Chapter

Introduction of Food

1
Microbiology
Introduction
Food microbiology is the study of
the microorganisms that inhabit, create,
• Introduction
• How Microorganisms
or contaminate food.
Cause Deterioration 1. This includes the study of microorganisms
• Preventing
Deterioration
causing food spoilage;
• The importance of 2. Pathogens that may cause disease
microorganism in food
• Good (Desirable)
3. Microbes used to produce fermented
• Bio preservation foods such as cheese, yogurt, bread, beer,
• Bio processing
• Probiotics
and wine; and
• Bad (Undesirable) 4. Microbes with other useful roles, such as
• Food borne
diseases
producing probiotics.
• Food spoilage Since our food supply primarily consists of plants
• Probiotics and Prebiotics
and animals or products derived from them, it is
common for microorganisms to interact with our
food. They often use food as a source of nutrients
for their own growth.
How Microorganisms Cause Deterioration:
These microorganisms alter the food in a few
ways.
5. They can synthesize new compounds that
produce strange flavours.
6. Break down (Enzymatic) existing compounds,
leading to off-putting tastes and odours.
Preventing Deterioration:
7. Minimize Contact
8. Eliminate Microorganisms: Use preservation
techniques such as pasteurization or canning to eliminate or
reduce the number of microorganisms in the food.
9. Understand Preservation
The importance of microorganism in food
Good (Desirable)
1. Food Bioprocessing: Microorganisms play a positive role in food
bioprocessing, where they are used for fermentation, production of various
foods and beverages (e.g., yogurt, cheese, bread), and enhancing the
flavour and nutritional value of food. Microbial enzymes are employed in
the production of food and food additives.

2. Food Bio preservation: Beneficial microorganisms, like certain lactic acid

bacteria, can be used for food preservation by producing organic acids and
other compounds that inhibit the growth of spoilage and pathogenic
microorganisms.

3. Probiotics: Certain microorganisms, known as probiotics, provide health

benefits when consumed as they support the gut microbiota, enhance
digestion, and boost the immune system.
• Probiotics are live microorganisms that are beneficial for human
health, particularly for digestive system and body’s natural immunity.
• Advantages:
1. Improve digestion system.
2. Support immune system.
3. Increase the absorption of calcium, in the prevention of
osteoporosis.
4. Preventing intestinal tract infections that are caused by candida
spp and Helicobacter pylori.
5. Cleaning the colon and improve constipation.
• Side effects of probiotics to human
• Diarrhea, Constipation, Bloating, Headache, Excessive drainage
syndrome.
Bad (Undesirable)
4. Foodborne Disease: Harmful microorganisms, such as bacteria, viruses,
and parasites, can lead to foodborne diseases when they contaminate food.
This can result in various illnesses, including gastroenteritis, food
poisoning, and more.
2. Food Spoilage: Microorganisms can cause food spoilage by breaking down
the components of food, resulting in undesirable changes in taste, texture,
and appearance. This can lead to food wastage
• Spoilage is the process by which food products become unsuitable for
consumption due to the growth of microorganisms or oxidation. This
can result in changes in the colour, texture, taste, and smell of the
food, as well as the production of toxins that can cause illness in
humans.
1. Microbial spoilage: This occurs when microorganisms such as
bacteria, yeast, or mould grow on the food product, leading to
changes in the colour, texture, taste, and smell of the food. This
can result in the production of toxins that can cause illness in
humans.
2. Chemical spoilage: This occurs when chemical reactions, such as
oxidation of fat or hydrolysis, occur in the food product, leading
to changes in the colour, texture, taste, and smell of the food.
This type of spoilage can be caused by exposure to light, heat, or
air.
3. Physical spoilage: This occurs when the food product is
physically damaged, leading to changes in the colour, texture,
taste, and smell of the food. This can be caused by bruising,
crushing, or cutting of the food product.
Sequence of events in food spoilage
• Microorganisms had to get into food from a source.
• Food environment should favour the growth of MO.
• Food need to store in a favourable condition.
• To allow sufficient number necessary to cause spoilage or change
in food
• To allow the produced enzyme to spoile the food
 Chapter

2
Food spoilage
Introduction
Food spoilage: Spoilage is the process by which food
• Introduction products become unsuitable for consumption due to the
• Factors affecting the growth of microorganisms or oxidation. This can result
development of
microorganisms in foods
in changes in the colour, texture, taste, and smell of the
• Microbial Spoilage of Foods food, as well as the production of toxins that can cause
• Putrefaction illness in humans.
• Fermentation
• Rancidity
Types of spoilage
• Fruits and vegetables 1. Physical spoilage: Due to physical damage.
• Microbial Spoilage of 2. Enzymatic spoilage: Break down of food by enzyme
Fruits and Vegetables
• Spoilage of Fruit and
3. Microbial spoilage
Vegetable Juices 4. Chemical spoilage
• Alcoholic Fermentation Factors affecting the development of microorganisms
of Fruit Juices
• Vegetable Juices
in foods
• Meat and meat products • Synergism: Synergism is a type of ecological
• Factors Affecting interaction where two or more species or organisms
Microbial Growth in
Meat:
work together, and their combined actions result in a
• General Types of Meat more beneficial outcome than if they were acting
Spoilage independently.
• Meat Spoilage Under
Aerobic Conditions by
• Antagonism: Antagonism is an ecological interaction
Bacteria, yeast and where two or more species or organisms actively
mould work against each other, resulting in negative effects
• Meat Spoilage Under
Anaerobic Conditions
on one or both parties.
• Poultry Meat Spoilage • Commensalism: Commensalism is an ecological
• Milk and milk product interaction where one species benefits from the
• Egg and egg products
relationship, while the other is neither significantly
harmed nor benefited.
 Intrinsic factor Extrinsic factor Implicit factor Processing factor

1. Nutrient 1. RH 1. Synergism 1. Irradiation

2. pH 2. Temperature 2. Antagonism 2. Washing
3. Water activity 3. Gasious 3. Commensalism 3. Slicing
4. OR-Potential atmosphare 4. Growth rate 4. Pasteurization
5. Ant-microbial 5. Packaging
constituents

Microbial Spoilage of Foods

Many different microorganisms can thrive in various types of foods, serving
as suitable growth media.
1. Putrefaction: In protein-rich foods, the action of proteolytic microbes
leads to the breakdown of proteins into amino acids, amines, ammonia,
and the production of hydrogen sulphide (H2S). This process is associated
with the decomposition and foul odours of protein-based foods.

2. Fermentation: In carbohydrate (CHO)-rich foods, saccharolytic microbes

break down carbohydrates, resulting in the production of organic acids,
alcohol, and gases. Fermentation is commonly used in the production of
various foods and beverages, such as bread, yogurt, and beer.

3. Rancidity: In fatty foods, lipolytic microbes can lead to the breakdown of

fats, resulting in the production of fatty acids and glycerol. This process
can cause fats and oils to become rancid, affecting taste and quality

Fruits and vegetables

Fruits and vegetables can deteriorate due to physical factors, the action of
their own enzymes, microbial action, or a combination of these factors.
Oxygen availability also plays a role in their deterioration.
Microbial Spoilage of Fruits and Vegetables (Cause)**
• Microbial spoilage of fruits and vegetables can be caused by various
microorganisms, including plant pathogens and saprophytic organisms that
act as secondary invaders, such as coliform bacteria.
• Fungal spoilage of vegetables can result in water-soaked mushy areas and
dry rots.
Spoilage of Fruit and Vegetable Juices
• Fruit and vegetable juices can be obtained by methods such as direct
squeezing, maceration or crushing, and water extraction. (How juice
extract)
• These juices may be used in natural concentration or concentrated by
evaporation or freezing. (Processing)
• Preservation methods for juices include canning, freezing, or drying.
(Preservation)
• Post-processing microorganisms, including moulds, bacteria, and yeasts,
can affect the quality and safety of juices.
• Yeasts are active in a temperature range of 15.6 to 35°C.
Alcoholic Fermentation of Fruit Juices
• Alcoholic fermentation of fruit juices can occur, with wild yeast species
like apiculate yeasts carrying out the first fermentation.
• The temperature range for yeast activity is 15.6 to 35°C.
• Above 32.2 to 35°C, lactobacilli can grow, and below 15.6°C, wild yeast
may grow.
• Fruit juices may undergo lactic acid fermentation of sugars by
heterofermentative and homofermentative lactic acid bacteria, as well as
other organic acid fermentations.
• Slime production by bacteria can occur in grape juice.
Vegetable Juices
• Vegetable juices generally have less sugar than fruit juices.
• Their pH ranges from 5.0 to 5.8, making them conducive to the growth of
fastidious lactic acid bacteria.
Meat and meat products
Raw meat can undergo changes due to various factors, including microbes
during slaughter, cutting, and processing, enzyme action, and fat oxidation.
Factors Affecting Microbial Growth in Meat:
• The invasion of tissues by microbes depends on factors such as the
microbial load in the gut of the animal, the physiological condition of the
animal immediately before slaughtering (pH levels), the method of
slaughtering, and the rate of cooling. (Invasion of tissue)
• Other factors affecting microbial growth in meat include Microbial
contamination, physical and chemical properties, the availability of
oxygen, temperature, and anaerobic conditions. (Growth)
General Types of Meat Spoilage
Meat can spoil under aerobic and anaerobic conditions, leading to various
types of spoilage.
Meat Spoilage Under Aerobic Conditions by Bacteria
Aerobic bacteria can cause surface slime, changes in meat colour (oxidizing
compounds), changes in fat, phosphorescence, various surface colours due to
pigmented bacteria, off odours, and off tastes.
Meat Spoilage Under Aerobic Conditions by Yeasts:
Yeasts can lead to sliminess, lipolysis, off odours, off tastes, and discolorations
(white, cream, pink, or brown).
Meat Spoilage Under Aerobic Conditions by Moulds
Moulds can cause stickiness, whiskers (white fuzzy growth), and various
coloured patches. They can also lead to the decomposition of fats and produce
off odours and off tastes.
Meat Spoilage Under Anaerobic Conditions (Facultative and Anaerobic
Bacteria):
• Anaerobic conditions can result in
• Souring, the action of meat enzymes during aging, anaerobic
production of fatty acids or lactic acid, proteolysis without
putrefaction (acid and gas formation by butyric Clostridium sp.),
• Putrefaction (foul-smelling compounds like hydrogen sulphide,
indole, skatole, ammonia, and amines by Clostridium sp.),
• Taint (souring or putrefaction).
Meat and meat products
Poultry Meat Spoilage
• Poultry meat can spoil for several reasons, including the action of bacteria
and enzymes.
• The primary source of spoilage bacteria is the intestine of the poultry.
• Surface odours become perceptible when the bacterial count on the skin
reaches about 2.5 million per square centimetre.
• Eviscerated poultry stored at temperatures of 10°C or below mainly spoils
due to Pseudomonas bacteria and yeasts.
• When stored above 10°C, Alcaligenes and Flavobacterium become more
involved in spoilage.
Milk and milk products
Factors Unfavourable for Milk spoilage
Milk and milk products can be unfavourably affected by factors like storage at
low temperatures, the destruction of lactic bacteria by heat, and neutralization
of acids.
Changes Produced by Proteolytic Microorganisms
Proteolytic microorganisms can bring about various changes in milk, including
acid proteolysis, sweet curdling, slow proteolysis by bacteria's enzymes.
Ropiness in milk
Nonbacterial ropiness (cream thickness, casein)
Bacterial ropiness (Alcaligens viscolactis)
Spoilage of Milk at Different Temperatures:
• Milk can spoil under different temperature conditions. At refrigerator
temperatures, psychrotropic bacteria like Pseudomonas can initiate
proteolysis, followed by the appearance of moulds.
• At room temperature, acid fermentation by bacteria occurs, along with the
presence of moulds and film yeasts.
Egg and egg products
Defects in Fresh Eggs:
• Cracks and leaks in eggshells can lead to spoilage.
• Loss of the natural protective "bloom" or gloss on the eggshell.
• Dirty spots on the exterior of the egg.
• Presence of blood clots in the interior of the egg.
• General bloodiness or translucent spots in the egg yolk.
Spoilage of Eggs Due to Microbial Causes:
• Microorganisms (MOs) can penetrate the pores of the eggshell to reach the
shell membrane.
• They then grow in the shell membrane and egg white.
• Eventually, they reach the egg yolk and lead to rapid spoilage.
Types of Bacterial Spoilage or Rots of Eggs:
• Bacterial spoilage of eggs can manifest in different types, including green
rots, colourless rots, black rots, red rots, pink rots, and off-flavours like
mustiness, earthy flavour, or fishy taste.
Fungal Spoilage of Eggs:
• Fungal spoilage of eggs can occur on the surface (superficial fungal spoilage) or lead to fungal rotting.
• Spoilage in fresh eggs can result from various factors, including physical defects and microbial contamination,
leading to changes in appearance, texture, and flavour.
Fish and other sea foods
Factors Affecting Spoilage:
• Type of Fish: The species of fish can influence its susceptibility to
spoilage.
• Condition When Caught: The freshness of the fish at the time of
capture is crucial.
• Contamination: The degree of contamination of fish flesh with
bacteria plays a role in spoilage.
• Temperature: Storage temperature significantly affects the rate of
spoilage.
• Discolorations: Different bacteria and microorganisms can cause
various discolorations in fish flesh.
Mechanism of Spoilage:
• Surface Growth: Bacteria, particularly Pseudomonas, initially grow on
the surface of the fish and then penetrate the flesh.
• Biochemical Changes: Fish flesh contains nitrogenous compounds
(amino acids, amines) and glucose. Bacteria metabolize these
compounds to produce trimethylamine, ammonia, amines, aldehydes,
and hydrogen sulphides, indicative of putrefaction.
• Odour and Taste: Musty or muddy odours or tastes in fish can result
from the growth of Streptomyces species in the environment, with the
flavour being absorbed by the fish.
 Chapter
Food as a Substrate for

3
Microorganisms
Introduction
1. Natural Interactions: Microorganisms play essential
roles in natural ecosystems, participating in various
• Introduction geochemical cycles. They help convert organic
• Intrinsic factor matter from dead plants and animals into forms that
• Hydrogen-ion plants can use, thereby contributing to nutrient
concentration (ph) cycling in nature.
• Moisture 2. Microorganisms and Food: Since our food supply
requirement: the primarily consists of plants and animals or products
concept of water derived from them, it is common for microorganisms
activity to interact with our food. They often use food as a
• Oxidation- source of nutrients for their own growth.
reduction 3. Food Spoilage: Microorganisms can cause the
potential deterioration of food by increasing their numbers,
• Nutritional utilizing nutrients, producing enzymatic changes,
requirement and generating off-flavours through breakdown or
• Antimicrobial synthesis of compounds. This is a normal
Substances consequence of their activity in nature.
• Extrinsic factor
• Temperature Factors of a food that influence microbial activity are
• Gases hydrogen-ion concentration, moisture, oxidation-
• RH reduction (O-R) potential, nutrients, and the presence of
inhibitory substances or barriers.
To prevent this we minimize the contact between
microorganisms and our foods (prevent contamination)
and also eliminate microorganisms from our foods, or at
least adjust conditions of storage to prevent their growth
(preservation). A knowledge of the factors that favour or
inhibit the growth of microorganism is essential to an
understanding of the principles of food spoilage and
preservation. The chief compositional
Hydrogen-ion concentration (ph)
• Every microorganism has a minimal, a maximal, and an optimal pH for
growth. Microbial cells are significantly affected by the pH of food
because they apparently have no mechanism for adjusting their internal
pH.
• Foods with low pH values (below 4.5) usually are not readily spoiled by
bacteria and are more susceptible to spoilage by yeasts and molds.
• A food with an inherently low pH would therefore tend to be more stable
microbiologically than a neutral food.
• Most fermentative yeasts are favored by a pH of about 4.0 to 4.5
• Most bacteria are favored by a pH near neutrality, although some, such as
the acid formers, are favored by moderate acidity
• The actively proteolytic bacteria, can grow in media with a high (alkaline)
pH, as found in the white of a stored egg.
• Vegetable juices have low buffering power, permitting an appreciable
decrease in pH with the production of only small amounts of acid by the
lactic acid bacteria.
• Milk, on the other hand, is fairly high in protein (a good buffer) and
therefore permits considerable growth and acid production by lactic acid
bacteria in the manufacture of fermented milks before growth of the starter
culture is finally suppressed.
• The inhibitory properties of many of the organic acids, including acetic,
benzoic, citric, lactic, proprionic, and sorbic acids, make them widely used
as acidulants or preservatives in foods. Also, changes in titratable acidity
are not always evident from pH measurements.
• Immediately after slaughter the pH of most beef muscles is 6.9 to 7.4 and
24hr. post-mortem is in the range of 5.6 to 6.4
• The pH of post slaughter chicken muscle is approximately 7.0
Moisture requirement: the concept of water activity
Microorganisms have an absolute demand for water, for without water no
growth can occur. As might be expected, the exact amount of water needed
for growth of microorganisms varies. This water requirement is best
expressed in terms of available water or water activity a w,

Water is made unavailable in various ways:

• Solutes and Ions: High concentrations of solutes (e.g., sugars, salts) tie up
water in a solution, reducing its availability. Osmosis can also cause water
to leave microbial cells when solute concentration is higher outside the
cells.
• Hydrophilic Colloids (Gels): Hydrophilic colloids, like agar, can limit
available moisture and hinder microbial growth even in small amounts.
• Water of Crystallization or Hydration: Water molecules in crystalline
structures (e.g., ice) are unavailable to microorganisms. As ice forms, the
water activity (aw) decreases due to increased solute concentration in the
unfrozen water. Lower aw inhibits microbial spoilage and chemical
reactions in food.
Most spoilage bacteria do not grow below aw of 0.91, while spoilage molds
can grow as low as 0.80. With respect to food poisoning bacteria,
Staphylococcus aureus has been found to grow as low as 0.86, while
Clostridium botulinum does not grow below 0.95.

Oxidation-reduction potential
O-R potential, or reducing and oxidizing power of the food itself, influence the
type of organisms which will grow and hence the changes produced in the
food.
• O-R Potential: The reducing and oxidizing power (O-R potential) of the
food itself is a crucial factor. It is determined by:
1. The inherent O-R potential of the original food.
2. The food's poising capacity, which reflects its resistance to changes in
O-R potential.
3. The oxygen tension in the surrounding atmosphere.
4. The degree to which the atmosphere can access the food.
• O-R Potential Influence:
1. A high O-R potential (oxidizing) favours the growth of aerobes but
can also permit facultative organisms to grow. Positive
2. A low O-R potential (reducing) is favourable for anaerobic or
facultative organisms. Negative
3. Growth of one organism can alter the O-R potential of the
environment, potentially inhibiting the growth of other organisms. For
example, anaerobes can reduce the O-R potential to levels that are
inhibitory to aerobes.
• Specific Microorganism Preferences
• Aerobic microorganisms like bacilli, micrococci, pseudomonads, and
Acinetobacter require positive Eh values or positive mV O-R
potentials.
• Anaerobic microorganisms like clostridia and Bacteroides thrive under
negative Eh values or negative mV O-R potentials.
• Aerobic Conditions Near the Surface: Fresh meat or whole fruits typically
have aerobic conditions near their surfaces, where oxygen is readily
available. This can support the growth of aerobic bacteria. However, deeper
layers may have anaerobic conditions, leading to putrefaction.
• Impact of Processing: Processing procedures, such as heating, can alter the
microbial environment within food:
1. Heating can reduce the O-R potential by destroying or altering
reducing and oxidizing substances.
2. It can also facilitate the diffusion of oxygen into the food, either by
destroying poising substances or by changing the food's physical
structure.
3. Processing may remove oxidizing or reducing substances. For
example, clear fruit juices lose reducing substances during extraction
and filtration, making them more favourable for the growth of yeasts.
• O-R Potential (Eh) in Foods
1. Plant foods, especially plant juices, tend to have positive Eh values
ranging from +300 to +400 mV, making them prone to spoilage by
aerobic bacteria and molds.
2. Solid meats have negative Eh values around -200 mV, while minced
meats typically have positive Eh values around +200 mV.
3. Cheeses of various types can have negative Eh values ranging from -
20 to around -200 mV.
Nutritional requirement
Microorganisms in food require various nutrients to grow and function
optimally. These include water, sugars (carbohydrates), proteins, fats, vitamins,
minerals, and related factors.
• Fats serve as an energy source when other options are scarce, with aerobic
microorganisms more involved in fat decomposition.
• Proteins and amino acids are used when better energy sources are lacking,
particularly by proteolytic organisms found in protein-rich foods like meats.
• pH plays a role, as proteolytic bacteria prefer neutral pH and are inhibited
by acidity.
• Microbes use more carbohydrates in the presence of sufficient nitrogen.
• Molds thrive in high sugar concentrations, yeasts in moderate levels, while
most bacteria prefer low sugar concentrations.
• Nutritional Differences:
1. Gram-positive bacteria often require accessory food substances or
vitamins for growth.
2. Gram-negative bacteria and molds can synthesize most or all of their
growth requirements.
• Specific Microbial Abilities:
1. Staphylococcus aureus, a Gram-positive bacterium, can synthesize
some accessory growth factors or vitamins.
2. Pseudomonas and E. coli, which are Gram-negative bacteria, can
synthesize nearly all the factors they need.
3. Lactic acid bacteria and many pathogens require external sources of
growth factors.
• Medium Quality and Growth Range:
1. The better the medium (nutritionally rich) for a microorganism, the
wider the range of temperature, pH, and water activity (aw) in which
it can grow.
• Food Composition:
1. Meats are rich in B vitamins, whereas fruits are lower in B vitamins
but high in ascorbic acid (vitamin C).
2. Fruits, due to their low B vitamin content, low pH, and positive
oxidation-reduction potential (En), tend to spoil more often due to
mold growth rather than bacterial growth.
• Egg White and Biotin:
• Egg white contains biotin but also has avidin, which binds to biotin
and makes it unavailable to microbes requiring biotin for growth.
• Impact of Food Processing and Storage:
1. Food processing can lead to a reduction in vitamin content.
2. Long-term storage, especially at elevated temperatures, may also
result in a decrease in the levels of accessory growth factors in foods.
Antimicrobial Substances: Foods naturally contain substances with
antimicrobial activity, which can inhibit the growth of microorganisms.
Some of these antimicrobial substances are inherent in the food, while others
may be intentionally added or produced during food processing and microbial
growth.
• Examples of Naturally Occurring Inhibitors:
1. Freshly drawn milk contains lactenins and anticoliform factors, which
have antimicrobial properties.
2. Eggs contain lysozyme, which possesses antimicrobial activity.
3. Goat milk has a higher lysozyme content compared to cow or sheep
milk.
4. Human milk has an exceptionally high lysozyme content, making it a
potent antimicrobial agent.
5. Spices like cloves (containing eugenols) and cinnamon (containing
cinnamaldehyde), as well as garlic (containing allicin), are rich in
antimicrobial constituents.
• Microbial Production of Inhibitory Compounds:
1. Microorganisms growing in food can produce substances that inhibit
the growth of other microorganisms, which can contribute to food
preservation.
2. Conversely, some microorganisms may degrade or destroy inhibitory
compounds in food.
• Impact of Heating and Food Processing:
1. Heating food can lead to the formation of inhibitory substances.
2. Heating lipids (fats) may accelerate auto oxidation, which can
generate antimicrobial compounds.
3. Concentrating sugar syrups can result in the production of furfural
and hydroxymethyl furfural compounds, which have inhibitory
effects on fermenting microorganisms.
 Chapter
Thermal processing of food

•
•
4Introduction
Commercially sterile food
• D-Value (Decimal
Introduction
Thermal processing of food involves the use of
heat to treat food products in order to achieve
specific objectives, such as preservation, pathogen
inactivation, or the alteration of food properties.
reduction time) Commercially sterile food
• Characteristics of D- Commercial sterilization is to attain a degree of
value sterility in the product being processed so that it
• D-value and slope
• Z-value (Thermal will not undergo spoilage and not be a public
resistance constant) health hazards.
• Thermal Death D-Value (Decimal reduction time): The D-value,
Time, F
• Expression for TDT
in the context of sterilization, represents the time
(F) or the equivalent conditions required to reduce the
• Spoilage Probability population of microorganisms in a product by a
• 12 D concept or Bot
factor of ten (or 90% reduction). (1 logarithmic
Cook
cycle).
• If a homogenious suspension of MO heated at
a constant temperature, the destruction of MO
commonly follows a logarithmic order of
death.
Number of MO

𝑇𝑖𝑚𝑒
So D value is the time required to reduce the population of MO at constant
temperature by
• A factor of 10
• A logarithmic cycle
• 90% of initial population
• 2D value is the time required to reduce the population of MO by 2
loagrithmic cycle or 99% reduction.
• 3D value is the time required to reduce the population of MO by 3
loagrithmic cycle or 99.9% reduction.
Characteristics of D-value:
• Indicates heat resistance of a MOs
• Remain the same for each log cycle
• It does not depends on the initial No of MOs
• Unit is time, like (Min)
• It is always 90% reduction in the existing MOS present in the system
• Time required to decrease MOS from 10,000 to 100, it will be 2D.
• D-value is 1 min at 85 0C; what does it mean? For each min processing at
85℃, MOs will reduce by 90% (1 log cycle)
D-value and slope
log(N)
7
6
f(x) = − 0.330573820989452 x + 6.03167249841905
5
4
3
2
1
0
0 2 4 6 8 10 12 14
Z-value (Thermal resistance constant)
Z value describe the influence of tmeperature on decimal reduction time or D
value.
• Z-value is the increase in tmeperature necessary to reduce the D value
by 90%.

Thermal Death Time, F

F value is the time required to cause a stated reduction in a population of MOs
or spores at a given temperature. It is also called Thermal Death Time (TDT).
• F0 is the no of minutes required to destroy a given no of microbial
population at 121.1 0C when z = 10 0C.
Expression for TDT (F)

TDT
𝐷𝑇
F is expressed as with 2
superscript and a subscript
10
D value

10 Z value
𝐷121.1

𝑇 121.1
Spoilage Probability
The spoilage probability is used to estimate the number of spoiled containers
within a total batch of processed product. We know that
• Thermal destruction/ inactivation is a logarithmic system.
• The survivor curve never reaches to zero.
Time N 2
10
0 102
101
𝐷
1D 101
10 0 𝑡
2D 100
−1
10
3D 10-1
10− 2
4D 10-2

From the right angle triangle

• Let r = No of can processed

• N0 = No. of spores/can
• r×N0 = total no of spores at the beginning of the process
• F = Total time of heating

If we want only one can at the end of heating to contain a spore, then

The ratio on the left side of Equation represents the total number of containers processed (r) and resulting in one
container with spoilage. The expression can be used to estimate the thermal death time required to accomplish a stated
spoilage probability, based on knowledge of the initial population and the decimal reduction time, D, for the microbial
population. It should be noted that the spoilage probability expression does assume that the survivor curve for the
spoilage microorganism follows a first-order model.
12 D concept or Bot Cook
Heat treatment is based on the most heat-resistant MOs related to health
hazards or spoilage. Thermal destruction is logarithmic and complete
destruction is impossible.
Probability of survival can be reduced to a small value. What should be the
level?

• Esty and Myer (1922) carried an experiment on spores/mL of Clostridium

botulinum
• At 121 0C reduced no to 1 survivor in 2.78 min
• Townsend (1938) corrected it to 2.45 min
For C. botulinum,
Z= 10 ℃, Temperature = 121 ℃; the reference F value ,

For Cl botulinum, a thermal process that is equivalent to F 0 is called Bot Cook.

1. The F value at 121.1◦C equivalent to 99.999% inactivation of astrain of
C.botulinum is 1.2 minutes. Calculate the D 0 value of this organism.
99.999% reduction is equal to 5 decimal reduction of C.botulinum.

Sterilizing Value or Lethality of a Process

S = number of decimal reduction =
1. Calculate F0 based on the 12D concept using the D0 value of C. botulinum
having D0 0.24 min and a most likely spore load in the product of 100.

2. The sterilizing value of a process has been calculated to be an F 0 of 2.88.

If each can contained spores of an organism having a D 0 value of 1.5 min,
Calculate the probability of spoilage from this organism. Assume the F 0
value was calculated using the same z value as the organism.

Since , then spoilage of 12 container in 100 can be expected.

3. The most probable spore load in a can food is 100 and the D 0 of the spore
is 1.5 minutes. Calculate a target F0 for a thermal process such that the
probability of spoilage is 1 in 100000. If under the same conditions C.
botulinum type B has a D0 of 0.2 min, would the target F0 value satisfy the
minimum 12D process for C. botulinum? Assume an initial spore load of
1 per can for C. botulinum.

Again for second condition

4. Estimate the spoilage probability of a 50-minute process at 113 0C when

D113 = 4 minutes and the initial microbial population is 10 4/container.

Since , then spoilage of one container in can be expected.

 Chapter
Indicator Organisms

•
5Microbial Quality &
Safety of Foods
• Quality Indicators
Microbial Quality & Safety of Foods
Microorganisms or their metabolites are used as
indicators to assess the microbial quality of foods.
The types and levels of these microorganisms can
provide insights into the overall quality of the
• Shelf-Life, and product.
Safety • Quality Indicators: The presence of specific
• Emphasis on Food microorganisms or their metabolic by-products
Safety
• Indicator vs index can serve as indicators of product quality. For
organisms example, the presence of certain lactic acid
• Criteria of Indicator bacteria in fermented foods like yogurt can
MOs
• Total and
indicate the quality of the fermentation
fecal coliform process.
• E. coli • Shelf-Life, and Safety: Microorganisms can
• Enteroibacteri
not only indicate existing quality but can also
aceae
• Coliform Criteria and help predict the shelf-life and safety of food
Standards products. For instance, the presence of certain
• Enterococci spoilage microorganisms might suggest a
• Relationship to
Sanitary Quality of
shorter shelf-life, while the absence of
Foods pathogenic microorganisms can indicate safety.
• Bifidobacterium • Emphasis on Food Safety: Microorganisms and
their metabolites are often employed more to
assess food safety and sanitation than to
evaluate quality.
Indicator vs index organisms
Aspect Indicator Microbes Index Microbes
Specific microorganisms
Specific microorganisms or groups chosen to represent the
Definition chosen to represent the general presence of ecologically
microbiological condition of food. similar pathogens or
conditions in food.

Used as surrogates to assess

Assess the general hygiene, processing,
food safety. Often more
Purpose and quality of food. Indicate whether
resistant or easily detected
sanitation standards are met.
than target pathogens.
Enterobacteriaceae for
Total coliforms, fecal coliforms, E.
Examples potential Salmonella
coli, Enterobacteriaceae, etc.
contamination, for instance.
The presence can indicate a
High levels may suggest poor
potential risk of pathogenic
sanitation practices or potential
Use in Food Safety contamination, and
contamination, raising quality and
appropriate measures can be
safety concerns.
taken to ensure food safety.

Criteria of Indicator MOs

• Present and detectable in desirable foods
• Growth and number should have a direct negative correlation with product
quality.
• Easily detectable
• Enumerable in short time
• Growth should not be affected by other flora in foods.
• Nonpathogenic
Examples of Indicator Mos
1. Total and Fecal Coliform
• Hygiene and Sanitation Indicator: Total and Fecal Coliforms are
classic indicator microorganisms used to assess the hygiene and
sanitation conditions in food processing environments.
• Historical Use in Water and Food: Initially, Total and Fecal Coliforms
were primarily used as indicators of fecal pollution in water sources.
2. E. coli
• Escherich (1885) identified a bacterium he initially named "Bacterium
coli commune." Today, this bacterium is known as Escherichia coli (E.
coli). E. coli has become a well-known indicator microorganism, and
certain strains can be used as a marker for fecal contamination and
sanitation issues in food.
3. Enterobacteriaceae
• Broad Family of Bacteria: Enterobacteriaceae is a large family of
bacteria that includes various species.
• Growth Conditions: Enterobacteriaceae species typically thrive under
conditions where E. coli can also grow.
• Detection Methods: Various detection methods are used to identify and
quantify Enterobacteriaceae in food.
Coliform Criteria and Standards
For Water:
• Lowest Possible and Feasible Number: Ideally, coliforms should be
absent in drinking water.
• Unsafe Level: Any presence of coliforms in drinking water is generally
considered unsafe.
For Dairy Products:
• Grade A Milk and Milk Products: Not over 10/mL.
• Certified Raw Milk: Not over 10/mL.
• Pasteurized Milk: Not over 1/mL.
• Precooked and Partially Cooked Frozen Foods: Not over 10/mL.
• Crabmeat: Not over 100/mL.
• Custard-Filled Items: Not over 100/mL
Limitation for food safety use
• Varied Suitability: Coliform tests are good indicators for milk and
water but may not be suitable for all food types.
• Dairy Industry: In dairy, coliform tests assess overall and plant
sanitation, not fecal contamination.
• Frozen Vegetables: E. coli presence in frozen blanched vegetables can
indicate processing issues but not necessarily pathogenic risk.
• Poultry: Coliforms are less useful in poultry due to the presence of
more concerning pathogens like salmonellae.
• Meat: Standard coliform tests may not be suitable for meat; specific criteria are often
needed.
• Shellfish: Coliform tests are widely used in shellfish sanitation to assess water quality and
potential fecal contamination.
Eneterococci
• Water Persistence: Enterococci persist in water, especially in low-
organic-matter conditions.
• Fecal Indicators: They indicate potential fecal contamination, often
used in water quality assessment.
• Comparison with E. coli: A Coli: Ent ratio of 4 or higher suggests
human waste contamination.
• Slow Die-Off Rate: Enterococci have a slower die-off rate in the
environment.
• Enterococci are hygiene and sanitation indicators in fermented dairy
products.
• They are gram-positive cocci and facultative anaerobic bacteria.
• Enterococci can tolerate a wide range of environmental conditions.
• Their use in cultured dairy products has limited research support.
Relationship to Sanitary Quality of Foods
• Good indicator of food sanitary quality for frozen foods
• Prior freezing, coliform is better indicator of sanitation
• Survival at freezing (-20 ℃) is higher of Enterococci than coliform
Bifidobacterium
• Classification: Bifidobacterium bifidum is a gram-positive, anaerobic,
catalase-negative, non-motile rod-shaped bacterium.
• Optimal pH: It thrives in environments with a pH range of 5 to 8.
• Metabolites: Bifidobacterium bifidum produces lactic and acetic acids as
metabolites.
• Indicative of Fecal Pollution: It is used as an indicator of fecal pollution,
particularly in water sources.
• Use in Fermented Foods: Some bifidobacteria are used in the production of
fermented foods, contributing to their flavour and texture.
• In hunan:10 to 100 times higher in human feces deposited in water than
coliform and enterococci
 Chapter
Food contamination

•
6Introduction
• Factors affecting
kinds and numbers
Introduction
We get foods from different sources. All foods
carry MOs from natural sources
Further contamination begins and continues
during when they handled and processed.
of microorganisms Factors affecting kinds and numbers of
in food microorganisms in food
• Cereal and Cereal Products • Source environment
• Microorganisms on
Harvested Grains • Microbes in the raw/unprocessed products
• Microorganism • Sanitary conditions during handling and
Families processing
• Microorganism
Counts in Wheat
• Adequacy of the packaging, handling and
Flour storage conditions
• Microorganisms in Contamination of Cereal and Cereal Products
Cornmeal, Flour,
Microorganisms on Harvested Grains: The
and Malts
• Mycotoxin exteriors of harvested grains can retain
• Sugars and Sugar Products microorganisms from their growth environment
• Vegetables and Fruit and may become contaminated by soil, insects,
• Meats and Meat Products
and other sources. Freshly harvested grains
typically contain thousands to millions of bacteria
per gram and various mold spores. Range (molds.
Yeast, Coliform)
Microorganism Families: The predominant
families of bacteria found on grains include
Pseudomonadaceae, Micrococcaceae,
Lactobacillaceae, and Bacillaceae. These
microorganisms can be partially removed during
grain processing but are mainly eliminated with
the outer portions of the grain during milling.
Microorganism Counts in Wheat Flour
• A few hundred to a few thousands bacteria per gram
• 20 – 30 bacillus spores per gram
• 50 -100 mold spores per gram
Microorganisms in Cornmeal, Flour, and Malts:
Cornmeal and flour contain several hundred to several thousand bacteria and
molds per gram. Fusarium and Penicillium species are the dominant molds.
Range ( molds; yeast; Coliform group )
Microorganisms in Baked Bread and Cakes: The surface of freshly baked
bread is usually free of viable microorganisms. However, it can be
contaminated by mold spores from the air during cooling and before
wrapping. Slicing and packaging can also introduce contamination. Some
spores of bacteria that can cause ropiness in bread survive the baking process.
Mycotoxin: Grains and cereal products are concerned of mycotoxin
Aspergillus flavus and A. parasiticus
Sugars and Sugar Products
Sucrose: The raw juice expressed from sugarcane may become high in
microbial content unless processing is prompt.
The relevant microorganisms in sugarcane are slime producers (Bacteria:
Leuconostoc and Bacillus) (Yeast: saccharomyces, Candida and Pichia)
Honey
Sources of Microorganisms in Honey
• The primary sources of microorganisms in honey are the nectar of
flowers and honeybees.
• Honey contains lysozyme, an enzyme with bacteriostatic and lytic
effects on most gram-positive bacteria.
Honey and C. botulinum Spores
• Honey is considered one of the suspected food sources for the
introduction of C. botulinum spores, which can cause infant botulism.
Microbial Content in Candies
• Candies from retail markets may contain varying amounts of bacteria,
ranging from 0 to 2 million bacteria per piece.
• Coliform bacteria are rarely found in candies.
Vegetables and Fruit
Harvesting and Transportation
• Contamination may occur during harvesting and transportation.
• Mechanical damage during transport can promote microbial growth.
• Precooling and refrigeration help slow this growth.
Washing and Processing
• Washing can distribute spoilage organisms.
• Recirculated water can add more microorganisms.
• Cleaning with detergents reduces microorganism numbers.
Retail Market
• Further contamination can happen in the retail market.
• Spraying with water and ice can add microorganisms and moisture.
Processing Plant
• Processing can reduce or add microorganisms, depending on
procedures.
Microbial Growth from Equipment
• Microbial growth on equipment influences food contamination.
• Adequate cleaning and sanitizing are essential to prevent this.
Microorganisms and range
Bacteria (Alcaligenes, Pseudomonuc, Micrococci, Bacillus)
Yeast (Fusarium, Penicillium)
Meats and Meat Products
Microorganism Sources in Meats:
• Healthy inner flesh of meats generally contains few or no
microorganisms.
• External contamination sources during slaughtering, handling, and
processing include the animal's exterior (hide, hooves, hair), intestinal
tract, and tools like knives.
Types of Contaminating Microorganisms
• Important genera of bacteria are: Moraxella, Alcaligenes,
Micrococcus, Streptococcus, Sarcina, Leuconostoc, Lactobacillus,
Proteus.
• Yeast: asporogenous
• Mold: Cladosporium, Sporotrichum, Geotrichum, Thamnidium,
Mucor, Penicillium
Fish and Other Seafoods
• Microbial Content: The microbial content in fish depends on the water
in which they live. Freshwater fish carry freshwater bacteria, while
saltwater fish may harbor saltwater bacteria.
• Outer Surface: The outer surface of fish often contains slime that can
be inhabited by bacteria like Pseudomonas, Moraxella, Alcaligenes,
among others.
• Intestinal Bacteria: Fish may have bacteria from both their skin and
the water in their intestines.
• Microbial Counts: The number of bacteria on the skin and slime of
fish can range from as low as 100 to several million per square
centimeter.
• Intestinal Flora: Intestinal fluid in fish can contain bacteria ranging
from 1,000 to 100 million per milliliter.
• Gill Tissue: Gill tissue may harbor bacteria at a concentration of 1,000
to 1 million per gram.
• Washing Effect: Washing fish can reduce the bacterial count on their
surfaces.