Bacteria

E.coli causes food poisoning and urinary tract infections. Mycobacterium tuberculosis causes
tuberculosis.

Viruses
Influenza virus causes 'flu'. Herpes simplex virus causes herpes.

Protozoa
Plasmodium causes malaria.

Fungi
Tinea causes ringwor

Epidemiology
Epidemiology is the study of the patterns, causes and prevention of human diseases. An
outbreak of a contagious disease that spreads rapidly and widely is called an epidemic. An
epidemic occurring over a very large area is called a pandemic. The term 'epidemiologic triangle'
is used to describe the intersection of host, agent and environment in analysing an outbreak.

The prevalence of a disease is the number of diseased individuals at any one time (point
prevalence) or over a given period (period prevalence). The incidence is the number of new
cases of a disease that occur within a defined population over an established period of time.

History of epidemics
The influenza (virus-caused) pandemic of 1918-1919 killed more people than World War I. It is
considered the most devastating epidemic in recorded world history. More people died of
influenza in a single year than in four years of the Black Death Bubonic Plague from 1347 to
1351.

Bubonic plague is caused by bacteria called Yersinia pestis. These bacteria live in fleas on rats.
Rats were abundant in medieval times due to very low hygiene standards, and the lack of sewage
systems and hot, running water. Bubonic plague killed about a quarter of the European
population of that time. Today this disease can be controlled by antibiotics but outbreaks sill occur
sometimes. See image 3.

Prevention of infections
The best way of fighting disease is to prevent it. The discovery and manufacture of antibiotics and
vaccines in the early part of the 20th century have changed the state of people's health forever.
There are three stages of prevention of infections: primary, secondary and tertiary.

The Primary stage - involves public education about infectious diseases.

The Secondary stage - involves treating the actual infection that has already occurred by
quarantining and/or vaccinating of infected individuals.

What pathogenic microorganisms may be found in milk?

Types of Microorganisms in Milk - The types of micro organisms found in milk
vary considerably, and are dependent upon the specific conditions associated
with a batch of milk. Bacteria, yeasts, moulds, and bacteriophages are commonly
encoun
How can you protect ourselves daily from contamination by pathoge...?
we can protect ourselves by washing our hands getting the proper needles and
using hand sanitizers we can protect ourselves by washing our hands getting the
proper needles and using hand sanitizers
The TWhat are the main diseases caused by waterborne pathogenic
microo...?
The majority of waterborne diseases arise as a result of the contamination of
water used for drinking with human or animal faeces. The pathogens contained
in the faeces of a diseased human or animal are distributed within the water body
and...

Should I have tests for these microorganisms, these pathogens??

Unless a health professional has some reason to search for a particular
pathogen we feel it is a waste of money and time looking for any specific invader
by the taking of blood tests or other traditional tests designed to find pathogens.
Ho...

Do the Sani-Check Dipslides identify specific microorganisms such...?

Not necessarily. The identification of specific microorganisms is more complex
and usually performed only within a microbiology laboratory.

Which are the most common wound pathogenic micro-organisms

which ...?
Staphylococcus aureus, haemolytic Streptococci, Escherichia coli, Klebsiella,
Enterococci, Pseudomonas, Enterococcus, ... Yeast: Candida albicans ( top )

Diseases Caused by Microorganisms

Pathogens attack and kill cells and tissues of the host directly or indirectly by producing
toxins. Microorganisms such as bacteria, fungi and protozoa can reproduce inside and
outside a host while viruses can only reproduce inside a living host. Specific diseases are
caused by specific microorganisms and show specific signs and symptoms.

Harmful Microorganisms
Some microorganisms are harmful to humans, animals and plants because they cause
diseases. They can also result in food turning bad and rotten. This may lead food
poisoning if the food is consumed. Microorganisms that cause diseases are called
pathogens. Pathogens consist of viruses, bacteria, fungi and protozoa.

Pathogens are classified as parasites or saprophytes. Parasites live inside a living

organism called the host, for example a human being or a cow. Saprophytes live on dead
tissues such as decaying plants and animals. Some bacteria turn food rotten and unsafe
for consumption. These microorganisms decompose food and release toxins (poison)
which result in food poisoning. Signs and symptoms of food poisoning are vomiting,
stomach ache and diarrhea.

Bacteria found in the mouth can cause tooth decay and gingivitis. Bacteria act on
carbohydrates (glucose) found in food remnants and change them into lactic acid which
dissolves the enamel layer of the teeth, causing small holes (caries). The action of
bacteria can also release a certain poison which causes the gums to swell and bleed.

Diseases Caused by Microorganisms

Pathogens attack and kill cells and tissues of the host directly or indirectly by producing
toxins. Microorganisms such as bacteria, fungi and protozoa can reproduce inside and
outside a host while viruses can only reproduce inside a living host. Specific diseases are
caused by specific microorganisms and show specific signs and symptoms.

Signs are external evidences, for example, swelling or fever due to a disease while
symptoms are internal and cause a change in the normal functions of the body.

Diseases Caused by Bacteria

Tuberculosis or commonly known as TB a) it is caused by the Bacillus bacteria which
enter the human body through the nose or mouth b) the bacteria attack and destroy the
lung tissues, lymph nodes, bones and joints of the patient c) the symptoms take time to
show. The patient first suffers from a dry cough and feels weak and tired. The cough is
continuous and becomes more severe. The patient then coughs out yellowish-green
sputum containing blood. The patient also loses weight because of a loss of appetite d) an
X-ray film of a tuberculosis patient’s chest shows black spots on the lungs

Cholera a) it is caused by the vibrios bacteria which contaminate drinking water and
food. Hence the bacteria enter the human body through the alimentary canal b) the
bacteria reproduce very fast inside the intestines and digest the mucous lining of the
intestinal walls c) the symptoms of this disease are vomiting, diarrhea and severe muscle
cramps d) this results in the patient suffering from dehydration due to the excessive loss
of water and this can cause death if not treated early.

Sexually Transmitted Diseases a) sexually transmitted diseases are diseases which

spread through sexual activities and infect both the male and female reproductive systems
b) two examples of such diseases are gonorrhea and syphilis.

c) Gonorrhea is caused by the cocci bacteria i) a male patient will suffer pain when
urinating. His penis will excrete a thick white liquid or pus. His testes become swollen. If
not treated, the person may become sterile ii) A female patient suffers from the
inflammation of the urethra, uterus and cervix. Sometimes the Fallopian tubes become
blocked and may lead to sterile.
d) Syphilis is caused by the spiral bacteria. The symptoms appear in three stages i) first
or early stage – ulcer and red spots appear around the vagina or penis ii) second or middle
stage – rashes occur on the skin of the body, ulcers appear in the mouth and throat, loss of
appetite and loss of weight as well as pain in the muscles, bones and joints iii) third or
final stage – the nervous system becomes infected and internal organs such as the heart
and brain may be damaged.

Diseases Caused by Viruses

Viruses are parasitic pathogens. They only reproduce inside a living host. Each type of
virus only infects the cells of a specific host. Examples of diseases caused by viruses are
influenza (the common cold), dengue fever, hepatitis and the Acquired Immune
Deficiency Syndrome (AIDS).

Influenza a) the virus which causes influenza or the common cold is found in the air b)
when a patient coughs or sneezes, the virus attacks the membrane lining of the respiratory
tract c) the patient suffers from blocked nasal cavities and excretes excessive mucus from
the nose d) the early symptoms of influenza are the abrupt onset of a fever, chills,
headache, pain in the muscles and joints, coughing and a sore throat d) influenza may
develop into pneumonia and other serious complications if it is not treated quickly and
effectively.

Dengue Fever a) this is dangerous and infectious disease caused by the dengue virus b) it
is spread by a vector, the Aides mosquitoes c) the symptoms are fever for about 5 to 7
days, severe pain throughout the body especially in the muscles, joints, bones and
eyeballs. There is often an intense headache and rashes on the skin d) in the more serious
hemorrhagic dengue fever there is bleeding in the nose, mouth and internal organs which
may result in death e) there is no antibiotic to treat dengue fever hence prevention is the
best way to control the disease.

Hepatitis a) there are two main types of hepatitis, hepatitis A and B which are caused by
different viruses b) the virus attacks liver cells. In severe cases, it leads to cirrhosis of the
liver as well as cancer of the liver c) the virus can be transmitted through contaminated
food or through the use of contaminated syringes. Table below shows a comparison
between the two types of hepatitis.

Diseases Hepatitis A Hepatitis B

Pathogen Hepatitis A virus Hepatitis B virus

Where the pathogen is found Urine and faces Saliva, blood and semen

Fever, rashes on skin,

Fever, headache, jaundice,
tiredness, loss of appetite,
Symptoms loss of appetite, tiredness and
pain in the joints, swollen
swollen liver.
liver, internal bleeding
Acquired Immune Deficiency Syndrome (AIDS) i) AIDS is caused by the human
immune –deficiency viruses (HIV-1 and HIV-2) ii) the virus attacks and kills white blood
cells and paralyses the immune system of the body iii) the virus is found in body fluids of
the patient like the saliva, blood, semen, mother’s milk and secretions from the vagina iv)
the incubation period for this disease varies from a few months to a few years before any
symptoms appear v) the symptoms are loss of appetite, fever, candidacies in the mouth
and throat as well as inflammation of the lymph nodes vi) since the patient’s immune
system is weak, the patient can easily contract other diseases such as pneumonia which
can lead to death.

Diseases Caused by Fungi

Fungi grow and multiply quickly in warm and moist environments. Fungi grow and
spread easily in the folds of human skin because it is warm and damp from sweat.
Examples of fungal diseases are tine and ringworm. Tine (Athlete’s foot) a) this disease
is caused by filamentous and round fungi b) the skin infected with tine appears bright
with white spots c) it is usually appears on the face, body, hands and legs.

Ringworm a) ringworm infection usually occurs in the gaps between the fingers and
toes, armpits and the area below the breasts b) the infected areas appear red in color.
Wounds and bleeding occur sometimes when the skin peels off c) if it grows on the head,
it may cause hair to drop off.

Diseases Caused by Pathogenic Protozoa

Protozoa which cause diseases are parasites which live inside the body of hosts. An
example of a disease caused by protozoa is malaria. Malaria a) it is caused by the
protozoan Plasmodium which is carried by the female Anopheles mosquitoes b)
Plasmodium enters the human body through mosquito bites and is then carried by the
bloodstream to the liver c) The parasite attacks the liver and red blood cells d) the
incubation period of this diseases varies from 12 to 30 days depending on the protozoan
species e) the disease begins with an ill feeling and a rapid rise in body temperature.
There is profuse sweating with intermittent fever. Headache and nausea are common f) in
advanced stages of the disease, there is enlargement of the spleen g) malaria can lead to
anemia.

Info Gallery
Malaria comes from the Italian word ‘Mala-aria’ which means bad air or air from marshy
land. A French army surgeon in Algeria in 1880 described the presence of the malarial
parasite in fresh human blood. Major Ronald Roses, a doctor in India, discovered the
malarial parasite in the stomach of the Anopheles mosquito in 1987. He was awarded the
Nobel Prize for his discovery which laid the foundation for the control o

Best Answer - Chosen by Voters

Types of Microorganisms in Milk - The types of micro organisms found in milk vary
considerably, and are dependent upon the specific conditions associated with a batch of
milk. Bacteria, yeasts, moulds, and bacteriophages are commonly encountered. Other
viruses and Protozoa are seldom observed in milk products, except as occasional
contaminants. Bacteria Bacteria are the most common, and probably the most numerous
of microorganisms with which the dairy processing industry is concerned. They belong to
four main groups: (1) cocci, usually gram positive, (2) gram positive non-sporeforming
rods, (3) gram positive sporeforming rods, and (4) gram negative non-spore forming rods.
Yeasts The yeasts most frequently encountered in milk and milk products act upon the
lactose to produce acid and carbon dioxide. Yeasts are more commonly found in raw,
cream during hot weather, but are potential contaminants throughout the year. Moulds
Moulds often grow in large concentrations and are visible as a fuzzy or fluffy growth.
They are sometimes observed on the surface, of butter, old cream, khoa, or cheese. They
are black, grey, green, blue or white. They discolour milk products and often produce
undesirable, at times repulsive, odours and flavours. Moulds are essential in production
of the certain kinds of cheese. Bacteriophages Bacteriophages are particularly
obnoxious in starter cultures used for making cultured milk, butter and cheese. Phages
will, kill the bacterial culture and the whole process of fermentation will be lost.
Source(s):
http://www.microbiologyprocedure.com/mic…

Dairy Microbiology

Basic Microbiology

• Microorganisms: Bacteria, Yeasts, Moulds

• Microbial Growth
• Detection and Enumeration

Microorganisms in Milk

• Spoilage Microorganisms
• Pathogenic Microorganisms

HACCP
Starter Cultures

• Mesophilic and Thermophilic

• Bacteriophages
• Starter Culture Preparation
Basic Microbiology
Microorganisms
Microorganisms are living organisms that are individually too small to see with the naked
eye. The unit of measurement used for microorganisms is the micrometer (µ m); 1 µ m =
0.001 millimeter; 1 nanometer (nm) = 0.001 µ m. Microorganisms are found everywhere
(ubiquitous) and are essential to many of our planets life processes. With regards to the
food industry, they can cause spoilage, prevent spoilage through fermentation, or can be
the cause of human illness.

There are several classes of microorganisms, of which bacteria and fungi (yeasts and
moulds) will be discussed in some detail. Another type of microorganism, the bacterial
viruses or bacteriophage, will be examined in a later section.

Bacteria
Bacteria are relatively simple single-celled organisms. One method of classification is by
shape or morphology:

• Cocci:

- spherical shape
- 0.4 - 1.5 µ m

Examples: staphylococci - form grape-like clusters; streptococci - form bead-like

chains

• Rods:

- 0.25 - 1.0 µ m width by 0.5 - 6.0 µ m long

Examples: bacilli - straight rod; spirilla - spiral rod

There exists a bacterial system of taxonomy, or classification system, that is

internationally recognized with family, genera and species divisions based on genetics.
Some bacteria have the ability to form resting cells known as endospores. The spore
forms in times of environmental stress, such as lack of nutrients and moisture needed for
growth, and thus is a survival strategy. Spores have no metabolism and can withstand
adverse conditions such as heat, disinfectants, and ultraviolet light. When the
environment becomes favourable, the spore germinates and giving rise to a single
vegetative bacterial cell. Some examples of spore-formers important to the food industry
are members of Bacillus and Clostridium generas.

Bacteria reproduce asexually by fission or simple division of the cell and its contents.
The doubling time, or generation time, can be as short as 20-20 min. Since each cell
grows and divides at the same rate as the parent cell, this could under favourable
conditions translate to an increase from one to 10 million cells in 11 hours! However,
bacterial growth in reality is limited by lack of nutrients, accumulation of toxins and
metabolic wastes, unfavourable temperatures and dessication. The maximum number of
bacteria is approximately 1 X 10e9 CFU/g or ml. Note: Bacterial populations are
expressed as colony forming units (CFU) per gram or millilitre.

Bacterial growth generally proceeds through a series of phases:

• Lag phase: time for microorganisms to become accustomed to their new

environment. There is little or no growth during this phase.
• Log phase: bacteria logarithmic, or exponential, growth begins; the rate of
multiplication is the most rapid and constant.
• Stationary phase: the rate of multiplication slows down due to lack of nutrients
and build-up of toxins. At the same time, bacteria are constantly dying so the
numbers actually remain constant.
• Death phase: cell numbers decrease as growth stops and existing cells die off.

The shape of the curve varies with temperature, nutrient supply, and other growth factors.
This exponential death curve is also used in modeling the heating destruction of
microorganisms.

Yeasts
Yeasts are members of a higher group of microorganisms called fungi . They are single-
cell organisms of spherical, elliptical or cylindrical shape. Their size varies greatly but
are generally larger than bacterial cells. Yeasts may be divided into two groups according
to their method of reproduction:

1. budding: called Fungi Imperfecti or false yeasts

2. budding and spore formation: called Ascomycetes or true yeasts

Unlike bacterial spores, yeast form spores as a method of reproduction.

Moulds
Moulds are filamentous, multi-celled fungi with an average size larger than both bacteria
and yeasts (10 X 40 µ m). Each filament is referred to as a hypha. The mass of hyphae
that can quickly spread over a food substrate is called the mycelium. Moulds may
reproduce either asexually or sexually, sometimes both within the same species. Asexual
Reproduction:

• fragmentation - hyphae separate into individual cells called arthropsores

• spore production - formed in the tip of a fruiting hyphae, called conidia, or in
swollen structures called sporangium

Sexual Reproduction: sexual spores are produced by nuclear fission in times of

unfavourable conditions to ensure survival.

Microbial Growth
There are a number of factors that affect the survival and growth of microorganisms in
food. The parameters that are inherent to the food, or intrinsic factors, include the
following:

• nutrient content
• moisture content
• pH
• available oxygen
• biological structures
• antimicrobial constituents

Nutrient Requirements: While the nutrient requirements are quite organism specific,
the microorganisms of importance in foods require the following:

• water
• energy source
• carbon/nitrogen source
• vitamins
• minerals

Milk and dairy products are generally very rich in nutrients which provides an ideal
growth environment for many microorganisms.
Moisture Content: All microorganisms require water but the amount necessary for
growth varies between species. The amount of water that is available in food is expressed
in terms of water activity (aw), where the aw of pure water is 1.0. Each microorganism
has a maximum, optimum, and minimum aw for growth and survival. Generally bacteria
dominate in foods with high aw (minimum approximately 0.90 aw) while yeasts and
moulds, which require less moisture, dominate in low aw foods ( minimum 0.70 aw). The
water activity of fluid milk is approximately 0.98 aw.

pH: Most microorganisms have approximately a neutral pH optimum (pH 6-7.5). Yeasts
are able to grow in a more acid environment compared to bacteria. Moulds can grow over
a wide pH range but prefer only slightly acid conditions. Milk has a pH of 6.6 which is
ideal for the growth of many microoorganisms.
Available Oxygen: Microorganisms can be classified according to their oxygen
requirements necessary for growth and survival:

• Obligate Aerobes: oxygen required

• Facultative: grow in the presence or absence of oxygen
• Microaerophilic: grow best at very low levels of oxygen
• Aerotolerant Anaerobes: oxygen not required for growth but not harmful if
present
• Obligate Anaerobes: grow only in complete absence of oxygen; if present it can
be lethal

Biological Structures: Physical barriers such as skin, rinds, feathers, etc. have provided
protection to plants and animals against the invasion of microorganisms. Milk, however,
is a fluid product with no barriers to the spreading of microorganisms throughout the
product.
Antimicrobial Constituents: As part of the natural protection against microorganisms,
many foods have antimicrobial factors. Milk has several nonimmunological proteins
which inhibit the growth and metabolism of many microorganisms including the
following most common:

1. lactoperoxidase
2. lactoferrin
3. lysozyme
4. xanthine

More information on these antimicrobials can be found in a chapter on dairy

microbiology and safety written by Vasavada and Cousin.
Where the intrinsic factors are related to the food properties, the extrinsic factors are
related to the storage environment. These would include temperature, relative humidity,
and gases that surround the food.

Temperature: As a group, microorganisms are capable of growth over an extremely

wide temperature range. However, in any particular environment, the types and numbers
of microorganisms will depend greatly on the temperature. According to temperature,
microorganisms can be placed into one of three broad groups:

• Psychrotrophs: optimum growth temperatures 20 to 30° capable of growth at

temperatures less than 7° C. Psychrotrophic organisms are specifically important
in the spoilage of refrigerated dairy products.
• Mesophiles: optimum growth temperatures 30 to 40° C; do not grow at
refrigeration temperatures
• Thermophiles: optimum growth between 55 and 65° C

It is important to note that for each group, the growth rate increases as the temperature
increases only up to an optimum, afterwhich it rapidly declines.
Detection and Enumeration of Microorganisms
There are several methods for detection and enumeration of microorganisms in food. The
method that is used depends on the purpose of the testing.
Direct Enumeration:

Using direct microscopic counts (DMC), Coulter counter etc. allows a rapid
estimation of all viable and nonviable cells. Identification through staining and
observation of morphology also possible with DMC.
Viable Enumeration:

The use of standard plate counts, most probable number (MPN), membrane
filtration, plate loop methos, spiral plating etc., allows the estimation of only
viable cells. As with direct enumeration, these methods can be used in the food
industry to enumerate fermentation, spoilage, pathogenic, and indicator
organisms.
Metabolic Activity Measurement:

An estimation of metabolic activity of the total cell population is possible using

dye reduction tests such as resazurin or methylene blue dye reduction, acid
production, electrical impedence etc. The level of bacterial activity can be used to
assess the keeping quality and freshness of milk. Toxin levels can also be
measured, indicating the presence of toxin producing pathogens.
Cellular Constituents Measurement:

Using the luciferase test to measure ATP is one example of the rapid and sensitive
tests available that will indicate the presence of even one pathogenic bacterial
cell.
Isolation of microorganisms is an important preliminary step in the identification
of most food spoilage and pathogenic organisms. This can be done using a simple
streak plate method.

Microorganisms in Milk
Milk is sterile at secretion in the udder but is contaminated by bacteria even
before it leaves the udder. Except in the case of mastisis, the bacteria at this point
are harmless and few in number. Further infection of the milk by microorganisms
can take place during milking, handling, storage, and other pre-processing
activities.
Lactic acid bacteria: this group of bacteria are able to ferment lactose to lactic
acid. They are normally present in the milk and are also used as starter cultures in
the production of cultured dairy products such as yogurt. Note: many lactic acid
bacteria have recently been reclassified; the older names will appear in brackets as
you will still find the older names used for convenience sake in a lot of literature.
Some examples in milk are:
 • lactococci
o L. delbrueckii subsp. lactis (Streptococcus lactis )
o Lactococcus lactis subsp. cremoris (Streptococcus
cremoris )

• lactobacilli
o Lactobacillus casei
o L.delbrueckii subsp. lactis (L. lactis )
o L. delbrueckii subsp. bulgaricus (Lactobacillus bulgaricus
)
• Leuconostoc

Coliforms: coliforms are facultative anaerobes with an optimum growth at 37° C.

Coliforms are indicator organisms; they are closely associated with the presence
of pathogens but not necessarily pathogenic themselves. They also can cause
rapid spoilage of milk because they are able to ferment lactose with the
production of acid and gas, and are able to degrade milk proteins. They are killed
by HTST treatment, therefore, their presence after treatment is indicative of
contamination.Escherichia coli is an example belonging to this group.

Significance of microorganisms in milk:

• Information on the microbial content of milk can be used to judge

its sanitary quality and the conditions of production
• If permitted to multiply, bacteria in milk can cause spoilage of the
product
• Milk is potentially susceptible to contamination with pathogenic
microorganisms. Precautions must be taken to minimize this possibility
and to destroy pathogens that may gain entrance
• Certain microorganisms produce chemical changes that are
desirable in the production of dairy products such as cheese, yogurt.

Spoilage Microorganisms in Milk

The microbial quality of raw milk is crucial for the production of quality dairy
foods. Spoilage is a term used to describe the deterioration of a foods' texture,
colour, odour or flavour to the point where it is unappetizing or unsuitable for
human consumption. Microbial spoilage of food often involves the degradation of
protein, carbohydrates, and fats by the microorganisms or their enzymes.
In milk, the microorganisms that are principally involved in spoilage are
psychrotrophic organisms. Most psychrotrophs are destroyed by pasteurization
temperatures, however, some like Pseudomonas fluorescens, Pseudomonas fragi
can produce proteolytic and lipolytic extracellular enzymes which are heat stable
and capable of causing spoilage.

Some species and strains of Bacillus, Clostridium, Cornebacterium, Arthrobacter,

Lactobacillus, Microbacterium, Micrococcus , and Streptococcus can survive
pasteurization and grow at refrigeration temperatures which can cause spoilage
problems.

Pathogenic Microorganisms in Milk

Hygienic milk production practices, proper handling and storage of milk, and
mandatory pasteurization has decreased the threat of milkborne diseases such as
tuberculosis, brucellosis, and typhoid fever. There have been a number of
foodborne illnesses resulting from the ingestion of raw milk, or dairy products
made with milk that was not properly pasteurized or was poorly handled causing
post-processing contamination. The following bacterial pathogens are still of
concern today in raw milk and other dairy products:

• Bacillus cereus
• Listeria monocytogenes
• Yersinia enterocolitica
• Salmonella spp.
• Escherichia coli O157:H7
• Campylobacter jejuni

It should also be noted that moulds, mainly of species of Aspergillus , Fusarium ,

and Penicillium can grow in milk and dairy products. If the conditions permit,
these moulds may produce mycotoxins which can be a health hazard.

HACCP
Raw and end-products may be tested for the presence, level, or absence of
microorganisms. Traditionally these practices were used to reduce manufacturing
defects in dairy products and ensure compliance with specifications and
regulations, however, they have many drawbacks:

1. destructive and time consuming

2. slow response
3. small sample size
4. delays in the release of the food

In the 1960's, the Pillsbury Company, the U.S. Army, and NASA introduced a
system for assuring pathogen-free foods for the space program. This system,
called Hazard Analysis and Critical Control Points (HACCP), is a focus on
critical food safety areas as part of total quality programs. It involves a critical
examination of the entire food manufacturing process to determine every step
where there is a possibility of physical, chemical, or microbiological
contamination of the food which would render it unsafe or unacceptable for
human consumption. These identified points are the critical control points (CCP).
There are seven prinicples to HACCP:
 1. analyze hazards
2. determine CCPs
3. establish critical limits
4. establish monitoring procedures
5. establish deviation procedures
6. establish verification procedures
7. establish record keeping procedures

Before these principles can be put into place, a prerequisite program and
preliminary setup is necessary. Prerequisite Program:

• premise control
• receiving and storage control
• equipment performance and maintenance control
• personnel training
• sanitation
• recall procedure

Preliminary Setup:

• assemble team
• describe the product
• identify intended use
• construct flow diagram and plant schematic
• verify the diagram on-site

Food Safety Enhancement Program-FSEP is The Canadian Food Inspection

Agency's HACCP initiative. There is extensive information at their Web site
regarding FSEP, including implementation manuals, HACCP curriculum
guidelines, and generic models.

Starter Cultures
Starter cultures are those microorganisms that are used in the production of
cultured dairy products such as yogurt and cheese. The natural microflora of the
milk is either inefficient, uncontrollable, and unpredictable, or is destroyed
altogether by the heat treatments given to the milk. A starter culture can provide
particular characteristics in a more controlled and predictable fermentation. The
primary function of lactic starters is the production of lactic acid from lactose.
Other functions of starter cultures may include the following:

• flavour, aroma, and alcohol production

• proteolytic and lipolytic activities
• inhibition of undesirable organisms
There are two groups of lactic starter cultures:

1. simple or defined: single strain, or more than one in which the

number is known
2. mixed or compound: more than one strain each providing its own
specific characteristics

Starter cultures may be categorized as mesophilic or thermophilic:

Mesophilic

 Lactococcus lactis subsp. cremoris

 L. delbrueckii subsp. lactis
 L. lactis subsp. lactis biovar diacetylactis
 Leuconostoc mesenteroides subsp. cremoris
Thermophilic

 Streptococcus salivarius subsp. thermophilus (S.thermophilus)

 Lactobacillus delbrueckii subsp. bulgaricus
 L. delbrueckii subsp. lactis
 L. casei
 L. helveticus
 L. plantarum
Mixtures of mesophilic and thermophilic microorganisms can also be used as in
the production of some cheeses.

Bacteriophage
Bacteriophages are viruses that require bacteria host cells for growth and
reproduction. Initially, the bacteriophage attaches itself to the bacteria cell wall
and injects nuclear substance into the cell. Inside the cell, the nuclear substance
produces shells, or phage coats, for the new bacteriophage which are quickly
filled with nucleic acid. The bacterial cell ruptures and dies as the new
bacteriophage are released. Bacteriophages are ubiquitous but generally enter the
milk processing plant with the farm milk. They can be inactivated heat treatments
of 30 min at 63 to 88° C, or by the use of chemical disinfectants. Bacteriophages
are of most concern in cheese making. They attack and destroy most of the lactic
acid bacteria which prevents normal ripening known as slow or dead vat.
Starter Culture Preparation
Commercial manufacturers provide starter cultures in lyophilized (freeze-dryed),
frozen or spray-dried forms. The dairy product manufacturers need to inoculate
the culture into milk or other suitable substrate. There are a number of steps
necessary for the propagation of starter culture ready for production:

1. Commercial culture
2. Mother culture - first inoculation; all cultures will originate from
this preparation
 3. Intermediate culture - in preparation of larger volumes of prepared
starter
4. Bulk starter culture - this stage is used in dairy product production

1) Spoilage occurs when microorganisms degrade the carbohydrates, proteins, fats of milk and
produce noxious, end products. Milk products as follows;

Spoilage type Oraganisms involved Signs of spoilage

Souring Lactobacillus sp. Streptococcus sp. Sour milk,Curd formation
Sweet curdling Bacillus sp. Alkaline pH

Proteus sp. Curd formation

Micrococcus sp.
Gas production Clastoridium sp. Explosion of curds

coliform bacteria
Ropiness Alcaligenes sp.,Klebsiella sp.,Enterobacter sp. Stringy or slimy milk
Red rot Serratia marcescens Red colaration
Gray rot clotridium sp. Gray colaration, Foul smell
Dairy mould Penicilium sp.,Geotrichum sp. Mouldy appearance

2) Sources of Microorganisms in Milk - There are several principal sources of

contamination of milk. From the time the milk leaves the udder, until it is dispensed into
containers, everything with which it comes into contact is a potential source of more
microorganisms. Milking performed under hygienic conditions, with strict attention to
sanitary practices, will reduce the entry of microorganisms into the milk. Naturally the
fewer the organisms that can get into the milk, the fewer have a chance to grow.

3)

Sources of Microorganisms - The Producing

Animal - Unless the producing animal is clean, and her
flanks, udder and teats given special sanitary care just
before milking, her body can be a source of considerable
contamination. The first few streams of milk from each
teat should, be collected, separated and discarded. This
flushes out the organisms that have entered the teat
through the teat opening. Milk from a cow with an
infected udder is likely to contain a large number of
organisms. The probability of diseases of the udder
contaminating the milk is very high. Mastitis, which is a
disease causing inflammation of the udder, contributes
considerable number of organisms, sometimes even
 blood cells, into the milk.

If all the milk is not drawn out of the udder but remains between milkings, it may spoil within the
udder itself and. thus increase the count considerably. Washing and massaging the cow's udder
with a warm detergent sanitizer solution before milking serves to clean the area. The hair of all
animals harbours organisms. The hair, dirt and dust often fall from the body into the milking pails
or the teat cups of milking machines. The modern practice is to keep the flanks clipped to minimize
contamination.

4)

Sources of Microorganisms - Milking Area - The

microbial content of air is greatly affected by many
conditions and practices. Dried dirt and filth is picked up
by all movements and carried about as dust in the
atmosphere. For this reason, dust may be the source of
almost evey kind of contamination.

The sprays which are sometimes used to cut down air contaniination are not very useful. However,
the main point of keeping the conditions clean and sanitary, is not to raise dust.

Sources of Microorganisms - Utensils and

Equipments- Utensils and equipments are known to be
the greatest sources of contamination. They may account
for as much as .100, 000 to a billion organisms per
milliliter. Pails, strainers, milking machines, cans, pipes,
bottles, and other equipment used for the handling of milk
are sometimes not properly washed and sanitized.

Organisms survive in the cracks, corners, crevices, dents, scratches, and other irregularities of the
utensils. Such neglect affords ideal conditions for the growth of microorganisms before the utensils
are used

5)

Sources of Microorganisms - The Personnel - All

persons involved in the milking process must be in good
health and must be careful in their personal cleanliness.
They should wash their hands, clean and rinse them
with an effective bactericidal solution, and dry them with
a clean towel before starting, and frequently during
work, and immediately following every rest stop. They
should keep finger nails free of dirt. Each person must
always carry a clean handkerchief and use it to prevent
the spraying of nasal and oral discharges into the
atmosphere, equipment or products.

They should, wear a neat and clean uniform. A surgical mask is an effective addition to the
uniform. Although workers may not contribute a large number of organisms, these are of
considerable importance since they may well be human pathogens. Probably the majority of milk
borne epidemics of disease were started by workers who were carriers, or who had mild cases, or
who were in close contact with others so affected. The importance of contamination contributed
by the workers is simply that it is the most dangerous.

6)

Sources of Microorganisms - Water is continuously

required for cleaning and processing operations in a
dairy. Special care or treatment must be taken to,
supply good quality water. Water quality will vary with
the source of supply. Water from surface supplies is
contaminated by dust, animals, plants, people, and
other agents.

The microbiological quality of water is tested before it is used in a dairy plant. The most common
test for water destined for domestic or industrial use, is the presumptive coliform test. This test
reveals faecal or sewage contamination. Chlorination of water is commonly practiced to assure
potability.

11) Milk Borne Diseases - The important diseases are, tuberculosis, brucellosis and Q fever.
Tuberculosis bacterium. Mycobacterium bovis is consumed in the milk and passes from human
intestine to the blood, from which it spreads to most organs. Brucellosis, a blood-disease is caused
by Gram-negative rod, Brucella abortus. When transmitted to man through cow milk, the bacterium
infect the blood -rich organs. Q fever, caused by rickettsia, Cxiella burnetii is also a milk borne
disease. Other important disorders associated with milk are primary atypical pneumonia,
toxoplasmosis, anthrax, streptococcal infections etc.

12) Pasterurization - The process was developed by Louis Pasteur in the 1860s to
eliminate bacteria in wines. The process for milk was adopted in 1895. Primary object
of this process is to eliminate disease-causing bacteria from milk, though the total number
of bacteria is also very much reduced during this process. It reduces the chances of
milk-spoilage.

13)

Temperature Response of Milk Microorganisms-

Microorganisms found in milk can also be described
according their optimum temperature for growth and
heat resistance. This is a very practical consideration
since milk is preserved by employing low temperatures
to prevent changes due to microbial activity and by high
temperatures to reduce microbial population and destroy
pathogens. All the four types of microorganisms i.e.
psychrophilic, mesophilic, thermophilic and thermoduric
 are found in milk.

Psychrophiles grow at temperatures just above freezing and at refrigeration tempratures. They
produce a wide variety of spoilage defects. The defects may result in the production bf many "off"
flavours and odours. The most commonly encountered psychrophilic bacteria are members of the
genera pseudomonas, Achromobacter, Vibrio, Flavobacterium and Alcaligenes, They arc killed in
the pasteurization process, but are sometimes found in pasteurized milk. The contamination
takes place after pasteurization from equipment, cans, bottles, and water.

14)

Milk Microorganisms Ability to Cause Infection and Diseases - Pathogenic organisms of both
bovine and human origin have been isolated from milk. Milk, therefore, can serve as a carrier of
diseases. Many serious epidemics were caused by the consumption of such products before this
fact was clearly recognized. However, this became less common as milk sanitation has improved
and pasteurization is being more widely practised. The disease organisms present in milk may be
derived from

(1) diseased animals or (2) persons collecting and handling milk: Thus the danger is due to the
inoculum and not to the growth of organisms in the milk. The health of animal is an important
factor. Several diseases of cattle including staphylococcal and streptococcal infections,
tuberculosis, brucellosis, salmonellosis, Q fever and Foot and mouth disease may be transmitted
to man. The organisms causing these diseases may get into the milk either directly from the
udder, or indirectly from infected body discharges, which may drop, splash, or be blown into the
milk.

Some of the important diseases of human origin that have been transmitted by milk are (1)
typhoid fever (2) diphtheria, (3) scarlet fever, (4) dysentery (5) septic sore throat and (6)
poliomyelitis. It is also possible for humans to infect animals. For example, mastitis may be
caused by a variety of organisms, including Staphylococcus aureus. The infecting organism, in
s9me cases, has been traced to humans.

15) Flash Method Pasteurization - This is modem method and also known as HTST
(high temperature, short time) method. Raw milk is first warmed using the heat of the
previously pasteurised milk. It then passes through a hot cylinder at 71.6°C for a period
of 15 to 17 seconds. The milk is then cooled rapidly in part by transferring its heat to the
incoming milk.

16)

Ultrapasteurization - It is also used in some dairy plants. Here the milk and milk products are
subjected to heat at 82°C for three seconds only. Pasteurisation destroys the pathogenic, bacteria
that may be transmitted by milk. The pasteurised milk is graded into different grades on the basis
of total bacterial counts. In different grades of milk, standards in respect of bacterial content per ml
of milk vary from localities to localities.
 Some bacteria are able to survive pasteurisation, causing milk spoilage. Streptococcus lactis,
Lactobacil1s casei and L. cremoris, produce acids, causing the protein to turn into curd. These
bacteria which are able to endure the heat of pasteurisation are called thermoduric. Pasteurisation
has no effect on thermophilic bacteria which grow well at 60°C to 70°C.

17)

PhosphateTest - An enzyme, phosphatase is normally found in milk. This enzyme in respect of

heat tolerance is similar to the rickettsia, Cxiella burnetii (Q. fever organism) or Mycobacterium
bovis (tuberculosis bacterium). It thus may be tested whether this enzyme is eliminated during
pasteurization, so that it could be assumed that these two microbes are also not present in the milk.

Milk sample is combined with disodium phenyl-phosphate. After few minutes of incubation, a
colour reagent (CQC) is added. If phosphatase is present the contents will become blue in colour.
If there is no colour change then it may be assumed that there is no phosphatase present

18)

Standard Plate Count-SPC - This determines the total number of bacteria in a specified amount
of milk, generally a milliliter (ml). This is used for grading of milk. Under aseptic conditions, one ml
of milk is added to 99 ml of distilled water or buffer. One ml and one-tenth ml samples are then
transferred to sterile Petri dishes.. These give 1: 100 and 1: 1000 dilutions of the milk respectively.
Other dilutions may also be prepared successively.

A growth medium like plate count agar or tryptone glucose yeast extract agar is then added, and
the milk samples are mixed with the medium. The dishes are incubated at 37°C for 24-48 hours.
The plates are then placed on a counting device like a Quebec colony counter and the number of
bacterial colonies is recorded. The colony count falling between 30 and 300 is selected and
multiplied by the reciprocal of the dilution factor to obtain the bacterial count per ml of milk. If 248
colonies appeared on the 1 : 100 plate and 16 colonies on ,the 1:1000 plate, 248 would be
selected and multiplied by 100, giving 24,800 total bacteria per ml of milk sample.

The 30 to 300 range is used because under 30 the chance of sampling error may be significant,
and over 300, due to overcrowding on the plate, there may be formed a single colony by more than
one bacteria.

19)

Reduction Test - This is a rapid test to find relative amount of bacteria in a milk sample. The
principle used is that the length of time for a colour change in a specific dye is proportional to the
number of bacteria in the sample. A sample of milk is mixed with the dye solution.(methylene blue
or resazurin) and this is incubated.

Due to growth of bacteria and subsequent fermentation of lactose of milk, the electrons released
are passed along to the dye molecules, reducing them. The enzyme involved is a reductase. On
reduction, methylene blue loses its colour and resazurin changes from normal slate blue to pink or
colorless form. Good quality milk normally shows minimal colour change even after five to six
hours, whereas contaminated milk shows color change rapidly within two hours or so.
20)

Breed Counting Method - This method does not distinguish between live and dead cells. It is a
direct count method in a known area of microscopic field. The sample is spread (about 0.0 I ml)
over I cm2 of a microscope slide. Milk smear is dried and stained with Newman - Lampert stain.
This stain fixes the smear, dissolves fat globules and stains bacteria with methylene blue. Slide is
then scanned under several oil immersion microscopic fields. Calculation, are made as follows:

Area of one microscope field = 0.02 mm2. Area over which milk sample is spread on the slide = 1
cm2 or 100 mm2 Then, no. of fields possible under the lens = 100 mm2/(or divided by) 0.02
mm2 = 5000 fields This number of field is for 0.01 ml of milk. However, final count is expressed
as bacterial number per ml of sample. Thus, factor is to be multiplied by 100. i.e. 5000 X 100 =
500,000

This is the microscope factor Le. 500,000 Suppose, 20 fields yielded an average of 0.4 bacteria
per field. Then, the final count would be 0.4 X500, 000 (microscope factor)= 200,000 bacteria per
ml of milk sample.

21) Antibiotic Detection Test- The test is performed to identify the presence of any antibiotic in
milk sample. It is likely that if cow is treated with antibiotics against some diseases as tuberculosis,
Q fever, brucellosis etc., the antibiotics may persist in the milk. A paper disc is dipped into the milk
and then applied to an agar plate inoculated with an organism like Bacillus subtilis. The presence
of an antibiotic in milk will be indicated by the development of a zone of inhibition around the disc.

22)

Sterilization of Milk - Boiling of milk is still widely used in our country. It kills all the vegetative
cells of microorganisms. It does not require refrigerati0n and has a relatively longer shelf life.
Commercial milk sterilization techniques are in the process of being developed.

Ultra pasteurization is a more recently perfected process in which milk is heated at 300F for 1 to 2
seconds. In addition the processing includes steps that eliminate any traces of cooked flavour.
The final product is comparable in flavour and nutritional quality to pasteurized milk.

23)

Grades of Milk - There are certain standards by which milk is judged. The grading of milk has
become an important factor in maintaining the health of the population. Grades of milk are based
upon regulations pertaining to production, processing, and distribution, e. g., sanitation,
pasteurization, and holding conditions, as well as upon bacterial standards.

The number of organisms permissible in different grades of milk vary some what, depending upon
standards set up by local public health authorities. However, the recommendations contained in
the U. S. Public Health Service publication "Milk Ordinance and Code".
24) Chemical and Other Standards for Grade A Milk and Other Milk Products -

Product chemical and other

Grade A raw milk for Antibiotics:less than 0.05 unit/ml by Bacillus subtilis method
pasteurization or equivalent
Grad A Pasturized milk products Phosphatasc:less than 1 mg/ml by Scharer rapid method
Grade A Pasteurized cultured
Phosphatasc:less than 1 mg/ml by Scharer rapid method
products

25) Bacterial Limit Standards for Grade A Milk and Other Milk Products -

Product Bacterial limits

Grade A raw milk
Individual producer milk not to exceed 100,000/ml prior to comming line
with other producer milk
for pasteurization
Grad A Pasturized
Milk and milk products:20,000/ml Coliform Limit:not exceeding 10/ml.
milk products
Grade A
Pasteurized
Exempt.

cultured products

26) Temperature Standards for Grade A Milk and Other Milk Products -

Product Temperature
Grade A raw milk
Cooled to 50F or less and maintained there at until proceesed.
for pasteurization
Grad A Pasturized
Cooled to 45F or less and maintained there at.
milk products
Grade A Pasteurized
Cooled to 45F or less and maintained there at.
cultured products

27) Dehydration of Milk - The milk can be preserved by dehydrating to various

degrees. The dehydrated milk has a longer "shelf-life" than the milk preserved by
pasteurization and refrigeration. Milk from which part or all of the water as been
withdrawn is termed concentrated or dried milk (milk powder).

28)

Concentrated Milk - Concentrated milk is the milk in which solids have been concentrated by
removal of water. Water may be removed by evaporation, by sublimation or by partial freezing
followed by removal of ice crystals. The first method is the most common. The higher
concentration of solids which results, provides the product some protection against spoilage by
certain chemical actions or microbial agents.

The degree of this protection is closely related to the extent of concentration; the greater the
proportion of the water removed, the more the protection. Whole milk, skim, or standardized milk
are used in the preparation of concentrated milks. Partially condensed milk or skim, sweetened or
unsweetened condensed milk or skim, evaporated milk or skim, and khoa are concentrated milks.

Prevention of Food Adulteration (P.F.A) Act, Government of India uses the term “evaporated" to
I4er to unsweetened condensed milk or skim.

29) Composition of Concentrated Milk

Product %of Milk fat %of Total milk solids %of Added cane sugar
Minimum Minimum
Min Max
Condenced milk 8.0 - 26.0 -
unsweetend or
evaporated milk
Condenced milk sweetend 9.0- 31.0 40.0
Condenced skim, -0.5 20.0 -
unsweetend,or
evaporated milk
Condenced skim, -0.5 26.0 40.0
sweetend
Khoa 20.0- - -

30)

Milk Powder - Powdered milk is made by the removal of most of the water from milk so that the
milk solids are accompanied by not more than 5%, perhaps as little as 2% -moisture. Water is
removed in two stages. In the first stage the milk is concentrated by vacuum evaporation to about
one-third of its original value. In the second stage the milk is dried, usually on a roll drier or in a
spray drier.

Three classes of milk powder are recognised by P. F. A. Act. They are milk powder, skimmed milk
powder, and partially, skimmed milk powder. The principal differences are in their final
composition, and are influenced by the class of milk which is dried.

31) Composition of Dried Milk

Powder %of fat %of Solid non- %of %of Ratio Fat / Solid non
fat Water fat
Whole milk 26.0 to 66.5 to 70.3 3.7 to 5.0 1:2:3 to1:2:7
28.5
Skim milk 1.3 to 1.6 93.5 to 94.9 3.5 to 5.0 1:10:2
Partially skimmed 8.5 to 24.0 71.0 to 86.5 4.2 to5.0 1:3 to 1:10:2
milk

32)

Fermented Milk - Long before microorganisms

had been discovered, butter milk and sour milk
were found palatable by some people in all
countries, Fermented milks are prepared by
allowing the normal lactic acid producing
organisms present to develop, or by inoculating
fresh milk with a portion of fermented milk of good
quality.

During fermentation, lactic acid thickens or curdles the milk and produces the desired sour
flavour. The nature of the product depends upon following factors: 1. The source of the milk
(cow, buffalo, goat, sheep, etc.) 2. The temperature to which milk is heated before inoculation.
3. The kinds of microorganisms in the starter. 4. The incubation temperature.

The market milk contains numerous species of

microorganisms in varying numbers. Their activity
is uncontrolled and, therefore, uniformity of
product is difficult to achieve. It is often
necessary and desirable to carry out controlled
fermentation with specific organisms. These
specific organisms have been identified. This has
led to the selection and propagation of known
cultures, sometimes referred to as starter cultures.
The principal organisms employed are species of
Streptococcus, Leuconostoc and Lactobacillus.

The starter culture may be a pure culture or mixture of known organisms. Each product requires
its own special starter and these contain different mixtures of organisms.The starter must be
capable of producing lactic acid at a rapid rate and outgrowing undesirable contaminants. Even
where only a single species is present in the starter, it is customary to use several different
varieties of the same species. This gives protection against culture being destroyed by a specific
bacteriophage. For these purposes lyophilized or other pure, stock cultures of desired organisms
are maintained by several commercial laboratories.

The consumption of fermented milk

preparations is widespread because of their
supposedly therapeutic value. Metchnikoff
(1908) developed the idea that the usual
Coliform and Clostridial bacteria in the intestine
of man form putrefactive products which are
distinctly injurious and the cause of definite
symptoms of disease. He even expressed the
opinion that these products are a common cause
of death attributed to old age. The remedy he
advocated was to prevent the activity of these
putrefiers by acidifying the intestinal contents
 with lactic acid.

This might be done by drinking fermented milk and introducing Lactobacillus bulgaricus which he
had isolated from Bulgarian fermented milk. This would prevent decomposition of proteins and thus
one might live for long periods as was the case with Bulgarian people. It was found later that L.
bulgaricus does not survive for very long in the intestine but L. acidophilus does and it also makes
an excellent tasting fermented milk. In recent years antibiotics are widely used which disturb the
normal balance of bacteria in the intestine. This has given a new impetus to the practice of
consuming Lactobacilli. This ensures the establishment of desirable microflora in the intestine during
or after antibiotic treatment. Fermented milks appear under various names. The names identify the
country or region where they

are produced and do not necessarily mean that they are fermented with different organisms. Some
preparations contain only one organism while others result from the combined action of two or more
organisms.

33)

Butter - Butter is made by churning

pasteurized sweet or sour cream, to separate fat
globules from the other constituents. The liquid
portion, buttermilk, drained off, and the granules
are further processed. Butter consists of about
80 percent fat, small percentages of lactose and
protein, and often 2 percent salt. The
remainder is water in the form of minute droplets
dispersed throughout the butterfat. The, salt is
dissolved in this water.

The cream from which the butter is churned may be soured naturally or by the addition of a
culture of organisms known as starter to improve the flavour. The advantages of first souring the
cream are: (l) the yield of butter is increased, owing to a better separation of fat, and (2) the
aroma and flavour may be greatly improved. Before a starter is added, the cream is pasteurized at
71° C for 30 minutes.

These eliminate undesirable organisms and

destroy milk lipase that would otherwise cause
rancidity during storage of butter. The starter
consists of two types of organism; 1. Those
producing a high acidity e. g. Streptococcus
lactis and S. cremoris and 2. Those imparting
characteristic aroma and flavour e. g.
Leuconostoc citrovorum and L. dextramicum.
The keeping quality of well made butter is good.
Although it has retained some of the bacteria of
the cream, it is low in sugar and protein. Since
the salt is all dissolved in aqueous fraction the
water present contains a high percentage of salt.

This prevents the growth of bacteria. Also, water droplets are so evenly distributed and so minute
that bacteria which are in these droplets, cannot multiply because of space limitation. Another
factor which restricts microbial growth is anaerobic condition except at the surface, where aerobic
microorganisms may develop. Undesirable changes which take place after butter has been
manufactured are produced largely by the growth of microorganisms. These are mainly aerobic
microorganisms which grow on the surface of the butter after its manufacture. The microorganisms
which cause rancidity, discoloration and off flavours are as follows:

1. Moulds of the genera Asrpergilhl8, Mucor,

Rhizopus, Clados porium, Alternaria. 2. Yeasts of
the genus Torula. 3. Bacteria of the genera
Pseudomonas and Achromobacter.
Microbiological tests carried out on butter are very
limited. Total bacterial count is not considered
significant because they are actually added as
starter. Yeast and moulds, when numerous, are
objectionable in butter.

Their presences indicate either improper pasteurization of cream or recontamination after

pasteurization. Samples with less than 10 yeasts and moulds per gram are classified as good. The
number of proteolytic bacteria is an indication of keeping quality and the count should be less than
50 per gram.

34)

Cheese - Cheese is basically a means of storing

milk and has been used for this purpose for at
least 2000 years. Cheese is the product made by
separating the casein of milk from the liquid or
whey. The butterfat often accompanies the
casein, but most of the lactose and other soluble
milk constituents remain in the whey. The
manufacture of all cheeses depends upon the
activities of selected microorganisms. Hundreds of
different kinds and varieties of cheeses are
produced by introducing many variables into the
manufacturing process. However, there are about
eighteen major types, the rest being variation of
these.

Cheese may be classified in several manners: on the basis of hardness, as nard, semi hard, soft
cheeses; on the basis of the principal ripening agents, as mould or bacterial cheeses; on the basis
of the method of coagulation, as rennet or acid curd cheese etc. Cheeses classified according to
their consistency and the use and nature of microbial ripening agents. Several kinds of cheese in
small quantities arc made in India. Panir is an ancient indigenous product. Other cheeses are
similar to common western types. Soft cheeses include Surti, Bandal and Dacc a. Hard cheeses are
the cheddar types.

Most cheese is made from either whole or

skimmed milk. Some varieties are prepared from
cream and whey. Milk from cows, goats, ewes
mares and other animals has been used for the
making of cheeses. Four steps are generally
followed in the preparation of cheese. These are
(I) inoculation of milk with starter culture and
curdling the milk, (2) processing of curd, (3)
salting, and (4) ripening. Milk inoculated with
starter culture ferments lactose with acid
production. The acid reacts with calcium to give
calcium lactate. When the content is reduced to a
certain low point, the casein precipitates with the
formation of firm curd. Starter culture consists of
Streptococcal lactis or S. cremoris if milk is held
below 38°C.

A starter culture for use at higher temperature (about 50°C) consists of S. thermophiles combined
with species of lactobacilli e.g., Lactobacillus lactis, L.Curd may also be prepared by adding the
enzyme renin to milk. The watery fluid which separates out during the formation of curd is called
Whey. It consists of 93 percent water, 5 percent lactose, and smaller amounts of minerals,
vitamins, lactalbumin, and fats.

The curd is separated from whey by various

methods depending on the amount of moisture
desired by various types of cheeses. Draining
of whey without pressure results in the
production of soft cheeses, which have high
moisture content. Hard cheeses require curd of
allow moisture content. This is achieved by heat,
pressure, and cutting of the curd. After the
moisture content has been reduced to the
desired limit, the curd is molded into a
characteristic shape and size.

Salt: (sodium chloride) is applied to practically all varieties of cheese. It is added by (1) floating
the cheese in a strong brine solution, or (2) rubbing the surface of the pressed cheese with dry salt.
The flavour of Taw cheese curd is very bland, and the cheese is rubbery in consistency. To be made
satisfactory for consumption, it must be set aside to ripen. During ripening, the flavour, aroma,
composition, and physical properties of the cheese undergo change due to the action of
microorganisms and their enzymes. The natures of the changes depend on the microorganisms
present or added and the control of temperature and relative humidity in the curing rooms,
chambers or caves. The development of microbial flora for ripening varies with the type of cheese.
In Roquefort and blue cheese, the curd is inoculated with spores of Penicillium roqueforti, In Swiss
cheese lactobacilli. Streptococci and propionibacteria arc added to the milk before the curdling
process. In. Limburger cheese the surface of the curd is allowed to be contaminated with desirable
organisms, mainly Brevibacterium linens, when placed in the ripening room. During the ripening
process, the degradation of lactose, protein, and fat is brought about by the activity of a mixed
population of microorganisms. Lactose is degraded to lactic acid and smaller amounts of volatile
acids, alcohol, and natural products. Bacteria of the lactic acid type are mall1ly responsible for the
flavour of hard cheeses. Propionibacteria ferments lactose, producing carbon dioxide. This results
in the formation of holes or "eyes" in Swiss cheese. Degradation of protein and fat is considerably
less in hard cheeses. In soft cheeses, protein is completely degraded to peptides and amino acids.
Cheese made from whole milk has considerable fat in the curd. Lipases elaborated by the mould
hydrolyse the fat, releasing butyric, caproic, caprylic, capric higher fatty acids. Some of these acids
are oxidized forming compounds contributing to flavour. Many undesirable microorganisms are
responsible for the spoilage of cheese. These may affect the flavour, aroma and texture of cheese.
Coliform species and certain yeasts ferment lactose with the production of acid and gas. Gas holes
are produced which give a swollen appearance to cheese. E. coli also imparts a bitter flavour to
cheese. The putrefactive organisms grow and become active when the acidity of cheese is reduced
during the ripening period. These organisms are responsible for putrid odours and flavours. The
discoloration of cheese is brought about by several pigment producing organisms. This may also
be caused by chemical reactions with metals such as copper and iron from the utensils. Several
epidemics have been traced to the consumption of cheese contaminated with pathogenic organisms.
The organisms most commonly associated with cheese born infections are Staphylococcus aureus,
Clostridium butylicum. Brucella melitensis and several species of salmonella.

35) Classification of Cheese -

Representative cheeses Distinctive organisms

Acid-curd

Soft :Cottage Not ripened

Cream
Rent-curd Ripened by surface growth
Soft :Limburger Streptococcus liquefaciens,Brevibacterium

Camembert Penicillium camemberti, Brevibacterium

Semihard :

Roquefort Penicillium roqueforti

Blue
Hard :Swiss Propionibacterium species

Cheddar Lactic group, Geotrichum

35)

Types of Microorganisms in Milk - The types

of micro organisms found in milk vary
considerably, and are dependent upon the specific
conditions associated with a batch of milk.
Bacteria, yeasts, moulds, and bacteriophages are
commonly encountered. Other viruses and
Protozoa are seldom observed in milk products,
except as occasional contaminants.

Bacteria Bacteria are the most common, and probably the most numerous of microorganisms
with which the dairy processing industry is concerned. They belong to four main groups: (1) cocci,
usually gram positive, (2) gram positive non-sporeforming rods, (3) gram positive sporeforming
rods, and (4) gram negative non-spore forming rods.

Yeasts The yeasts most frequently encountered

in milk and milk products act upon the lactose to
produce acid and carbon dioxide. Yeasts are
more commonly found in raw, cream during hot
weather, but are potential contaminants
throughout the year.

Moulds Moulds often grow in large concentrations and are visible as a fuzzy or fluffy growth. They
are sometimes observed on the surface, of butter, old cream, khoa, or cheese. They are black,
grey, green, blue or white. They discolour milk products and often produce undesirable, at times
repulsive, odours and flavours. Moulds are essential in production of the certain kinds of cheese.

Bacteriophages Bacteriophages are particularly

obnoxious in starter cultures used for making
cultured milk, butter and cheese. Phages will,
kill the bacterial culture and the whole process of
fermentation will be lost.

Microorganisms found in milk can also be described on the basis of the following characteristics; 1.
Biochemical activities. 2. Temperature response. 3. Ability to cause infection and disease.
Biochemical activities If allowed to stand under condition that permit bacterial growth, raw milk
of a good sanitary quality will rapidly undergo a series of chemical changes. The principal change is
lactose fermentation to lactic acid. This change is brought about by acid uric lactic organisms,
especially Strepotococcus lactis and certain lactobacilli. These include two distinct biochemical
types, homo-and heterofermentative. In homofermentation lactic acid is the major product of
lactose fermentation. Heterofermentative organisms, however, produce lactic, acetic, propionic,
and some other acids, and some alcohols and gases such as CO2 and H2 Organisms continue to
form lactic acid until the concentration of acid is itself too great for the organisms to remain live.
Microbacteria, micrococci, coliforn18, etc. also ferment lactose to lactic acid and other products.
Many Clostriifiul1J species and, some yeasts such as Torula lactic, and Torula cremoris ferment
lactose with acid and gas production. As the acidity continues to increase and reaches a pH of 4.7,
it eventually causes a precipitation of casein. Organisms capable of metabolizing lactic and other
acids develop especially acid uric, yeasts and moulds. The acidity of milk is diminished and the
alkaline products of protein decomposition such as amines, ammonia and the like are produced.
This is accomplished by many species of the genera Bacillus, Clostridium, Pseudomonas, Proteus
and numerous other forms. The action of microorganisms does not involve fat as readily as it does
lactose and protein. Lipolysis results from the action of lipase produced by bacteria such as
Pseudomonas, Achromobacter and by some yeasts and moulds. Fat is hydrolysed to glycerol and
fatty acids. Some of the fatty acids, for example, butyric and caproic acid give milk products,
distinctive and usually rancid, odours and flavours. Several microorganisms also bring about certain
objection able changes in the milk which may not be deleterious to health. Rapines in milk is
sometimes encountered. The milk become ropy or slimy and may be pulled out into long threads.
It is produced by several organisms but the most important species is Alcaligenes viscolactis. A
rapid fermentation of lactose in milk is sometimes observed and is known as stormy fermentation.
This is brought about by Clostridium perfringens. The curd become torn to shreds by the vigorous
fermentation and gas production. Several organisms have been isolated from milk which impart
brilliant colours. Pseudomonas syncyanea imparts blue colour, pseudomonas synxantha yellow
colour and Serratia marcescens red colour to the milk.

Temperature response Microorganisms found in milk can also be described according their
optimum temperature for growth and heat resistance. This is a very practical consideration since
milk is preserved by employing low temperatures to prevent changes due to microbial activi1y'and
by high temperatures to reduce microbial population and destroy pathogens. All the four types of
microorganisms i.e. psychrophilic, mesophilic, thermophilic and thermoduric are found in milk.
Psychrophiles grow at temperatures just above freezing and at refrigeration tempratures. They
produce a wide variety of spoilage defects. The defects may result in the production bf many "off"
flavours and odours. The most commonly encountered psychrophilic bacteria are members of the
genera pseudomonas, Achromobacter, Vibrio, Flavobacterium and Alcaligenes, They arc killed in
the pasteurization process, but are sometimes found in pasteurized milk. The contamination takes
place after pasteurization from equipment, cans, bottles, and water. The most important mesophilic
bacteria are streptococci, lactobacilli and coliforms, which produce acid and gas and off flavours.
They are killed in the pasteurization process Thermophilic bacteria grow well at the temperature
used in pasteurization, specially when the low temperature holding method is followed. Most
thermophilic forms are found in two genera, Bacillus and Clostridium. Thermoduric organisms are
regarded as those which survive pasteurization but do not grow at pasteurization temperatures. The
most common thermoduric bacteria are found in the genera Microbacterium, Corynebacterium,
Micrococcus, Streptococcus and Bacillus. Excessive numbers of thermoduric bacteria in milk make it
difficult to meet the grading standard. Ability to cause infection and disease Pathogenic
organisms of both bovine and human origin have been isolated from milk. Milk, therefore, can serve
as a carrier of diseases. Many serious epidemics were caused by the consumption of such products
before this fact was clearly recognized. However, this became less common as milk sanitation has
improved and pasteurization is being more widely practised. The disease organisms present in milk
may be derived from (1) diseased animals or (2) persons collecting and handling milk: Thus the
danger is due to the inoculum and not to the growth of organisms in the milk. The health of animal
is an important factor. Several diseases of cattle including staphylococcal and streptococcal
infections, tuberculosis, brucellosis, salmonellosis, Q fever and Foot and mouth disease may be
transmitted to man. The organisms causing these diseases may get into the milk either directly
from the udder, or indirectly from infected body discharges, which may drop, splash, or be blown
into the milk. Some of the important diseases of human origin that have been transmitted by milk
are (1) typhoid fever (2) diphtheria, (3) scarlet fever, (4) dysentery (5) septic sore throat and (6)
poliomyelitis. It is also possible for humans to infect animals. For example, mastitis may be caused
by a variety of organisms, including Staphylococcus aureus. The infecting organism, in s9me cases,
has been traced to humans.

36)

Milk Spoilage - Spoilage occurs when microorganisms degrade the carbohydrates,

proteins, fats of milk and produce noxious, end products. It may be seen that
Lactobacillus or Streptococcus species ferment the lactose to lactic acid and acetic acids
turning the mi1k sour. They may produce enough acid to curdle the protein and form
sour curd.

Attack of milk protein by Micrococcus, Bacillus or Proteus results into sweet curdling.
There is little acid formation. If milk becomes contaminated with Gram-negative rods-of
coliform group of bacteria, such as E. coli or Enterobacter aerogenes, or Clostridium sp.,
there is .acid and gas formation from the lactose.

This stormy fermentation causes the explosion of curds. Ropiness, like bread develops
from Alcaligenes, Klebsiella and Enterobacter. Serratia marcescens causes the
development of a red pigment.

37)PATHOGENIC ORG:
The previous chapter mentioned that different types of microorganisms interact with human
bodies on a regular basis. They can be harmless, harmful or beneficial. Harmful microorganims
are also called pathogenic. This chapter looks at disease- causing bacteria and viruses.

Communicable diseases
The ability of a microorganism to cause disease is called pathogenicity. There are several
pathogens that can cause serious harm or even immediate death.

Invasion and multiplication of pathogenic microorganisms in the body is called an infection. When
we are infected by pathogens we become sick, which means that our bodies stop functioning
properly. Infectious agents, such as bacteria, a virus, fungi or protozoa cause communicable
diseases. Communicable diseases can be spread from one person to another.

Infection transmission
All living organisms have a natural or acquired resistance mechanism called immunity. When we
get sick, for example, we use different body cells and chemicals to fight bacteria. Bacteria in their
turn use different chemicals to fight us. That is why infection is sometimes referred to as a race
between pathogen and host organism. The infection can be transmitted by direct or indirect
contact.

Direct contact transmission - involves any direct contact with an infected individual. Infection
can be passed in water droplets through a sneeze, cough, laugh or exhalation and through bodily
fluids. Most communicable diseases like colds, influenza, tuberculosis and HIV are spread from
person to person through infected fluid droplets.

Indirect contact transmission - is a method of spreading infection from person to person that
involves contact with a contaminated object. Objects can become contaminated when touched by
someone with an infection. The infected object is called a fomite.

Another form of infection transmission through indirect contact is through the oral-feacal route,
which usually involves ingesting contaminated water.

A third method of indirect contact involves vector-borne diseases, which are carried by animals
and insects. A vector is an organism that serves as a 'go-between' in the transmission of a host-
to-host disease.

Examples of diseases that can be transmitted via indirect contact are cholera, Salmonellosis and
dysentery. See image 1.

Types of pathogens
See image 2.

Natural or human-triggered changes in the environment might upset the natural balance between
living organisms. These new environmental conditions may encourage pathogens, allowing them
to multiply rapidly and increase the risk of exposing humans who share that environment. Here
are the main groups of human pathogens with some examples of the diseases they cause.

Bacteria
E.coli causes food poisoning and urinary tract infections. Mycobacterium tuberculosis causes
tuberculosis.

Viruses
Influenza virus causes 'flu'. Herpes simplex virus causes herpes.

Protozoa
Plasmodium causes malaria.

Fungi
Tinea causes ringworm.

See animation 1.

Epidemiology
Epidemiology is the study of the patterns, causes and prevention of human diseases. An
outbreak of a contagious disease that spreads rapidly and widely is called an epidemic. An
epidemic occurring over a very large area is called a pandemic. The term 'epidemiologic triangle'
is used to describe the intersection of host, agent and environment in analysing an outbreak.

The prevalence of a disease is the number of diseased individuals at any one time (point
prevalence) or over a given period (period prevalence). The incidence is the number of new
cases of a disease that occur within a defined population over an established period of time.

History of epidemics
The influenza (virus-caused) pandemic of 1918-1919 killed more people than World War I. It is
considered the most devastating epidemic in recorded world history. More people died of
influenza in a single year than in four years of the Black Death Bubonic Plague from 1347 to
1351.

Bubonic plague is caused by bacteria called Yersinia pestis. These bacteria live in fleas on rats.
Rats were abundant in medieval times due to very low hygiene standards, and the lack of sewage
systems and hot, running water. Bubonic plague killed about a quarter of the European
population of that time. Today this disease can be controlled by antibiotics but outbreaks sill occur
sometimes. See image 3.

Prevention of infections
The best way of fighting disease is to prevent it. The discovery and manufacture of antibiotics and
vaccines in the early part of the 20th century have changed the state of people's health forever.
There are three stages of prevention of infections: primary, secondary and tertiary.

The Primary stage - involves public education about infectious diseases.

The Secondary stage - involves treating the actual infection that has already occurred by
quarantining and/or vaccinating of infected individuals.

The Tertiary stage - involves the recovery from illness.

Bioterrorism
Bioterrorism is the deliberate use of microorganisms, or their toxins, to cause death or disease.
Biological and chemical agents that could be used include anthrax, small pox, West Nile virus and
cholera. Biosecurity refers to the policies and measures taken for protecting a nation's food
supply and agricultural resources from both accidental contamination and deliberate attacks of
bioterrorism.