UNIT 1

Introduction
Dairy microbiology is a branch of microbiology that focuses on the microorganisms
associated with milk and dairy products. It studies the beneficial, spoilage, and pathogenic
microorganisms that influence the quality, safety, and shelf life of dairy products. Dairy
microbiology plays a crucial role in ensuring the hygienic production, processing, and
distribution of milk and its derivatives.

SIGNIFICANCE OF DAIRY MICROBIOLOGY

Milk is a highly nutritious food, serving as an ideal medium for microbial growth due
to its rich composition of carbohydrates, proteins, fats, vitamins, and minerals. Understanding
the microbiology of milk helps in controlling microbial contamination, ensuring product
safety, and improving production techniques for fermented dairy products.

Ta
Source Percent composition
ble :
(Animal) Total Protein Casein Whey protein Fat Carbohydrate
Buffalo 4.0 3.5 0.5 7.5 4.8
Cow 3.5 2.8 0.7 3.7 4.8
Goat 3.6 2.7 0.9 4.1 4.7
Sheep 5.8 4.9 0.9 7.9 4.5
Percent composition of milk of different animal origins

1. Ensuring Milk Safety

 Milk is a nutrient-rich medium, making it prone to contamination by harmful

microorganisms like Escherichia coli, Salmonella spp., and Listeria monocytogenes.

 Dairy microbiology ensures the application of pasteurization, sterilization, and other

safety measures to eliminate pathogenic microbes, safeguarding public health.

2. Quality Control
 The microbial load significantly affects the sensory attributes of dairy products,
including taste, texture, and odor.

 Routine microbiological testing ensures that dairy products meet the required
standards and remain of high quality.

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3. Spoilage Prevention
 Spoilage microorganisms like Pseudomonas spp. and Bacillus spp. degrade milk and
reduce its shelf life.

 By implementing proper microbial management and storage techniques, spoilage can

be minimized.

4. Production of Fermented Dairy Products

 Beneficial microorganisms such as Lactobacillus and Streptococcus species are vital
for producing yogurt, cheese, butter, and other fermented products.

 These microbes enhance the flavor, texture, and nutritional value of the products,
while also acting as natural preservatives.

5. Development of Probiotics
 Probiotic bacteria like Lactobacillus acidophilus and Bifidobacterium are used in
functional foods to improve gut health and boost immunity.

 Dairy microbiology enables the development and optimization of probiotic-rich dairy

products.

6. Improved Shelf Life

 Innovations in packaging, combined with microbiological expertise, help in extending
the shelf life of dairy products.

 Techniques such as aseptic processing and modified atmosphere packaging prevent

microbial contamination.

7. Economic Impact
 Microbial control in dairy processing reduces wastage due to spoilage, saving
resources and increasing profitability.

 Fermentation and probiotics add value to dairy products, creating new revenue
streams for the industry.

8. Public Health Contribution

 Dairy microbiology ensures that milk and its derivatives are free from harmful
residues, toxins, and pathogens.

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  Monitoring antibiotic resistance in dairy products helps maintain safe consumption
standards.

9. Hygiene Monitoring in the Dairy Industry

 Dairy microbiology plays a critical role in monitoring hygiene during milk
production, storage, and processing.

 It ensures compliance with food safety regulations, fostering consumer trust.

10. Research and Development

 Advances in dairy microbiology lead to the discovery of novel starter cultures and
functional foods.

 It also supports innovation in dairy preservation and bioactive product development.

SOURCES OF CONTAMINATION IN MILK

Milk is a highly nutritious and perishable food that is susceptible to contamination by

various microorganisms. These contaminants can originate from multiple sources, including
the animal, environment, equipment, and human handling. Microbial contamination of milk
can be from the internal and/ or external sources

1. Contamination from the Udder

a) Infected Udder (Mastitis)
Animals suffering from mastitis (inflammation of the udder) often shed pathogenic
bacteria like Staphylococcus aureus, Streptococcus agalactiae, and E. coli into milk. Such
infections not only affect milk quality but can also pose health risks to consumers. Varying
numbers of bacteria are found in aseptically drawn milk with the reported counts of <100-
10,000 CFU/ml from normal udder.

b) External Contamination of Udder

 Dirt, dust, manure, and mud on the udder’s surface can enter milk during the milking
process.

 Improper cleaning of the udder before milking increases this risk.

 The environmental conditions such as soil, manure, mud, feed or bedding; determines
what kind of microbes will dominate in milk.

 Predominantly micrococci and coagulase negative staphylococci exist.

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  Aerobic thermoduric organisms are entirely Bacillus spp. The more frequent are B.
licheniformis, B. subtilis,

2. Contamination During Milking

a) Milking Equipment
 Milk comes into direct contact with milking machines, pipelines, and containers. If
these are not cleaned or sterilized properly, they can harbor microorganisms like
Pseudomonas, Bacillus, and yeast.

b) Milker’s Hands
 The hands of the person milking the animal can introduce bacteria like E. coli and
Salmonella if proper hygiene is not maintained.

 Dirty gloves or nails can further add contaminants.

3. Contamination from the Animal’s Environment

a) Bedding and Manure
Bedding material such as straw and manure in the milking area can transfer
microorganisms to the udder or milk. Milk produced on farms with poor hygiene practices
may undergo significant spoilage and have a shorter shelf- life, when compared to milk
produced under hygienic conditions. Microbes associated with the bedding materials
include:

 Coliforms
 Spore-formers
 Staphylococci
 Streptococci
 Clostridium spores from soil or manure can enter the milk, leading to spoilage or
health issues.

4. Contamination During Storage and Transport

a) Improper Cooling
 Milk should be cooled immediately to below 4°C after milking to prevent bacterial
growth. Failure to do so allows rapid multiplication of spoilage organisms like
Pseudomonas.

b) Storage Containers
 Using dirty, rusted, or non-food-grade storage containers can introduce bacteria and
chemicals into milk.

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c) Transport Vehicles
 If milk transport tanks are not properly cleaned and sanitized, they can harbor
pathogens and spoilage microorganisms.

5. Contamination from Feed and Water

a) Contaminated Feed
 Animal feed that is contaminated with fungi, bacteria, or chemicals can indirectly
affect the microbial quality of milk.
 For example, molds in feed can produce toxins that may pass into milk.
b) Contaminated Drinking Water
 Non-potable water consumed by animals can introduce pathogens like Salmonella or
Cryptosporidium into milk.

6. Contamination by Air and Dust

 Airborne particles in the milking environment can carry microorganisms such as
spores of Bacillus, Clostridium, or molds.

 Dust from the barn or surrounding areas can settle in open milk containers,
introducing contaminants.

7. Contamination from Human Handling

a) Lack of Hygiene
 Milkers or workers who do not wash their hands or wear clean clothes can introduce
pathogens.
 Skin infections or respiratory droplets from workers can also contaminate milk.

b) Sick Workers
 Workers suffering from infections such as Salmonella or Shigella can transfer these
pathogens into milk.

8. Contamination from Adulteration

a) Addition of Water
 Adding water to milk to increase volume is a common practice, but if the water is
contaminated, it can introduce harmful microorganisms.

b) Chemical Adulterants
 Some chemicals added to milk as preservatives can encourage microbial growth if not
properly regulated.

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9. Contamination During Processing
a) Unclean Processing Units
 Processing plants that are not properly sanitized can become breeding grounds for
microorganisms like Listeria monocytogenes and E. coli.

b) Incomplete Pasteurization
 Improper or incomplete pasteurization can fail to kill all pathogenic bacteria, leaving
some to survive and multiply.

10. Contamination from Packaging

a) Non-Sterile Packaging Material
 Packaging materials like plastic bags or bottles can introduce microorganisms if not
sterilized before use.

b) Improper Sealing
 Poorly sealed packages can allow air and microorganisms to enter, leading to
contamination.

Common Microorganisms Found in Contaminated Milk

1. Pathogens: E. coli, Salmonella spp., Listeria monocytogenes, Staphylococcus aureus.
2. Spoilage Microbes: Pseudomonas spp., Bacillus spp., yeasts, and molds.
3. Toxin-Producing Microbes: Clostridium botulinum and Aspergillus (aflatoxin).

HYGIENIC MILK PRODUCTION

Hygienic milk production refers to the methods and practices used to ensure milk is
produced, handled, and processed in a way that minimizes contamination, maintains quality,
and ensures safety for consumers. This process involves strict adherence to cleanliness and
sanitation at every stage, from the farm to the consumer.

Principles of Hygienic Milk Production

1. Producing milk free from contaminants.
2. Minimizing microbial growth during and after milking.
3. Preserving milk’s nutritional and sensory qualities.

Steps in Hygienic Milk Production

1. Animal Health and Hygiene
 Healthy Animals:

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 o Only healthy animals should be milked to ensure the milk is free from
infections like mastitis.
o Regular veterinary checks should be conducted to monitor animal health.
 Clean Animals:
o Animals should be kept clean, especially the udder and teats.
o Regular grooming and proper bedding should be maintained to prevent
contamination from dirt, manure, and bedding material.

2. Milking Area Hygiene

 Clean Environment:
o The milking area should be clean, well-ventilated, and free from dust, insects,
and rodents.
 Proper Lighting and Drainage:
o Good lighting helps identify dirt or contamination. Proper drainage prevents
water stagnation, which can harbor bacteria.

3. Udder and Teat Cleaning

 Wash the udder and teats with clean water or a sanitizing solution before milking.
 Dry the teats using a clean, dry towel to avoid water droplets that may carry bacteria
into the milk.

4. Milking Practices
 Hand Milking:
o Hands should be thoroughly washed with soap and water before milking.
o Use disposable gloves to minimize contamination.

 Machine Milking:
o Milking machines should be sanitized before and after every use.
o Ensure no residues remain in pipelines or containers.

Cleanliness
 Milker should always wear neat and clean clothing and caps.
 Milker should also maintain adequate personal cleanliness ( i.e. haircut, trimming of
beard and cutting the nail regularly) to avoid any microbial contaminations
 Cuts, blisters or boils, if any must be covered with clean dressing
 Washing of hands with detergents and drying with clean towel is must prior to hand
milking
 Never smoke, eat drink or spit while milking
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  Avoid coughing and sneezing while milking

5. Milking Equipment Hygiene

 Use stainless steel or food-grade plastic equipment as they are easier to clean and
sanitize.
 Clean equipment with hot water, detergent, and a disinfectant after each milking
session.
 Store equipment in a clean, dry area to prevent microbial growth.
Hygiene of milking utensils
 The milking utensils should be of uniform size. Having small mouths to avoid
external contamination. These should be preferably made up of non-rusting and non-
absorbent materials (i.e. aluminium or galvanized iron). Stainless steel is ideal, but
costly. All the utensils should be free from dents, cracks and crevices. The utensils
should be scrubbed and cleaned before and after each milking.

6. Milk Handling and Storage

 Filtering:
o Filter milk immediately after milking to remove visible impurities.
 Cooling:
o Cool milk to 4°C or below as soon as possible to prevent microbial growth.
o Use refrigeration or ice tanks for bulk milk cooling.
 Storage Containers:
o Use clean, sealed, and food-grade containers to store milk. Avoid open
containers.
7. Transport Hygiene
 Use clean and sanitized milk tanks or cans for transport.
 Transport milk in refrigerated vehicles to maintain the cold chain.
 Avoid delays in transporting milk to processing plants.

8. Worker Hygiene and Training

 Workers involved in milk production should:
o Maintain personal hygiene by wearing clean clothes, gloves, and head covers.
o Wash hands thoroughly before and after handling milk.
o Be free from infectious diseases to avoid contaminating the milk.
 Regular training should be provided to workers on best practices for hygienic milk
production.

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9. Water Quality
 Use potable water for cleaning animals, equipment, and milking areas.
 Test water regularly for microbial and chemical safety.

10. Pest Control

 Keep milking and storage areas free from pests such as flies, rodents, and insects,
which can contaminate milk.
 Use pest control measures that do not harm the milk or environment.

Benefits of Hygienic Milk Production

1. Hygienic practices ensure milk is free from off-flavors, sediments, and microbial
contamination.
2. Reduces the risk of milkborne diseases caused by pathogens like E. coli, Salmonella,
and Listeria.
3. Proper handling slows microbial growth, extending the milk’s shelf life.
4. Reduced spoilage and waste lead to better profitability for farmers and processors.
5. Hygienic milk production ensures adherence to food safety standards set by
government and international bodies.
Challenges in Hygienic Milk Production
1. Lack of awareness among farmers about hygiene practices.
2. High costs of advanced milking and cooling equipment.
3. Limited access to veterinary services in rural areas.
4. Inadequate infrastructure for cold storage and transport.

MICROBIAL CHANGES IN MILK DURING PRODUCTION AND PROCESSING

Milk is an ideal medium for microbial growth due to its rich composition of proteins,
fats, lactose, and vitamins. These nutrients not only support beneficial microorganisms but
also enable the growth of spoilage and pathogenic microbes if proper precautions are not
taken. The microbial changes during production and processing are critical to understand,
particularly for microbiology students, to ensure milk quality, safety, and shelf life.

During Milking
Microbial contamination begins during milking. The udder, especially in animals with
mastitis, serves as a primary source, introducing pathogens like Staphylococcus aureus,
Streptococcus agalactiae, and E. coli. External contaminants from dust, manure, or the
milking area can add to the microbial load. Equipment used in milking, if not properly
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cleaned and sanitized, can harbor bacteria like Pseudomonas, Bacillus, and Clostridium.
Additionally, improper handling by milkers with unwashed hands or gloves increases the risk
of contamination.

During Storage at the Farm

Immediately after milking, milk should be cooled to below 4°C to slow bacterial
growth. Failure to do so allows rapid multiplication of mesophilic bacteria such as
Lactococcus lactis, leading to souring. Psychrotrophic bacteria like Pseudomonas fluorescens
thrive at low temperatures and produce heat-stable enzymes, such as lipases and proteases,
that degrade fats and proteins even after pasteurization. These enzymatic activities result in
spoilage characterized by rancidity and bitterness. Additionally, spore-forming bacteria like
Clostridium can survive in dormant forms and germinate under favorable conditions.

During Transportation
Transporting milk in not sanitized containers or tanks often leads to secondary
contamination by microbes like Salmonella and Enterobacter. Delays in transportation or
failure to maintain a cold chain (keeping milk consistently below 4°C) can allow
psychrotrophic and thermophilic bacteria to proliferate, particularly during warmer
conditions. These bacteria may produce toxins or enzymes that compromise milk quality and
safety.

During Processing
Processing, particularly pasteurization, plays a key role in reducing microbial content.
Pasteurization effectively destroys most pathogens, including Salmonella spp., Listeria
monocytogenes, and Mycobacterium tuberculosis. However, heat-resistant bacteria such as
Bacillus cereus and Clostridium botulinum spores may survive if not handled correctly.
Ultra-high temperature (UHT) processing eliminates these bacteria, but improper aseptic
packaging can reintroduce contaminants. Post-pasteurization contamination is also a concern,
especially in plants where equipment is not thoroughly cleaned or where human handling is
involved.

Post-Processing and Retail Storage

After processing, microbial growth in milk depends on storage conditions. If stored
above the recommended temperature, psychrotrophic bacteria can grow, leading to spoilage.
Molds and yeasts may also develop if packaging materials or seals are compromised. Biofilm
formation by bacteria like Pseudomonas on equipment or packaging can act as a persistent
source of contamination, leading to reduced shelf life.

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Key Factors Influencing Microbial Changes
1. Intrinsic Factors:
o Nutrients: Milk’s rich composition supports diverse microbial populations.
o pH: The near-neutral pH of milk (6.4–6.8) provides an optimal environment
for microbial growth.

2. Extrinsic Factors:
o Temperature: Improper cooling accelerates microbial multiplication.
o Hygiene: Cleanliness of milking areas, equipment, and handlers significantly
impacts contamination levels.

3. Microbial Interactions:
o Some microorganisms inhibit others through competition or the production of
antimicrobial compounds like bacteriocins. For instance, lactic acid bacteria
(LAB) produce organic acids that lower pH and inhibit spoilage organisms.

4. Spore-Forming Bacteria:
o Dormant spores of bacteria like Clostridium and Bacillus resist heat treatment
and may germinate under favorable storage conditions, causing spoilage and
safety issues.

Spoilage Microorganisms in Milk

1. Psychrotrophs:
o Pseudomonas fluorescens – Produces enzymes that cause rancidity.
2. Lactic Acid Bacteria (LAB):
o Lactobacillus and Lactococcus – Cause souring by fermenting lactose to lactic
acid.
3. Spore-Formers:
o Bacillus cereus – Causes sweet curdling and flat sour spoilage.
4. Yeasts and Molds:
o Grow under low pH conditions, especially in fermented milk products.

Impact of Microbial Changes

1. Quality Issues:
o Off-flavors, discoloration, and curdling are common signs of microbial
spoilage.
2. Safety Concerns:

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 o Pathogenic bacteria like Salmonella, E. coli O157 and Listeria monocytogenes
can lead to foodborne illnesses.
3. Economic Losses:
o Spoiled milk leads to reduced profitability for producers and processors.
4. Health Benefits (in controlled settings):
o Controlled microbial activity is desirable in producing fermented milk
products like yogurt and cheese.

MASTITIS
Mastitis is an inflammation of the mammary gland (udder) in dairy cows, primarily
caused by infections with bacteria, although viruses and fungi can also play a role. It is one of
the most common and costly diseases in dairy farming, affecting milk production and quality,
as well as the overall health of the cow. The disease can be caused by a wide range of
pathogens, including environmental and contagious bacteria, such as Escherichia coli and
Staphylococcus aureus, as well as less commonly by fungi or viruses

Life Cycle of Mastitis in Cows

The life cycle of mastitis in cows can be broken down into stages based on the
infection's initiation, progression, immune response, and eventual recovery or resolution.
Mastitis is primarily caused by bacterial pathogens that invade the cow's mammary gland.

1. Exposure to Pathogens
Mastitis begins when pathogenic microorganisms (bacteria, viruses, or fungi) come into
contact with the cow’s udder. The most common cause of mastitis is bacterial infection,
which is often transmitted from the environment or during milking. The primary sources of
these pathogens include:

 Contaminated Milking Equipment: Bacteria can enter the udder when milking
equipment, such as the teat cups, liners, or milk collection systems, is not properly
sanitized.

 Cow’s Environment: Dirty bedding, manure, or wet conditions can introduce

bacteria like E. coli, Staphylococcus aureus, & Streptococcus agalactiae into the
udder.

 Teat Injury: Injuries to the teat, such as from rough handling or faulty milking
equipment, can provide an entry point for bacteria. Pathogens are usually introduced
through the teat canal, which serves as the gateway for bacteria to enter the mammary
gland.
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2. Infection and Initial Response
Once the bacteria enter the udder through the teat canal, they begin to multiply in the
milk-producing tissues of the mammary gland. The bacteria thrive in the nutrient-rich
environment of the udder, where they can cause localized infection and inflammation.

 Bacterial Growth and Tissue Damage:

The bacteria multiply, causing tissue damage by releasing toxins and enzymes.
These enzymes break down the structural integrity of the udder tissue, leading to
inflammation.

 Immune System Activation:

The cow’s immune system detects the presence of pathogens and begins an
inflammatory response. White blood cells (such as neutrophils and macrophages)
migrate to the site of infection to fight the invading bacteria. This response leads to
the classic signs of mastitis, including swelling, heat, and redness in the affected
udder.

 Increase in Somatic Cell Count (SCC):

As part of the immune response, somatic cells (mainly leukocytes, such as
neutrophils) flood the udder to combat infection. An elevated SCC is a key indicator
of mastitis, with higher counts signifying more severe infections.

3. Progression of Infection
The progression of mastitis can be categorized into clinical mastitis and subclinical
mastitis based on the severity of symptoms.

 Clinical Mastitis:
This type is characterized by visible signs of infection. The udder becomes
swollen, hot, and painful, and the milk may appear discolored or contain pus, blood,
or clots. In more severe cases, the cow may exhibit fever, reduced milk yield, and
general malaise. Clinical mastitis is usually the result of more aggressive bacterial
strains and can quickly progress to severe forms of infection if not treated.

 Subclinical Mastitis:
In this type, there are no visible changes in the udder or milk, but there are
internal changes. The somatic cell count increases, which indicates that the immune
system is fighting off the infection. Subclinical mastitis is often harder to detect, as
the cow does not show obvious signs. However, it can still lead to reduced milk
quality and yield over time.
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4. Recovery
Spontaneous Healing:
In some cases of mild mastitis, particularly those caused by environmental
bacteria, the cow’s immune response is sufficient to resolve the infection without the
need for antibiotics or other interventions.

Treatment and Veterinary Intervention:

Antibiotics, anti-inflammatory drugs, and proper milking practices can help
accelerate recovery. However, some cows may require long-term treatment, especially
if the infection is chronic. In some instances, repeated or systemic antibiotic therapy
may be necessary to clear the infection.

UNIT 2

MORPHOLOGY AND CLASSIFICATION OF DAIRY BACTERIA: LACTIC ACID

BACTERIA (LAB)
Lactic acid bacteria (LAB) are a group of bacteria commonly found in dairy products,
where they play a crucial role in fermentation, milk preservation, and flavor development.
These bacteria are known for their ability to produce lactic acid as the major metabolic
product from carbohydrates, particularly lactose, and they are essential in the production of
various dairy products like yogurt, cheese, and kefir. LAB also contribute to the preservation
of dairy products by lowering the pH, which inhibits the growth of spoilage organisms and
pathogens.

Morphology of Lactic Acid Bacteria (LAB)

Lactic acid bacteria are typically Gram-positive, non-spore-forming, and anaerobic or
facultative anaerobic microorganisms. Their morphology can vary depending on the species,
but some common characteristics include:
1. Shape:
LAB can exhibit a wide range of shapes:
o Cocci (spherical): These are round-shaped bacteria, often seen in pairs or chains.
Examples include Lactococcus and Enterococcus species.
o Rods (bacilli): These are elongated, rod-shaped bacteria, such as Lactobacillus and
Pediococcus species.
o Branching filaments: In some species like Bifidobacterium, the bacteria form
branched structures.

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 2. Size:
LAB cells are generally small, ranging from 0.5 to 1 micrometer in diameter
for cocci and up to 4-5 micrometers in length for rod-shaped bacteria.
3. Gram Staining:
Lactic acid bacteria are Gram-positive, meaning they retain the crystal violet
stain and appear purple under a microscope due to their thick peptidoglycan cell wall.
4. Oxygen Requirements:
o Anaerobic: Some LAB, such as Lactobacillus and Bifidobacterium, are obligate
anaerobes, meaning they grow only in the absence of oxygen.

o Facultative Anaerobic: Others, like Lactococcus and Streptococcus, can grow in

both the presence and absence of oxygen.

5. Catalase Activity:
LAB are catalase-negative, meaning they do not produce the enzyme catalase,
which breaks down hydrogen peroxide.
6. Endospore Formation:
LAB are generally non-spore-forming, although some species, like
Pediococcus, can form heat-resistant cells that resemble spores under certain
conditions.

Classification of Lactic Acid Bacteria (LAB)

Lactic acid bacteria are classified based on their ability to ferment carbohydrates
(mainly sugars), their morphological characteristics, and their biochemical properties. The
main families and genera of LAB in dairy are:

1. Family: Lactobacillaceae
o Genera: Lactobacillus, Pediococcus, Leuconostoc, Lactococcus,
Streptococcus
o Lactobacillus: This is one of the most important genera in dairy fermentation.
These bacteria are typically rod-shaped and are found in a wide range of
fermented dairy products. They can be classified further into different species
such as Lactobacillus acidophilus (used in yogurt production), Lactobacillus
bulgaricus (used in yogurt and cheese), and Lactobacillus casei (found in
cheese and probiotic products).

o Lactococcus: Cocci-shaped bacteria, Lactococcus species, such as

Lactococcus lactis, are used in the production of cheese and buttermilk. They
are important for milk fermentation and acid production.

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 o Streptococcus: Streptococcus thermophilus is commonly used in yogurt and
cheese production. These cocci bacteria thrive in warm environments and help
in the fermentation process by producing lactic acid.

o Leuconostoc: These bacteria are involved in the fermentation of dairy

products, but also vegetables and other foods. Leuconostoc mesenteroides is
known for its role in butter and fermented vegetable production.

2. Family: Bifidobacteriaceae
o Genera: Bifidobacterium
Bifidobacterium species, such as Bifidobacterium bifidum and Bifidobacterium lactis,
are often found in probiotic dairy products. They are typically branched rods and are
involved in the fermentation of lactose to produce lactic acid. These bacteria are
considered beneficial for gut health.

3. Family: Enterococcaceae
o Genera: Enterococcus
Enterococcus species, such as Enterococcus faecalis and Enterococcus
faecium, are commonly found in dairy products. These bacteria are resilient to harsh
conditions such as high salt concentrations and can contribute to fermentation
processes. They can also play a role in ripening some cheeses.

4. Family: Carnobacteriaceae
o Genera: Carnobacterium
While less common in dairy products, some Carnobacterium species can be
involved in fermentation processes, particularly in vacuum-packed or preserved dairy
products. They produce lactic acid and are often found in fermented foods.

Metabolism of Lactic Acid Bacteria

Lactic acid bacteria are heterofermentative or homofermentative, based on their fermentation
pathways:
 Homofermentative LAB: These bacteria produce mainly lactic acid from sugars like
glucose or lactose. They are the primary organisms used in dairy fermentation. For
example, Lactococcus lactis and Streptococcus thermophilus are homofermentative
and produce primarily lactic acid.
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  Heterofermentative LAB: These bacteria produce not only lactic acid but also other
by-products such as carbon dioxide and ethanol, depending on the fermentation
pathway. For example, Leuconostoc mesenteroides produces lactic acid along with
carbon dioxide, contributing to gas formation in some fermented products.

Significance of Lactic Acid Bacteria in Dairy

1. Fermentation:
LAB are essential for the fermentation of milk and other dairy products. They
convert lactose (milk sugar) into lactic acid, which lowers the pH of the milk, aiding
in coagulation and flavor development. This is crucial for products like yogurt,
cheese, and kefir.

2. Probiotic Effects:
Certain LAB strains, such as Lactobacillus acidophilus and Bifidobacterium
bifidum, have beneficial effects on gut health, making them popular in probiotic dairy
products.
3. Preservation:
By producing lactic acid, LAB inhibit the growth of spoilage organisms and
pathogens, extending the shelf life of dairy products.

4. Flavor Development:
LAB contribute to the unique flavors and textures of fermented dairy products.
They produce organic acids, diacetyl (which imparts a buttery flavor), and other
volatile compounds that are important for the taste profile of cheese, yogurt, and other
fermented dairy items.

MORPHOLOGY AND CLASSIFICATION OF DAIRY BACTERIA:

LACTOCOCCUS
Lactococcus is a genus of lactic acid bacteria (LAB) that plays a crucial role in dairy
fermentation. These bacteria are widely used in the production of fermented dairy products
like cheese, buttermilk, and sour cream. The genus Lactococcus is essential for the dairy
industry due to its ability to ferment lactose, the primary sugar in milk, and produce lactic
acid, which helps in coagulating milk proteins and creating the characteristic texture and
flavor of dairy products.
Morphology of Lactococcus
Lactococcus species share several common morphological characteristics, which
include:

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 1. Shape:
o Lactococcus bacteria are typically cocci (spherical) in shape. They are usually
small, ranging from 0.5 to 1 micrometer in diameter.
o These cocci can occur in pairs or chains. When observed under a microscope,
Lactococcus typically forms short chains or diplococci (pairs of cells) rather
than large clusters, which helps differentiate it from other types of bacteria.

2. Gram Staining:
o Lactococcus species are Gram-positive, meaning they retain the crystal violet
dye during the Gram-staining procedure. This gives them a purple appearance
under the microscope. This characteristic is due to their thick peptidoglycan
layer in the cell wall.

3. Oxygen Requirements:
o Lactococcus bacteria are facultatively anaerobic, meaning they can grow
both in the presence and absence of oxygen. However, they typically grow
better under anaerobic conditions (without oxygen), which is ideal for
fermentation.

4. Catalase Activity:
o Lactococcus is catalase-negative, meaning they do not produce the enzyme
catalase, which breaks down hydrogen peroxide. This feature is used as a
distinguishing characteristic in laboratory identification.

5. Non-Spore-Forming:
o Like many other LAB, Lactococcus species are non-spore-forming, meaning
they do not form spores to survive harsh conditions. They rely on their ability
to grow in the presence of food sources like milk for survival.

6. Size and Arrangement:

o The size of Lactococcus cells ranges from 0.5 to 1 micrometer in diameter.
The cells are arranged in pairs or chains, which is a common feature of the
genus.

Classification of Lactococcus
Lactococcus is classified within the family Lactobacillaceae, which includes other
well-known LAB genera such as Lactobacillus and Streptococcus. The classification of
Lactococcus is based on its genetic, metabolic, and biochemical properties. Below is an
overview of the classification:
1. Domain:
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 o Bacteria
2. Phylum:
o Firmicutes
This phylum includes Gram-positive bacteria with a high proportion of
peptidoglycan in their cell walls.
3. Class:
o Bacilli
This class includes many bacteria used in fermentation, including LAB.
4. Order:
o Lactobacillales
This order includes various genera of LAB, which are important for dairy
fermentation.
5. Family:
o Lactobacillaceae
The family includes other LAB genera like Lactobacillus, Pediococcus,
Leuconostoc, and Streptococcus.
6. Genus:
o Lactococcus
This genus includes several species that are widely used in dairy fermentation.

Species of Lactococcus
The genus Lactococcus includes several important species, the two most commonly
used in dairy fermentation are:
1. Lactococcus lactis
o This is the most important species in dairy production, especially in the
fermentation of milk to produce products like cheese, butter, and buttermilk.
o Lactococcus lactis is subdivided into two main subspecies:
 Lactococcus lactis subsp. lactis: Used mainly for cheese making,
such as in the production of cheddar and other hard cheeses.

 Lactococcus lactis subsp. cremoris: Often used in the production of

cultured dairy products such as sour cream and yogurt.
2. Lactococcus garvieae
o This species is less commonly used in dairy fermentation but can be found in
some dairy environments and has been associated with cases of mastitis in
dairy cows.

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Metabolism of Lactococcus
Lactococcus bacteria are homofermentative, meaning they primarily produce lactic
acid from the fermentation of sugars (such as lactose, the sugar in milk). This process helps
to reduce the pH of milk, leading to the coagulation of casein, a protein in milk that forms the
curds in cheese-making.
 Fermentation Products: In addition to lactic acid, some species of Lactococcus can
also produce small amounts of other by-products, such as carbon dioxide or ethanol,
although lactic acid is the major product.

 Lactose Utilization: Lactococcus species are capable of fermenting lactose, the

primary sugar in milk, into lactic acid. This is essential for the fermentation of dairy
products and for lowering the pH to preserve milk.

Significance of Lactococcus in Dairy Products

1. Fermentation of Dairy Products:
o Lactococcus species are key players in the fermentation of dairy products.
They are used in the production of a wide range of fermented products,
including:
 Cheese: Lactococcus lactis is essential in the production of many
cheeses, including cheddar, mozzarella, and cottage cheese.
 Yogurt: Certain strains of Lactococcus are used to ferment milk into
yogurt, producing the characteristic texture and tart flavor.
 Buttermilk and Sour Cream: Lactococcus species are also used in
the production of cultured dairy products like buttermilk and sour
cream.
2. Milk Preservation:
o By producing lactic acid during fermentation, Lactococcus lowers the pH of
milk, which helps preserve the product by inhibiting the growth of spoilage
bacteria and pathogens.
3. Flavor Development:
o The fermentation process, carried out by Lactococcus, contributes to the
development of unique flavors in dairy products. The lactic acid produced
during fermentation also acts as a natural preservative.
4. Probiotic Potential:

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 o Some strains of Lactococcus are considered beneficial to human health and are
used in probiotic dairy products, although their probiotic effects are not as
well-studied as those of other LAB genera, such as Lactobacillus.
MORPHOLOGY AND CLASSIFICATION OF DAIRY BACTERIA:
STREPTOCOCCUS
Streptococcus is a genus of Gram-positive bacteria that is widely found in the environment,
including in dairy products. Streptococci play a significant role in the dairy industry,
particularly in the fermentation processes involved in the production of various dairy
products such as cheese, yogurt, and butter. They are known for their ability to ferment
lactose and produce lactic acid, which contributes to the preservation and texture
development in dairy items.

Morphology of Streptococcus
1. Shape:
o Streptococcus bacteria are cocci (spherical) in shape. These cocci often
appear in chains or pairs when viewed under a microscope. This chain
formation is a key characteristic of the genus and distinguishes them from
other bacterial genera.

o The size of individual cells ranges from 0.5 to 1 micrometer in diameter.

2. Gram Staining:
o Streptococcus species are Gram-positive, meaning they retain the crystal
violet stain used in the Gram staining process, resulting in a purple color
when viewed under a microscope. This is due to their thick peptidoglycan
layer in the cell wall.

3. Catalase Activity:
o Streptococcus species are catalase-negative, which means they do not
produce the enzyme catalase. This is an important differentiating feature, as
catalase-positive bacteria (like Staphylococcus) can break down hydrogen
peroxide, whereas Streptococcus cannot.

4. Oxygen Requirements:
o Streptococcus bacteria are facultatively anaerobic, meaning they can grow
both in the presence and absence of oxygen. However, they prefer
environments with low oxygen for optimal growth, making them well-suited
for fermentation processes in dairy production.

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 5. Non-Spore-Forming:
o Streptococcus species do not form spores, which means they are more
vulnerable to environmental stress than spore-forming bacteria. However, they
can survive in various dairy environments as they are adapted to thrive in
nutrient-rich conditions such as milk.

Classification of Streptococcus
The genus Streptococcus is classified within the family Streptococcaceae, which is
part of the order Lactobacillales. The classification of Streptococcus is based on genetic,
biochemical, and metabolic characteristics, including their ability to ferment different sugars
and their hemolytic activity. The major species of Streptococcus found in dairy include:

1. Domain:
o Bacteria
2. Phylum:
o Firmicutes
This phylum includes Gram-positive bacteria with a thick peptidoglycan cell
wall.
3. Class:
o Bacilli
A class that includes various lactic acid bacteria, including Streptococcus.
4. Order:
o Lactobacillales
This order includes several important genera involved in dairy fermentation,
including Lactobacillus, Lactococcus, and Streptococcus.
5. Family:
o Streptococcaceae
This family includes all the species of Streptococcus.
6. Genus:
o Streptococcus

Important Species of Streptococcus in Dairy Production

Several species of Streptococcus are important in dairy production due to their ability
to ferment milk and contribute to the production of dairy products. Some of the key species
include:
1. Streptococcus thermophilus

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 o This is one of the most important species of Streptococcus in the dairy
industry, particularly for yogurt and cheese production.

o Streptococcus thermophilus is a thermophilic bacterium, meaning it thrives at

higher temperatures, typically around 42-45°C, which makes it ideal for
yogurt fermentation. It is often used in combination with Lactobacillus
bulgaricus to ferment milk and produce yogurt.

o It ferments lactose to produce lactic acid, which lowers the pH of milk,

causing it to coagulate and form yogurt. It also contributes to the development
of the characteristic flavor and texture of yogurt.

2. Streptococcus lactis
o Streptococcus lactis is used in the production of cheese, particularly in the
early stages of fermentation.

o It is involved in acid production and contributes to the curd formation process

by converting lactose into lactic acid.

3. Streptococcus cremoris
o This species is commonly used in cheese production, especially in the
production of cheddar and other semi-soft cheeses.

o Streptococcus cremoris is known for its ability to produce lactic acid quickly,
which helps in curdling milk and starting the cheese-making process.

4. Streptococcus salivarius
o While this species is primarily associated with the human oral cavity, certain
strains of S. salivarius have been used in the production of fermented dairy
products such as probiotic yogurt.

Metabolism of Streptococcus
Streptococcus species are homofermentative, meaning they primarily produce lactic
acid from the fermentation of carbohydrates, particularly lactose. This production of lactic
acid is central to their role in dairy fermentation, as it lowers the pH of milk, which helps to
coagulate the proteins and form curds, essential for cheese-making and yogurt production.

1. Fermentation Pathways:
o Streptococcus species primarily rely on homofermentative fermentation,
where they convert sugars like lactose into lactic acid, with very little or no

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 production of other by-products. This is different from heterofermentative
LAB, which can produce other by-products such as ethanol or carbon dioxide.

2. By-products of Fermentation:
o The main by-product of Streptococcus fermentation is lactic acid, which
contributes to the acidity and preservation of fermented dairy products.
o In some cases, certain Streptococcus species may also produce small amounts
of other volatile compounds that influence the flavor profile of fermented
products.

Significance of Streptococcus in Dairy Products

1. Fermentation of Dairy Products:
o Streptococcus species, particularly Streptococcus thermophilus, are essential
for the fermentation of dairy products. They play a key role in converting milk
into products like yogurt, cheese, and buttermilk.
o Streptococcus thermophilus is typically used in combination with other LAB
like Lactobacillus bulgaricus in the production of yogurt.

2. Milk Preservation:
o The production of lactic acid by Streptococcus lowers the pH of the milk,
which helps to preserve the product by creating an acidic environment that
inhibits the growth of spoilage microorganisms and pathogens.

3. Flavor Development:
o Streptococcus species contribute to the development of characteristic flavors
in dairy products. In yogurt and cheese, they are involved in the production of
lactic acid, which gives the product its tart flavor, and other compounds that
contribute to the final taste and texture.

4. Probiotic Potential:
o Some species of Streptococcus, like Streptococcus thermophilus, are used in
probiotic dairy products. These strains are beneficial for gut health as they can
survive in the gastrointestinal tract and may provide various health benefits,
such as improving digestion and supporting the immune system.

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 MORPHOLOGY AND CLASSIFICATION OF DAIRY BACTERIA:
LACTOBACILLUS
Lactobacillus is one of the most important genera of lactic acid bacteria (LAB),
playing a central role in the fermentation of dairy products. These bacteria are widely used in
the production of yogurt, cheese, sauerkraut, pickles, and other fermented foods. Lactobacilli
are known for their ability to ferment sugars, such as lactose, into lactic acid, which lowers
the pH of the food, preserving it and contributing to the development of characteristic flavors.

Morphology of Lactobacillus
1. Shape:
o Lactobacillus bacteria are rod-shaped (bacilli), and their size typically ranges from
0.5 to 1 micrometer in diameter and 2 to 9 micrometers in length. The rods can
appear as straight or slightly curved, depending on the species.
o They may occur as single cells, pairs, or chains, though chains are less common than
with other LAB genera, such as Streptococcus.

2. Gram Staining:
o Lactobacillus species are Gram-positive, meaning they retain the crystal violet dye
during the Gram staining process, which gives them a purple appearance. This is due
to their thick peptidoglycan layer in the cell wall, a characteristic of Gram-positive
bacteria.

3. Catalase Activity:
o Lactobacillus species are catalase-negative, meaning they do not produce the
enzyme catalase, which breaks down hydrogen peroxide. This is an important
characteristic used in differentiating them from other Gram-positive bacteria like
Staphylococcus.

4. Oxygen Requirements:
o Lactobacillus species are facultatively anaerobic, meaning they can grow in both the
presence and absence of oxygen. However, they typically grow better in low-oxygen
or anaerobic environments, which is ideal for their role in fermentation processes.
o Some species of Lactobacillus are aerotolerant anaerobes, meaning they can
tolerate oxygen but do not use it for growth.

5. Non-Spore-Forming:
o Lactobacillus is non-spore-forming, meaning they do not form spores to withstand
extreme environmental conditions. They depend on the availability of nutrients for
survival and reproduction, particularly in dairy products.

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Classification of Lactobacillus
The genus Lactobacillus belongs to the family Lactobacillaceae, which is part of the
order Lactobacillales. The classification of Lactobacillus is based on various factors,
including its metabolic properties, ability to ferment sugars, and the production of lactic acid.
1. Domain:
o Bacteria

2. Phylum:
o Firmicutes
The phylum Firmicutes includes Gram-positive bacteria with a high content of
peptidoglycan in their cell walls.

3. Class:
o Bacilli
This class includes rod-shaped bacteria, many of which are involved in fermentation,
including Lactobacillus.

4. Order:
o Lactobacillales
This order includes several genera of LAB, including Lactobacillus, Streptococcus,
and Lactococcus.

5. Family:
o Lactobacillaceae
This family includes the genus Lactobacillus and other LAB that are important in
fermentation.

6. Genus:
o Lactobacillus

Important Species of Lactobacillus in Dairy Production

Several species of Lactobacillus are widely used in dairy fermentation due to their
ability to convert lactose into lactic acid, a process essential for the production of dairy
products such as yogurt, cheese, and kefir. Some key species include:
1. Lactobacillus bulgaricus
o Lactobacillus bulgaricus is one of the primary bacteria used in the production of
yogurt. It is used in combination with Streptococcus thermophilus to ferment milk.

o This species is thermophilic, meaning it thrives at higher temperatures, typically

between 42-45°C. It ferments lactose to produce lactic acid, which helps in the
coagulation of milk proteins and the development of the yogurt’s texture and flavor.

2. Lactobacillus acidophilus
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 o Lactobacillus acidophilus is commonly found in probiotic dairy products, such as
yogurt and fermented milk drinks.

o This species is considered a probiotic because it can survive in the human

gastrointestinal tract and may provide health benefits, including improving digestion
and supporting immune function.

o Like other Lactobacillus species, it ferments lactose into lactic acid, lowering the pH
of the milk and contributing to the flavor and preservation of the dairy product.

3. Lactobacillus casei
o Lactobacillus casei is used in the production of cheese, particularly in the ripening
process of cheeses like cheddar, gouda, and Swiss. It contributes to the development
of texture and flavor during the aging process.

o It is capable of fermenting a wide range of sugars, including lactose, and can tolerate
a relatively broad range of pH and temperature conditions.

4. Lactobacillus helveticus
o Lactobacillus helveticus is involved in the production of Swiss cheese, Parmesan,
and other aged cheeses.

o This species produces lactic acid and other metabolites that contribute to the
characteristic flavor and texture of aged cheeses. It also plays a role in the breakdown
of milk proteins during the ripening process.
Metabolism of Lactobacillus
Lactobacillus species are primarily homofermentative, meaning they produce lactic
acid as the main end-product from the fermentation of carbohydrates, particularly lactose.
This production of lactic acid is essential for the fermentation of dairy products, as it lowers
the pH and causes the milk proteins to coagulate, leading to the formation of curds in cheese
and the thickening of milk in yogurt.
 Fermentation Pathways: Lactobacillus species mostly follow the
homofermentative pathway, where they convert sugars like lactose directly into
lactic acid. Some species can also follow heterofermentative pathways, where they
produce other by-products such as carbon dioxide or ethanol, although this is less
common in dairy-related species.
 By-products of Fermentation: The primary by-product of Lactobacillus
fermentation is lactic acid. This acidification of milk helps in the preservation of dairy
products and is key to their flavor. In some cases, other volatile compounds like
acetaldehyde or diacetyl may also contribute to the flavor profile of the product.

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Significance of Lactobacillus in Dairy Products
1. Fermentation of Dairy Products:
o Lactobacillus species are integral to the production of several dairy products. They
are particularly important in the production of yogurt, cheese, kefir, and other
fermented milk products.

o In yogurt production, Lactobacillus bulgaricus works in symbiosis with

Streptococcus thermophilus to ferment milk and produce the characteristic tangy
flavor and texture.

2. Milk Preservation:
o Through the production of lactic acid, Lactobacillus helps to lower the pH of milk,
which prevents the growth of spoilage bacteria and pathogens, thus extending the
shelf life of dairy products.

3. Flavor Development:
o The lactic acid produced during fermentation by Lactobacillus contributes to the tart
and slightly sour flavor of products like yogurt and kefir. In cheese production,
Lactobacillus species help in the breakdown of proteins and fats during ripening,
which enhances the complexity of the flavor.

4. Probiotic Potential:
o Certain species of Lactobacillus, such as Lactobacillus acidophilus, are considered
probiotics due to their beneficial effects on human health. They can survive in the
digestive tract, improve gut health, and boost the immune system, making them
valuable in the production of functional foods.

MORPHOLOGY AND CLASSIFICATION OF DAIRY BACTERIA:

BIFIDOBACTERIUM
Bifidobacterium is a genus of Gram-positive, anaerobic bacteria commonly found in
the gastrointestinal tract of humans and animals, as well as in fermented dairy products.
These bacteria are especially known for their beneficial role as probiotics, contributing to gut
health and digestion. Bifidobacteria are also important in the dairy industry for their
involvement in the fermentation of dairy products, such as yogurt and other probiotic-rich
foods.

Morphology of Bifidobacterium
1. Shape:
o Bifidobacterium species are Gram-positive, Y-shaped or branching rod-
shaped bacteria. The characteristic bifurcated or "Y" shape distinguishes them
from other rod-shaped bacteria.
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 o They can also appear as curved or branched rods, giving them a distinct
appearance when observed under the microscope.

2. Size:
o The size of Bifidobacterium cells generally ranges from 0.5 to 1.2
micrometers in diameter and 1 to 5 micrometers in length, depending on the
species and growth conditions.

3. Gram Staining:
o Bifidobacterium species are Gram-positive, meaning they retain the crystal
violet stain and appear purple under a microscope due to their thick
peptidoglycan cell wall.

4. Catalase Activity:
o These bacteria are catalase-negative, meaning they do not produce the
enzyme catalase, which breaks down hydrogen peroxide. This trait is similar
to other lactic acid bacteria (LAB) such as Lactobacillus and Streptococcus.

5. Oxygen Requirements:
o Bifidobacterium species are strict anaerobes, meaning they thrive in
environments that lack oxygen. They can survive in the absence of oxygen but
do not grow in the presence of air.
o Some species are also microaerophilic, meaning they can tolerate very low
oxygen concentrations.

6. Non-Spore-Forming:
o Bifidobacterium species do not produce spores, which makes them more
sensitive to environmental stress compared to spore-forming bacteria.
However, they are quite resilient in the right environments, particularly in the
human gut and dairy products.

Classification of Bifidobacterium
The genus Bifidobacterium belongs to the family Bifidobacteriaceae, which is part
of the order Bifidobacteriales. The classification of Bifidobacterium is based on genetic,
biochemical, and morphological characteristics. Here is the taxonomic classification:
1. Domain:
o Bacteria
2. Phylum:

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 o Actinobacteria
Bifidobacterium belongs to this phylum, which is a group of Gram-positive
bacteria known for their high G+C content in DNA.
3. Class:
o Actinobacteria
This class includes a variety of important bacteria, including many that are
involved in human health and fermentation.
4. Order:
o Bifidobacteriales
This order includes only a few genera, with Bifidobacterium being the most
well-known and significant in dairy products and probiotics.
5. Family:
o Bifidobacteriaceae
This family includes Bifidobacterium and other closely related genera.
6. Genus:
o Bifidobacterium

Important Species of Bifidobacterium in Dairy Production

Several species of Bifidobacterium are used in the dairy industry, particularly in the
production of probiotic dairy products such as yogurt, kefir, and fermented milk. Some
notable species include:
1. Bifidobacterium bifidum
o This species is one of the most common Bifidobacterium species found in
fermented dairy products. It is often used as a probiotic in yogurt and other
dairy products due to its beneficial effects on human gut health.

o Bifidobacterium bifidum is believed to support the digestive system, enhance

immune function, and prevent the growth of harmful bacteria in the intestines.

2. Bifidobacterium animalis
o Bifidobacterium animalis is another important species in the dairy industry,
frequently used in probiotic yogurts and fermented milk drinks.

o This species is associated with improving gut health, enhancing the immune
response, and reducing gastrointestinal disorders such as constipation and
irritable bowel syndrome (IBS).

3. Bifidobacterium lactis

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 o Bifidobacterium lactis is a well-known species used in the fermentation of
dairy products such as yogurt, particularly in products marketed as "probiotic"
due to its health-promoting properties.

o Bifidobacterium lactis is used to enhance digestive health, improve the

absorption of nutrients, and boost the immune system. It is often added to
infant formulas and other probiotic-rich foods.

4. Bifidobacterium breve
o This species is involved in the fermentation of certain types of dairy,
especially in products aimed at promoting gut health.

o Bifidobacterium breve can help balance the intestinal flora and has been
shown to support the immune system and prevent gastrointestinal diseases.

Metabolism of Bifidobacterium
Bifidobacterium species are primarily homofermentative lactic acid bacteria,
meaning they primarily produce lactic acid from the fermentation of carbohydrates, such as
lactose. However, they can also produce small amounts of acetic acid and other by-products,
which can contribute to the flavor of fermented dairy products.
1. Fermentation Pathways:
o Bifidobacterium species primarily ferment sugars like lactose, galactose, and
fructose, converting them into lactic acid. In addition to lactic acid, some
species can produce acetic acid, which also plays a role in maintaining the
acidic environment of dairy products and contributing to their preservation.

2. By-products of Fermentation:
o The primary by-product of Bifidobacterium fermentation is lactic acid, which
helps acidify the product and preserves it. In some cases, Bifidobacterium also
produces acetic acid, which further lowers the pH and inhibits the growth of
harmful microbes.

o Some species of Bifidobacterium may also produce beneficial compounds,

such as bacteriocins, which can inhibit the growth of pathogenic bacteria.

Significance of Bifidobacterium in Dairy Products

1. Probiotic Functionality:
o Bifidobacterium species are widely recognized for their probiotic properties.
They are believed to support gut health by promoting the growth of beneficial
bacteria and inhibiting the growth of harmful microbes in the digestive tract.
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 o These bacteria can help in the fermentation of lactose in individuals who are
lactose intolerant, contributing to the production of dairy products that are
easier to digest.

2. Fermentation of Dairy Products:

o Bifidobacterium species play an important role in the fermentation of dairy
products like yogurt, kefir, and certain cheeses. Their fermentation of lactose
helps in the production of lactic acid, which causes milk to coagulate and form
the characteristic texture of yogurt and other fermented dairy products.

3. Health Benefits:
o Bifidobacterium has been shown to improve digestive health by regulating the
intestinal flora. It is also linked to immune system support and the prevention
of gastrointestinal disorders such as diarrhea, constipation, and irritable bowel
syndrome (IBS).

o Some studies suggest that Bifidobacterium may also have a role in preventing
infections and enhancing overall gut function, making it a valuable ingredient
in functional foods.

4. Milk Preservation:
o Through the production of lactic acid, Bifidobacterium helps to preserve dairy
products by lowering their pH, thus preventing the growth of spoilage
microorganisms and extending shelf life.

ROLE OF PSYCHROTROPHIC BACTERIA IN SPOILAGE OF MILK

Psychrotrophic bacteria can grow at refrigeration temperatures (typically 4°C) but
may grow more slowly than mesophilic or thermophilic bacteria. They are capable of
multiplying in the cool storage conditions typical of milk and dairy products. Psychrotrophic
bacteria are often capable of producing extracellular enzymes such as proteases (which
break down proteins) and lipases (which break down fats). These enzymes are key
contributors to the spoilage of milk and other dairy products.
1. Enzyme Production and Its Impact on Milk Spoilage
Psychrotrophic bacteria are notorious for producing various extracellular enzymes,
such as lipases (enzymes that break down fats) and proteases (enzymes that break down
proteins). These enzymes have a significant role in milk spoilage and contribute to the
following effects:

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a. Breakdown of Milk Proteins (Proteolysis)
 Proteases produced by psychrotrophic bacteria break down milk proteins, particularly
casein, which is the primary protein in milk.

 The breakdown of casein leads to the formation of peptides and amino acids. These
by-products are often bitter or off-flavored, contributing to the undesirable taste of
spoiled milk.

 In addition to taste changes, the breakdown of proteins can cause milk to become
more acidic. This acidification leads to coagulation or curdling of the milk, altering
its texture and making it less desirable for consumption.

 Peptides and amino acids can also act as nutrients for other microorganisms,
facilitating further microbial growth and accelerating the spoilage process.

b. Breakdown of Milk Fats (Lipolysis)

 Lipases produced by psychrotrophic bacteria target the fat molecules in milk,
breaking them down into free fatty acids and glycerol.

 The release of free fatty acids causes the milk to develop a rancid or soapy taste.
This rancidity is often considered one of the most noticeable signs of milk spoilage.

 Lipolysis not only impacts the flavor of milk but also changes its texture. As fats are
broken down, the consistency of milk may become greasy or slimy, further
contributing to the loss of product quality.

2. Sourness and Acidification

Psychrotrophic bacteria are often capable of fermenting lactose, the main sugar in
milk, into lactic acid. The production of lactic acid causes several spoilage effects:
a. Souring of Milk
 As the bacteria ferment lactose, they produce lactic acid as a byproduct. The
acidification of milk causes a sharp, sour taste, which is an obvious sign of
spoilage.

 Although lactic acid production is desirable in controlled dairy fermentation (e.g., for
yogurt or cheese), uncontrolled acidification by psychrotrophic bacteria results in an
unpleasant, overly sour flavor.

 The souring process also leads to a decrease in milk’s pH, causing the proteins to
denature and coagulate, leading to curd formation or separation of curds and whey.

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 This texture change is undesirable for fresh milk but may be acceptable in certain
fermented dairy products when controlled.

3. Off-Flavors and Odors

Psychrotrophic bacteria contribute to the development of off-flavors and off-odors in
milk, which result from their metabolic activities:
a. Bitter and Rancid Tastes
 As mentioned, proteases and lipases produced by psychrotrophic bacteria break
down proteins and fats, respectively, into bitter peptides and free fatty acids. These
components contribute to the bitter and rancid flavors in spoiled milk.

 In addition, the release of volatile fatty acids and aldehydes contributes to a strong,
unpleasant odor, often described as sour or putrid, which makes milk unappetizing.

b. Other Unpleasant Odors

 Certain psychrotrophic bacteria, such as Pseudomonas species, are capable of
producing volatile sulfur compounds (such as hydrogen sulfide), which contribute
to a rotten egg smell in milk.

 The development of these off-odors is often a sign that psychrotrophic bacteria have
grown to high numbers and have significantly degraded the quality of the milk.

4. Impact on Shelf Life

The activity of psychrotrophic bacteria significantly reduces the shelf life of milk.
Even when milk is refrigerated, these bacteria can multiply slowly, leading to early spoilage.
The following points summarize their impact on shelf life:
a. Continued Growth at Low Temperatures
 Unlike mesophilic bacteria (which prefer warmer temperatures), psychrotrophic
bacteria can continue to multiply at refrigeration temperatures (4°C), allowing them to
spoil milk even under controlled conditions.

 Even though refrigeration slows bacterial growth, it does not completely stop it.
Therefore, the presence of psychrotrophic bacteria in milk can still result in spoilage
within a short period, particularly if the milk has been improperly handled, stored, or
exposed to contamination.

b. Shorter Storage Period

 Milk and other dairy products that are not handled or stored properly (e.g., not kept at
the correct temperature) are more susceptible to spoilage by psychrotrophic bacteria.

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 These bacteria can rapidly reduce the milk’s shelf life, making it unsuitable for
consumption.

 Retailers and consumers often face losses due to milk spoiling prematurely, even
though it is stored in refrigerators or chillers.

5. Contribution to Milk Texture Changes

The breakdown of milk proteins and fats by psychrotrophic bacteria also leads to
texture changes in the milk. This includes:
a. Curdling or Coagulation
 As proteolysis (protein breakdown) occurs, the proteins in milk, especially casein,
begin to denature and form clumps or curds. This results in curdling or coagulation,
which is an undesirable change in the texture of milk, particularly for fresh milk.

 This is often accompanied by whey separation, which further contributes to a poor

texture and makes the milk appear grainy or watery.

b. Sliminess
 Some psychrotrophic bacteria, particularly those in the Pseudomonas group, can
produce exopolysaccharides (EPS), leading to the formation of slimy or viscous
milk. This slime formation can be a clear indication of microbial contamination and
spoilage.

ROLE OF MESOPHILIC BACTERIA IN SPOILAGE OF MILK

Mesophilic bacteria are microorganisms that thrive in moderate temperatures,
typically between 20°C to 45°C. In the context of milk, they can cause spoilage under both
ambient and refrigerated conditions if the milk is not properly handled. These bacteria are
commonly found in milk during the milking process, storage, and transportation stages.
Unlike psychrotrophic bacteria, which grow best at lower temperatures, mesophilic bacteria
tend to grow more rapidly at room temperature or slightly above.
These bacteria play a significant role in milk spoilage, especially when milk is
improperly stored or handled, leading to various forms of deterioration such as souring,
curdling, and texture changes. Mesophilic bacteria can contribute to both uncontrolled
acidification of milk and the breakdown of key milk components, causing undesirable flavors
and reducing the milk's shelf life.

1. Souring of Milk
One of the most common forms of spoilage caused by mesophilic bacteria in milk is
souring, which is the result of excessive lactic acid production.

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Mechanism of Souring:
 Mesophilic lactic acid bacteria (LAB), such as Lactococcus, Lactobacillus, and
Streptococcus species, consume lactose (the primary sugar in milk) and convert it
into lactic acid.

 The accumulation of lactic acid leads to a decrease in pH, making the milk
increasingly acidic.

 As the pH drops, milk proteins (especially casein) begin to denature, causing the
milk to curdle or form lumps. This is especially common when milk is stored at
room temperature for extended periods.

The resulting sour taste is an obvious sign of spoilage. This souring is more pronounced
when milk is stored improperly at temperatures that favor mesophilic bacterial growth.

2. Curdling and Coagulation

Mesophilic bacteria are also responsible for curdling or coagulation of milk, a
process that occurs due to the acidification caused by the bacterial breakdown of lactose.
Mechanism of Curdling:
 As lactic acid accumulates in the milk, the pH level drops. This acidic environment
causes the casein proteins in milk to lose their solubility and form curds.

 In the presence of certain mesophilic bacteria, the production of proteases (enzymes

that break down proteins) can enhance the breakdown of casein, leading to
coagulation or the formation of curds, resulting in a clumpy texture.

While curdling is a natural part of dairy fermentation (e.g., in cheese production),

uncontrolled coagulation by mesophilic bacteria in fresh milk is undesirable, as it indicates
spoilage. This is particularly a problem in milk that has been improperly stored at ambient
temperatures for too long.

3. Off-Flavors and Odors

The metabolic activities of mesophilic bacteria can lead to the production of
unpleasant flavors and odors in milk, which are a result of their breakdown of milk
proteins, fats, and sugars.
a. Production of Off-Flavors:
 The proteolysis (breakdown of proteins) by mesophilic bacteria releases amino acids
and peptides, some of which can have a bitter or fishy taste.

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  The breakdown of milk fats by mesophilic lipases (fat-degrading enzymes) can result
in the production of free fatty acids, leading to a rancid or soapy taste.
b. Production of Unpleasant Odors:
 In some cases, mesophilic bacteria such as Enterobacter and Clostridium species
produce volatile sulfur compounds (such as hydrogen sulfide), which give milk a
rotten egg smell.

 The accumulation of these metabolites further reduces the milk's quality, making it
unappealing for consumption.
4. Gas Production
Certain mesophilic bacteria, especially Enterobacter and Clostridium species, can
ferment milk sugars and produce gases such as carbon dioxide and hydrogen. The presence
of these gases leads to several negative effects:
a. Swelling or Bulging of Containers:
 Gas production causes swelling or bulging of milk containers, especially in sealed
packaging. This is a common indicator of spoilage due to bacterial fermentation.

 In extreme cases, the production of gas can lead to rupture of milk containers or
cause an unpleasant foamy texture.

b. Off-Flavor Production:
 The accumulation of gases in milk can also contribute to the development of off-
flavors, as some gases (e.g., hydrogen sulfide) are foul-smelling.

5. Reduced Shelf Life

Mesophilic bacteria are one of the main contributors to the shortening of milk's shelf
life. Their ability to grow rapidly at room temperature or slightly higher temperatures means
that unrefrigerated milk will spoil much faster. This is especially a concern when milk is
stored improperly or handled incorrectly during transportation or retail.
a. Rapid Growth at Ambient Temperatures:
 Mesophilic bacteria grow best at temperatures ranging from 20°C to 45°C. If milk is
left at room temperature for extended periods, these bacteria can multiply quickly,
leading to rapid spoilage.

 In milk that is improperly stored at higher temperatures, mesophilic bacteria can also
outcompete other microbes, causing an accelerated spoilage process.

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b. Spoilage in Processed Milk:
 Even pasteurized milk, which is typically heated to kill harmful bacteria, can still be
susceptible to contamination from mesophilic bacteria if it is not stored properly after
processing.

 Contamination may occur during packaging or through exposure to contaminated

equipment, leading to faster spoilage and shortened shelf life.

6. Contribution to Milk Texture Changes

Mesophilic bacteria contribute to texture changes in milk, primarily through the
breakdown of proteins and fats:
a. Sliminess and Viscosity:
 Certain mesophilic bacteria, such as Enterobacter and Lactobacillus, produce
exopolysaccharides (EPS), which are long-chain sugar molecules. These EPS can
make milk appear slimy or viscous, which is a clear sign of bacterial contamination.

 The increase in viscosity makes the milk feel thicker and less desirable to drink, often
leading to the milk being discarded.

ROLE OF THERMOPHILIC BACTERIA IN SPOILAGE OF MILK

Thermophilic bacteria are microorganisms that thrive at high temperatures, typically

between 45°C to 75°C, and sometimes even higher. These bacteria are particularly
concerning in milk and dairy products because they can survive not only at ambient
temperatures but also at pasteurization temperatures (around 60°C to 72°C). While
pasteurization is designed to kill most harmful bacteria, some thermophilic bacteria are
heat-resistant and can cause spoilage in milk, especially if it is improperly stored or handled
after pasteurization.

Thermophilic bacteria contribute to spoilage by producing enzymes that break down

proteins, fats, and sugars in milk, leading to changes in taste, texture, and overall quality.
They can also lead to off-flavors, curdling, and gas production in milk. Understanding the
role of thermophilic bacteria in spoilage is crucial for the dairy industry to improve milk shelf
life and maintain product quality.

1. Heat Resistance and Spoilage

Thermophilic bacteria are able to survive pasteurization due to their heat-resistant
spores or their ability to endure the high temperatures used in the pasteurization process.

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a. Surviving Pasteurization:
 Pasteurization is a process used to heat milk to a specific temperature for a set period
to kill harmful bacteria and extend shelf life. However, some thermophilic bacteria
can produce spores that allow them to withstand the heat treatment.

 After pasteurization, if milk is improperly handled or stored, these heat-resistant

thermophiles can germinate and grow, causing spoilage even in milk that has been
heated.
b. Common Thermophilic Bacteria in Milk:
 Bacillus species (e.g., Bacillus stearothermophilus) and Clostridium species (e.g.,
Clostridium butyricum) are common thermophilic bacteria found in milk. They are
known to form heat-resistant spores that can survive pasteurization and grow under
favorable conditions.

2. Production of Off-Flavors
Thermophilic bacteria contribute to spoilage in milk by producing off-flavors, often
due to the breakdown of milk proteins and fats. These bacteria produce enzymes such as
proteases and lipases that break down milk's components, resulting in rancid or bitter
flavors.

a. Proteolysis and Off-Flavors:

 Proteases break down milk proteins (especially casein), producing peptides and amino
acids, some of which can result in bitter or unpleasant tastes.
 In some cases, the breakdown of proteins can lead to the production of ammonia,
which imparts an off-putting, fishy odor to the milk.

b. Lipolysis and Rancid Tastes:

 Lipases produced by thermophilic bacteria break down milk fats into free fatty acids
and glycerol. The free fatty acids, particularly butyric acid, contribute to the
development of a rancid or soapy taste in milk.

3. Curdling and Coagulation

The enzymes produced by thermophilic bacteria also contribute to the curdling and
coagulation of milk, which leads to changes in its texture and appearance.

a. Acidification and Curdling:

 Thermophilic bacteria, like Lactobacillus thermophilus and Streptococcus
thermophilus, produce lactic acid from the fermentation of lactose. This

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 acidification leads to a decrease in the pH of the milk, causing casein proteins to
denature and form curds.

 While curdling is desirable in some dairy products (e.g., cheese), in fresh milk, it is an
undesirable sign of spoilage caused by excessive acid production.

b. Protein Breakdown and Texture Changes:

 Proteolytic enzymes produced by thermophilic bacteria can also directly break down
casein, leading to coagulation or separation of the milk into solid curds and liquid
whey.
 This process results in texture changes such as the milk becoming grainy or clumpy,
which is not desirable for fresh milk consumption.

4. Gas Production
Some thermophilic bacteria are capable of fermenting sugars in milk, such as
lactose, and producing gases like carbon dioxide and hydrogen.

a. Gas Formation and Container Bulging:

 The production of gases by thermophilic bacteria leads to swelling or bulging of milk
containers, especially in sealed packaging. This is an indication that microbial
fermentation is taking place and the milk is spoiling.

 Gas production can also result in the formation of bubbles or foam in milk, altering
its texture and making it undesirable for consumption.

b. Off-Odors Associated with Gas:

 In some cases, the gases produced during fermentation by thermophilic bacteria may
contribute to unpleasant odors such as sulfur or rotten egg smells, which are
characteristic of spoilage caused by Clostridium species.

5. Impact on Shelf Life

Thermophilic bacteria contribute to the reduction of milk shelf life due to their
ability to grow and spoil milk at higher temperatures. These bacteria can cause rapid
spoilage of milk, especially if it is not stored or processed properly after pasteurization.

a. Spoilage After Pasteurization:

 Although pasteurization is designed to kill most harmful microorganisms,
thermophilic bacteria that produce heat-resistant spores can survive and proliferate,
causing spoilage even in pasteurized milk.

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  This is especially true if milk is stored at temperatures that favor the growth of these
thermophiles (e.g., at room temperature or higher).

b. Shortened Shelf Life of Dairy Products:

 Products like yogurt, cheese, and butter are particularly susceptible to thermophilic
spoilage if the milk used in their production is contaminated with thermophilic
bacteria.

 Even when stored in refrigeration, thermophilic bacteria can continue to grow, leading
to shorter shelf life for dairy products.

ROLE OF THERMODURIC BACTERIA IN SPOILAGE OF MILK

Thermoduric bacteria are microorganisms that can survive and grow in milk at
high temperatures but do not necessarily thrive at these temperatures. They are capable of
withstanding heat treatments, such as pasteurization, which are meant to kill or reduce
harmful microbial populations in milk. These bacteria can survive at temperatures above
the normal pasteurization range (typically 60°C to 72°C) but will grow optimally at lower
temperatures.

While thermoduric bacteria are less likely to cause immediate spoilage during
pasteurization, they are still responsible for spoilage during storage and post-
pasteurization if milk is not handled or stored correctly. These bacteria play a significant
role in reducing milk's shelf life and can lead to undesirable changes in the milk's taste,
texture, and overall quality.

1. Heat Resistance and Post-Pasteurization Growth

Thermoduric bacteria can survive pasteurization temperatures because they have
developed the ability to endure brief exposure to heat, unlike other bacteria that are
eliminated during pasteurization.

a. Surviving Pasteurization:
 Pasteurization is a heat treatment designed to kill most harmful bacteria in milk.
However, thermoduric bacteria can survive this process due to their ability to
withstand high temperatures.

 After pasteurization, these bacteria can remain dormant in the milk but may begin to
grow and multiply if the milk is not stored correctly, especially if it is stored at
refrigeration temperatures (around 4°C).

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b. Common Thermoduric Bacteria:
 Common thermoduric bacteria found in milk include species like Bacillus and
Clostridium (in spore form), and some lactic acid bacteria (e.g., Lactococcus,
Lactobacillus, and Streptococcus species). These can survive pasteurization but may
not necessarily multiply unless environmental conditions are conducive.

2. Contribution to Spoilage
Thermoduric bacteria contribute to spoilage by fermenting lactose and breaking
down proteins and fats, leading to off-flavors, curdling, and changes in texture. While
thermoduric bacteria are typically less aggressive in spoilage compared to psychrotrophic or
mesophilic bacteria, their long-term effects can still cause significant deterioration.

a. Acid Production and Souring:

 Some thermoduric bacteria, such as lactic acid bacteria (LAB) (Lactococcus,
Streptococcus), can ferment the lactose in milk to produce lactic acid. This
acidification leads to a lowered pH, which may cause souring of milk.

 The lowered pH also causes casein (milk proteins) to precipitate, leading to

curdling and a sour taste.

b. Breakdown of Milk Fats (Lipolysis):

 Thermoduric bacteria like Bacillus and Clostridium species can produce lipases,
enzymes that break down milk fats into free fatty acids and glycerol. These fatty
acids, particularly butyric acid, result in a rancid or soapy taste, an undesirable
feature in milk.

 The breakdown of milk fats can cause the development of off-flavors, particularly
in milk that has been stored for a long period.

c. Proteolysis and Texture Changes:

 Some thermoduric bacteria also produce proteases, enzymes that break down milk
proteins (e.g., casein), leading to a bitter or off taste. This breakdown may also
result in undesirable changes in the texture of the milk, such as graininess or a
clumpy appearance.

3. Gas Production and Container Bulging

In addition to the production of lactic acid and lipids, some thermoduric bacteria,
particularly Clostridium species, produce gas as a metabolic byproduct during the

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fermentation of sugars. This gas production can contribute to milk spoilage in the following
ways:

a. Gas Formation:
 Gas production by thermoduric bacteria leads to the formation of carbon dioxide and
sometimes hydrogen. The accumulation of these gases can cause swelling or bulging
of milk containers, especially those that are sealed. This is a common sign of spoilage.
b. Off-Odors from Gas:
 Gas production, particularly from Clostridium species, can result in unpleasant
odors, such as sulfur or rotten egg smells. These odors contribute significantly to the
unpalatability of the milk.

4. Impact on Shelf Life

Thermoduric bacteria contribute to the shortened shelf life of milk by continuing to
grow and multiply after pasteurization, especially when milk is not stored properly. Their
heat resistance allows them to survive pasteurization and persist in milk during storage,
leading to spoilage over time.

a. Post-Pasteurization Growth:
 Milk that is stored at improper temperatures (e.g., above 4°C) can allow thermoduric
bacteria to multiply. These bacteria continue to break down proteins, fats, and
sugars in milk, leading to spoilage even after pasteurization.

b. Reduced Shelf Life in Dairy Products:

 The presence of thermoduric bacteria in milk affects not only fresh milk but also
processed dairy products. In products like cheese, yogurt, and cream, thermoduric
bacteria may still lead to spoilage by producing off-flavors, curdling, and gas
production, reducing the product's overall shelf life.

EFFECT OF PROCESSING ON MICROORGANISMS IN MILK

Milk processing plays a pivotal role in determining the microbial content of the
product, with the primary objective of ensuring food safety, preserving quality, and
extending shelf life. The methods employed in milk processing—such as pasteurization,
homogenization, fermentation, ultra-high temperature (UHT) treatment, and cooling—
are designed to reduce the microbial load, control spoilage, and prevent the growth of
harmful microorganisms. However, some microorganisms may survive certain processes and

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affect the quality of the milk. Below is a detailed explanation of the effects of these common
processing techniques on the microorganisms present in milk.

1. Pasteurization
Pasteurization is a heat treatment method applied to milk to eliminate pathogens and
spoilage-causing microorganisms without altering the taste, texture, and nutritional quality of
milk. This process involves heating milk to a specific temperature for a set duration and then
cooling it rapidly to prevent microbial growth.
Effect on Microorganisms:
 Destruction of Pathogens: Pasteurization effectively destroys many harmful
microorganisms that cause foodborne illnesses, such as Salmonella, E. coli, Listeria,
and Mycobacterium tuberculosis. The treatment ensures that milk is safe for
consumption by eliminating these pathogens.

 Reduction of Spoilage Microorganisms: Besides pathogens, pasteurization also

targets spoilage-causing microorganisms like Lactobacillus and Streptococcus. These
bacteria contribute to souring, curdling, and off-flavors. However, thermoduric
bacteria (heat-resistant microorganisms like Bacillus and Clostridium) may survive
pasteurization and later cause spoilage during storage.

 Types of Pasteurization: There are different methods of pasteurization:

o High-Temperature Short-Time (HTST): In this method, milk is heated to
72°C for 15-20 seconds, which kills most spoilage organisms and pathogens.

o Ultra-Pasteurization (UP): This involves heating milk to 135°C for 2-5

seconds to eliminate nearly all microorganisms, extending the shelf life of
milk.

2. Homogenization
Homogenization is a mechanical process in which milk is forced through small
openings at high pressure to break down the fat globules, thereby creating a smooth and
uniform texture. It is usually performed after pasteurization.
Effect on Microorganisms:
 Homogenization itself does not kill microorganisms. However, it helps in improving
the texture and consistency of milk by ensuring that the fat is evenly distributed.
This process does not have a direct impact on the microbial content.

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  By breaking down fat globules into smaller particles, homogenization increases the
surface area for microbial attachment. This may potentially make the milk more
susceptible to contamination if not handled properly, as microorganisms can adhere
more easily to the milk's surface.
3. Fermentation (for Yogurt, Cheese, and Other Dairy Products)
Fermentation is the process by which specific starter cultures of microorganisms
(typically lactic acid bacteria) are added to milk to ferment lactose into lactic acid, creating
fermented dairy products like yogurt, cheese, and buttermilk.
Effect on Microorganisms:
 Dominance of Beneficial Microorganisms: During fermentation, lactic acid
bacteria (such as Lactococcus, Lactobacillus, and Streptococcus) proliferate and
produce lactic acid. The production of lactic acid lowers the pH of the milk, creating
an acidic environment that inhibits the growth of pathogens and spoilage bacteria.

 Inhibition of Pathogens: The acidification process helps to preserve the product by

reducing the growth of harmful microorganisms like E. coli and Salmonella.
Additionally, other byproducts of fermentation, such as bacteriocins, produced by
lactic acid bacteria, help further suppress the growth of undesirable organisms.

 Survival of Non-Spoiling Bacteria: Some thermophilic lactic acid bacteria, such as

Streptococcus thermophilus, can survive fermentation and contribute to the flavor and
texture of the final product.

4. Ultra-High Temperature (UHT) Treatment

Ultra-High Temperature (UHT) treatment involves heating milk to extremely high
temperatures, typically between 135°C and 150°C, for a very short period (2-5 seconds),
followed by rapid cooling. UHT is commonly used to increase the shelf life of milk.
Effect on Microorganisms:
 Destruction of Pathogens and Spoilage Microorganisms: UHT treatment
effectively eliminates almost all microorganisms, including pathogens and most
spoilage bacteria. This process ensures that the milk is safe to drink even without
refrigeration until it is opened.

 Thermoduric and Thermophilic Bacteria: Despite its effectiveness in destroying

most microorganisms, thermoduric and thermophilic bacteria (such as certain strains
of Bacillus and Clostridium) can survive UHT treatment. These heat-resistant
microorganisms may cause spoilage if milk is not stored properly after processing.

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  Extended Shelf Life: UHT milk has a significantly longer shelf life, often lasting up
to 6-9 months without refrigeration (before opening), making it ideal for distribution
in regions with limited access to refrigeration.

5. Cooling and Storage

After milk has been processed through methods like pasteurization or UHT treatment,
it is rapidly cooled to refrigeration temperature (below 4°C). Proper storage conditions are
essential to maintain the safety and quality of milk.
Effect on Microorganisms:
 Inhibiting Growth: Cooling milk quickly and storing it at 4°C or lower helps to
slow down the growth of psychrotrophic bacteria that can still be present after
pasteurization. These bacteria, although less active at cold temperatures, can still
multiply if milk is not handled or stored correctly.

 Risk of Contamination: Improper handling or contamination during storage can

lead to the growth of spoilage microorganisms. For example, if the milk is stored in
dirty containers or at temperatures above 4°C, it becomes vulnerable to the growth of
psychrotrophic or thermoduric bacteria, leading to spoilage and contamination.

6. Filtration and Separation

Filtration and separation are common methods used to remove contaminants, such as
dirt, somatic cells, and larger microorganisms from milk. Separation is also used to divide
milk into its components, such as cream and skim milk.
Effect on Microorganisms:
 Removal of Larger Contaminants: Filtration helps to remove larger contaminants,
including some bacteria and debris. However, this process is not sufficient to
eliminate all microorganisms, especially smaller bacteria that may still be present in
the milk.

 Concentration of Microorganisms in Fat: In the separation process, the cream

portion of milk may concentrate microorganisms, particularly lipolytic bacteria that
thrive in fat-rich environments. These bacteria can lead to spoilage, particularly by
producing off-flavors and rancidity in milk and cream.

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7. Milk Powdering (Spray Drying)
Milk powdering, or spray drying, is a process that involves removing the moisture
from milk to produce powdered milk. The milk is first evaporated and then sprayed into hot
air, which evaporates the remaining water, leaving dry milk particles.
Effect on Microorganisms:
 The low moisture content in milk powder limits the growth of most microorganisms
by reducing water activity. As a result, powdered milk has a much longer shelf life
compared to liquid milk.
 Some heat-resistant microorganisms, including spore-forming bacteria like
Bacillus and Clostridium, may survive the spray-drying process. However, these
bacteria are generally unable to grow in the dry environment unless the powder is
rehydrated.
 The drying process helps preserve milk for long periods, but once reconstituted with
water, the milk becomes susceptible to microbial growth if not handled properly.

Conclusion
The processing of milk through various methods like pasteurization, UHT treatment,
fermentation, and homogenization has a profound impact on the microbial content of milk.
These methods are designed to kill or suppress harmful microorganisms, improve the shelf
life of milk, and ensure safety for consumption. However, certain microorganisms, especially
thermoduric and thermophilic bacteria, may survive these processes and contribute to
spoilage if milk is not stored properly. Thus, milk handling and storage practices are equally
important to maintain milk quality and prevent spoilage. Proper cooling, hygiene, and storage
are essential to ensuring that milk remains safe, fresh, and free from microbial contamination.

UNIT – 3
Microbiological Methods of Milk Testing
Milk is a perishable product that can be contaminated with various microorganisms,
including pathogens and spoilage organisms. Testing the microbiological quality of milk is
essential for ensuring its safety, quality, and compliance with food standards. Several
microbiological methods are used to detect, enumerate, and identify these microorganisms,
which include bacteria, yeasts, molds, and spores. These tests help monitor the sanitation of
milk production processes and ensure that the milk is safe for human consumption.
1. Standard Plate Count (SPC)
The Standard Plate Count (SPC), also known as Total Viable Count (TVC), is one
of the most common methods used to assess the overall microbial load in milk. This method

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involves diluting a milk sample and plating it on a suitable agar medium to count the number
of viable microorganisms that can grow and form colonies.
Procedure:
 A sample of milk is serially diluted and spread onto agar plates (usually nutrient agar
or Plate Count Agar).
 The plates are incubated at an appropriate temperature (typically 30-37°C) for 24-48
hours.
 The colonies that form are counted and expressed as colony-forming units (CFU) per
milliliter of milk.
Significance:
 The SPC provides an indication of the general microbial quality of milk, although it
does not differentiate between types of microorganisms. It is often used to evaluate
sanitation during the milk production and handling processes.
 High SPC values indicate poor hygiene and possible contamination during milking,
storage, or transportation.
2. Coliform Count
The Coliform Count is a test used to detect the presence of coliform bacteria,
particularly Escherichia coli, which is an indicator of fecal contamination in milk. Coliform
bacteria are a group of microorganisms found in the intestines of warm-blooded animals and
are often used as an indicator of unsanitary conditions during milk production or handling.
Procedure:
 A sample of milk is inoculated into selective media (such as MacConkey agar),
which allows the growth of coliform bacteria while inhibiting other microorganisms.
 The milk is incubated at 35-37°C for 24-48 hours, and the number of colony-forming
units (CFUs) is counted.
 A positive coliform count indicates poor hygiene practices and potential
contamination with fecal material.
Significance:
 Coliforms, especially E. coli, are used as an indicator of possible pathogenic
contamination in milk, as they can cause foodborne illnesses if present in large
numbers.
 The absence of coliforms is an indication of proper hygiene and cleanliness during
milking and milk handling.

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3. Somatic Cell Count (SCC)
The Somatic Cell Count (SCC) test measures the number of somatic cells (primarily
white blood cells) in milk. A high SCC is an indicator of mastitis, an inflammation of the
udder caused by infection. The SCC is an important parameter used to assess the quality and
health of the milk and the cow's udder.
Procedure:
 Milk samples are collected from individual cows and analyzed using an electronic
counting device or microscopic methods to determine the number of somatic cells
per milliliter of milk.
 The SCC is reported as cells per milliliter (cells/mL).
Significance:
 An elevated SCC (above 200,000 cells/mL) is often a sign of mastitis and can affect
the taste, texture, and nutritional quality of milk. It also indicates the need for
veterinary treatment for the affected cow.
 Lower SCC values are desirable and indicate healthy cows and high-quality milk.

4. Detection of Pathogens (e.g., Salmonella, Listeria, Staphylococcus aureus)

The detection of specific pathogenic bacteria such as Salmonella, Listeria
monocytogenes, and Staphylococcus aureus is critical for ensuring the safety of milk and
milk products.
Procedure:

 Selective media and enrichment broths are used to isolate pathogens from milk
samples. For example:
o Salmonella can be detected using Selenite F broth for enrichment and XLD
agar for isolation.
o Listeria can be isolated using PALCAM agar.
o Staphylococcus aureus can be identified using Mannitol Salt Agar or
Baird-Parker agar.

Significance:
 The presence of pathogens like Salmonella, Listeria, and S. aureus in milk is a serious
health risk, as they can cause foodborne illnesses. These tests are crucial for ensuring
that milk is free from harmful microorganisms that can affect consumer health.

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5. Petrifilm™ Method
The Petrifilm™ method is a rapid and convenient test for detecting specific types of
bacteria in milk, such as Coliforms and Enterobacteriaceae. This method uses pre-prepared,
ready-to-use agar film plates for microbial detection.
Procedure:
 A small amount of milk is applied to the Petrifilm™ plate, which contains a special
nutrient medium that supports the growth of target microorganisms.
 The plate is incubated at the appropriate temperature, and colonies that grow are
identified based on their appearance on the film.
 The number of colonies is counted, and the results are compared to the standard.
Significance:
 The Petrifilm™ method offers rapid results compared to traditional plating
techniques. It is commonly used for routine milk quality testing and pathogen
detection.
 The simplicity and speed of the test make it ideal for quality control in dairy
production facilities.

6. Yeast and Mold Count

The Yeast and Mold Count is a method used to detect the presence of yeasts and
molds in milk, which can cause spoilage and affect the sensory properties of dairy products.
Procedure:
 A sample of milk is inoculated on a Sabouraud Dextrose Agar plate, which supports
the growth of yeasts and molds.
 The plates are incubated at 25-30°C for several days, and the number of colonies is
counted.
Significance:
 High counts of yeasts and molds indicate improper handling, contamination, or
storage conditions. These microorganisms can lead to fermentation, off-flavors, and
spoilage of milk and dairy products.
 Yeast and mold testing is particularly important for products like cream, cheese, and
yogurt.

7. Biochemical Tests (e.g., Catalase, Coagulase)

Biochemical tests are used to identify specific bacteria based on their enzymatic
activities. For instance, the catalase test detects the enzyme catalase produced by certain

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bacteria, while the coagulase test identifies Staphylococcus aureus, which produces the
enzyme coagulase.
Procedure:
 The catalase test involves adding hydrogen peroxide to a bacterial culture. If bubbles
form, the bacteria are catalase-positive (e.g., Staphylococcus species).

 The coagulase test involves mixing a bacterial culture with rabbit plasma. If the
plasma clots, the bacteria are coagulase-positive (e.g., Staphylococcus aureus).

Significance:
 These tests help in the identification of specific pathogens that can contaminate milk.
Identifying such pathogens is crucial for ensuring the safety of milk and dairy
products.

MILK-BORNE INFECTIONS
Milk is a highly nutritious substance that can become a source of infection if
contaminated with harmful microorganisms. These milk-borne infections can pose significant
health risks, particularly when milk is consumed in its raw or improperly processed form. The
pathogens responsible for milk-borne infections can include bacteria, viruses, and parasites.
Infections can result from poor hygiene, contaminated water sources, or diseases in cows, and
the infection can be transmitted to humans through the consumption of contaminated milk.
To prevent such infections, it is essential to ensure proper milk handling, sanitation, and
pasteurization.

Bacterial Milk-Borne Infections

1. Salmonella spp.
Salmonella is a major pathogen that can cause gastrointestinal infections when
transmitted through contaminated milk. This bacterium can infect cows, especially those with
poor health, and can be shed in their milk. Milk can also become contaminated if proper
sanitation and milking practices are not followed. Additionally, environmental contamination
by feces can be a significant source of Salmonella in milk.
Symptoms of Salmonella infection include severe abdominal cramps, diarrhea (which
may be bloody), nausea, and vomiting. In some cases, the infection can become more severe,
leading to septicemia or dehydration if left untreated. To reduce the risk of Salmonella
transmission, proper hygiene during milking and pasteurization of milk are critical.
2. Mycobacterium tuberculosis (Tuberculosis)
Tuberculosis (TB) caused by Mycobacterium tuberculosis can be transmitted through
the consumption of raw milk from infected cows. TB in cows is rare, but it can still occur in
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areas with poor animal health management. Although milk is not the most common source of
TB transmission, it remains a risk in regions where pasteurization is not commonly practiced.
Symptoms of bovine tuberculosis in humans include chronic cough, fever, weight
loss, and night sweats. Infected individuals can also experience respiratory distress and
pleuritic pain. TB can cause lung infection, and in severe cases, it can be fatal if not treated
promptly with antibiotics. Pasteurization of milk effectively eliminates the Mycobacterium
tuberculosis bacteria, thus preventing its transmission to humans.

3. Escherichia coli (E. coli)

Escherichia coli is a bacterium that is naturally present in the intestines of humans
and animals, but certain strains, like E. coli O157, can cause severe foodborne illness when
consumed in contaminated milk. This pathogen is particularly concerning because it can
cause hemolytic uremic syndrome (HUS), a severe condition that affects the kidneys and can
lead to kidney failure.
Milk can become contaminated with E. coli through fecal contamination of milk
during milking, storage, or transportation. Improper handling or unclean equipment used in
milking and processing can facilitate the growth and spread of this pathogen.
Symptoms of an E. coli infection include severe abdominal cramps, bloody diarrhea,
vomiting, and sometimes fever. Young children, elderly individuals, and those with
weakened immune systems are particularly vulnerable to the more serious effects of E. coli
infection. Pasteurization is an effective way to kill E. coli bacteria and reduce the risk of
infection.
4. Brucella spp. (Brucellosis)
Brucellosis, caused by Brucella bacteria, is a zoonotic disease that can be transmitted
to humans through the consumption of unpasteurized milk from infected animals, especially
cattle, goats, sheep, and buffalo. Infected animals excrete Brucella bacteria in their milk,
which can be ingested by humans if the milk is not properly treated.
Symptoms of brucellosis include fever, joint pain, fatigue, sweating, and headaches. The
disease can also cause miscarriages in pregnant women and infertility in men. Brucellosis can
lead to chronic conditions such as arthritis or endocarditis if left untreated. Pasteurization of
milk is the best method to prevent the transmission of Brucella bacteria.
5. Listeria monocytogenes
Listeria monocytogenes is a bacterium that can cause listeriosis, an infection that can
result from consuming contaminated dairy products, including milk. Listeria can thrive in
refrigerated conditions, making it a unique threat to dairy products that are not consumed
immediately. The infection can occur when raw milk from infected cows is not properly
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pasteurized or when milk comes into contact with contaminated equipment during
processing.
Symptoms of listeriosis in healthy individuals include fever, muscle aches, and
gastrointestinal symptoms. However, the infection can be more serious in vulnerable
populations such as pregnant women, newborns, elderly individuals, and those with
weakened immune systems. In pregnant women, listeriosis can lead to miscarriages,
stillbirths, or premature birth. For newborns, it can cause meningitis or sepsis. Pasteurization
effectively eliminates Listeria, making it a vital process for preventing infection.

6. Staphylococcus aureus
Staphylococcus aureus is a bacteria that is commonly associated with mastitis in
cows. This infection results in the production of milk that is contaminated with
Staphylococcus aureus, which can lead to foodborne illness if the milk is consumed without
proper pasteurization. This pathogen can survive in milk even if it is improperly stored,
making it a significant concern for the dairy industry.
Symptoms of infection from S. aureus in humans include vomiting, diarrhea, and
abdominal cramps. In more severe cases, it can lead to toxic shock syndrome or food
poisoning. To minimize the risk of contamination, milk should be pasteurized, and proper
milking and storage hygiene practices must be followed to prevent bacterial growth.

Prevention of Milk-Borne Infections

To reduce the risk of milk-borne infections, several preventive measures should be
followed:
1. Pasteurization: The process of heating milk to a specific temperature for a set period
of time kills harmful pathogens without significantly altering the taste or nutritional
value of the milk. Pasteurization is the most effective way to prevent many of the
pathogens discussed above.
2. Good Milking Practices: Hygiene is crucial during milking. Farmers should ensure
that cows' udders are clean, and milking equipment is sanitized regularly to prevent
contamination.
3. Proper Storage: Milk should be stored at cool temperatures (refrigeration at or
below 4°C) to slow down the growth of any remaining bacteria and prevent spoilage.
4. Health Monitoring of Animals: Regular veterinary checks should be conducted to
monitor for diseases like mastitis and tuberculosis, which can lead to infected milk.
5. Milk Quality Testing: Regular microbiological testing of milk helps identify
contamination and ensure the safety of milk before it reaches consumers.
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 MILK TOXIC INFECTION

Milk toxic infections occur when toxins produced by bacteria contaminate milk, either
during the milking process, improper handling, or insufficient pasteurization. Unlike
infections caused by live pathogens, which can multiply in the body after ingestion, toxic
infections result from the consumption of toxins produced by bacteria in the milk. These
toxins are capable of causing serious illnesses and in some cases can be fatal. To understand
milk toxic infections, it's crucial to examine the bacterial pathogens that produce these toxins,
the symptoms they cause, and the preventive measures required to protect milk quality and
public health.

Causes of Milk Toxic Infections

Milk can be contaminated by toxin-producing bacteria that either directly produce
toxins in the milk or enter the milk through poor hygiene practices, infection in cows, or
improper milk storage. The major bacterial pathogens responsible for milk toxic infections
include:
1. Staphylococcus aureus and Enterotoxins
Staphylococcus aureus is a gram-positive bacterium that commonly contaminates
milk through mastitis in cows. Mastitis, an inflammation of the udder, is one of the most
frequent causes of milk contamination with S. aureus. This bacterium is known to produce
enterotoxins, which are heat-stable and can survive even after the milk is pasteurized. When
milk contaminated with these toxins is consumed, it can lead to food poisoning.
 Symptoms: The enterotoxins produced by S. aureus are among the most common
causes of staphylococcal food poisoning. Symptoms of milk toxic infections caused
by S. aureus include:
o Vomiting
o Diarrhea
o Abdominal cramps
o Nausea
o In more severe cases, it can cause toxic shock syndrome, which can be life-
threatening if not treated promptly.
The heat stability of these toxins means that even pasteurization, which typically kills the
bacteria, may not fully eliminate the toxins once they have been produced. Therefore, milk
that has been contaminated with S. aureus should be handled and stored properly to prevent
toxin formation.
<

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2. Clostridium botulinum and Botulinum Toxin
Clostridium botulinum is a spore-forming, anaerobic bacterium that produces
botulinum toxin, one of the most potent neurotoxins known to humans. The spores of this
bacterium can survive in the environment and can contaminate milk if cows are fed
contaminated feed or if milk is stored improperly. Botulinum toxin can be present in dairy
products, including raw milk or fermented milk products, particularly if they are stored at low
temperatures that allow the spores to germinate and produce toxins.
 Symptoms: Botulinum toxin causes botulism, a rare but potentially fatal condition.
Symptoms of botulism from contaminated milk or dairy products include:
o Weakness
o Difficulty swallowing
o Double vision
o Muscle paralysis
o Respiratory failure
o In severe cases, death due to respiratory failure.
This disease is caused when botulinum toxin interferes with neuromuscular junctions, leading
to paralysis.
3. Escherichia coli and Shiga Toxins
Certain strains of Escherichia coli, particularly E. coli O157, can be present in fecally
contaminated milk. E. coli can enter milk if it is improperly handled or if milking equipment
is contaminated. Some strains of E. coli produce Shiga toxin, which can cause severe illness
in humans.
 Symptoms: Infection with E. coli O157 can lead to gastrointestinal illness
characterized by:
o Severe abdominal cramps
o Bloody diarrhea
o Vomiting
o Fever (in some cases)
In rare but severe cases, E. coli O157 infection can lead to hemolytic uremic syndrome
(HUS), a condition that can cause acute kidney failure and can be fatal. The Shiga toxin
produced by this strain damages the blood vessels and can lead to thrombocytopenia (low
platelet count), hemolytic anemia, and renal failure.
E. coli is easily killed by pasteurization, but if milk is consumed raw, contamination can lead
to these potentially life-threatening conditions.

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Symptoms of Milk Toxic Infections
The symptoms of milk toxic infections are usually gastrointestinal and neurological in
nature, depending on the type of toxin involved. The severity of symptoms can vary from
mild to life-threatening, and in some cases, infections can lead to long-term complications.
Common symptoms of milk toxic infections caused by bacterial toxins include:
 Gastrointestinal symptoms:
o Vomiting
o Diarrhea (often watery or bloody)
o Abdominal cramps
o Nausea
 Neurological symptoms (in cases of botulism):
o Muscle weakness
o Difficulty swallowing or breathing
o Blurred or double vision
o Respiratory failure
 Systemic symptoms:
o Fever (in some cases)
o Dehydration (due to excessive fluid loss from vomiting and diarrhea)
o Headaches
o Muscle aches
In the most severe cases, milk toxic infections can lead to death, especially if caused by
potent neurotoxins like botulinum toxin or Shiga toxin.

Prevention of Milk Toxic Infections

The best way to prevent milk toxic infections is through a combination of good
agricultural practices, proper milk handling, and pasteurization. Below are the main
preventive measures:
1. Pasteurization
Pasteurization is the most effective method to kill harmful bacteria and inactivate
toxins in milk. The process involves heating milk to a specific temperature (typically 72°C
for 15 seconds) to kill pathogenic microorganisms while preserving the nutritional value of
the milk. Pasteurization is especially important for preventing the growth of Staphylococcus
aureus, Clostridium botulinum, and E. coli.

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2. Good Milking Practices
Proper hygiene during the milking process is essential in preventing contamination of
milk. This includes:
 Cleaning the udder thoroughly before milking.
 Sanitizing milking equipment after each use.
 Using gloves and ensuring that the environment is clean and free from fecal
contamination.
3. Milk Storage and Handling
Milk should be stored at low temperatures (below 4°C) to slow down the growth of
bacteria and to prevent the formation of toxins. Milk that is stored improperly (at room
temperature or above) can quickly become a breeding ground for bacteria like
Staphylococcus aureus, which can produce enterotoxins.
4. Regular Health Monitoring of Cows
Regular veterinary checks are crucial to ensure that cows are free from mastitis and
other diseases that can result in the contamination of milk with bacteria and toxins. Cows
should be treated promptly if infected to prevent the spread of pathogens into the milk supply.
5. Testing for Pathogens
Regular microbiological testing of milk helps identify contamination with harmful
bacteria or toxins. Milk producers should test for specific pathogens such as E. coli, Listeria
monocytogenes, and Salmonella, ensuring that the milk is safe for consumption.

PATHOGENS ASSOCIATED WITH MILK AND MILK PRODUCTS AND THEIR

PRESERVATION
Milk and milk products, due to their nutrient-rich nature, provide an excellent medium
for the growth of a wide variety of microorganisms. While many of these microorganisms are
harmless, some can cause foodborne illnesses. The major pathogens associated with milk and
milk products are bacteria, viruses, and parasites that can be introduced during the milking
process, from infected cows, or during improper handling and storage. Understanding these
pathogens and the methods of preservation is crucial in maintaining milk safety and quality.

1. Pathogens Associated with Milk

a. Bacterial Pathogens
Bacteria are the most common type of pathogen found in milk and milk products.
These pathogens can cause a range of illnesses, from mild gastroenteritis to severe, life-
threatening conditions. Key bacterial pathogens include:

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i. Escherichia coli (E. coli)
Escherichia coli is a fecal indicator bacterium, which can enter milk through
contaminated water, fecal matter, or improper milking techniques. Some strains of E. coli,
such as E. coli O157, produce Shiga toxins, which can cause severe foodborne illness,
including hemolytic uremic syndrome (HUS). This can lead to kidney failure and even death,
especially in vulnerable populations such as children, the elderly, and immunocompromised
individuals.
 Symptoms: Diarrhea (often bloody), abdominal cramps, vomiting, and in severe
cases, kidney failure.
ii. Listeria monocytogenes
Listeria monocytogenes is a psychrotrophic bacterium that can grow at low temperatures
and is responsible for listeriosis, a serious infection that can lead to meningitis and
septicemia. It is commonly found in raw milk or unpasteurized dairy products. Pregnant
women, newborns, elderly, and immunocompromised individuals are particularly at risk.
 Symptoms: Fever, muscle aches, nausea, and diarrhea. In severe cases, it can cause
meningitis and miscarriage.
iii. Staphylococcus aureus
Staphylococcus aureus is known to produce enterotoxins that cause staphylococcal
food poisoning. It can contaminate milk during improper milking or mastitis in cows. The
enterotoxins produced by this bacterium are heat-stable, which means they can survive
pasteurization if present in milk before it is processed.
 Symptoms: Vomiting, diarrhea, abdominal cramps, and nausea. Symptoms typically
begin within a few hours of consuming contaminated milk.
iv. Salmonella spp.
Salmonella bacteria can be found in raw milk, dairy products, and egg products.
Salmonella infection, known as salmonellosis, results from consumption of contaminated
milk, and the bacteria often survive even in refrigerated milk if it is not handled correctly.
 Symptoms: Diarrhea, fever, abdominal cramps, and vomiting. In severe cases, it can
lead to sepsis or reactive arthritis.
v. Mycobacterium bovis
Mycobacterium bovis, the causative agent of bovine tuberculosis, can be transmitted
to humans through consumption of raw milk or unpasteurized dairy products. The bacterium
is capable of surviving in milk for a long period.
 Symptoms: Tuberculosis in humans, leading to chronic coughing, weight loss, fever,
and night sweats. It can also affect the lungs, bones, and gastrointestinal system.
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b. Viral Pathogens
Though viruses do not multiply in milk, they can still be transmitted through milk
consumption if the cow is infected. Common viral pathogens include:
i. Hepatitis A Virus
Hepatitis A can be transmitted through contaminated milk or dairy products when
proper hygiene and sanitation practices are not followed. Hepatitis A affects the liver and can
cause symptoms ranging from mild to severe.
 Symptoms: Jaundice, fatigue, abdominal pain, nausea, and vomiting.
ii. Foot-and-Mouth Disease Virus (FMDV)
Foot-and-Mouth Disease primarily affects livestock (such as cattle, sheep, and goats),
the virus can be excreted in the milk of infected animals. Though humans are not directly
affected by FMD, it can lead to economic losses in the dairy industry.

c. Parasitic Pathogens
While parasitic infections in milk are less common, certain protozoa and helminths
can be transmitted through milk, especially when cows are infected with parasites such as
Giardia or Toxoplasma.
i. Toxoplasma gondii
Toxoplasmosis is caused by the Toxoplasma gondii parasite, which can be transmitted
to humans through consumption of raw milk or unpasteurized cheese. It is particularly
dangerous for pregnant women as it can lead to miscarriage or birth defects.
 Symptoms: Flu-like symptoms, muscle aches, and in severe cases, neurological
damage in immunocompromised individuals or infants.

2. Preservation of Milk and Milk Products

The preservation of milk is critical to ensure its safety and quality. Several methods are used
to control the growth of microorganisms, ensuring that milk is free from harmful pathogens.
Common milk preservation techniques include:
a. Pasteurization
Pasteurization is the most effective and widely used method to destroy harmful
bacteria in milk. During pasteurization, milk is heated to a specific temperature for a
specified period (e.g., 72°C for 15 seconds for high-temperature short-time
pasteurization) to kill pathogenic microorganisms while preserving the flavor and nutritional
value of the milk. This process does not sterilize milk but significantly reduces the number of
viable pathogens.

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  Effectiveness: Pasteurization kills many harmful bacteria such as Salmonella, E. coli,
and Listeria, but heat-resistant toxins produced by bacteria like Staphylococcus
aureus may remain.
b. Ultrahigh Temperature (UHT) Treatment
UHT treatment involves heating milk to a temperature of 135°C for 2-5 seconds.
This process kills almost all pathogens and extends the shelf life of milk without
refrigeration, making it convenient for long-term storage. UHT milk, often packaged in
aseptic containers, can be stored at room temperature until opened.
c. Refrigeration
Refrigeration is essential for slowing down the growth of microorganisms in milk.
Milk should be stored at 4°C or lower immediately after milking. Cold storage is especially
important for raw milk or milk that has been processed to a lesser degree (such as pasteurized
milk).
d. Fermentation
Fermentation is a preservation method that involves the use of lactic acid bacteria (LAB)
to convert lactose into lactic acid, thereby lowering the pH and creating an acidic
environment that inhibits the growth of harmful bacteria. This method is used in the
production of yogurt, cheese, and other fermented dairy products.
 Lactic Acid Bacteria: Common LABs used in dairy fermentation include
Lactobacillus, Streptococcus, and Lactococcus. These bacteria acidify the product and
improve its shelf life while imparting desirable flavors.
e. Canning
Canning involves sealing milk in airtight containers and then heating it to a high
temperature to kill bacteria and other microorganisms. This process is used for evaporated
milk and condensed milk and provides long shelf life even without refrigeration.

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 UNIT – 4
STARTER CULTURES OF FERMENTED MILKS: DAHI

Dahi is a traditional fermented milk product popular in South Asia. It is made by

fermenting milk using specific starter cultures that contain lactic acid bacteria (LAB). These
bacteria are responsible for the fermentation process, which converts lactose (milk sugar) into
lactic acid, lowering the pH and giving dahi its characteristic tangy flavor and thick
consistency. The starter culture used for making dahi is essential for controlling the
fermentation process and ensuring the desired texture, flavor, and safety of the product.

Starter Cultures in Dahi Production

The starter cultures used in dahi generally consist of lactic acid bacteria, primarily species
of the genera Lactobacillus and Streptococcus. These microorganisms not only help in
fermentation but also contribute to the health benefits of dahi, including probiotics that
support gut health. The specific starter cultures used for dahi include:
1. Streptococcus thermophilus: This bacterium is often used as a primary culture in
dahi-making. It is a thermophilic bacterium, meaning it thrives at higher
temperatures (around 40°C to 45°C). Streptococcus thermophilus is responsible for
the initial acidification of milk, contributing to the characteristic tart flavor of dahi.
2. Lactobacillus bulgaricus: Another important bacterium in the dahi starter culture,
Lactobacillus bulgaricus works synergistically with Streptococcus thermophilus. It
helps in further acid production and contributes to the texture and aroma of the
product. This bacterium also plays a role in increasing the viscosity of dahi and
enhancing its probiotic properties.
3. Lactobacillus acidophilus: Although not always included in all dahi cultures,
Lactobacillus acidophilus is sometimes added to enhance the health benefits of dahi,
particularly its probiotic effects on gut health.
4. Bifidobacterium species: Bifidobacterium is another type of probiotic bacteria that
may be included in dahi starter cultures, particularly for its beneficial effects on
digestion and immune system function. It helps in the fermentation process and
contributes to the overall health benefits of the product.
Fermentation Process
To make dahi, fresh milk is heated to about 85°C to denature the proteins and then
cooled to around 45°C, which is the optimal temperature for the starter culture to grow. A
small amount of the starter culture, containing the lactic acid bacteria, is then added to the

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milk. The milk is kept at this temperature for several hours, during which the bacteria ferment
the lactose into lactic acid. As the pH drops, the milk coagulates, thickens, and forms dahi.
The fermentation time can vary, but typically, dahi takes about 6-8 hours to set at a
temperature of around 40-45°C. After this, it is cooled and stored under refrigeration to
prevent further fermentation and to maintain its texture and taste.
Importance of Starter Cultures in Dahi
 Flavor Development: The specific combination of bacteria in the starter culture is
responsible for developing the characteristic flavor of dahi, which is mildly sour but
not as sharp as yogurt.
 Texture and Consistency: The bacteria in the starter culture help to thicken the milk
and give dahi its smooth, creamy texture. The acid produced during fermentation
helps the proteins in the milk (particularly casein) to coagulate, forming the desired
consistency.
 Probiotic Properties: The lactic acid bacteria used in the starter culture are beneficial
for gut health. They help in maintaining a healthy balance of intestinal flora and may
aid in digestion. Dahi is also considered a good source of probiotics, which are
beneficial for overall digestive health.
STARTER CULTURES IN LASSI PRODUCTION

Lassi is typically made by blending yogurt or dahi with water, but the starter cultures
used in the fermentation of yogurt play a crucial role in shaping the final product. The main
microorganisms used in the production of yogurt, and consequently lassi, include:

1. Streptococcus thermophilus: This thermophilic bacterium is commonly used in the

production of yogurt and lassi. It helps in the fermentation process by converting
lactose into lactic acid, which lowers the pH of the milk. This acidification leads to
the characteristic tart flavor of yogurt, and consequently, of lassi.
2. Lactobacillus bulgaricus: This bacterium works symbiotically with Streptococcus
thermophilus to further acidify the milk and enhance the texture of the product.
Lactobacillus bulgaricus is also responsible for creating the smooth and creamy
consistency of yogurt and lassi. It contributes to the mild, tangy flavor that defines
lassi.
3. Lactobacillus acidophilus: This bacterium is sometimes added to starter cultures for
making yogurt and lassi to enhance the probiotic content. Lactobacillus acidophilus is
beneficial for gut health and digestion. It aids in the fermentation process and helps
improve the texture and health benefits of lassi.

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 4. Bifidobacterium species: Similar to Lactobacillus acidophilus, Bifidobacterium
species are probiotics that can be added to the starter cultures for their health-
promoting benefits. They support digestion and the immune system and contribute to
the overall probiotic effect of the fermented product.
5. Lactococcus lactis: Sometimes used as part of starter cultures, Lactococcus lactis is a
mesophilic bacterium that is capable of fermenting milk at moderate temperatures. It
is commonly used in the production of various fermented dairy products and may
contribute to the flavor and texture of lassi when included in the starter culture.
Fermentation Process of Lassi
The process of making lassi typically begins with preparing yogurt or curd. The steps for
making lassi are as follows:
1. Preparation of Yogurt (Dahi): Fresh milk is heated and then cooled to a temperature
of about 45°C, which is optimal for the growth of starter cultures. A small amount of
pre-existing yogurt (which contains the starter cultures) is added to the milk. The milk
is then kept at this temperature for several hours to allow fermentation. The bacteria
in the starter culture convert lactose into lactic acid, which thickens the milk and gives
it the characteristic tangy flavor of yogurt.
2. Blending with Water and Spices: Once the yogurt has fermented and set, it is mixed
with cold water to form lassi. The amount of water can vary, depending on the desired
thickness of the beverage. Additional ingredients such as salt, sugar, mint, cumin, and
fruit may be added to enhance flavor.
3. Fermentation of Lassi (Optional): In some traditional recipes, lassi is allowed to
ferment for a short period after being mixed with water and spices, particularly if the
intention is to achieve a more pronounced tartness and flavor profile.
Importance of Starter Cultures in Lassi Production
 Flavor Development: The starter cultures used in lassi production are responsible for
developing its characteristic tangy and slightly sour flavor. Streptococcus
thermophilus and Lactobacillus bulgaricus are the main contributors to the acidic
taste, while Lactobacillus acidophilus and Bifidobacterium species contribute to a
smoother, more balanced flavor and health benefits.
 Texture and Consistency: The lactic acid bacteria in the starter culture help to
coagulate the proteins in milk (casein), resulting in a thick and creamy texture. This is
important in both yogurt and lassi. While lassi is typically diluted with water, the
fermentation process still impacts the final texture, making it smooth and palatable.

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  Probiotic Properties: One of the key benefits of lassi is its probiotic content. The live
bacteria in the starter culture, particularly Lactobacillus and Bifidobacterium species,
promote digestive health by maintaining a healthy balance of gut bacteria. These
probiotics can help with lactose digestion, improve immune function, and support
overall gut health.
 Health Benefits: The starter cultures used in lassi production not only help with
flavor and texture but also contribute to the health-promoting properties of the
product. The probiotics in lassi help improve gut flora, prevent constipation, and may
reduce the risk of certain gastrointestinal diseases. Additionally, lassi is a good source
of calcium, proteins, and vitamins such as B12 and riboflavin.

STARTER CULTURES OF FERMENTED MILKS: YOGURT

Yogurt is one of the most popular fermented dairy products globally, known for its
creamy texture, tangy taste, and numerous health benefits. The transformation of milk into
yogurt is driven by the action of specific microorganisms known as starter cultures. These
cultures consist primarily of lactic acid bacteria (LAB) that ferment the milk sugar (lactose)
into lactic acid, which lowers the pH of the milk, causing it to coagulate into a thick, smooth,
and flavorful product. The quality, flavor, and probiotic content of yogurt largely depend on
the selection of these starter cultures.

Starter Cultures Used in Yogurt Production

The key microorganisms used in yogurt production are lactic acid bacteria—mainly
Streptococcus thermophilus and Lactobacillus bulgaricus. Other strains may be used to
enhance flavor, texture, and health benefits. Below are the main bacteria that make up the
starter culture for yogurt:
1. Streptococcus thermophilus:
o Streptococcus thermophilus is a thermophilic bacterium that thrives at high
temperatures (about 40°C to 45°C), making it an ideal microorganism for
yogurt fermentation. It is responsible for the initial stage of fermentation by
rapidly converting lactose into lactic acid, which helps to acidify the milk and
create the tangy taste typical of yogurt.

o This bacterium also contributes to the soft, smooth texture of yogurt, as the
acidification process allows proteins, particularly casein, to coagulate and
form a gel-like structure.

2. Lactobacillus bulgaricus:

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 o Lactobacillus bulgaricus works in synergy with Streptococcus thermophilus to
produce yogurt. This bacterium also ferments lactose to produce lactic acid but
at a slower rate than Streptococcus thermophilus. It contributes to the
development of the characteristic flavor and aroma of yogurt.

o Lactobacillus bulgaricus is also essential for the thickening process in yogurt.

It produces enzymes that help break down milk proteins, contributing to the
creamy texture and enhancing the sensory characteristics of the yogurt.

3. Lactobacillus acidophilus:
o Lactobacillus acidophilus is a probiotic bacterium that is often added to
yogurt starter cultures for its beneficial effects on gut health. This bacterium
ferments lactose to produce lactic acid and can enhance the texture of yogurt,
giving it a smoother, more creamy consistency.

o Lactobacillus acidophilus is well-known for its ability to survive in the human

digestive system and provide health benefits, such as improving digestion and
boosting immunity.

4. Bifidobacterium species:
o Bifidobacterium is another group of probiotics commonly included in yogurt
cultures to increase the product’s health benefits. These bacteria are believed
to promote gut health by maintaining the balance of good bacteria in the
intestines and may reduce symptoms of irritable bowel syndrome,
constipation, and other gastrointestinal disorders.
o Bifidobacteria contribute to the acidification of milk and may also play a role
in improving the texture and stability of yogurt.

5. Lactococcus lactis:
o Lactococcus lactis is another species used in yogurt starter cultures,
particularly in certain regions or types of yogurt. It is a mesophilic bacterium,
meaning it can grow at moderate temperatures (around 30°C). It is typically
used in combination with other LAB strains to improve the fermentation
process and flavor profile of yogurt.
Fermentation Process of Yogurt
The process of making yogurt begins by heating fresh milk to around 85°C to
denature the proteins and eliminate any potential pathogens. The milk is then cooled to about
43°C to create an optimal environment for the starter cultures to grow. A small amount of the

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starter culture is added to the milk, typically around 1-2% of the total volume. The milk is
then incubated at this temperature for about 4-6 hours to allow the bacteria to ferment the
lactose into lactic acid. This acidification process causes the milk proteins to coagulate,
thickening the mixture and forming yogurt.
Once the yogurt has set, it is cooled to stop further fermentation, and then it is stored
in refrigerators to maintain its consistency, taste, and freshness. If the yogurt is not
refrigerated properly, fermentation will continue, potentially causing the product to become
overly sour or thin.
Importance of Starter Cultures in Yogurt Production
1. Flavor Development:
o The combination of Streptococcus thermophilus and Lactobacillus bulgaricus
is responsible for creating the characteristic tangy flavor of yogurt. These
bacteria generate lactic acid as they ferment lactose, which gives yogurt its
sour taste. The specific balance and ratio of these bacteria in the starter culture
directly affect the flavor profile of the final product.
2. Texture and Consistency:
o The acidification caused by the bacteria leads to the coagulation of casein
proteins in milk, which thickens the liquid into yogurt. Lactobacillus
bulgaricus also produces enzymes that help break down proteins and
contribute to the smooth, creamy texture of the yogurt.
3. Probiotic Benefits:
o Yogurt is known for its probiotic content, which includes live beneficial
bacteria such as Lactobacillus acidophilus and Bifidobacterium species. These
probiotics offer numerous health benefits, including supporting digestion,
enhancing gut health, and boosting the immune system. The presence of
probiotics also makes yogurt a beneficial food for people with lactose
intolerance, as the bacteria partially digest lactose during fermentation.
4. Health Benefits:
o In addition to providing probiotics, yogurt is a good source of essential
nutrients such as calcium, protein, vitamins (B12 and riboflavin), and
minerals. The fermentation process enhances the bioavailability of some
nutrients, making them easier for the body to absorb.

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 STARTER CULTURES OF FERMENTED MILKS: ACIDOPHILUS MILK

Acidophilus milk is a fermented dairy product that is produced by fermenting milk

with the bacterium Lactobacillus acidophilus, a probiotic microorganism known for its
health-promoting properties. This milk is particularly beneficial for individuals with lactose
intolerance and those looking to improve their gut health, as Lactobacillus acidophilus aids in
the digestion of lactose and supports a healthy digestive system.

Starter Cultures in Acidophilus Milk

The primary starter culture used in the production of acidophilus milk is Lactobacillus
acidophilus, although other bacterial strains may also be included to enhance the
fermentation process and improve the flavor and texture of the product.
1. Lactobacillus acidophilus:
o Lactobacillus acidophilus is the main bacterium responsible for the
fermentation of milk into acidophilus milk. This bacterium is a probiotic that
ferments lactose into lactic acid, which lowers the pH of the milk, leading to
the formation of a gel-like consistency. The lactic acid also contributes to the
characteristic tangy flavor of acidophilus milk.

o Lactobacillus acidophilus helps in breaking down lactose, making the milk

more digestible for individuals with lactose intolerance. It is also beneficial for
gut health as it encourages the growth of good bacteria in the intestines,
improves digestion, and can help balance the gut microbiota.

2. Streptococcus thermophilus (Optional):

o In some cases, Streptococcus thermophilus, a thermophilic bacterium, is
added to the starter culture. This bacterium works symbiotically with
Lactobacillus acidophilus in yogurt and other dairy products to ferment milk.
It helps in acid production and contributes to the overall texture and
consistency of the product. It also plays a role in the initial fermentation phase.

3. Bifidobacterium species (Optional):

o Some formulations of acidophilus milk may also contain Bifidobacterium
species, which are probiotics that support digestive health and contribute to
the health benefits of the milk. These bacteria can further enhance the gut
health-promoting properties of acidophilus milk, and help in improving the
digestive system’s overall balance.

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Fermentation Process of Acidophilus Milk
The process of making acidophilus milk involves adding Lactobacillus acidophilus
(and other optional strains) to pasteurized milk. The milk is then incubated at a temperature
of around 40°C to 45°C for several hours, allowing the bacteria to ferment the milk. During
fermentation, Lactobacillus acidophilus converts lactose into lactic acid, which results in the
thickening of the milk and gives it a characteristic tangy taste. The milk also develops a
smooth, creamy texture due to the coagulation of casein proteins.
Once the milk has fermented and reached the desired consistency, it is cooled to stop
further fermentation and then stored under refrigeration to maintain its flavor, texture, and
probiotic content.

STARTER CULTURES OF FERMENTED MILKS: CULTURED BUTTERMILK

Cultured buttermilk is a popular fermented dairy product made from the liquid that
remains after churning cream to make butter. Traditionally, buttermilk was the liquid left
behind from churning curd, but today, it is often produced by fermenting low-fat milk or
skim milk with specific starter cultures. The fermentation process imparts a tangy taste,
enhances digestibility, and provides several health benefits, especially due to its probiotic
content.

Starter Cultures in Cultured Buttermilk

The primary microorganisms used to produce cultured buttermilk are lactic acid bacteria
(LAB), which ferment the lactose in milk into lactic acid, lowering the pH and leading to the
formation of the characteristic sour flavor and thicker texture. The typical starter cultures
used in cultured buttermilk include:
1. Lactococcus lactis subsp. lactis:
o Lactococcus lactis is one of the main bacterial strains used in the production
of cultured buttermilk. This bacterium is responsible for fermenting the lactose
in the milk, producing lactic acid and contributing to the tangy taste and slight
thickening of the buttermilk. It is often used as a starter culture in the dairy
industry for various fermented products, including cultured buttermilk.

2. Lactococcus lactis subsp. cremoris:

o Lactococcus lactis subsp. cremoris is another strain frequently used in
cultured buttermilk production. It ferments lactose to produce lactic acid, and
its activity aids in the coagulation and souring of the milk. This strain is also

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 responsible for contributing to the smooth texture and mild, slightly tangy
flavor of buttermilk.

3. Leuconostoc mesenteroides:
o Leuconostoc mesenteroides is often included in cultured buttermilk starter
cultures to enhance flavor and promote the fermentation process. This
bacterium produces lactic acid and other flavor compounds, which give
cultured buttermilk its characteristic sourness. It also contributes to the smooth
and slightly creamy texture of buttermilk.

4. Streptococcus thermophilus (Occasionally used):

o Streptococcus thermophilus is a thermophilic bacterium used in combination
with other LAB strains to produce a well-balanced cultured buttermilk. While
it is more commonly used in yogurt production, it may be added to cultured
buttermilk starter cultures to improve fermentation efficiency and contribute to
the flavor profile.

5. Lactobacillus species (Occasionally used):

o Lactobacillus species, such as Lactobacillus acidophilus and Lactobacillus
bulgaricus, may be used as secondary cultures in cultured buttermilk. These
bacteria are known for their ability to promote the growth of beneficial gut
flora, providing health benefits such as aiding digestion and supporting
immune function.
Fermentation Process of Cultured Buttermilk
The process of making cultured buttermilk involves inoculating milk or cream with a
starter culture containing the appropriate strains of lactic acid bacteria. The milk is typically
pasteurized to eliminate any harmful microorganisms, and then cooled to the optimal
fermentation temperature of around 30°C to 35°C. Once the starter culture is added, the milk
is allowed to ferment for several hours.
During fermentation, the lactic acid bacteria begin to metabolize the lactose in the
milk, producing lactic acid. This acidification process lowers the pH of the milk, causing the
proteins (such as casein) to coagulate and form a gel-like structure, which thickens the liquid.
As the bacteria continue to produce lactic acid, the milk develops a tangy, sour flavor and a
thicker, creamier texture, characteristic of cultured buttermilk.
Once the fermentation process is complete, the cultured buttermilk is cooled to stop
further acidification and stored in a refrigerated environment to maintain its quality, flavor,
and probiotic content.
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 STARTER CULTURES OF CHEESES

Cheese production involves the fermentation of milk by specific microorganisms

known as starter cultures. These cultures are essential for the cheese-making process as they
help in coagulating milk proteins, producing lactic acid, and contributing to the texture,
flavor, and preservation of the cheese. The types of starter cultures used vary depending on
the type of cheese being produced. The main role of these cultures is to convert lactose into
lactic acid, which lowers the pH of the milk, leading to the coagulation of casein proteins and
the formation of curds. The starter cultures also help in developing the characteristic flavor
and aroma of different cheese varieties.

Types of Starter Cultures Used in Cheese Making

Starter cultures for cheese typically consist of lactic acid bacteria (LAB), which can
be classified into mesophilic, thermophilic, and proprietary cultures based on their
temperature preferences. Some cheeses also include secondary cultures, such as molds or
yeasts, to enhance flavor development and create distinctive textures. Below are the major
categories of starter cultures used in cheese-making:
1. Mesophilic Starter Cultures
Mesophilic bacteria are those that grow best at moderate temperatures, usually around
20°C to 30°C. These cultures are used for cheeses that are made at lower temperatures, like
cheddar, cream cheese, Brie, and Colby.
 Lactococcus lactis subsp. lactis:
o This is one of the primary bacteria used in mesophilic starter cultures. It is
responsible for the initial fermentation of lactose into lactic acid, helping to
curdle the milk. This strain is widely used in cheeses like cheddar and cream
cheese.

 Lactococcus lactis subsp. cremoris:

o Another important bacterium in mesophilic cultures, Lactococcus cremoris
helps in acidifying the milk and contributes to the smooth texture and mild
flavor of cheeses.

 Leuconostoc species:
o Leuconostoc mesenteroides and other strains are used to enhance the flavor
profile and the aroma of cheeses. They are also involved in the production of

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 gases that contribute to the texture and the formation of eye holes in cheeses
like Swiss cheese.

 Enterococcus faecalis:
o Sometimes used in mesophilic cultures, Enterococcus faecalis contributes to
the development of flavor and texture in certain cheeses like cheddar and
gouda.

2. Thermophilic Starter Cultures

Thermophilic bacteria thrive at higher temperatures, typically around 40°C to 45°C.
These bacteria are used for cheeses that require higher temperature fermentation and are
involved in producing cheeses like Parmesan, Mozzarella, Swiss, and Provolone.
 Streptococcus thermophilus:
o This is a key bacterium used in thermophilic starter cultures. It is particularly
important in the production of yogurt as well as Mozzarella and Provolone.
Streptococcus thermophilus works synergistically with other bacteria like
Lactobacillus to produce lactic acid and contribute to the firm texture of the
cheese.

 Lactobacillus delbrueckii subsp. bulgaricus:

o While commonly associated with yogurt production, this bacterium is
sometimes used in combination with Streptococcus thermophilus for cheeses
like Provolone and Parmesan. It helps to lower the pH and develop the
characteristic flavor.

 Lactobacillus helveticus:
o Used for cheeses like Swiss and Gruyère, Lactobacillus helveticus plays a key
role in developing the flavor profile and texture, especially in aged cheeses. It
helps in the breakdown of proteins during aging, enhancing the flavor
complexity.

3. Probiotic and Secondary Starter Cultures

In addition to the primary cultures, some cheeses are made with probiotic cultures or
secondary cultures, which can introduce additional beneficial microorganisms for flavor
development and health benefits. These are added during or after the primary fermentation
stage.
 Bifidobacterium species:

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 o Sometimes added to cheeses to enhance their probiotic properties,
Bifidobacterium species can improve gut health and support the immune
system. These are commonly found in functional cheeses or cheeses marketed
as having added health benefits.

 Penicillium molds:
o Used in blue cheeses like Roquefort, Gorgonzola, and Stilton, Penicillium
roqueforti and Penicillium glaucum are secondary cultures responsible for
creating the distinctive blue veins and pungent flavor. Penicillium camemberti
is used in Camembert and Brie, contributing to their creamy texture and
bloomy rind.

 Geotrichum candidum:
o Used in cheeses like Brie and Camembert, this yeast contributes to the
development of the characteristic soft and creamy texture of the cheese and
aids in the formation of the rind.
Role of Starter Cultures in Cheese Production
1. Acidification:
o Starter cultures are responsible for acidifying the milk by converting lactose
into lactic acid. This acidification lowers the pH of the milk, which helps
coagulate the milk proteins and creates the curds. This is the first essential step
in cheese-making.

2. Flavor Development:
o The bacteria and molds involved in the fermentation process break down
proteins and fats in the milk into smaller compounds, such as amino acids and
fatty acids, which contribute to the flavor profile of the cheese. For example,
Lactobacillus helveticus plays a major role in the development of aged cheese
flavors.

3. Texture Formation:
o The fermentation process also affects the texture of the cheese. Starter cultures
influence the curd texture by producing lactic acid, which causes casein
proteins to coagulate. Different cultures and fermentation times result in
different textures, from soft and creamy to firm and crumbly.

4. Preservation:

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 o The acidification of the milk by starter cultures helps in preserving the cheese
by lowering the pH, creating an environment that inhibits the growth of
harmful bacteria. Additionally, the production of antimicrobial compounds by
certain bacteria helps extend the shelf life of the cheese.

MICROBIOLOGY OF FRESH AND RIPENED CHEESES

Cheese is a fermented dairy product that is produced through the action of

microorganisms. These microorganisms, which include lactic acid bacteria (LAB), molds,
and yeasts, play crucial roles in the production, texture, flavor, and safety of cheese. The
microbiological process involved in the making of fresh cheeses (such as paneer, ricotta, and
cream cheese) differs significantly from that of ripened cheeses (such as cheddar, gouda, and
brie). This difference is largely due to the variations in the type of microorganisms used, the
fermentation conditions, and the length of time the cheese undergoes fermentation and aging.

Microbiology of Fresh Cheeses

Fresh cheeses are those that are typically not aged or undergo minimal aging. They
have a soft, moist texture and a mild flavor due to a shorter fermentation period. The
microbiological processes in fresh cheeses mainly involve lactic acid bacteria (LAB), which
contribute to the acidification of milk and curd formation.
Microorganisms in Fresh Cheeses
1. Lactic Acid Bacteria (LAB):
o Lactococcus lactis and Lactococcus cremoris are commonly used in the
production of fresh cheeses. These bacteria convert lactose into lactic acid,
which lowers the pH of the milk and causes the casein proteins to coagulate,
forming curds.

o Other strains of Streptococcus thermophilus and Lactobacillus species may

also be used, depending on the specific type of fresh cheese being made.
These bacteria also contribute to the flavor and texture by producing small
amounts of flavor compounds such as diacetyl and acetaldehyde.

2. Yeasts:
o Yeasts such as Saccharomyces cerevisiae and Candida species may be
present in small quantities in fresh cheeses. They do not play a major role in
fermentation but can contribute to flavor complexity and help in preventing
spoilage by outcompeting harmful microorganisms.
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 3. Molds:
o Fresh cheeses generally do not contain molds, but some types, like fresh
ricotta or cream cheese, may occasionally have molds if they are made under
unsanitary conditions. These molds are not intentionally added and can lead to
spoilage if they grow.
Fermentation and Texture Formation
 The primary biochemical process in the production of fresh cheese is lactic
fermentation, where lactose is converted into lactic acid by LAB. This lowers the pH
and causes the milk proteins (mainly casein) to coagulate, forming curds.

 The texture of fresh cheese remains soft and moist because the curds are not subjected
to a long aging process. In some cases, salt is added to the curd to enhance flavor and
act as a preservative.

 Fresh cheeses are typically consumed shortly after production, and their shelf life is
relatively short, as they do not undergo the preservation effects of aging or drying.
Microbiology of Ripened Cheeses
Ripened cheeses (also known as aged cheeses) undergo a fermentation and
maturation process that significantly changes their texture, flavor, and microbiological
composition. The ripening of cheese involves enzymatic activity and the action of various
microorganisms, which break down proteins, fats, and carbohydrates over time.
Microorganisms in Ripened Cheeses
1. Lactic Acid Bacteria (LAB):
o LAB play a vital role in both the initial fermentation and the ripening of
cheese. In the early stages of ripening, bacteria like Lactococcus lactis and
Streptococcus thermophilus are involved in the fermentation of lactose into
lactic acid, which lowers the pH and forms the curds.

o During the aging process, Lactobacillus species (e.g., Lactobacillus

helveticus, Lactobacillus casei) break down the proteins in the curd into
smaller peptides and amino acids, which are responsible for the development
of complex flavors. The breakdown of fats by LAB also produces fatty acids,
contributing to the texture and flavor of the cheese.

2. Lipolytic Bacteria:
o Some ripened cheeses, such as Brie and Camembert, may have specific
lipolytic bacteria (bacteria that break down fats), such as Pseudomonas and

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 Corynebacterium, which contribute to the development of the characteristic
flavor profile.

o Lipolysis leads to the formation of free fatty acids that give certain cheeses
their rich, tangy, and sometimes rancid flavor notes.
3. Molds:
o Penicillium species are among the most well-known molds involved in cheese
ripening. Molds such as Penicillium roqueforti and Penicillium glaucum are
used in the production of blue cheeses like Roquefort, Gorgonzola, and
Stilton. These molds contribute to the formation of blue veins and the
distinctive strong flavor of these cheeses by releasing enzymes that break
down proteins and fats.

o Penicillium camemberti is used for the surface ripening of soft cheeses like
Brie and Camembert. This mold grows on the surface, creating a white,
bloomy rind and producing enzymes that help in the breakdown of proteins
and fats, contributing to the creamy texture and mild flavor of these cheeses.
4. Yeasts:
o In some aged cheeses, yeasts (e.g., Geotrichum candidum) may be involved in
the early stages of ripening. These yeasts help create the rind and contribute to
the flavor development by breaking down fats and proteins. Yeasts are
especially important in cheeses like Camembert and Brie, where they are used
to help form the characteristic soft, creamy texture.
Fermentation and Aging Process in Ripened Cheeses
 Initial Fermentation: Just like in fresh cheeses, lactic acid bacteria (LAB) play a
crucial role in the initial acidification of the milk. However, the difference in ripened
cheeses is that the fermentation continues during aging. After the curd is formed and
salted, the cheese is stored in controlled temperature and humidity conditions,
allowing microbial activity to continue.
 Proteolysis and Lipolysis: The proteolysis (breakdown of proteins) and lipolysis
(breakdown of fats) processes that occur during aging are essential for developing the
texture and flavor of ripened cheeses. The breakdown of proteins results in the
formation of amino acids, some of which contribute to the umami and savory
flavors in cheeses like Parmesan and Gruyère. Lipolysis leads to the formation of
free fatty acids that create characteristic rich, nutty, or sharp flavors in cheeses like
Brie and cheddar.

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  Microbial Rind Formation: In certain ripened cheeses, such as washed rind cheeses
like Munster and Limburger, specific bacteria such as Brevibacterium linens are
responsible for the rind formation. These bacteria are known for their ability to break
down amino acids, which results in the production of ammonia and the development
of the strong-smelling rind.
Microbiology of Accelerated Cheese Ripening
Cheese ripening, or aging, is a complex process where microorganisms play a
significant role in developing the cheese's texture, flavor, and aroma. Traditional cheese
ripening takes months or even years, but with modern techniques, the process can be
accelerated through specific environmental conditions and the manipulation of microbial
activity. This accelerated ripening is important in producing cheeses more quickly for
commercial purposes while maintaining desirable qualities. The microbiology of accelerated
cheese ripening involves the use of different microorganisms, temperature, humidity, and
enzymatic activity to fast-track the usual biochemical changes that occur during aging.
Key Factors in Accelerated Cheese Ripening
Accelerating the ripening of cheese involves manipulating several factors to speed up the
natural fermentation processes that typically occur over long periods. These factors include:
1. Increased Temperature:
o One of the main ways to accelerate ripening is by increasing the temperature
at which the cheese is stored. Typically, ripened cheeses are aged at 10°C to
15°C for several months. In accelerated ripening, temperatures may be raised
to 20°C to 30°C, which stimulates microbial activity and enzyme production,
leading to quicker changes in texture and flavor.

2. Modified Humidity:
o Adjusting the humidity levels during aging helps manage the moisture content
of the cheese, affecting the texture and microbial growth. Higher humidity can
speed up the enzymatic breakdown of fats and proteins, influencing the speed
of flavor development.

3. Use of Active Starter and Secondary Cultures:

o Lactic acid bacteria (LAB) are still fundamental to accelerated ripening, but
a more concentrated or active form of these bacteria may be used to initiate
fermentation faster. Additionally, the use of probiotic cultures and lipolytic
(fat-degrading) bacteria speeds up the breakdown of fats and proteins, leading
to quicker flavor formation.

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 o Certain molds such as Penicillium species (e.g., Penicillium camemberti or
Penicillium roqueforti) can be applied on the surface or incorporated into the
cheese for enhanced ripening. These molds break down fats and proteins,
contributing to characteristic flavors and textures in cheeses like Brie and Blue
cheese.
4. Ultrafiltration and Microfiltration:
o Microfiltration or ultrafiltration can be used in cheese production to
concentrate milk proteins and reduce moisture content. This modification can
speed up the ripening process by promoting quicker acidification and curd
formation, reducing the time needed for the cheese to mature.
Microorganisms Involved in Accelerated Cheese Ripening
The use of specific microorganisms or the enhancement of natural microbial activity is
key to accelerating the cheese ripening process. These microorganisms work in synergy with
each other to break down milk components like proteins, fats, and carbohydrates into smaller
molecules that contribute to flavor and texture.
1. Lactic Acid Bacteria (LAB):
o LAB play a pivotal role in both the initial fermentation and ripening of cheese.
In accelerated ripening, strains of Lactococcus lactis, Streptococcus
thermophilus, and Lactobacillus species are often used in higher
concentrations to quickly acidify the milk, which helps in curd formation and
also initiates the breakdown of proteins.

o Lactobacillus helveticus, commonly used in Swiss or Gruyère cheese, is

particularly effective in protein breakdown, producing small peptides and
amino acids that contribute to the flavor.

o Lactobacillus casei and Lactobacillus rhamnosus are also employed to

improve the flavor profile and contribute to the faster acidification process.

2. Lipolytic Bacteria:
o To speed up the breakdown of fats, lipolytic bacteria such as
Corynebacterium species or Pseudomonas species are introduced. These
bacteria break down fats into free fatty acids, which are crucial for developing
the characteristic aroma and flavor of many cheeses. They also influence the
cheese's texture by enhancing its softness and creaminess.
3. Molds:

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 o Penicillium species, such as Penicillium roqueforti, which are used in blue
cheeses like Gorgonzola and Roquefort, can also be part of the accelerated
ripening process. These molds produce enzymes that break down both proteins
and fats, creating strong flavors and distinctive textures much more quickly
than natural ripening.

o In surface-ripened cheeses like Brie and Camembert, Penicillium camemberti

molds are used to break down the cheese's surface and interior, contributing to
the soft, creamy texture and the characteristic aroma.

4. Yeasts:
o Yeasts, such as Geotrichum candidum, play a role in some cheeses like Brie
and Camembert, where they help develop the rind and contribute to the
breakdown of fats. They can also accelerate the ripening process by promoting
faster growth on the surface of the cheese and aiding in flavor development.

MICROBIAL PRODUCTION OF CHEESE

The production of cheese is a complex biochemical process that relies heavily on the
action of microorganisms, particularly bacteria, molds, and yeasts. These microorganisms
play a vital role in fermenting the milk, coagulating the proteins, developing flavors, and
forming the characteristic texture of cheese. The microbial production of cheese involves
several key steps: milk preparation, fermentation, curd formation, whey drainage, salting, and
ripening.

Steps in Microbial Cheese Production

1. Milk Preparation and Pasteurization
The first step in cheese making is the preparation of milk. Fresh, high-quality milk is
used to ensure that the cheese has the desired characteristics. Milk is typically pasteurized by
heating it to around 72°C for 15 seconds. This process kills harmful bacteria and pathogens,
ensuring the safety of the final product. After pasteurization, the milk is cooled to an optimal
temperature for fermentation.

2. Addition of Starter Cultures

Once the milk is cooled, starter cultures of lactic acid bacteria (LAB) are added.
These microorganisms are responsible for fermenting lactose into lactic acid, lowering the pH
of the milk, and initiating curd formation. The starter cultures used depend on the type of
cheese being made:
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  Mesophilic cultures (e.g., Lactococcus lactis) are used for cheeses like cheddar and
brie, which ripen at lower temperatures.
 Thermophilic cultures (e.g., Streptococcus thermophilus and Lactobacillus species)
are used for cheeses like mozzarella and Swiss cheese, which require higher
fermentation temperatures.
In addition to LAB, secondary cultures (e.g., molds and yeasts) may be added depending on
the desired characteristics of the cheese.
3. Coagulation and Curd Formation
After adding the starter cultures, rennet (an enzyme) is introduced to coagulate the
milk proteins. Rennet contains chymosin, which acts on the milk protein casein, causing the
milk to coagulate and form curds. This process is known as coagulation.
In some cheeses, the curds are cut to facilitate whey expulsion. The size of the curds affects
the texture of the cheese—smaller curds result in firmer cheeses, while larger curds result in
softer cheeses.
4. Curd Cooking and Whey Drainage
Once the curds have formed, they are cooked at varying temperatures depending on
the cheese type. For example, cheddar curds are heated to about 40°C, while mozzarella
curds are heated to 60°C.
Heating the curds helps expel more whey, making the curds firmer. The whey, which
contains water, lactose, soluble proteins, and other nutrients, is drained off. The amount of
whey left in the curds influences the final texture of the cheese.

5. Salting
After whey drainage, the cheese curds are salted. Salt plays several roles in cheese
production:
 Flavor enhancement.
 Preservation by inhibiting the growth of spoilage microorganisms.
 Texture development, as salt draws moisture from the curds, helping to firm them.
Salting can be done in several ways:
 Dry salting: Salt is directly applied to the curd.
 Brining: The cheese is soaked in a saltwater solution.

6. Molding and Pressing

The curds are then placed into molds to give the cheese its final shape. Depending on
the type of cheese, the curds may be pressed to remove additional whey and compact the

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curds together. The pressing process also helps to enhance the texture and develop the desired
consistency.

7. Ripening (Aging)
The final step in cheese production is ripening or aging, during which the texture,
flavor, and aroma of the cheese develop. Microorganisms, particularly lactic acid bacteria and
molds, continue to break down proteins, fats, and carbohydrates in the cheese, leading to the
formation of flavor compounds such as amino acids, fatty acids, and peptides.
The ripening process can vary greatly depending on the type of cheese. For example:
 Soft cheeses like Brie or Camembert ripen quickly and have a creamy texture due to
the action of molds and yeasts.
 Hard cheeses like cheddar or Parmesan require longer ripening, often for several
months, where the proteins are extensively broken down, and complex flavors
develop.
PRODUCTION OF YOGURT

Yogurt is a popular dairy product made by fermenting milk with specific bacterial
cultures. The fermentation process involves the conversion of lactose (milk sugar) into lactic
acid, which causes the milk proteins to coagulate and form a thickened, tangy product. The
microbial production of yogurt is a controlled process, requiring specific strains of bacteria
and careful temperature management. Below is a detailed explanation of the yogurt
production process.

Microorganisms in Yogurt Production

The production of yogurt involves the use of starter cultures, which are specific strains
of bacteria that ferment the milk to produce yogurt. The two primary bacterial species used in
yogurt making are:
1. Lactobacillus delbrueckii subsp. bulgaricus
2. Streptococcus thermophilus
These bacteria work symbiotically to acidify the milk, producing the characteristic flavor,
texture, and consistency of yogurt.
 Lactobacillus delbrueckii subsp. bulgaricus is responsible for the production of
flavor compounds and contributes to the thick texture of the yogurt.

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  Streptococcus thermophilus is the first bacterium to start the fermentation process
by converting lactose into lactic acid, lowering the pH and helping the milk proteins
coagulate.
In addition to these primary bacteria, other beneficial bacteria, such as Lactobacillus
acidophilus and Bifidobacterium bifidum, may be added to enhance the probiotic content
of the yogurt.
Steps in Yogurt Production
1. Milk Preparation
The first step in yogurt production is selecting fresh, high-quality milk. The milk can
be whole milk, skim milk, or partially skimmed milk, depending on the desired final
product. The milk is then pasteurized to kill any harmful bacteria and other microorganisms.
Pasteurization typically involves heating the milk to about 85°C for 30 minutes or 90°C for 5
minutes. This step ensures that the milk is free from pathogens and also denatures the milk
proteins, helping the yogurt achieve the desired texture.
After pasteurization, the milk is cooled to about 42°C-45°C (optimal temperature for
bacterial growth and fermentation).
2. Inoculation with Starter Cultures
Once the milk is cooled to the appropriate temperature, the starter cultures of
Lactobacillus bulgaricus and Streptococcus thermophilus are added. These cultures are
usually in the form of freeze-dried powder or liquid culture. The bacteria are added in a
specific ratio to ensure proper fermentation. The addition of these bacteria starts the
fermentation process, during which they convert lactose into lactic acid.
In some cases, additional bacteria such as Lactobacillus acidophilus or
Bifidobacterium may be added to create probiotic yogurt, which provides additional health
benefits.

3. Fermentation Process
After the cultures are added, the milk is maintained at a constant temperature of
around 42°C-45°C for about 4-6 hours. During this period, the bacteria ferment the lactose
into lactic acid. The acidification of the milk lowers the pH, causing the casein proteins
(milk proteins) to coagulate and form a gel-like structure. As the pH drops, the yogurt
becomes thicker and takes on its characteristic tart flavor.
The fermentation time can be adjusted based on the desired texture and taste of the
yogurt. Longer fermentation results in a thicker, more acidic yogurt, while shorter
fermentation produces a milder taste and thinner consistency.

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4. Cooling and Stirring
Once the desired pH and consistency are achieved, the yogurt is cooled rapidly to
stop the fermentation process. This is typically done by lowering the temperature to about
4°C. After cooling, the yogurt may be stirred to break up the curd structure, resulting in a
smooth texture. This is especially common for set yogurt (where yogurt is allowed to
ferment in individual containers) versus stirred yogurt, which is mixed after fermentation.
At this stage, optional ingredients such as flavors, sweeteners, fruit, or
preservatives can be added, depending on the type of yogurt being produced (e.g., fruit
yogurt or flavored yogurt).
5. Packaging and Storage
Once the yogurt is cooled, stirred, and mixed (if necessary), it is packaged into
containers. These containers are then sealed and stored under refrigerated conditions to
maintain the freshness and prevent the growth of unwanted microorganisms.
Yogurt is generally stored at 4°C to 7°C to preserve its quality. During storage, the
yogurt continues to undergo slow fermentation, which can improve its flavor profile.

UNIT – 5

MICROBIOLOGICAL STANDARDS AND QUALITY OF DAIRY PRODUCTS

Cream
Microbial Quality
 Cream is a dairy product made by separating the fat from milk. It has a high-fat
content and provides an ideal environment for microbial growth, especially if
improperly handled or stored.

 The main contaminants in cream include lactic acid bacteria (LAB), yeasts, molds,
and pathogens like Staphylococcus aureus and Salmonella spp..

Microbiological Standards
 Total Plate Count (TPC): This is a general indicator of the microbial load in cream.
The acceptable limit is generally less than 50,000 CFU/g in high-quality cream.

 Coliform Count: Coliforms are indicator organisms for fecal contamination. The
limit for coliforms is typically <10 CFU/g in cream.

 Pathogens: No pathogens, particularly Listeria monocytogenes, Salmonella, and

Escherichia coli, should be present in cream. These are harmful to health and are
regulated to zero tolerance levels.

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  Yeasts and Molds: These microorganisms can lead to spoilage and fermentation of
cream, causing off-flavors. Their permissible level is usually <10 CFU/g.

Butter
Microbial Quality
 Butter is made by churning cream to separate butterfat from buttermilk. It is a
perishable product due to its fat content, which can support the growth of
microorganisms.

 Lactic acid bacteria, including Lactococcus and Leuconostoc, are predominant in

butter. These bacteria contribute to the characteristic flavor of fermented butter.

 Pathogenic bacteria like Staphylococcus aureus, Salmonella, and Clostridium

botulinum can also be present in butter, especially if produced under unsanitary
conditions.

Microbiological Standards
 Total Plate Count (TPC): The acceptable level for TPC in butter is typically
<100,000 CFU/g for high-quality butter.

 Coliforms: Coliform bacteria should be absent or <10 CFU/g in butter.

 Yeasts and Molds: The permissible level for yeasts and molds in butter is generally
<10 CFU/g, as these can cause spoilage.

 Pathogens: There should be no presence of harmful pathogens such as Listeria

monocytogenes and Salmonella spp. in butter. These pathogens must be absent in all
dairy products to ensure consumer safety.

Dried and Evaporated Milk

Microbial Quality
 Dried milk (also known as powdered milk) and evaporated milk are both milk
products that have been concentrated by removing most of the water content. These
processes help in extending shelf life but do not completely eliminate the risk of
microbial contamination.

 Lactic acid bacteria, enteric pathogens, and spore-forming bacteria (such as Bacillus
cereus) are common contaminants in dried and evaporated milk.

 Salmonella, E. coli, and Campylobacter can also be found in these products if proper
sanitation measures are not followed during production.

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Microbiological Standards
 Total Plate Count (TPC): The acceptable TPC in dried milk is typically <50,000
CFU/g. For evaporated milk, the standard is generally <10,000 CFU/g.

 Coliforms: Coliform bacteria should be absent or <10 CFU/g in both dried and
evaporated milk.

 Pathogens: Both Salmonella and E. coli should be absent in dried and evaporated
milk. The standards for these pathogens are typically zero tolerance.

 Spore-forming Bacteria: Spore-forming bacteria such as Bacillus cereus should be

absent, as they can survive the drying and heating processes and lead to spoilage.

Sweetened Condensed Milk

Microbial Quality
 Sweetened condensed milk is milk that has been evaporated and then sweetened with
a large amount of sugar. The high sugar content in sweetened condensed milk inhibits
the growth of many microorganisms; however, some pathogens may still survive if
the product is not processed correctly.

 Lactic acid bacteria and yeasts can still be present, though they do not grow
effectively due to the high sugar concentration.

 The major concern with sweetened condensed milk is the presence of Clostridium
botulinum, as the low acidity and sealed container can provide an anaerobic
environment that favors its growth.

Microbiological Standards

 Total Plate Count (TPC): The acceptable TPC for sweetened condensed milk is
usually <50,000 CFU/g.

 Coliforms: Coliform bacteria should be absent or <10 CFU/g in sweetened

condensed milk.

 Pathogens: The most important standard for sweetened condensed milk is zero
tolerance for Clostridium botulinum. Other pathogens like Salmonella, E. coli, and
Listeria should also be absent.

 Yeasts and Molds: These microorganisms should also be absent or present in very
low amounts (usually <10 CFU/g), as they can cause fermentation and spoilage.

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 MICROBIOLOGICAL SPOILAGE OF MILK PRODUCTS
Milk products, such as milk, cream, cheese, butter, yogurt, and ice cream, are highly
nutritious and perishable. Due to their high moisture content, they provide an ideal
environment for microbial growth. Microbial spoilage of milk products can affect their taste,
texture, appearance, and safety, leading to reduced product quality and potential health risks.
The primary microorganisms involved in spoilage are bacteria, yeasts, and molds. This
spoilage is influenced by factors such as the type of product, storage conditions, and the
presence of contaminants during production.

Types of Microbial Spoilage

1. Bacterial Spoilage
Bacteria are the primary agents responsible for the spoilage of most milk products. The type
of bacteria involved can vary depending on the conditions of the product and its processing
history.
a) Lactic Acid Bacteria (LAB)
 Common Genera: Lactobacillus, Lactococcus, Streptococcus, Leuconostoc.
 Spoilage Effects: Lactic acid bacteria are typically associated with the souring of
milk and dairy products. They ferment lactose, producing lactic acid, which lowers
the pH and results in an undesirable sour flavor. Over time, this fermentation can also
lead to changes in texture (e.g., curdling or thickening) and the production of gases
that may cause bulging of packaging.
 Products Affected: Milk, cream, yogurt, and other fermented dairy products.
b) Pathogenic Bacteria
 Common Pathogens: Escherichia coli, Salmonella, Listeria monocytogenes,
Staphylococcus aureus.
 Spoilage Effects: While these bacteria may not always cause spoilage in terms of
sensory changes, their presence poses serious health risks. They can multiply if milk
or dairy products are not pasteurized properly or stored under appropriate conditions.
These pathogens can cause foodborne illnesses like diarrhea, vomiting, and more
severe infections in susceptible populations.
 Products Affected: Raw milk, cream, butter, cheese, and yogurt.
c) Psychrotrophic Bacteria
 Common Genera: Pseudomonas, Aeromonas, Flavobacterium.
 Spoilage Effects: These bacteria can grow at refrigeration temperatures, making them
a major concern in refrigerated milk products. They produce enzymes that break

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 down fats and proteins, leading to off-flavors, rancidity, and spoilage. Rancidity due
to lipolysis (fat breakdown) often leads to a bitter, unpleasant taste.
 Products Affected: Milk, cream, butter, and ice cream.

2. Yeast Spoilage
Yeasts are microorganisms that thrive in high-sugar or high-moisture environments
and can cause fermentation in milk products. Yeast spoilage often leads to off-flavors and
changes in texture.
 Common Genera: Candida, Saccharomyces, Rhodotorula.
 Spoilage Effects: Yeasts can ferment lactose and other sugars in milk, producing
carbon dioxide and ethanol, which leads to gas formation, bubbling, and souring. This
can result in undesirable flavors, such as alcoholic or fruity notes, and changes in
texture, like a slimy or overly viscous consistency.
 Products Affected: Cream, yogurt, and kefir, particularly when storage conditions
are not optimal or when the product is contaminated during handling.

3. Mold Spoilage
Molds are fungi that grow in moist environments and can cause both visible and
invisible spoilage in milk products. While most molds are not pathogenic, some can produce
mycotoxins, which are toxic and harmful to health.
 Common Genera: Penicillium, Aspergillus, Cladosporium.
 Spoilage Effects: Molds typically grow on the surface of dairy products, forming
visible colonies that can affect the appearance and texture. They produce enzymes
that degrade proteins and fats, leading to the breakdown of the product. Molds can
also cause changes in flavor, producing musty, earthy, or bitter notes.
 Products Affected: Cheese, particularly those with high moisture content, such as
soft cheeses (brie, camembert), as well as cream and yogurt.

Factors Contributing to Microbial Spoilage

1. Temperature: Dairy products are highly susceptible to microbial growth, and
improper storage temperature is one of the most common causes of spoilage. For
example, refrigeration at temperatures above 4°C can encourage the growth of
psychrotrophic bacteria, while improper pasteurization may allow pathogens to
survive and proliferate.
2. Processing Conditions: Insufficient heat treatment (e.g., underpasteurization) or
contamination during production can lead to an increased microbial load in the final

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 product. Pasteurization, sterilization, and fermentation are critical steps in controlling
microbial populations in dairy products.
3. Packaging: Improper packaging or exposure to air can introduce or allow
microorganisms to grow in dairy products. For example, poorly sealed packaging can
lead to contamination with air-borne yeasts or molds, while damaged packaging can
allow bacteria to enter and spoil the product.
4. Storage Duration: The longer dairy products are stored, the higher the risk of
microbial spoilage. This is especially true for products like milk and cream, which
have relatively short shelf lives. Even when refrigerated, the microbial load can
increase over time, leading to spoilage.
5. Sanitation Practices: Poor hygiene during milk collection, handling, and processing
can introduce microorganisms into the product. Contaminated equipment, utensils, or
workers' hands can all contribute to microbial spoilage.

Common Milk Products and Their Microbial Spoilage

1. Milk
 Microbial spoilage: Caused by lactic acid bacteria, psychrotrophic bacteria, and
pathogens like E. coli or Salmonella. These can lead to souring, curdling, and the
potential for foodborne illnesses.
 Signs of spoilage: Sour odor, off-flavors, and curdling.

2. Cream
 Microbial spoilage: Lactic acid bacteria, yeasts, and molds can lead to souring, off-
flavors, and texture changes.
 Signs of spoilage: Sour taste, off-flavors, mold growth on the surface.

3. Butter
 Microbial spoilage: Psychrotrophic bacteria and molds can lead to rancidity and
changes in flavor and texture.
 Signs of spoilage: Bitter or rancid taste, off-flavors.

4. Cheese
 Microbial spoilage: Molds and spoilage bacteria, like Pseudomonas or
Brevibacterium, can alter texture, flavor, and cause the product to deteriorate.
 Signs of spoilage: Molds on the surface, sliminess, ammonia-like smell.

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5. Yogurt
 Microbial spoilage: Yeasts, molds, and spoilage bacteria can cause fermentation or
souring.
 Signs of spoilage: Separation of whey, yeast-like odors, and changes in texture (e.g.,
sliminess).

Prevention of Microbial Spoilage

To prevent microbial spoilage of milk products, several practices can be implemented:
1. Proper Pasteurization
2. Refrigeration
3. Good Manufacturing Practices (GMP)
4. Proper Packaging

MICROBIAL SAFETY OF FERMENTED DAIRY PRODUCTS

Fermented dairy products are produced through the action of beneficial
microorganisms, primarily lactic acid bacteria (LAB), which help ferment milk sugars
(lactose) to produce lactic acid. This process lowers the pH of the product, creating an
environment that inhibits the growth of many harmful microorganisms. However, like all
food products, fermented dairy products can still pose safety risks if contamination occurs
during production, processing, or storage.

Factors Affecting the Safety of Fermented Dairy Products

Several factors influence the microbiological safety of fermented dairy products, including:
1. Pasteurization: The pasteurization of milk before fermentation is a critical step in
ensuring the safety of fermented dairy products. Pasteurization kills most pathogens
that may be present in raw milk, such as Salmonella, E. coli, and Listeria. However,
pasteurization does not guarantee the absence of all microorganisms, so proper
handling after pasteurization is also essential.
2. Fermentation Conditions: The temperature, time, and conditions under which
fermentation occurs play a significant role in the safety of the final product.
Inadequate fermentation may not sufficiently lower the pH to inhibit the growth of
pathogenic organisms. Fermented dairy products should be allowed to ferment for the
appropriate amount of time at the correct temperature to ensure the growth of
beneficial microorganisms and the suppression of harmful bacteria.

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 3. Storage Conditions: Fermented dairy products are highly perishable. Storage at
improper temperatures can promote the growth of spoilage microorganisms and
pathogens. Refrigeration is crucial for maintaining the safety of fermented dairy
products. Products like yogurt, kefir, and cheese should be stored at temperatures
below 4°C to limit microbial growth.
4. Hygiene and Sanitation: Proper hygiene practices during milk collection,
fermentation, and packaging are essential to prevent contamination with harmful
bacteria. Contaminated equipment, improper handling, or exposure to air-borne
pathogens can introduce harmful microorganisms into the product.

Safety Measures for Fermented Dairy Products

To ensure the safety of fermented dairy products, various measures must be taken
during their production, storage, and consumption:

1. Pasteurization and Heat Treatment

 Proper pasteurization (heating milk to 72°C for 15 seconds or equivalent) is essential
to kill most pathogens while preserving the quality of the milk.
 In some products, ultra-pasteurization (higher temperature for shorter periods) may
be used for longer shelf life.

2. Temperature Control
 Refrigeration is critical for maintaining the safety and quality of fermented dairy
products. Products should be stored at or below 4°C to prevent the growth of harmful
bacteria and ensure their freshness.
 Fermented dairy products should not be left out at room temperature for extended
periods, as this may allow pathogenic bacteria to grow.

3. Starter Culture Control

 The use of starter cultures (specific strains of LAB) helps ensure that only beneficial
bacteria proliferate during fermentation, outcompeting harmful microorganisms.
These cultures should be carefully chosen and maintained to ensure consistent product
quality and safety.

4. Hygiene and Good Manufacturing Practices (GMP)

 Cleanliness during milk handling, fermentation, and packaging is crucial. Regular
cleaning and sanitizing of equipment help prevent contamination.
 Adhering to good manufacturing practices (GMP) ensures that the production
environment remains free from contaminants.
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5. Packaging
 Proper packaging can help protect fermented dairy products from microbial
contamination. Packaging should be airtight and clean to prevent exposure to air-
borne contaminants, such as molds or yeasts.

Regulatory Standards and Quality Assurance

To ensure the safety of fermented dairy products, various regulatory bodies have set
standards for their production, labeling, and sale. These standards typically focus on:
 Microbiological limits for pathogens such as Listeria monocytogenes, Salmonella,
and E. coli.

 Limits for spoilage organisms, including lactic acid bacteria, yeasts, and molds, to
ensure the product remains safe and of high quality.

 Labeling regulations to inform consumers about product ingredients, storage

requirements, and expiration dates.

MECHANISM OF ACTION OF PROBIOTICS AND THEIR HEALTH BENEFITS

Probiotics are live microorganisms, which when administered in adequate amounts,

confer a health benefit on the host, typically by improving or restoring the gut microbiota
balance. The health-promoting effects of probiotics are primarily attributed to their
interaction with the gut microbiome and immune system. Here is a detailed look at the
mechanisms of action of probiotics and their health benefits:

Mechanisms of Action
1. Restoration of Gut Microflora Balance
o Colonization of the Gut: Probiotics help restore the natural balance of gut
bacteria, which can be disrupted by factors such as illness, antibiotics, or a
poor diet. Beneficial probiotic strains, such as Lactobacillus and
Bifidobacterium, can colonize the gut and inhibit the growth of harmful
pathogens, preventing dysbiosis (imbalance of gut microbiota).

2. Production of Antimicrobial Substances

o Antibacterial Activity: Many probiotics produce antimicrobial substances
like lactic acid, hydrogen peroxide, and bacteriocins, which lower the pH of
the gut, creating an unfavorable environment for pathogenic microorganisms.

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 These antimicrobial compounds help reduce the colonization of harmful
pathogens like Clostridium difficile, Salmonella, and Escherichia coli.

3. Immune System Modulation

o Immune Enhancement: Probiotics can interact with gut-associated lymphoid
tissue (GALT) and influence the production of cytokines, antibodies, and other
immune responses. By enhancing the immune function, probiotics can help
protect against infections and support overall immune health.

o Regulation of Inflammatory Responses: Probiotics may help to regulate

inflammatory pathways by decreasing the levels of pro-inflammatory
cytokines, such as tumor necrosis factor (TNF), and increasing anti-
inflammatory cytokines. This can be particularly beneficial in conditions like
irritable bowel syndrome (IBS), Crohn’s disease, and ulcerative colitis.

4. Inhibition of Pathogen Adhesion

o Competition for Adhesion Sites: Probiotics can prevent harmful pathogens
from adhering to the intestinal walls by competing for the same binding sites.
This physical barrier reduces the chance of infection and colonization by
pathogenic microbes.

5. Enhancement of Gut Barrier Function

o Strengthening the Intestinal Mucosal Barrier: Probiotics support the
integrity of the intestinal barrier by promoting the production of mucus and
enhancing the expression of tight junction proteins that seal the intestinal cells
together. This reduces the permeability of the gut lining, preventing harmful
substances and pathogens from entering the bloodstream.

Health Benefits of Probiotics

1. Gut Health and Digestion
o Relief from Gastrointestinal Disorders: Probiotics are particularly beneficial
for people suffering from gastrointestinal disorders like IBS, inflammatory
bowel diseases (IBD), diarrhea, and constipation. They help improve gut
motility, reduce bloating, and alleviate abdominal discomfort.

o Prevention of Antibiotic-Associated Diarrhea: Probiotics can prevent

diarrhea associated with antibiotic use by restoring the balance of gut
microbiota disrupted by antibiotics.

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 o Prevention and Treatment of Infections: Probiotics have been shown to
reduce the incidence of Clostridium difficile-associated diarrhea and
gastroenteritis caused by pathogenic organisms like Rotavirus.

2. Immune System Support

o Boosting Immune Response: Probiotics can enhance both innate and
adaptive immune responses, leading to improved resistance to infections. They
stimulate the production of antibodies and modulate immune cells like
macrophages and T lymphocytes, making the body more efficient in fighting
off pathogens.

o Reduction of Inflammation: Probiotics may help in reducing systemic

inflammation associated with chronic conditions like rheumatoid arthritis,
obesity, and cardiovascular diseases.

3. Mental Health Benefits

o Gut-Brain Axis: There is growing evidence suggesting that probiotics may
benefit mental health through the gut-brain axis. Probiotics can influence the
production of neurotransmitters like serotonin and gamma-aminobutyric acid
(GABA), which can reduce symptoms of depression, anxiety, and stress.

4. Improved Lactose Digestion

o Lactose Intolerance: Probiotics can help individuals with lactose intolerance
by enhancing the breakdown of lactose in the gut. Certain probiotic strains,
such as Lactobacillus acidophilus, produce lactase, the enzyme required for
lactose digestion.

5. Allergy Management
o Modulation of Allergic Responses: Probiotics have been shown to reduce the
symptoms of allergies, including eczema and hay fever, by modulating
immune responses and enhancing the gut’s defense mechanisms. This is
particularly beneficial in children with a predisposition to allergies.

6. Cholesterol Reduction
o Lowering Cholesterol Levels: Certain probiotic strains, such as
Lactobacillus reuteri, can help lower blood cholesterol levels by breaking
down bile salts in the intestine, which reduces the reabsorption of cholesterol
into the bloodstream.

7. Enhancement of Nutrient Absorption

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 o Increased Nutrient Bioavailability: Probiotics may enhance the absorption
of essential nutrients, including minerals like calcium, magnesium, and iron,
thus supporting bone health and reducing the risk of deficiencies.

REGULATIONS AND SAFETY OF PROBIOTICS

The safety and efficacy of probiotics are of great concern, as they are consumed as
functional foods or dietary supplements. Regulatory frameworks for probiotics vary from
region to region, but they generally involve the following principles:

1. Regulatory Bodies and Guidelines

 United States (FDA): In the U.S., probiotics are generally considered dietary
supplements and are not subject to the same strict regulations as drugs. However, the
FDA monitors claims made by manufacturers and ensures that products are safe and
meet labeling requirements. Probiotics are not required to prove their efficacy before
being sold, but manufacturers must ensure their products do not cause harm.
 European Union (EFSA): In Europe, probiotics are regulated more rigorously. The
European Food Safety Authority (EFSA) evaluates health claims related to
probiotics and requires scientific evidence to substantiate claims of health benefits.
Products claiming health benefits must undergo safety evaluations and clinical studies
to prove their efficacy.
 World Health Organization (WHO) and FAO: The WHO and the FAO have set
guidelines for the use of probiotics, including criteria for safety, such as the
requirement that probiotic strains must be non-pathogenic, stable, and capable of
survival in the gastrointestinal tract.

2. Safety Standards for Probiotics

 Strain-Specific Safety: Only specific strains of microorganisms can be classified as
probiotics. Strains should be well-characterized, and their safety should be established
through clinical trials, focusing on their ability to not cause infections or produce
harmful byproducts.
 Monitoring and Reporting: Adverse effects, if any, must be reported by consumers
and healthcare providers. Common issues might include bloating, gas, or mild
digestive disturbances, but severe side effects are rare.

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3. Health Claims and Marketing
 Health Claims: Regulatory bodies require that any health claims made by probiotic
manufacturers must be substantiated by scientific evidence. For example, claims that
probiotics can treat or prevent diseases such as irritable bowel syndrome or infections
must be backed by clinical studies. False claims are subject to penalties and removal
from the market.
 Labeling: Probiotic products must be clearly labeled with the strain(s) used, the
dosage, and storage requirements. Some regions also require that the product clearly
state whether it is a supplement or a food product.

4. Safety Concerns and Quality Control

 Quality Control: To ensure the safety and efficacy of probiotics, manufacturers must
adhere to Good Manufacturing Practices (GMP), ensuring that products are
consistently produced, stored, and handled to maintain quality.
 Potential Risks: While probiotics are generally safe for most people, they may not be
suitable for individuals with compromised immune systems (e.g., those with HIV,
cancer, or organ transplants). In such cases, the use of probiotics should be supervised
by a healthcare provider.

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