered by different ways depending upon the
technique of manufacture.
15.4.2. Applications in Food Fermentation

Pure or mixed microbial cultures may be added for development of dairy products
like fermented milks, butter, cheese, bread making, malt beverages, wines and
vinegar. No microbial culture is added if the concerned organisms are known to be
present in sufficient number in the original raw material e.g. in the preparation of
sauerkraut, fermented pickles, processing of cocoa, coffee, etc. The fermentation
does not only preserve the food but also support distinctive, new and desired food
productions that is carried out by molds, yeast, bacteria or their combinations.
Mainly bacteria is used in the production of fermented milks, molds in cheese
varieties and oriental foods, while yeast is used in bread, wine, distilled liquor and
beer productions.
15.4.3. Microorganisms as a Food or Source of Enzymes

Microorganisms are used as a food (protein, fat, acids and polysaccharides) for
human beings or as a source of enzymes to be used in the processing of foods.
During World War II, Germans produced yeast and molds in some quantity as food
source when there was shortage of protein and vitamins. After the war, the British
established a plant in Jamaica for the production of food yeast. But now increasing
quantities of microbes are being grown directly for such purposes. From this,
concept of Single Cell Protein (SCP) was originated. It is a term applied to the
microbial cells grown and harvested for humans or animal food. Research on SCP
has been inspired by a concern over the food shortage or food crisis that arises due
to increased population of any specified region. Various yeast, molds, algae and
bacteria are possible microbes for SCP production. Scenedesmusa cutus (267-3A)
and Spirulina maxima have been used in culture ponds with some limitations.
Yeasts Candida utilis (torula yeast) are the widely accepted and used
microorganism for SCP production. For fat production Candida pulcherrima,
Torulopsis lipofera, Saccharomyces cerevisiae and Rhodotorula glutinis among
yeast, Trichosporon pullulans among the yeast like and Geotrichum candidum of
mold strains have been studied. Enzymes as biocatalyst are being used by living
cells and responsible for the numerous metabolic processes of the cell. Microbial
enzymes, sources and their usage are described below in Table 15.1 (Frazier and
Westhoff 2008).
Table 15.1 Microbial enzymes, sources and their application

Enzyme Source Industry Application
Amylase Aspergillus niger Baking Flour supplement
Aspergillus oryzae Brewing Mashing
Cellulase Aspergillus niger Food Preparation of liquid-
coffee concentrates
Glucose oxidase Aspergillus niger Food Glucose removal from
egg solids
Lactase S. fragils Dairy Hydrolysis of lactose
Protease Aspergillus niger Food Prevents chill haze in
beer, bread
Source: (Frazier and Westhoff 2008)
15.5. Application of Food Microbiology

As a discipline, the application of food microbiology to systems grasps the methods
involved in the assessment of microbial food safety and the advantageous use of
microorganisms in following of food fermentation, Probiotics development, food
spoilage and food borne diseases etc. as below:
15.5.1. Food Fermentation

Fermentation in food processing is the conversion of carbohydrates to alcohols and
carbon dioxide or organic acids using yeasts, bacteria, or a combination under
anaerobic conditions. Fermentation usually implies that the action of
microorganisms is desirable. The science of fermentation is also known as
zymology or zymurgy. The term "fermentation" is sometimes used to specifically
refer to the chemical conversion of sugars into ethanol, a process which is used to
produce alcoholic beverages such as wine, beer, and cider. Fermentation is also
employed in the leavening of bread (CO2 produced by yeast activity); in
preservation techniques to produce lactic acid in sour foods such as sauerkraut, dry
sausages, kimchi, and yogurt; and in pickling of foods with vinegar (acetic acid)
It involves genetically modified strain development and over production of desired
metabolic activities, sequencing genome of developed strains for better
understanding of their characteristics and effective methods to use microbial
metabolites in food processing.
15.5.2. Probiotics Development

According to the 2001 definition by the World Health Organization (WHO),
probiotics are “live microorganisms which, when administered in adequate
amounts, confer a health benefit on the host.”
Probiotics are live bacteria and yeasts that are thought to be beneficial in preventing
several health conditions. They are usually consumed as supplements and are also
referred to as “good bacteria.” Probiotics are said to restore the balance of bacteria
in the gut when it has become disrupted through long-term antibiotic use or
gastrointestinal disease.
Introduction of recent methods for better use of lactic acid bacteria for the
production of immunity proteins, gene sequencing for better understanding of their
characteristics and uses in food applications and determining antimicrobial
metabolites secreted by probiotics.
15.5.3. Food Spoilage

Spoilage is the process in which food deteriorates to the point in which it is not
edible to humans or its quality of edibility becomes reduced. Various external
forces are responsible for the spoilage of food. Food that is capable of spoiling is
referred to as perishable food.
Food microbiology involves the development of new techniques for prevention of
microbial spoilage, enzymatic spoilage by bacteria in frozen and refrigerated foods,
using nanotechnology to identify harmful bacteria and expected shelf life of foods
and importance of environmental factors on the response of harmful bacteria to
antimicrobial agents.
15.5.4. Food Borne Diseases

Foodborne illness (also foodborne disease and colloquially referred to as food
poisoning) is any illness resulting from the food spoilage of contaminated food,
pathogenic bacteria, viruses, or parasites that contaminate food, as well as chemical
or natural toxins such as poisonous mushrooms and various species of beans. Rapid
finding of disease causing bacteria in food and surroundings by using recent
techniques of biotechnology and nanotechnology and relationship of environmental
factors with identification and destruction of pathogenic bacteria, viruses and
parasites are also desired areas of foodborne disease factor.
15.5.5. Miscellaneous
Nevertheless, application of microbiology in combination with Hazard Analysis
Critical Control Point for food production, processing and preservation,
Assessment of microbiology of raw and lower temperature processed ready to eat
foods, Total quality management and control from farm to fork and Food safety
legislations and its enforcement criteria
15.6. Role of Food Microbiologist

Food Microbiology is a practical science and the major role of food microbiologist
is to ensure nutritious and safe food to the consumer. For efficient and reliable
production of food, a thorough understanding of microbial ecosystem is necessary.
The food microbiologist controls the exterior and interior biochemical reactions of
microbes in the food systems that can cause spoilage, address public health issues
and fermentation of food with full command in areas of a) microbial quality of
foods and its ingredients by using applicable methods, b) Knowledge about food
rules and laws (state, federal and international), c) Determining microbes with their
origin that cause spoilage and health issues, d) Adaptation of recent techniques in
food processing, their issues and solutions, e) Basic pathogenesis mechanism of
food borne microbes, f) Methods of identification and isolation of harmful
microbes from food and control measure g) Design operative sanitation techniques
to prevent decomposition and harmful micro-organisms issues in food processing
facilities, h) Knowledge about of starter cultures productions for fermentation and
prebiotics, i) Recognition of microbiological issues of imported foods world wide
and j) Educating food microbiology and its importance to various stakeholders.
15.7. Significance of Microorganisms in Food

Foods free of microorganisms are rare and their population in food depends upon
the entry points/status of microbes, growth pattern, survival rate and compatibility
with food. Microorganisms can enter into food through raw material or during
harvesting/slaughter, processing, storage and supply chains. The population and
survival of different microorganisms is estimated by food characteristics, its storage
conditions, characteristics of microbes and the processing. In most cases, the
presence of microbes in food is not considered as harmful. In some occasions
though, microbes show their occurrence and importance in several ways like food
spoilage, diseases spread caused by microbial contaminated foods and food
preparations that are more reliable through food fermentation
15.8. Environmental Microbiology of Food

Generally, environment components are atmosphere, lithosphere, hydrosphere and
mesosphere each with different share of microbes that depend upon geographical
location and environmental conditions of the components. Natural micro flora
maintains ecological balance with their hosts. Different kinds of microorganisms
can contaminate food through environmental components such as soil, air, water,
human activities, food elements, equipment, and miscellaneous sources. Entrance
and population of microbes in food system depends upon the sanitation level of
food system during handling and processing. For the control of microbial
contamination, one should know the source of microbes in food, its killing
techniques, microbiological standards, specifications and quality of food. Most
foods materials are not germ free may have microbes naturally and can also be
contaminated through environment. For safe and longer storage life of foods,
destruction of food spoiling microbes and creation of unfavourable conditions for

their survival and growth should be ensured (Frazier and Westhoff 2008).
15.8.1. Atmospheric Microbiology

Dust and moisture droplets in air are the main sources of microorganisms in
atmosphere, hence microbial population depends upon humidity, dust particles size,
temperature, air speed and their drying resistance. Commonly, the air contains
spores of molds, Clostridium spp, Bacillus spp, few Gram-positive bacteria (e.g.,
Sarcina spp, Micrococcus spp.) and yeasts. Air is the main transmitting agent of
harmful microbes from contaminated surroundings. Entry of microbes by air in the
food could be prevented by sterilizing the environment, filtering particles of dust in
the air, adjusting air pressure, increasing dryness, concentration and UV rays (Ray
and Bhunia 2013).
15.8.2. Lithospheric Microbiology

Agricultural soils and the soils under livestock may have several types and high
populations (billions/g) of microorganisms due to favourable multiplication and
growth medium (soil). Numerous kinds of molds, bacteria and yeast contaminate
foods through soil. Harmful bacteria and viruses enter into the food through soil
particles contaminated with faecal materials. Fish harvesting areas and soil are also
the source and entry points of pathogenic microbes and parasites in the food
systems. Issue of microbial contamination through soil can be solved by removing
the soil from foods through washing and cleaning. Thus shielding the food from
soil particles is commonly acceptable to reduce the hazard of food contamination
through soil (Ray and Bhunia 2013).
15.8.3. Hydrospheric Microbiology

Hydrosphere (fresh and sea water) is the largest part of the biosphere containing
huge number of microorganisms. Water play a key role in many processed foods,
agriculture, drinking water, aquaculture, marine products, cleaning food
commodities, food preservation and equipment washing. Thus water quality has
pronounced impact on microbial quality of food. Pathogenic bacteria, viruses, and
parasites can enter into food through polluted water used in food processing. Water
contains spoilage causing bacteria like Alcaligenes, Pseudomonas and
Flavobacterium. Untreated water may also be contaminated with coliforms and
pathogens (mainly enteric types). To overcome the problems, water free of
microorganisms should be used in food processing.
Cyanobacteria from the unicellular and the dinoflagellates from the multicellular
organisms have the aquatic origin and badly affect food quality and safety by
producing toxins in the food. These toxic foods, if ingested, can cause illness such
as paralytic shellfish poisoning (Ray and Bhunia 2013).
15.8.4. Equipment Microbiology

Equipment may be contaminated by various microorganisms from air, raw foods,
water and workers handling the equipment. Microrganisms under favourable
environmental conditions and time, rapidly grow and contaminate bulk of food,
multiply and are continuously transferred by the contaminated equipments that are
being used longer periods.
In some equipment, from uncleaned and unsanitized parts, harmful microorganisms
can enter into the food. Due to improper cleaning of tools and equipments like
knives, cutting boards, spoons, and similar articles microbes can enter into food
commodities. Pseudomonas, Listeria, Escherichia, Enterococcus, Salmonella,
Lactobacillus, Leuconostoc, Clostridium, Bacillus spp., yeasts and molds can get
entry into the food from equipments (Ray and Bhunia 2013).To decrease the
microbial levels in the foods, instruments design should be such that they are easy
to clean and sanitize and their cleaning at short intervals is recommended.
15.8.5. Miscellaneous
Packaging materials should be free of pathogenic microorganisms, vermins, flies,
rodents and birds during food processing and in storage places. Harmful
microorganisms in food deteriorate the quality of food, therefore, appropriate
precautions should be adapted to prevent contamination of food from these sources.
15.9. Contamination of Foods

Food having undesirable characters with respect to impurity is unfit for use and is
termed as contaminated. Contamination implies the unintentionally addition of
things in food other than the product and microbes. Physical contaminants are
materials that become part of a food and may or may not change the quality of the
food itself. For instance, metal filings or broken pieces of glass have occasionally
gotten into foods causing injury if swallowed. It usually includes pieces of glass,
wood, packaging material, rodent, insects and their droppings. Insecticides and
herbicides used to control the insects and weeds in crops for better yield could be
food contaminant. Pesticide residues on food may enter the food while deteriorating
the quality. Pathogens are harmful microorganisms and cause of human diseases. A
change in a food that makes it unfit or undesirable for consumption is called as
food spoilage, a foodborne illness caused by a toxin produced by microbes in food
is called food intoxication whereas, a foodborne illness caused by ingestion of
foodborne pathogens is called as food infection.
15.9.1. Major Sources of Contamination

Many sources can contaminate the food including antibiotics and hormones
production in meat and dairy foodstuff. Food may also be contaminated with
chemicals which come from pesticides, during industrial transportations, or during
use of chemicals in packing of food.
A number of contaminants can accumulate in many kinds of animals and are

frequently found in meat, poultry, fish and dairy products. Pesticides found in
fruits, vegetables and other agricultural supplies are also the source of
contamination. The children under age of five years are the most vulnerable to
these infections (Abraham et al. 2012).
15.9.1.1. Air
The air is an important potential source of microorganisms, including pathogens.
Air filter should be used in high risk environment to avoid microbial contamination
of food. Such production practices should be adopted that minimizes aerosol
generation within the high-risk area. Tuberculosis, measles and mumps could be
transmitted by coughing or sneezing during processing and handling of foods.
Infinite number of microbes can infect the surroundings or persons by a single
sneeze.
15.9.1.2. Water
Water contaminated with human or animal faeces contains harmful microbes and it
causes several diseases. Water borne diseases can be overcome by drinking safe
and pure water.
15.9.1.3. Soil
Contamination through soil, plants and water can cause a variety of diseases. Soil is
the loving habitat for most of fungal agents (mycoses) in addition to bacterial
source. Histoplasmosis causing pathogens live in high organic and bird droppings
loaded soil.
15.9.1.4. Animals
Animal can also act as infectious agents in the same mode as human infect others.
They may be cause of severe clinical cases or transporters exposed to the diseases
and phases of such transmissions.
15.9.2. Contamination through Traditional Marketing Systems

Local or long transported food used by most of the communities internationally is
now recognized as contaminated by environmental factors. Multiple agents can
contaminate the same food commodity. Major issues of evaluation, risk assessment
and management of food contamination through environment have been studied.
Diet varies according to sex, age and society differences. Foods in Pakistan are
shortened of nutrients like vitamin A, calcium, folic acid and dietary fibre. In a
study, it was observed that concentration of zinc, iron and potassium were higher
while concentration of sodium, fat, saturated fat and sucrose were lower when only
conventional foods were used instead of market foods. In this population, the move
away from conventional food towards a diet composed completely of market food.
It was characterized by an upturn in absolute energy intake and an increase in the
relative contributions of carbohydrate (particularly sucrose), fat and saturated fat.
This type of dietary change from conventional to market based foods can produce
potential negative health consequences so it is very necessary to document the

current health situation in the country (Receveur et al. 1997).
15.9.3. Contamination of Milk and Meat in Pakistani Market

System
Many foods like dairy products, processed meats and seafood may cause human
infection. Tuberculosis and brucellosis are not the main diseases but milk-borne
illness of salmonellosis and campylobacter enteritis is spreading day by day due to
use of untreated milk. Milk can be disinfected by heating it. It is awful news that
the continued sale of raw milk is being allowed in Great Britain (Galbraith et al.
1982). Milk and meat may serve as important carrier for dissemination of the
pathogenic bacteria of animal origin like E. coli to consumers or humans. Cattle are
a major reservoir of pathogenic E. coli and faecal contamination in the animal meat
occurs during its handling, slaughtering and processing. Traditional ways of
slaughtering and processing of meat in Pakistan also help to increase the occurrence
of E. coli to many folds. E. coli was first recognized as a cause of illness in 1982
during an outbreak of severe bloody diarrhoea that was traced to contaminated
hamburgers. In meat, if E. coli is present, it will cause economic losses to the
farmers. Milk, on the other hand is commonly contaminated with aflatoxin M1.
15.9.4. Contamination of Microbes through Workers /Services

Provider
Contact of microbial infected hands of the workers to the food products imparts a
large contamination to food. To avoid this, hands should be washed properly.
Infected equipment and contaminated raw materials causes production of
contaminated products and even workers families through contact means if do not
adapt proper hygienic practices. In many cases infection in factory has been spread
by faeces of infected person. Foods composed of multi-ingredient with limited
descriptions served in restaurants or hotels, at schools and at catered events are
vulnerable to contamination. Readymade foods including baked commodities,
beverages and products of animal origin are identified as contaminated. In some
cases, it is hard to announce the worker as cause or victim of diseases. But mostly
workers have role in spreading disease although they do not accept that they are ill
for a variety of reasons, but subsequent tests and examination provid indication of
infection (Todd et al. 2007).
15.9.5. Mode of Microbial Transmission

Transmission of microbes occurs from reservoir to the host in several ways, the two
basic modes are direct mode and indirect approach. Direct transfer of microbes
occurs when infectious and susceptible person are in direct contact with each other.
Indirect transmission of microbes happens when susceptible are infected through
spores or harmful microbes present in the air or environment. Exact recognition of
the transfer method can enable for upgraded anticipation and diseases control. By
knowing the mode of microbe’s transmission, one can illustrate biological impacts
of pathogens. Black band disease in corals, avian influenza, severe wasting disease
in deer, are few of the examples. Knowledge of the transmission corridor is also
necessary for understanding the evolution of environmentally significant
pathogenic species (Cortez and Weitz 2013).
15.10. Control of Microorganisms in Food

Food can be preserved by simply manipulating physical and chemical factors that
interrupt microbial growth. For studying preservation, the key objective is to learn
the impact of various preservation methods on the microbial population.
15.10.1. Physical Methods

15.10.1.1. Use of High Temperature
High temperature i.e., above room temperature is used to kill the microbes for food
preservation. In food preservation, there are two temperature categories that are
commonly used: pasteurization and sterilization.
i) Pasteurization
In the Pasteurization treatment, high temperature is applied for the destruction of
disease causing microorganisms and for the killing or reducing the spoilage
microorganisms in specific foods, milk is the main example of pasteurization. Milk
processing for microbial control is achieved by heating at different temperatures for
corresponding time periods such as: 65°C for half an hour, 72°C for 15 seconds etc.
These temperature and time ranges are enough to kill most of the vegetative /
pathogenic molds, yeasts and bacteria. In brewing industry pasteurization (to
destroy spoilage organisms) of bear is usually done at 60°C for 7 to 15 minutes.
ii) Sterilization
Sterilization is the destruction of all kind of microorganisms including their spores
in a material or on an object when measured by any appropriate enumeration
method. Sterilization is done to make the foods or objects sterile. Sometimes,
canned foods are labelled as commercially sterile to identify that either the number
of organisms that survive are very low or no organism can be detected during any
cultural method and they are insignificant under different storage conditions.
Micro-organisms may present in canned foods but the can’s inner conditions e.g.
pH, temperature etc. may not support for microbial growth (Jelen 1982). The
sterilisation is normally done at 121°C for 15 minutes.
iii) Ultra High Temperatures (UHT)
Ultra high temperature or flash pasteurization is used for processing of many
products. The fruit juice is subjected to 116°C for few moments before filling. The
basic features of the ultra-high temperature include its continuous nature, aseptic
handling of product from the sterilizer, aseptic storage, very high temperatures of
140-150°C and usually little time for a few seconds essential to attain commercially
sterile product. Consumers like ultra-high temperature processed milks than
conventional heated pasteurized milk or milk products. The sterilized produces can
be kept at ambient temperature without flavor changes up to 8 weeks. Higher

temperatures for short time are effective in killing the microorganisms without
bringing adverse changes in product (Burton 1985).
iv) Dry Heat
It is used to sterilize materials and surfaces which are highly heat resistant and
containing no liquids. Penetration rate of dry heat is very slow than moist heat so
higher temperatures of 160-170°C or more for longer time periods (several hours)
are required, depending on the load and distribution of materials in the oven (Bone
1973).
v) Steam
The autoclave is the main example of moist heat treatment which is a sophisticated
version of the pressure cooker that is used in canning and bottling of foods. Moist
heat penetrates at faster rate than dry heat and it is used to sterilize agar
preparations and culture solutions. A main reason for pressurized steam is to
destroy bacterial spores that can survive during boiling process. In autoclave,
typically a pressure of 15 psi is sufficient to create steam at a high enough
temperature of 121°C to kill the endospores.
15.10.1.2. Use of Low Temperature
Food spoilage is the result of different chemical reactions. Microorganisms and
endogenous enzymes are responsible for these chemical reactions. The rate of many
chemical reactions varies with temperature changes; as temperature is decreased the
degree of reaction slows down. Use of low temperature, actually slows down the
chemical reactions that are responsible for food spoilage.
i) Freezing
Freezing is very significant method of food preservation because of restoration of
freshness and nutrient quality of food for a longer time period. Food material
begins to freeze at temperature between -0.5 to -3°C. During freezing the water
changes into ice and the concentration of solute increases in the unfrozen water
which decreases its freezing point, even at very low temperature of -60°C.
Temperature below -18°C is used for freezing the food commodities. At low
temperature, microbial growth is hindered, but residual microorganisms and
activity of some enzymes such as lipase can cause spoilage of food products.
Blanching of fruits and vegetables should be done before freezing to avoid
enzymatic activity which can cause the discoloration of food product during
storage.
ii) Chill Storage
In chill storage, food products are kept above their freezing temperature which is
normally in the range of 0-5°C. Chill storage delays the process of spoilage as well
as changes the nature of spoilage. Low temperature changes the spoilage
characteristics as low temperature stop the growth of mesophilic and thermophilic
microbes and leads to a micro flora dominated by psychrophilic bacteria. Low
temperature can also alter the physiology of microorganisms that changes the
spoilage characteristics. Psychrophilic bacteria can grow at chilling temperature but

their activity is so negligible that the onset of spoilage is delayed. The growth of
microorganisms at low temperature is connected to the composition of the cell
membrane. At low temperature, phase transition takes place in the structure of cell
membrane and it changes from crystalline liquid form to rigid gel form. By this
phenomena transportation of solute is limited. So chilling process will result in a
cold shock phenomenon which leads to cause the death and injury of microbial
population (Ronsivalli and Charm 1975).
15.10.1.3. Pascalization (High Pressure Processing)
Application of high hydrostatic pressure technique contains pressure container,
fluid for the transfer of pressure and pressure generator pump. High pressure
processing (HPP) maintain the quality of product. Nutritious value, flavor,
appearance, color and texture of HPP foods resemble closely to fresh food
products.
High hydrostatic pressure affects the non-covalent bonds like hydrogen,
hydrophobic and ionic bonds. It can damage the structure of proteins by affecting
the hydrogen, hydrophobic and ionic forces. Proteins of eggs, meat and soya form
gels while other proteins are not affected and this can have the adverse effects on
food quality when the enzymatic activity decreases the shelf life of product. For
example in orange juice pectin esterase must be inactivated to preserve the desired
cloudiness of the product. High pressure may affect the non-protein macro
molecules such as high pressure processed starch products have sweet taste because
of alterations in the starch properties which permit greater access to the amylase
(Alpas 2000).
15.10.1.4. Hyper Filtration
Hyper filtration technique is used to make food and products free from bacteria and
fungi. Viruses could not be excluded from food products due to its very small size
in nature. The technique is mainly used for the sterilization of fluids. Microbes can
also be destroyed by other methods such as heat processing and irradiation in
combination to Hyper filtration, although the technique is not economical and not
used for bulk food preservation.
15.10.1.5. Hyperosmotic
Hyperosmotic condition preserves food because of squeezing water out of bacteria
and fungi so that they cannot grow further. Jams and pickles are good examples of
its application because of high solute loads that are hyperosmotic to the cytoplasm
of bacteria which removes water from the cells by osmosis. But some microbes rise
in hyperosmotic conditions, for instance some yeast in brine pickles and surface
molds in jams.
15.10.1.6. Modification of Atmosphere
Modified atmosphere prevents the growth of anaerobes, that would otherwise spoil
perishable food products, while improving nutritional and keeping quality of food
products although it is less efficient in killing spoilage causing microbes. The
practice is useful if combined with refrigeration temperature. Three different
measures are being used to modify the atmosphere: vacuum packaging, modified-
atmosphere packing or gas blushing and controlled atmosphere (Farber 1991).
15.10.1.7. Control of Water Activity (aw)
The water activity of a food may be lowered physically by extraction of liquid /
water by drying or freezing methods. It can be dropped by using high sugar and
salts. Drying enhances the shelf stability of food because of restricted water
availability to microbes in dried food products. Sometimes drying just stops the
growth of microorganisms and does not kill the microbes. Freeze drying is very
operative to hinder the growth of microorganisms. In freeze-drying, the product is
subjected to freezing temperature and then water is removed under vacuum.
Growth of microbes is inhibited at 0.6 water activity level. Intermediary moisture
foods (IMF) are generally described as those foods with aw in the range 0.6 to 0.85.
Moisture content in the range of 15-50% prevents the growth of bacteria, yeast and
molds with extension of the shelf-life of foods at room temperature.
Fire drying gives a smoked flavor to the desired products as well it increases shelf
life. In warm environments fruits, grains, coffee and fish are dried in direct sun
light however, this method offers best chances of adulteration by rodents, insects,
birds and dust etc. Solar drying is economical but less efficient than machine drying
methods. In the mechanical methods like drum drying, product is sheeted over
rotating drums with temperature (150oC) by steam generation method. As drum
rotates gradually the film dries and the product is scrapped off in the form of sheet
by a fixed blade. Evaporation of some moisture concentrates the soluble solids and
thus decreases the water activity. Tomato paste is the example in which the
evaporation of moisture is common technique of food preservation (Troller and
Christian 1978).
15.10.1.8. Use of Irradiation
Radiations are the release of energy from the space or material medium.
Electromagnetic waves are applied in food processing. The electromagnetic
spectrum is further sub divided according to the use of radiations in food
processing: X-rays, gamma rays, ultraviolet rays and microwaves. Ionizing
radiations are of primary interest in food processing.
i) Ultraviolet Light
Ultraviolet light is very effective against bacterial population having the
wavelength of 2600°A. Proteins and nucleic acid absorb it, where it is responsible
for photochemical changes in cell and ultimately causes the cell death. The UV
light produces lethal mutation in bacterial cells by disturbing nucleic acids. The
application of UV light to food products is limited due to its poor penetrative
capacities. The main problem of UV light is that it may cause oxidative changes in
food that leads to rancidity and discoloration. Ultra violet light is also used for
treatment of baked products before wrapping.
ii) Ionizing Radiation
Ionizing radiations are high energy (>10eV) gamma and x-rays. These rays have
the high energy due to which they hit electrons away from the molecules and ionize
them. When these ionizing radiations pass through the microbial cells they create
hydrogen ions, hydroxide ions and some types of peroxides which causes
intracellular damages (Thayer et al. 1991).
a. Gamma Rays
Gamma rays are highly speeded electrons used for food preservation
which are formed from the highly excited nucleus of cobalt 60 and cesium
137. These are the most inexpensive radiations. The source elements are
recovered from waste products of atomic reactions. These radiations are
having very high dissemination capabilities in contrast to Beta rays.
b. X-rays
X-rays are generated in evacuated tube by the collision of high velocity
electrons on the heavy metals. Their effects on food preservation are
similar to that as gamma rays.
iii) Microwaves
Microwaves generate an electromagnetic field in which food material is located.
Food material is electrically neutral. When electromagnetic field is created,
molecules move from one side to another side. In this process, molecules bring into
line themselves along with the changing electromagnetic field. As the molecules
move fastly to reach at +ive and -ive pools, friction between molecules causes
heating of molecules. As a result, microwave energy is produced. Two frequencies
915 and 2450 megacycles are frequently researched by scientists. At 915
megacycles, molecules of food material swing forth and back 915 million times per
second.
iv) Radicidation, Radappertization and Radurization of Foods
a. Radicidation is same as the pasteurization of milk. It is application of
radiation to food products to enhance their shelf life by lowering the
number of live non-spore forming harmful microbes. The irradiation
dose for this process is 2.5-10 kGy.
b. Radappertization is same to commercial sterilization radiation
treatments and is applied to food to kill all pathogens and most food
spoilage bacteria. Level of radiations is typically between 30-40 kGy.
c. Radurization is considered equal to pasteurization. It is the application
of radiation to the food product in order to enhance their keeping
quality by reduction in number of viable specific food spoilage
microorganisms. Typical level to achieve this is 0.75-2.5 kGy. This
technique is applicable for cereal grains, fruits, vegetables, meat,
poultry and seafood.
15.10.2. Chemical Methods

Chemical compounds which hinder or help to prevent the growth of
microorganisms or deterioration are called preservatives. Preservatives could be
such compounds that kill the specific target organisms or they may only inhibit
their growth.
15.10.2.1. Organic Acids and Esters
Organic acids are produced by microbes and also by using chemical methods of
production such as food-grade acetic acid derived from chemical origin sometimes
act as a substitute to vinegar. Organic acids are added as element in fermented
products such as sauces and pickles, or they may be produced in situ in the vast
variety of lactic-fermented products. Benzoic acid which is naturally present in tea,
greengage plums, cherry bark and cranberries but are also commercially
manufactured for use in food. Undissociated organic acids act as antimicrobial
agents therefore, used to inhibit growth of yeast and molds. Organic acids do their
best job in non-acid foods. Their activity against microbes rises as ester group
carbon chain length increases hence lowers water solubility and become less
efficient in fat based products.
15.10.2.2. Sulfur Dioxide
Sulfur dioxide is used to prevent oxidation and browning reactions in many food
commodities. SO2 is colorless and readily soluble in water. It is effective to control
spoilage causing microbes in food. Gram positive bacteria are very resistant against
this gas. Acceptable level of sulfite application in fresh sausage is 450mg/kg in
England. In wine making, the tolerance of the wine yeast Saccharomyces cerevisiae
to SO2 levels around 100mg/kg is exploited to inhibit the population of undesired
yeast and bacteria. Fruits over supplies can be stored for longer duration by
application SO2 (Davidson and Branen 1993).
15.10.2.3. Nitrite
In the beginning of curing processes, nitrite was manufactured by the bacterial
lessening of nitrate present as an impurity in the crude salt used, but now nitrate, or
more commonly sodium or potassium salts of nitrite are added. Nitrite is very
effective against bacterial population. The most important is the ability of nitrite to
hinder the growth of Clostridium botulinum which is spore forming bacteria and
heat resistant. To attain its killing in meat, initial dose of 100mg/kg of nitrite is
applied. Nitrous acid is the active agent because when pH is decreased the bacterial
inhibition increases. Nitrite is also effective against the spores of microbes. It acts
as inhibitor of spore’s germination (Woods et al. 1981).
15.10.2.4. Natamycin
Natamycin is produced by bacterium Streptomyces natalensis. It was previously
known as pimaricin which is a polyene macrolide antibiotic. It is very effective
against the fungal sterol and ergo sterol. Its mode of action is through rupturing of
the fungal cell membrane. As the consequence, solutes from the cytoplasm are
leached outside and cell dies and therefore it is effective antifungal agent. Its
solubility in water is low so it is used in suspension form to inhibit fungal
population. It has some rewards in competition to sorbate, as it does not penetrate
into product. It is also effective at elevated pH. It does not kill the beneficial
bacteria (Holley 1981).
15.10.3. Bio-Preservation of Food

Bio-preservation is defined as the use of natural or controlled microbiota or
antimicrobials for the preservation of food and shelf life extension. This is
performed by using beneficial bacteria that produces fermentation products. The
process is used to limit and control the spoilage also disable the pathogens present
in food. In current scenario, lactic acid bacterial (LAB) antibiotics (bacteriocins)
are used as an important part of hurdle technology. They efficiently control
spoilage bacteria and other pathogens when used in combination with other
preservative techniques. They also hinder the activities of a wide range of
organisms like gram negative bacteria which are naturally resistant.
15.10.3.1. Bacteriocins in Food Application
These are bacterial ribosomal produced proteins or peptides containing
antimicrobial characteristics. Bacteriocins comprise of a very heterogeneous group
with respect to their composition, physical, chemical properties and primary
structure. Food and feed fermentation of lactic acid bacteria are basics on which
bacteriocins functions as bio-preservation depends. Lactic acid bacteria and their
bacteriocins have been consumed from long times and their use in food is safe.
Large numbers of bacteriocins have role in the manufacturing of basic food
materials in various fields such as in agriculture, veterinary and aquaculture fields.
Lacticin and nisin have been presented into different prophylactic measures against
the treatment of mastitis. In aquaculture colicin like bacteriocins have activity
against many gram negative pathogens for the objective of biocontrol. The major
field of application of bacteriocins to inhibit the pathogenic or spoilage bacteria is
during food manufacturing and preservation. Principally bacteriocins are used in
three different arrangements, 1) In situ production by protective or/and starter
cultures, 2) a fermenter ingredient of a bacteriocins strain, 3) additive component in
purified preparation. Dairy starters that produce nisin are developed specifically to
stop the growth of Clostridium tyrobutyricum in semi hard and Staphylococcus
aureus in acid coagulated cheeses. Protective cultures have been used to enhance
the quality of meat and fish products, which do not contribute in the sensory
characteristics of food (Schillinger et al. 1996).
15.10.3.2. Bacteriophage in Food Applications
Bacteriophages are capable of, 1) as phage therapy to lessen or prevent the
colonization and diseases in animals, 2) as bio-control to increase the storage
period of perishable foods products, 3) as bio-sanitizer to eliminate the
contaminants from the carcasses and other products that are in raw form and to
disinfect contact surfaces. Bacteriophages have been used to decrease the pathogen
burden in animal farming, during milking and after slaughtering of animals. Phages
decrease the number of foodborne pathogens e.g. Salmonella and Escherichia coli
0157:H7 on fresh poultry and Listeria monocytogenes on ripened cheeses. When
chickens are treated with bacteriophages against Salmonella and Campylobacter,
significant lowering of bacterial load is detected after the treatment with
bacteriophage especially when applied before slaughtering of the animal (Atterbury
et al. 2003). Bio-sanitation with phage treatment has been used to lessen formation
of biofilm or reduce / eliminate the Staphylococcus aureus skin or nasal

colonization to the persons who handle the food. The phage specificity to host
sometimes restricted only to a few number of strains and they form a burden to
their use as food preservatives or bio control. When phages are familiarized into
milk which is contaminated with Salmonella in cheddar cheese production,
bacterial cells decrease after storage. In the same way, growth of Staphylococcus
aureus in milk and during the production of curd is hindered by phages, its
inhibition continues during storage and ripening of the semi hard and acid
coagulated cheeses. Other examples of bio-preservation with the use of
bacteriophages include the prevention of Salmonella typhimurium on meat
frankfurters and Entero bactersakazakii in rebuilt infant formula. They are also
used for the recognition of foodborne pathogens (Goode et al. 2003).
15.10.3.3. Endolysins in Food Applications
Endolysins are powerful antimicrobials and have role in the cure of infections from
bacteria in animal system. Large numbers of endolysins have activity against
various foodborne pathogens. Basically in the food production or processing
operations, biocontrol of pathogens by the use of endolysins has been used in dairy
industry. The purified endolysins are used in milk pasteurization treatment to
abolish Staphylococcus aureus. Recombinant lactic acid bacteria have capability to
secrete Listeria endolysins but their activity as antagonistic in milk and other food
systems have not been verified (Zimmer et al. 2002).
15.11. Food Borne Illnesses

Food borne diseases has been internationally defined as; “Any infectious or toxic
natured disease produced by or thought to be caused by, the ingesting of food or
water”. According to Dorland's Medical Dictionary and CDC food poisoning guide,
foodborne illness (also foodborne disease and colloquially referred to as food
poisoning) is consequential from the ingestion of food polluted by disease
causing microorganisms (Adams and Moss 2008).
Food is essential to the conservation of life but it may spread the diseases. Harmful
compounds can be present in the food. Furthermore, the hazards created by natural
toxins, foods may also act as the source by which an exogenous harmful agent may
be ingested. The agents causing human illness conveyed by food are bacteria,
viruses, fungi, parasites, toxins produced by microbes, chemicals and toxins
naturally present in plants and animals. Many experiments about hazards linked
with food have resulted that microbes are of supreme significance (Ray and Bhunia
2013).
Various foods take part in food borne illnesses but meat and meat products are the
source for communication of a major share of food borne illnesses. Meat can be
contaminated during its preparation. Milk can be a source of antibiotic resistant
zoonotic pathogens (S. aureus, Strept. agalactiae, E. coli, K. pneumoniae, S.
Typhimurium and Serratia marescens). Furthermore, relying fish may be fabricated
into smaller cuts at the local level with the
The most commonly recognized food borne pathogens include bacteria (e.g. E.coli,
Campylobacter, Salmonella, L. monocytogenes, Shigella, C. botulinum, C.
perfringens, S. aureus, B. cereus); viruses (Hepatitis A virus, Norovirus.
Rotavirus); parasites (Taeniasaginata, Echinococcus, Ascarislum bricoides,
Trichinell aspiralis, Fasciola, Cryptosporidium parvum, Entamoeba histolytica,
Toxoplasma gondii Taeniasolium,); mycotoxins and marine biotoxins (Hassanain
et al. 2013).
15.11.1. Food Intoxication

Bacteria and other microbes produce toxins in food. If that food is ingested, then
food intoxication may occur causing diseases. Toxins may or may not change the
sensory properties like odor and flavor of food. Main toxin producing bacteria are
Clostridium botulinum and Staphylococcus aureus (Ray and Bhunia 2013). The
toxins are resistant against high and freezing temperatures. Bacteria introduced
during harvesting, handling, processing and preparation may responsible to
contaminate food. When food contaminated with exotoxins is ingested, the person
can be diseased like vomiting, nausea, and diarrhoea. Numbness and tingling
around the lips can also be detected. The symptoms usually appear quickly because
the toxins act speedily in the body. People with food intoxication get sick
immediately. Some important food intoxication is described here:
15.11.1.1. Botulism
It is caused by botulinum neurotoxin contaminated foods. Main symptoms are
muscle paralysis exhibited by slurred speech, double vision, drooping eyelids,
blurred vision, and difficulty in swallowing, dry mouth and muscle weakness. If not
properlt treated, symptoms can lead to paralysis of the legs, arms, trunk and
respiratory muscles. Symptoms can appear from 6 hours of ingestion up to 10 days
(CDC 2008). An outbreak of type E food borne botulism has happened in Egypt in
1991 and the patients were cured with both heptavalent botulism immune globulin
provided to the Egyptian Ministry of Health by the U.S. Army and commercially
available trivalent antitoxins. Possible serum illness in hospital was verified for half
patients who were receiving commercial antitoxins (Hassanain et al. 2013).
15.11.1.2. Staphylococcal Intoxication
Staphylococcal food poisoning (SFP) or intoxication is one of the common
categories of food-borne disease which is produced by the ingestion of foods
containing thermo stable toxins formed by strains of the bacterium Staphylococcus
aureus. It is extensively dispersed in water, air, dust, sewage, foods, humans and
animals. It can cause infections and diseases like boils, carbuncles, bullous
impetigo, toxic shock syndrome, scalded skin syndrome, enterocolitis,
osteomyelitis and food poisoning. It is usually transmitted to the foods and food
products due to their poor handling. The growth of the bacterium is ideal in protein
rich foods with high salt content. Foods that are often implicated in staphylococcal
food intoxication include meat, poultry, fish, salads, milk/ milk products and
cream-filled bakery products.
Communal indications include abdominal cramps, nausea, vomiting, diarrhoea,

dehydration, prostration, weakness and subnormal body temperature.
Staphylococcal food intoxication is seldom fatal and recovery usually occurs within
24-48 hours. Numerous antigenically distinct types of staphylococcal enterotoxins
are produced by strains of S. aureus which are named as A, B, C1, C2, C3, D, E, G,
H, I and J. Among these, staphylococcal enterotoxin A is very noxious and
concerned in most of the food intoxication (Singh and Panday 2012).
15.11.1.3. Clostridium Perfringens Intoxication
It occurs 6 to 1 day after consumption of spoiled food. Abdominal pain and
diarrhoea are its signs. Sometimes vomiting and fever are also happened. In very
few cases it is lethal. In much infected form involve "Type C" strains of the
organism causes ulcers of stomach. (Hassanain et al. 2013).
15.11.2. Food Infections

Food infection happens as a consequence of intake of contaminated food and water
with pathogenic microbes. For bacteria to cause food infection, it is required for
them to remain viable during the consumption of food (Ray and Bhunia 2013).
Ingestion of V. parahaemolyticus in raw or undercooked seafood, typically oysters,
causes serious gastroenteritis. The incubation period of 1 day is tailed by severe
watery diarrhoea, vomiting, nausea, abdominal cramps and fever (Hassanain et al.
2013).
15.11.2.1. Campylobacteriosis
Human gets infected with Campylobacter by absorption of contaminated food
(chicken meat). Most strains of C. jejuni produce a toxic material (cytodistending
toxin) that delays the cell division. The sites of tissue injury include the jejunum,
ileum and colon. Campylobacteriosis causes inflammation, bloody diarrhoea,
dysentery, cramps, pain and fever. C. jejuni can lead to secondary Guillain-Barré
syndrome (Hassanain et al. 2013).
15.11.2.2. Food Borne Salmonellosis
Salmonella typhimurium and S. enteritidis are the main causing agents of
salmonellosis. Improperly cooked meat, infected eggs and milk, reptiles, pet
rodents, spoiled fruits and vegetables are the sources of these microbes. Diarrhoea,
fever, vomiting and abdominal cramps are signs of salmonellosis and these signs
occurs within three days of infection. Food net stated two infectious diseases
involving contaminated tahini from Egypt resulting in an outbreak of S. Montevideo
infection (Hassanain et al. 2013).
15.11.3. Mycotoxicoses
Many strains of molds, while developing in appropriate environment, yield toxic
materials denoted as mycotoxins. Ingesting of food containing these toxins causes
mycotoxicoses. In this disease, symptoms are very potential and hard to identify.
Elevated humidity and temperature supports its spreading. High moisture grains are
the mostly spoiled with these microbes.
15.11.3.1. Aflatoxins
Aspergillus parasiticus and Aspergillus flavus are the production agents of
aflatoxins. Tree nuts, maize, peanuts, oil seeds including corn and cotton are also
infected by aflatoxins. Aflatoxins suppress the immune system, causes mutation,
teratogenic and carcinogenic compounds. They mainly effect the liver. Aflatoxins
cause necrosis, cirrhosis and cancer of the liver (Hassanain et al. 2013).
15.11.3.2. Altertoxins
Altertoxins are originated from Alternaria spp. Wheat and tomatoes contain some
quantity of the toxins. The toxins easily contaminate the grain products. So during
grains processing and storage, hygienic practices should be adapted to avoid
contamination.
15.11.3.3. Fumonisins
Corn can be simply contaminated by Fusarium moniliforme. Fumonisins B1 and
B2 are more toxic and infectious than others (B3, B4, A1 and A2) Fumonisin B1
causes oesophageal cancer in humans.
15.11.3.4. Ochratoxin
Penicillium and Aspergillus species are the causing agents of these toxins.
Aspergillus ochraceus can contaminate alcoholic beverages. Aspergillus
carbonarius produces toxic compounds in fruit juices. Ochratoxin A has been
considered as cancer causing agent in the nephrons and urinary tract (Shephard
2008)
15.11.3.5. Zearalenone
Fusarium graminearum and related species are its production sources. Wheat,
barley and sorghum are more affected by these toxins. This toxin creates
reproductive tract problems in animal species (vaginal prolapse, infertility,
vulvaloedema, and mammary hypertrophy in females and feminization of males,
atrophy of testes and enlargement of mammary glands).
Severe mycotoxicoses can lead to death. Proper treatment should be given if some
persons get involved in the mycotoxicoses. Symptoms appearance in infected
persons are delayed after ingestion of contaminated food. Safe food quality should
be maintained at homes and industry to avoid these infections (Hassanain et al.
2013).
15.11.4. Viral infections

15.11.4.1. Hepatitis A Virus (HAV)
Hepatitis A virus (HAV) is a critical contagious disease of the liver. Its virus is
conveyed by the food or water contaminated with faeces or oral secretions. Fish
living in polluted water may cause the infection. Zaher et al. (2008) in Egypt
proved the existence of HAV in products of meat and milk. HAV vaccine has been
proven effective against this infection (Hassanain et al. 2013).
15.11.4.2. Hepatitis E
Hepatitis E is not a severe disease and may be treated with mild medication. Its
chances of lethality are 2%. It is severe in pregnancy phase, connected with a
syndrome called "fulminant hepatic failure" with death chances of around 20%.
Contamination of water with faecal residues is its major causing source. It is not an
infectious disease.
15.11.4.3. Rotavirus
Rotavirus causes the stomach diseases and flu in humans. Nearly every child in the
world has been infested with rotavirus at least once but adults are infrequently
diseased. Zaher et al. (2008) revealed the occurrence of Rotavirus in meat and milk
products intended for human consumption in different localities in Egypt. Rotavirus
is transferred by faeces and oral secretions. It infects the cells of small intestine and
generates a toxin, which brings gastroenteritis, leading to severe diarrhoea and
dehydration leading to death (Hassanain et al. 2013)
15.11.5. Current Status of Food Borne Pathogenicity in Pakistan

Abdominal pain and dysentery are mainly caused by Campylobacter infections.
Studies showed that meat and milk in Pakistan are contaminated with
Campylobacters. In a study 1636 samples of meat, milk and vegetables from three
major cities of Pakistan were collected and examined. Study confirmed that 48%
samples of milk and meat and 40.9% of vegetables were contaminated with
Campylobacters. Shigellosis is more acute and hazardous than Campylobacter
infections. Shigella Spp. are more resistant to killing methods and they cause a lot
of diseases now a day. Shigellosis is spreaded by deprived hygienic environments,
polluted water and inaccessibility of better health care services. Hence, this disease
is common in poor population of Pakistan. Vegetables consumed in Pakistan serve
as a main source of nutrients like vitamins, minerals, bioactive compounds and
fiber. Listeria monocytogenesis commonly present in uncleaned vegetables, thus
more care has to be exercised to get advantages from their nutrients (Soofi et al.
2011; Akhtar 2013). Diarrhoea is the most prevalent of the food borne illness
dominant in Pakistan and is primary health issue in Pakistani children and infants
(Soofi et al. 2011).
Foods mismanagement is major causing agent of diseases caused by food. Fruit
chats were found heavily contaminated with Enterobacter spp., E. coli, Klebsiella
spp., Salmonella spp., S.aureus and S. Epidermidis. The arrangement of fruit chats
without cover is the major cause of contamination from surroundings. Khoya and
burfi, are also contaminated from S. aureus, E. coli and Klebsiella spp. (Farzana
et al. 2011; Akhtar 2013). Aflatoxins are present in Pakistani foods because of the
warm and humid climate (Mobeen et al. 2011).
S. aureus, C. perfringens and Bacillus cereus may contaminate the home made
foods in Pakistan. Another study verified higher percentage of the tested samples of
ice cream to be highly polluted with Enterobacter (En). Aerogenes (34%), S.

aureus (26%), Proteus spp., (16%), Streptococcus (St.) faecalis (12%),
Citrobacterspp (10%) and B. cereus (4%) confirming that commercially prepared
ice-cream was not suitable for consumption (Akhtar 2013).
15.11.6. Role of Government Agencies in Controlling Food Borne

Pathogens
According to the report of World Health Organization, foodborne illnesses results
in economic loss by affecting the economic productivity at individual as well as
community level. The situation gets worse with no investigation and monitoring of
food borne diseases at any level in Pakistan. In developed world, food safety has
great interest. Food safety is at 11th number priority of World Health Organization
(Akhtar 2013).
In modern worlds, investigation of diseases caused by food is basic module of food
safety systems. Investigation details are helpful for planning, applying and
evaluating public health policies. So investigation systems should be made
applicable to control foodborne diseases.
In the United States multiple investigation systems are in use to deliver information
of foodborne illness. Many health agencies also support for collection of
information. These systems emphasis on pathogens, their mode of transmission
through food and have been used broadly for many years (Foodborne Disease
Surveillance, 1982).
In Pakistan and other developing countries where centralized monitoring and
investigation systems are not working efficiently, small researches have been done
to investigate these diseases. But there is not a one standard approach and the
resulting variation, make this investigation data even more susceptible to bias
(Younus et al. 2008). Numerous key international bodies are operational on food
safety. Several agencies have run different programs for safe food supplies and
management that includes: World Health Organization, Department of Food Safety
and Zoonoses; Food and Agriculture Organisation, Nutrition And Consumer
Protection Division; World Organization for Animal Health, Working Group on
Animal Production Food Safety; European Commission, DG Health and Consumer
Protection; United States Food and Drug Administration, Centre for Food Safety
and Nutrition.
15.12. Conclusion
Conclusively, food microbiology is an applied science that deals with the study of
microorganisms related to the food materials whether they are harmful or
beneficial. Microorganisms cause food spoilage making food unfit for human
consumption and it may be a root for different food borne diseases in human body,
if ingested. To avoid spoilage, foods are preserved through various techniques like
drying, freezing, chemical preservatives, water activity control, irradiation or
through bio-preservation. Microorganisms are beneficially important in food
fermentations, culture preparations and for enzyme productions. Major sources of

microbial contamination are water, air, soil, animals and workers. In the present
era, microbial food borne diseases (Food infection and food intoxication) are very
common leading to the death if not properly treated. To avoid the microbial
contamination of foods, proper hand washing, cross contamination, safe cooking
and storage temperature, safe preparatory operations and proper public training
programs for personnel hygiene must be taken into considerations. Furthermore,
strategies should be adopted regarding awareness programs on hygiene and
sanitation at the door step level.
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Chapter 15

Muhammad Saeed and Tahir Zahoor*

15.1. Introduction to Food Microbiology

Managing editors: Iqrar Ahmad Khan and Muhammad Farooq

15.2. Scope of Food Microbiology

15.2.1. Food Spoilage

15.2.2. Food Preservation

15.3. Origin of Microorganisms

themselves accordingly to survive in nature. Multicellular organisms grown in the

15.4. Importance of Food Microorganisms

15.4.1. Production of Cultures for Food Application

15.4.2. Applications in Food Fermentation

15.4.3. Microorganisms as a Food or Source of Enzymes

Table 15.1 Microbial enzymes, sources and their application

15.5. Application of Food Microbiology

15.5.1. Food Fermentation

15.5.2. Probiotics Development

15.5.3. Food Spoilage

15.5.4. Food Borne Diseases

15.6. Role of Food Microbiologist

15.7. Significance of Microorganisms in Food

15.8. Environmental Microbiology of Food

destruction of food spoiling microbes and creation of unfavourable conditions for

15.8.1. Atmospheric Microbiology

15.8.2. Lithospheric Microbiology

15.8.3. Hydrospheric Microbiology

15.8.4. Equipment Microbiology

15.9. Contamination of Foods

15.9.1. Major Sources of Contamination

A number of contaminants can accumulate in many kinds of animals and are

15.9.2. Contamination through Traditional Marketing Systems

potential negative health consequences so it is very necessary to document the

15.9.3. Contamination of Milk and Meat in Pakistani Market

15.9.4. Contamination of Microbes through Workers /Services

15.9.5. Mode of Microbial Transmission

15.10. Control of Microorganisms in Food

15.10.1. Physical Methods

be kept at ambient temperature without flavor changes up to 8 weeks. Higher

spoilage characteristics. Psychrophilic bacteria can grow at chilling temperature but

15.10.2. Chemical Methods

15.10.3. Bio-Preservation of Food

of biofilm or reduce / eliminate the Staphylococcus aureus skin or nasal

15.11. Food Borne Illnesses

15.11.1. Food Intoxication

Communal indications include abdominal cramps, nausea, vomiting, diarrhoea,

15.11.2. Food Infections

15.11.4. Viral infections

15.11.5. Current Status of Food Borne Pathogenicity in Pakistan

ice cream to be highly polluted with Enterobacter (En). Aerogenes (34%), S.

15.11.6. Role of Government Agencies in Controlling Food Borne

fermentations, culture preparations and for enzyme productions. Major sources of

Factors contributing to outbreaks and description of outbreak categories. J.

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