FPT 112: General Food Microbiology

Module 1

Week 6: October 2-6, 2023 | 1st Semester, S.Y. 2023-2024

Introduction
Bacteria are prokaryotic organisms that do not contain chlorophyll. They are
unicellular and do not show true branching. They differ from eukaryotes in not
having a nuclear membrane, a nucleolus, and cell organelles like mitochondria,
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Golgi apparatus and endoplasmic reticulum. They have a single circular
chromosome.

This chapter will deal with the nutrition, growth, multiplication of bacteria,
and how to control their growth.
Intended Learning Outcomes
After reading this lesson, you will be able to:

Discuss salient aspects of nutrition, gaseous requirements, temperature

requirements and other physiological requirements for growth;

describe how bacteria grow and multiply;

List the salient features of the bacterial growth curve

Topic 3: Microbial Nutrition and Growth

3.2.1 Bacterial nutrition

The bacterial cell has the same general chemical pattern as the cells of other
organisms. The bacterial cell contains water (80% of total weight), proteins,
polysaccharides, lipids, nucleic acids, mucopeptides and low molecular weight
compounds. For growth and nutrition of bacteria, the minimum nutritional
requirements are
water, a source of carbon, a source of nitrogen and some inorganic salts. Water
is the vehicle of entry of all nutrients into the cell and for the elimination of waste
products.

Nutrition is a process in which living organisms

acquire energy and nutrition through their surroundings to carry out various
cellular processes. All living organisms have different nutritional requirements
for growth and development. Bacteria are prokaryotic organisms that need energy
and nutrients to undergo activities like growth, reproduction, and metabolism.

Nutrient Sources in Bacteria

Bacteria necessarily require macronutrients (such as carbon, nitrogen, hydrogen,

oxygen, phosphorus, and sulfur) to build important biomolecules (proteins and
nucleic acids) that constitute most cellular components.
 Carbon is the essential macronutrient that constitutes a significant portion of
the cell. It builds biomolecules like carbohydrates, proteins, nucleic acids,
lipids, etc.
 Nitrogen is necessary to build proteins and nucleic acids. Bacteria may utilize
 atmospheric nitrogen or other inorganic forms.
 Hydrogen and oxygen constitute many organic compounds, and they are part
of water.
 Phosphorus is necessary to synthesize nucleotides and phospholipids.
 Sulfur is a part of amino acids (like cysteine, methionine).
 Besides all of the above, potassium, magnesium, calcium and sodium are
essential to maintain the bacterial cell structure and function.
Bacteria need micronutrients (like iron, boron, chromium, and manganese) in
trace or minute amounts to perform electron transport chain reactions and other
processes.
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 Bacteria need iron for the functioning of cytochromes and to undergo
electron transport reactions.
 Some bacteria utilize elements
like boron, chromium and manganese as they function as primary
cofactors.

Energy Sources in Bacteria

They obtain energy from direct light and the oxidation-reduction reactions of
different chemical compounds present in our environment. Depending upon the
energy sources, the nutrition in bacteria is mainly autotrophic and heterotrophic.
Phototrophic bacteria possess bacteriochlorophyll pigments to synthesize food,
while heterotrophic bacteria rely on another host for food and energy.

Nutritional Types of Bacteria

Depending upon the energy, electron and carbon sources, bacteria can be classified
into different groups, as mentioned in the diagram below. Phototrophs and
chemotrophs are the two groups of bacteria that obtain energy through direct
sunlight and chemicals.

Then, lithotrophs and organotrophs represent a bacterial group that use reduced
inorganic carbon and organic compounds as electron donors, respectively. At last,
bacteria utilizing carbon sources are grouped into autotrophs and heterotrophs.
Photoautotrophic and chemoautotrophic bacteria exhibit autotrophic nutrition,
whereas photoheterophic and chemoheterotrophic bacteria exhibit heterotrophic
nutrition. Chemoheterotrophs either undergo saprophytic, symbiotic and parasitic
modes of nutrition.
 Figure 1. Types of Bacteria according to Nutrition

Based on Energy Source

1. Chemotrophs includes organisms that rely on chemical energy obtained

through the oxidation of chemical compounds. Such organisms use
a chemosynthesis mechanism where they use the energy released by the
oxidation of inorganic molecules to make organic substances. Based on the
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cellular carbon source, chemoautotrophs and chemoheterotrophs are the two
common types. Conversely, chemoorganotrophs and chemolithotrophs are the
two groups of chemotrophs based on the origin of a hydrogen donor.
2. Phototrophs include organisms that utilize light energy or capture energy
photons. Such organisms use a photosynthesis mechanism where they use light
energy to make organic molecules. Based on the cellular carbon source,
photoautotrophs and photoheterotrophs are the two common types.

Table 1. Types of bacteria based on different energy sources.

Nutritional Nutritional Types of Energy Electron Carbon

Examples
Class Bacteria Source Source Source
CO2 or
Phototrophs Photolithotrophs Light Inorganic Thiospirillum, Chromatiium etc.
organic
Photoorganotrophs Light Organic Organic Rhodospirillum rubrum
Inorganic Cyanobacteria, Green sulfur
Photoautotrophs Light CO2
or organic bacteria and Purple bacteria etc.
Green nonsulfur bacteria, Purple
Photoheterotrophs Light Organic Organic
nonsulfur bacteria etc.
CO2 or Hydrogen, Iron and Sulfur
Chemolithotrophs Chemical Inorganic
organic oxidizing bacteria etc.
Chemotrophs
Chemoorganotrophs Chemical Organic Organic Psuedomonas pseudoflova
Inorganic or
Chemoautotrophs Chemical
organic
CO2 Sulfobolus
Chemoheterotrophs Chemical Organic Organic Clostridium
Based on Electron Source

1. Lithotrophs use reduced organic compounds as electron donors and

thereby using a lithotrophic mode of nutrition. They can be further
categorized into chemolithotrophs and photolithotrophs.
 Chemo-lithotrophic bacteria gain energy from the reduced
inorganic compounds. For example, Nitrosomonas use NH3 as a source of an
electron.
 Photo-lithotrophic bacteria gain energy from light and use reduced
inorganic compounds. For example, Chromatium okeinii uses H2S as an
electron source.
2. Organotrophs use organic compounds as electron donors and thereby
undergoing the organotrophic mode of nutrition. They can be further
classified into chemoorganotrophs and photoorganotrophs.
 Photo-organotrophic bacteria capture light energy and utilize
organic compounds. For example, Rhodospirillum uses succinate as a source
of an electron.
 Chemo-organotrophic bacteria solely obtain energy from organic
compounds. For example, Pseudomonas pseudoflora use glucose and amino
acids as a source of an electron.

Based on Carbon Source

 Autotrophs

They typically make food from inorganic sources (H 2O, CO2, H2S salts). The
autotrophic bacteria are of two types:
1. Photoautotrophs capture the photons of sunlight through
bacteriochlorophylls and convert them into chemical energy. They generally
reduce CO2 to carbohydrates. Inorganic substances serve as hydrogen donors.
2. Unlike photoautotrophs, chemoautotrophs do not require light and
chlorophyll pigments. These bacteria undergo oxidation-reduction reactions of
certain inorganic substances. As a result of this, the energy is released or the
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reaction is exothermic. Chemoautotrophic bacteria derive this energy to carry
out the synthetic processes of the cell.

Heterotrophs

They obtain their-ready made food from organic substances, living or dead.
These are of two types:

1. Photoheterotrophs harness light energy but do not utilize CO 2 as the sole

carbon source. They fulfil carbon and electron requirements through organic
compounds. Such bacteria possess bacteriochlorophyll pigment.
2. Chemoheterotrophs derive energy from organic sources like
carbohydrates, lipids and proteins. Depending upon the mechanism of how
chemoheterotrophs get their organic nutrients, there are three main categories
listed below.
 Saprophytic bacteria obtain nutrition from dead organic matter such as
decomposed leaves and vegetables. These bacteria secrete enzymes to degrade
complex food, such as carbohydrates and protein, into simple absorbable
compounds. Examples are Bacillus mycoides, B. ramosus, Acetobacter etc.
 Parasitic bacteria obtain their nutrition from the living host tissues. They
may be harmless or deleterious that may cause severe illnesses and diseases.
Examples are Rickettsia, Chlamydia, Bacillus typhosus, Vibrio
cholerae, Pseudomonas citri etc.
 Symbiotic bacteria are closely associated with some organisms called
symbionts. They are beneficial to organisms. The common examples are the
nitrogen-fixing bacteria (like Rhizobium, Clostridium etc.), those living inside
the roots of leguminous plants. Nitrifiers perform a primary role in fixing free
atmospheric nitrogen into nitrogenous compounds, which plants utilize. In
return, the plant harbors the bacteria by giving essential nutrients and a
favorable environment for growth.

Therefore, we can conclude that the nutritional types of bacteria rely upon
different energy sources like light energy and chemical energy to undergo various
cellular processes.

3.2.2. Factors Affecting Microbial Growth

Intrinsic factors
Factors intrinsic refer to aspects of the composition of the food itself. They
include:
1.) Nutrients are available to microbes, including sugars, carbohydrates,
proteins, and others
 2.) Acidity/alkalinity or pH of the food
3.) The amount of water available in the food for microbes (water activity
or aw)
Whether the food contains natural antimicrobial chemicals or features that
limit growth
4.) How much natural oxygen is in the food (redox potential)

Extrinsic factors
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Factors extrinsic to the food are essentially the environmental surroundings of the
food. They include:
1.) The temperature of the food during preparation and storage. Most
microbes prefer warm conditions. If temperature is not managed and controlled,
microbes can grow.
2.) The time available for microbes present in food to grow, from the time
of manufacture to the time of consumption. This includes shelf life considerations
for the food as well.
3.) The atmosphere for the food during preparation and storage. This is a
very important concept—food packaging that changes the normal atmospheric
conditions of oxygen, nitrogen, and carbon dioxide is called “reduced oxygen
packaging”. Some bacteria survive best in normal aerobic conditions, while others,
such as C. botulinum, prefer an anaerobic environment.
4.) Relative humidity. This is the level of moisture around the food—is it
dry or wet?

Processing factors
Food processing is often overlooked when foods are evaluated, but it is essential to
understanding risk. Activities during processing can increase risk. For example, the
act of slicing a tomato increases the risk of Salmonella growth. When a fruit is cut,
such as a tomato or a melon, the outer skin and protective covering of the fruit is
damaged. If the interior flesh of the fruit can support bacterial growth, when its
outer skin (tomato) or rind (melon) is damaged, there is a risk that bacteria can
enter the fruit and grow. This may occur if the skin is soiled (containing dirt and
bacteria), or if the knife or utensil used to cut into the fruit is soiled. Processing can
also decrease risk, such as when raw milk is pasteurized, which kills microbes
during the heating step.

Microbe factors
Foods—especially fresh foods—rarely have only one microbe present. A good
example of this is fresh cabbage, which has likely hundreds of microbe varieties
attached to the outer surface of the leaves, stalk, and roots. When fresh cabbage is
submersed in a salty fluid and fermented, fast-growing populations of lactic acid
bacteria change the environment to a more acidic, friendly one in which they can
grow rapidly.

Intrinsic factors: Nutrients, pH, a , antimicrobials, and redox potential

w
Intrinsic factors refer to things that are naturally occurring in the food. These
include: the available nutrients in the food; the natural acidity in the food; naturally
occurring chemical compounds that may give the food natural resistance to other
microbes, such as spoilage organisms; water availability for reactions; and the
 energy potential of the food.

Available nutrients
The most important condition for microbial growth is a supply of suitable nutrients
that can be used as metabolic fuel for growth and reproduction. All living cells
need sources of carbon, oxygen, hydrogen, nitrogen, sulphur, phosphorous, and
other elements. If the supply of one of more nutrients is lacking in the composition
of the food, microbes will have no fuel to maintain growth and reproduction.
Bacteria can be specialized in terms of the types of nutrients they can consume.
For example, different types of bacteria will consume different types of sugars,
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such as sucrose, lactose, glucose, and sorbitol.

Acidity/alkalinity or pH of foods
The acidity or alkalinity of a food is given by its pH. pH is a measurement the
number of hydrogen ions (H+) in water-soluble substances (pH stands for
“potential of hydrogen”). Greater numbers of hydrogen ions makes foods more
acidic. pH values are by numbers on a scale from 1 to 14. Substances pH 7, the
middle value, are neither acidic or alkaline (basic), and are said to have a neutral
pH. Substances with pH values below 7 are acidic. Acidity increases as the number
decreases, with pH 1 being the most acidic. Substances with pH values above 7 are
alkaline. Alkalinity increases as the number increases, with pH 14 being the most
alkaline.

Some foods are acidic. For example, lemons contain natural citric acid and have a
pH 3 to 4. Some foods are only slightly acidic or neutral, such as cabbage, which
has a pH of 6.5 to 7.0. The pH of a food can be measured with paper strips called
pH strips or indicator paper, which containing chemicals that change color based
on the amount of hydrogen ions in the food. Figure 5 shows the relative
acidity/alkalinity of some common foods and substances, using the appropriate
colors of indicator paper.

Figure 2. pH range of foods and other substances

Food microbiologists and food-safety specialists consider most foods with pH

above 4.6 to be potentially hazardous. Most foodborne pathogens thrive in slightly
acidic conditions, from pH above 4.6 to a slightly alkaline pH of 7.5. Most foods
we eat also have a pH in this range. Many spoilage organisms, such as yeasts and
molds, tolerate pH values lower than 4.6 and are often the first microbes to start
spoiling foods such as strawberries and tomatoes. Because lemons contain citric
acid, they take a longer time to spoil; but will eventually also grow mold and
deteriorate. Foodborne pathogens can survive, even if they don’t grow, at higher
and lower pH values as well.

Water activity (aw or Aw)

Because nutrients for microbes are predominantly in a soluble form, the
availability of water is another factor that affects microbial growth. Enzymes
(those specialized proteins that serve as tools to cells) use water to break down
carbohydrates and other sources of food through a process called hydrolysis. Water
is an essential ingredient for these activities. The availability of water in a food is
 referred to as water activity (aw). You might also see water activity given as Aw.
While it is generally true that moist foods have greater water activity than dry
foods, there are some exceptions.
Some foods are preserved by lowering their water activity. The use of salt to
preserve foods does two things: it draws water away from cells, making it
unavailable to microbes, and it binds excess water molecules, lowering the
available water in the food substrate. These actions lower aw and limit the growth
of all microbes, for example, pathogenic and spoilage-types of yeasts, moulds and
bacteria. Sugar solutions can also limit the availability of water. Meat jerky is an
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example of a food that is preserved by lowering its water activity. The water
activity of this food product usually ranges from 0.7 to 0.8, well below the level
most bacteria can survive, but not below the level spoilage molds can grow. Meat
jerky is preserved through a combination of drying and addition of chemical
preservatives (e.g., nitrite) to give this product a long shelf life. Drying is one of
the oldest methods of food preservation. Lacking available water, meat jerky is an
inhospitable substrate for bacteria. Bacteria generally require more water (i.e. have
higher aw requirements) than some fungi, such as the spoilage molds that grow on
the outside of dried meats.

The following terms are used to describe microbes according to the water activity
environments they can grow in:
Halotolerant microbes grow in high concentrations of salt
Osmotolerant microbes grow in high concentrations of organics (e.g.,
sugars)
Xerotolerant microbes grow on dry foods

Figure 3. Shows the water activity (aw) levels of various foods and microbes that can grow at those
levels.

Water activity is not the same as percent moisture of a food. Although water
activity can be expressed mathematically, this will not be covered in this course.
Food with the same water activity can vary wildly in percent moisture. For
example, each of the following foods may have a water activity of approximately
0.7, but very different percent moistures (Adams & Moss, 2000):
Oil-rich nuts have a percent moisture of 4 to 9 %
Protein-rich legumes have a percent moisture of 9 to 13 %
Sucrose-rich dried fruits have a percent moisture of 18 to 25 %
 One definition of a potentially hazardous food is that its water activity is above
0.85. Because their water activity is 0.7, the three foods above would not be
considered potentially hazardous—even though their percent moisture varies
widely. The cut-off for most microbial action (including fungi) is a water activity
of around 0.6.

Based on the parameters of pH and water activity (a w), the defining characteristics for a
potentially hazardous food is a pH above 4.6 and a water activity above 0.85
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Figure 4. pH and water activity levels that make foods potentially hazardous

Antimicrobials
Many foods have naturally occurred compounds that suppress the growth of
microbes, called antimicrobials. Vegetables, fruits, and spices contain chemicals
that prevent and suppress fungal and bacterial growth as a protective mechanism to
plant injury. Plant injury can occur when insects or weather cause bruising or
cutting of plant stems, roots, or leaves. Naturally occurring antimicrobials help to
prevent infection by spoilage and pathogenic microbes. Many plants we use as
spices contain antimicrobials, such as cloves(eugenol), oregano (carvacrol and
thymol), thyme (thymol), garlic (allicin), and ginger (olioresin), along with many
others (Liu et al., 2017). These compounds are used in the modern food industry
during food production and food packaging to limit growth of undesirable
microbes (Liu et al., 2017). Another example of naturally occurring antimicrobials
may be found in eggs. Egg whites (albumen) contain several protein compounds,
including the antimicrobial enzyme lysozyme (Clay, Lock, Dolman, Radford, &
Board, 2003). This enzyme makes up 3.5 % of the albumen contents and protects
the egg whites from bacterial growth by damaging the outer cell walls of bacteria
(Clay et al., 2003). Other compounds in eggs raise the pH of the egg white to a
more inhospitable alkaline pH that makes bacterial growth more difficult (Clay et
al., 2003).
Although foods may contain intrinsic antimicrobial properties, these are not
enough to overcome all pathogenic bacteria, especially when foods are not
properly handled and temperature-controlled. For example, in BC, lack of proper
temperature control (temperature abuse) has caused both botulism and
salmonellosis (from growth of Clostridium botulinum and Salmonella
respectively). Garlic-in-oil stored at room temperature caused thirty-six people to
become severely ill from botulism (St Louis et al., 1988). Cases of salmonellosis
were linked to temperature abuse and improper cooking of eggs in bakery products
(McIntyre, Jorgenson, & Ritson, 2017; Strauss et al., 2000).

Redox potential
Redox potential (Eh) refers to the likelihood that chemical components of a food
(for e.g., vitamins and proteins that are part of the food) will accept or lose
(transfer) electrons, and is determined based on a chemical assessment of a food.
 Electron transfers take place during oxidation-reduction reactions (or redox
reactions). Transfer of electrons drives many chemical activities in microbes, such
as the actions of enzymes and various living functions known as metabolic
activities. The redox potential of a food can to some degree inform what microbes
can survive in a food and what microbial activity has already occurred in a food.
To explain this term in more detail is beyond what we wish to teach in this course.
The key concepts associated with this intrinsic factor are:
Microbial growth in a food will reduce its redox potential
Increasing air (oxygen) will increase the redox potential
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Increasing acidity will increase the redox potential

Of importance to the food industry is the use of redox dyes that measure microbial
growth and activity in foods. For example, methylene blue is used in the brewing
industry to tell whether yeast cells are alive and active (their cytoplasm does not
stain blue) or inactive and dead (they do stain blue).

Figure 5.Yeast (Saccharomyces sp.) stained with methylene blue dye

Extrinsic factors: Temperature, time, atmosphere and relative humidity

Extrinsic factors refer to conditions present outside of the food, similar to the
weather when we wake up: is it wet and muggy or dry and hot? For example, the
potatoes you dug out of your garden will keep better if they are kept dry and in a
cool basement, rather than stored wet in a hot room.
Temperature
Temperature affects the rate of microbial growth. In this section, we will
concentrate on the effect of temperature on bacterial growth. All bacteria have an
optimum, maximum, and minimum temperature for growth. Temperature
variations of only a few degrees may favour the growth of a completely different
species of bacteria, so not only can a change in temperature alter the rate of growth
of bacterial colonies, it can impact which species thrive. The danger zone is a
term used to describe the temperatures between 4°C and 60°C (Figure 9). This is
the optimal temperature range in which most food pathogens and spoilage bacteria
grow.
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Figure 6. Growth of bacteria in the danger zone

 Bacteria and other microbes can be classified into four groups, depending
on the temperature zone in which they grow.
 Psychrophiles grow in cold temperatures between −20°C and 20°C,
preferring temperatures in the range of 0°C to 10°C. These microbes will
grow in refrigerators—and even on frozen foods!
 Mesophiles grow in moderate temperature between 10°C and 50°C,
preferring temperatures between 20°C and 40°C. Most human pathogens
are mesophiles and prefer body temperatures, which range from 35°C to
37°C.
 Thermophiles grow in hot temperatures between 20°C up to 120°C!
Thermophiles prefer temperatures between 40°C and 80°C.
 Hyperthermophiles (a subset of the thermophiles group) have an optimum
temperature greater than 75°C. This class of microorganisms tolerates the
highest temperatures of any known organisms (some living at temperatures
greater than 100°C and up to 120°C).

While freezing will not kill most bacteria, there is a defined thermal death point
(TDP) above which bacteria can no longer survive. Food scientists use this value
to calculate how much heat is required to cook a food to a safe end-point. For
example, E. coli is a mesophile. Found in the human gut, these bacteria prefer a
body temperature of 37 °C. While E. coli will not grow or multiply at refrigeration
temperatures of 4°C or lower, they will not die. Instead, these bacteria will lie
dormant until conditions become more favorable for growth. E. coli survives well
on meat surfaces. Once meat is taken out of the freezer or refrigerator, the E. coli
bacteria have an opportunity to begin growing. Cooking the meat to the thermal
death point of the E. coli bacteria will ensure these bacteria are destroyed. We will
learn more about microbial reduction later in this lesson when we talk about
thermalization of microbes. Thermalization describes the heating process and
 population reduction (or logarithmic reduction) of microbes, and in layman terms,
may be thought of as the cook step.

Time
When considering time as a factor in food microbiology, consider the importance
of time with respect to food hazards, food processes, and food shelf life.
1. For food hazards, time is most often linked to how long it takes bacteria to
grow and double in numbers. Time is also needed for chemical hazards to
develop, such as putrescine formation during decomposition (spoilage).
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2. Food processes are often described in terms of temperature and time. Recipes
and food-safety plans need to specify the temperature and the time—how long
—the food is held at that temperature.
3. Food shelf life is described as a best-before date. This is the time given by a
manufacturer for when the food will retain its best quality attributes (taste,
texture, and smell).

Food hazards and food processes may be considered in seconds, minutes, hours
and months, while shelf life is generally considered in hours, months, and in the
case of canned foods, years. Time must be considered within the context of all of
above examples, and many others.

Atmospheric conditions
While many animals, plants, and bacteria need oxygen, it can be toxic and fatal to
some species. Life on Earth had to adapt to the levels of oxygen in the air. The air
we breathe is composed of 21 % oxygen, 78 % nitrogen, and 0.3 % carbon dioxide
and other gases.
Different bacteria have a different tolerance for the presence or absence of oxygen,
and are adapted to thrive in many different oxygen conditions. The following video
explains the oxygen preferences (requirements) of various bacteria.

Relative humidity
Relative humidity is the amount of moisture in air or packaging. It is affected by
temperature and pressure. For the food microbiologist, relative humidity is
important as it can affect the water activity, aw, of foods. A high relative humidity
will allow water to be absorbed by food, while a low relative humidity may cause
the food to lose water and shrivel (desiccate). High humidity can also cause food
spoilage, as the excess moisture on the surface of foods allows growth of molds,
other fungi and microbes. Low humidity can change the effectiveness of cooking
processes—without enough moisture, higher temperatures and longer times may
be required to adequately cook foods (dry heat versus wet heat).

3.2.3 Growth and Multiplication of Bacteria

Bacteria divide by binary fission. When a bacterial cell reaches a certain size, it
divides to form two daughter cells. Nuclear division is followed by cell division.
The interval of time between two cell divisions, or the time required for a
bacterium to give rise to two daughter cells under optimum conditions, is called
the generation time or the population doubling time. In Escherichia coli and many
other medically important bacteria, the generation time is about 20 minutes. Some
bacteria are slow-growing. The generation time in tubercle bacilli is about 20
hours.
 In Lepra bacilli, it is as long as about 20 days. When bacteria are grown in a vessel
of liquid medium, multiplication is arrested after a few cell divisions due to
depletion of nutrients or accumulation of toxic products. This is a batch culture.
By the use of special devices for replenishing nutrients and removing bacterial
cells (chemostat or turbidostat), it is possible to maintain a continuous culture of
bacteria for industrial or research purposes.

When bacteria multiply in host tissues, the situation may be intermediate between
a batch culture and a continuous culture. The source of nutrients may be
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inexhaustible but the bacteria have to fight the defense mechanisms of the host.
Bacteria growing on solid media (for example blood agar, MacConkey agar) form
colonies. Each colony represents a cluster of cells derived from a parent cell. In
liquid media, growth is diffuse.

Bacterial growth may be considered at two levels: increase in the size of the
bacterial cell and increase in the number of cells. Growth in numbers can be
studied by bacterial counts. Two types of bacterial counts can be made: total count
and viable count. The total count gives the total number of cells in the sample,
irrespective of whether they are living or not. It can be done by various methods,
for example direct counting under the microscope using counting chambers. The
viable count measures the number of living cells, that is, cells capable of
multiplication. Viable counts are obtained by dilution or plating methods. In the
dilution method, the suspension, whose cell count is to be determined, is serially
diluted. The dilutions are made to the point beyond which unit quantities do not
yield growth when inoculated into suitable liquid media. Each dilution is
inoculated into the respective tubes containing liquid media. The viable count is
statistically evaluated from the number of tubes showing growth. This method is
not accurate but is used for the estimation of “presumptive coliform count” in
drinking water. The presumptive coliform count is a method of estimating the level
of pollution of drinking water. In the plating method, appropriate dilutions are
inoculated on solid media, either on the surface of plates or as pour plates. The
number of colonies that develop after incubation gives an estimate of the viable
count. The method commonly employed is that described by Miles and Misra
(1938) in which serial dilutions are dropped on the surface of dried plates and
colony counts obtained.

3.2.6 Bacterial growth curve

When a bacterium is seeded into a suitable liquid medium and incubated, its
growth follows a definite course. If bacterial counts are made at intervals after
inoculation and plotted in relation to time, a growth curve is obtained. The curve
shows the following phases:

Lag phase: Immediately following the seeding of a culture medium, there is no

appreciable increase in number, though there may be an increase in the size of the
cells. This initial period is the time required for adaptation to the new environment.
The necessary enzymes and metabolic intermediates are built up in adequate
quantities for multiplication to proceed. The maximum cell size is obtained
towards the end of lag phase. The duration of the lag phase varies with the species,
size of the inoculum, nature of the culture medium and environmental factors such
as temperature.

Log (logarithmic) or exponential phase: Following the lag phase, the cells start
 dividing and their numbers increase exponentially or by geometric progression
with time. If the logarithm of the viable count is plotted against time, a straight line
will be obtained. In this phase, cells are smaller and stain uniformly.

Stationary phase: After a varying period of exponential growth, cell division

stops due to depletion of nutrients and accumulation of toxic products. The number
of new cells formed is just enough to replace the number of cells that die.
Equilibrium exists between the dying cells and the newly formed cells. So, the
viable count remains stationary. In this phase, cells are frequently gram variable
and show irregular staining. Sporulation occurs at this stage.
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Phase of decline: This is the phase when the population decreases due to cell
death. Besides nutritional exhaustion and toxic accumulation, cell death may also
be caused by autolytic enzymes.

Topic 4: An Overview to Control of Microorganisms

Basic terms used in discussing the control of microorganisms include:
1. Sterilization
Sterilization is the process of destroying all living organisms and viruses. A
sterile object is one free of all life forms, including bacterial endospores, as
well as viruses.
2. Disinfection
Disinfection is the elimination of microorganisms, but not necessarily
endospores, from inanimate objects or surfaces.
3. Decontamination
Decontamination is the treatment of an object or inanimate surface to make it
safe to handle.
4. Disinfectant
A disinfectant is an agents used to disinfect inanimate objects but generally to
toxic to use on human tissues.
5. Antiseptic
An antiseptic is an agent that kills or inhibits growth of microbes but is safe to
use on human tissue.
6. Sanitizer
A sanitizer is an agent that reduces microbial numbers to a safe level.
7. Antibiotic
An antibiotic is a metabolic product produced by one microorganism that
inhibits or kills other microorganisms.
8. Chemotherapeutic synthetic drugs
Synthetic chemicals that can be used therapeutically.
9. Cidal
An agent that is cidal in action will kill microorganisms and viruses.
10. Static
An agent that is static in action will inhibit the growth of microorganisms.

Control of microorganisms is essential in order to prevent the transmission of

diseases and infection, stop decomposition and spoilage, and prevent unwanted
 microbial contamination. Microorganisms are controlled by means of physical
agents and chemical agents. Physical agents include such methods of control as
high or low temperature, desiccation, osmotic pressure, radiation, and filtration.
Control by chemical agents refers to the use of disinfectants, antiseptics,
antibiotics, and chemotherapeutic antimicrobial chemicals.
The basis of chemotherapeutic control of bacteria is selective toxicity. Selective
toxicity means that the chemical being used should inhibit or kill the intended
pathogen without seriously harming the host. A broad spectrum agent is one
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generally effective against a variety of Gram-positive and Gram-negative
bacteria; a narrow spectrum agent generally works against just Gram-positives,
Gram- negatives, or only a few bacteria. As mentioned above, such agents may
be cidal or static in their action. A cidal agent kills the organism while a static
agent inhibits the organism's growth long enough for body defenses to remove
it.
There are two categories of antimicrobial chemotherapeutic agents: antibiotics
and synthetic drugs. Antibiotics are metabolic products of one microorganism
that inhibit or kill other microorganisms. Chemotherapeutic synthetic drugs
are antimicrobial drugs synthesized by chemical procedures in the laboratory.
Many of today's antibiotics are now actually semi-synthetic and some are even
made synthetically.
Antibiotics are metabolic products of one microorganism that inhibit or kill
other microorganisms. Why then do bacteria produce antibiotics? There is
growing support for multiple actions for microbial antibiotic production:
o If produced in large enough amounts, antibiotics may be used as a weapon to inhibit or kill
other microbes in the vicinity to reduce competition for food.
o Antibiotics produced in sublethal quantities may function as interspecies quorum sensing
molecules enabling a number of different bacteria to form within a common biofilm where
metabolic end products of one organism may serve as a substrate for another. All the
organisms are protected within the same biofilm.
o Antibiotics produced in sublethal quantities may function as interspecies quorum sensing
molecules enabling some bacteria to manipulate others to become motile and swim away thus
reducing the competition for food.
o Antibiotics action may result in the degradation of bacterial cell walls or DNA and these
products can act as cues that trigger other bacteria to produce a protective biofilm.
o Antibiotics produced in sublethal quantities may trigger intraspecies quorum sensing.
Exposure to low concentrations of an antibiotic may trigger bacteria to produce quorum
sensing molecules that trigger the population to produce a protective biofilm. The biofilm then
protects the population from greater concentrations of the antibiotic.

Summary
2. Physical control includes such methods of control as high or low
temperature, desiccation, osmotic pressure, radiation, and filtration.
3. Chemical control refers to the use of disinfectants, antiseptics, antibiotics, and
chemotherapeutic antimicrobial chemicals.
4. Sterilization is the process of destroying all living organisms and viruses.
5. Disinfection is the elimination of microorganisms, but not necessarily
endospores, from inanimate objects or surfaces.
6. Decontamination is the treatment of an object or inanimate surface to make it
safe to handle.
 7. A disinfectant is an agents used to disinfect inanimate objects but generally to
toxic to use on human tissues.
8. An antiseptic is an agent that kills or inhibits growth of microbes but is safe to
use on human tissue.
9. A sanitizer is an agent that reduces microbial numbers to a safe level.
10. An antibiotic is a metabolic product produced by one microorganism that
inhibits or kills other microorganisms.
11. Synthetic chemicals that can be used therapeutically.
12. An agent that is cidal in action kills microorganisms.
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13. An agent that is static in action inhibits the growth of microorganisms.
14. Selective toxicity means that the chemical being used should inhibit or kill
the intended pathogen without seriously harming the host.
15. A broad-spectrum agent is one generally effective against a variety of Gram-
positive and Gram-negative bacteria.
16. A narrow spectrum agent generally works against just Gram-positives, Gram-
negatives, or only a few bacteria.
In conclusion
Like other life forms, microbes require energy to grow and reproduce. They
have evolved over millennia, and now inhabit diverse and sometimes extreme
environments. Control of microorganisms is essential in order to prevent the
transmission of diseases and infection, stop decomposition and spoilage, and
prevent unwanted microbial contamination.

References
Books
 Doyle, M. P., Diez-Gonzalez, F., & Hill, C. (Eds.). (2020). Food
microbiology: fundamentals and frontiers. John Wiley & Sons.
 Engelkirk, P. G., Duben-Engelkirk, J., & Fader, R. C. (2020). Burton's
microbiology for the health sciences. Jones & Bartlett Publishers.
 Matthews, K. R., Kniel, K. E., & Montville, T. J. (2017). Food
microbiology: an introduction. John Wiley & Sons