Nutrition and Growth of Bacteria MODULE

Microbiology

3
Notes
NUTRITION AND GROWTH OF
BACTERIA

3.1 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,
golgi apparatus and endoplasmic reticulum. They have a single circular
chromosome.
This chapter will deal with the growth and multiplication of bacteria and their
requirements for the same. It will deal with the energy requirements and their
ability to synthesise essential metabolites.

OBJECTIVES
After reading this lesson, you will be able to:
z discuss salient aspects of nutrition, gaseous requirements, temperature
requirements and other physiological requirements for growth
z describe how bacteria grow and multiply
z list the salient features of the bacterial growth curve

3.2 GROWTH OF BACTERIA

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.

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 MODULE Nutrition and Growth of Bacteria

Microbiology 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.
Bacteria can be classified nutritionally based on their energy requirements and
on their ability to synthesise essential metabolites. Bacteria which derive energy
Notes from sunlight are called phototrophs. Those that obtain energy from chemical
reactions are called chemotrophs. Bacteria that can synthesise all their organic
compounds are called autotrophs. They are able to use atmospheric carbon
dioxide and nitrogen. They are capable of independent existence in water and
soil. They are of no medical importance. Some bacteria are unable to synthesise
their own metabolites. They depend on preformed organic compounds. They are
called heterotrophs. These bacteria are unable to grow with carbon dioxide as
the sole source of carbon. Their nutritional requirements vary widely. Some may
require only a single organic substance like glucose. Others may need a large
number of different compounds like amino acids, nucleotides, lipids, carbohydrates
and coenzymes.
Bacteria require a supply of inorganic salts. They require anions like phosphate
and sulphate anions and cations like sodium, potassium, magnesium, iron and
calcium. Some ions like cobalt may be required in trace amounts.
Some bacteria require certain organic compounds in minute quantities. These
are called growth factors or bacterial vitamins. Growth factors are called
essential when growth does not occur in their absence. Accessory growth factors
are those which enhance growth without being absolutely necessary for it. In
many cases, bacterial vitamins are same as vitamins necessary for nutrition of
mammals, for example, B group vitamins – thiamine, riboflavin, pyridoxine,
nicotinic acid, folic acid and vitamin B12.

3.2.2 Gaseous Requirements

Depending on the influence of oxygen on growth and survival, bacteria are
divided into aerobes and anaerobes. Aerobic bacteria require oxygen for
growth. They may be obligate aerobes or facultative anaerobes. Obligate
aerobes grow only in the presence of oxygen, for eg. Cholera bacillus.
Facultative anaerobes are ordinarily aerobic but can grow in the absence of
oxygen, though less abundantly. Most bacteria of medical importance are
facultative anaerobes. Anaerobic bacteria, such as clostridia grow in the
absence of oxygen. Obligate anaerobes may even die on exposure to oxygen.
Microaerophilic bacteria are those that grow best in the presence of low oxygen
tension.

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Nutrition and Growth of Bacteria MODULE
In case of aerobes, atmospheric oxygen is the final electron acceptor in the Microbiology
process of respiration (aerobic respiration). In this case, the carbon and energy
source may be completely oxidised to carbon dioxide and water. Energy is
provided by the production of energy-rich phosphate bonds and the conversion
of adenosine diphosphate (ADP) to adenosine triphosphate (ATP). This process
is called oxidative phosphorylation.
Anaerobic bacteria use compounds like nitrates or sulphates instead of oxygen
as final electron acceptors in the process of respiration (anaerobic respiration). Notes
A more common process used by these bacteria in anaerobic metabolism is
fermentation. It is defined as the process by which complex organic compounds,
such as glucose, are broken down by the action of enzymes into simpler
compounds without the use of oxygen. This process leads to the formation of
several organic end products such as organic acids and alcohols, as well as of
gas (carbon dioxide and hydrogen). For example, Escherichia coli ferments
glucose with the production of acid and gas. It also ferments lactose. During the
process of fermentation, energy-rich phosphate bonds are produced by the
introduction of organic phosphate into intermediate metabolites. This process
is known as substrate-level phosphorylation. The energy-rich phosphate
groups so formed are used for conversion of ADP to ATP.
All bacteria require some amounts of carbon dioxide for growth. This is obtained
from the atmosphere or from the cellular metabolism of the bacterial cell. Some
bacteria like Brucella abortus require much higher levels of carbon dioxide (5-
10%) for growth. They are called capnophilic.

3.2.3 Temperature Requirements

Bacteria vary in their requirement of temperature for growth. For each species,
there is a “temperature range”, and growth does not occur above the maximum
or below the minimum of this range. The temperature at which growth occurs
best is known as the “optimum temperature”. In the case of most pathogenic
bacteria, the optimum temperature is 37ºC.
Bacteria which grow best at temperatures of 25-40ºC are called mesophilic, for
example Escherichia coli. Psychrophilic bacteria are those that grow best at
temperatures below 20ºC. They are soil and water saprophytes and may cause
spoilage of refrigerated food. Thermophilic bacteria are those which grow best
at high temperatures, 55-80ºC. They may cause spoilage of underprocessed
canned food. Some thermophiles, for example Geobacillus stearothermophilus,
form spores that are highly thermoresistant.

3.2.4 Other physiological Requirements

Moisture and drying, hydrogen ion concentration, light, osmotic effect and
mechanical and sonic stress may also influence the growth and multiplication
of bacteria.

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 MODULE Nutrition and Growth of Bacteria

Microbiology 3.2.5 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
Notes 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 inexhaustible but the bacteria have to fight the defence 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.

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Nutrition and Growth of Bacteria MODULE
3.2.6 Bacterial growth curve Microbiology

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 Notes
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.
Phase of decline: This is the phase when the population decreases due to cell
death. Besides nutritional exhaustion and toxic accumulation, cel death may also
be caused by autolytic enzymes.

INTEXT QUESTIONS 3.1

1. Bacteria that can synthesise all their organic compounds are called:
A. Phototrophs B. Chemotrophs
C. Autotrophs D. heterotrophs
2. Organisms that are ordinarily aerobic but can grow in the absence of oxygen
are called:
A. Aerobes B. Facultative anaerobes
C. Anaerobes D. Obligate anaerobes

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 MODULE Nutrition and Growth of Bacteria

Microbiology 3. The process by which complex organic compounds, such as glucose, are
broken down by the action of enzymes into simpler compounds without the
use of oxygen is called:
A. Fermentation B. Substrate-level phosphorylation
C. Oxidative phosphorylation D. Photosynthesis
4. Organisms which are mesophilic grow at:
Notes
A. Temperature of 0-20ºC B. Temperature of 50-60ºC.
C. Temperature of 25-40ºC D. Concentration of 5-10% CO2
5. In the bacterial growth curve, the phase in which there is an exponential
increase in the number of cells is:
A. Lag phase B. Log phase
C. Stationary phase D. Phase of decline

WHAT YOU HAVE LEARNT

z Bacteria are unicellular organisms (prokaryotes) which lack nuclear
membrane, nucleolus and other cell organelles like mitochondria, endoplasmic
reticulum and golgi apparatus.
z Bacterial cell contains water (80% of total weight), proteins, polysaccharides,
lipids, nucleic acids, mucopeptides and low molecular weight compounds.
z For growth and multiplication of bacteria, the mimimal nutritional
requirements are water, a source of carbon, a source of nitrogen and some
inorganic salts.
z Phototrophs are bacteria that derive energy from sunlight while chemotrophs
derive theirs from chemical reactions.
z Autotrophs are bacteria that synthesise all their organic compounds while
heterotrophs are unable to do so.
z Heterotrophs depend on preformed organic compounds.
z Aerobic bacteria require oxygen for growth and may be obligate aerobes
or facultative anaerobes.
z Obligate aerobes grow only in the presence of oxygen.
z Facultative anaerobes grow in the presence or absence of oxygen.
z Anaerobic bacteria grow in the absence of oxygen.
z Obligate anaerobes may even die on exposure to oxygen.
z Microaerophilic bacteria grow best in the presence of low oxygen tension.

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Nutrition and Growth of Bacteria MODULE
z Depending on requirements of temperature for growth, bacteria can be Microbiology
classified as mesophilic (25-40ºC), psychrophilic (below 20ºC) and
thermophilic (55-80ºC).
z Moisture and drying, hydrogen ion concentration, light, osmotic effect and
mechanical and sonic stress may also influence the growth and multiplication
of bacteria.
z Bacteria divide by binary fission.
Notes
z When the bacterial cell reaches a certain size, it divides to form two daughter
cells.
z Nuclear division is followed by cell division.
z The time interval between two cell divisions is the generation time or the
population doubling time. It may vary from 20 minutes (coliform bacilli)
to 20 hours (tubercle bacilli) to 20 days (lepra bacilli).
z The bacterial growth curve consists of a lag phase, a log phase, a stationary
phase and a decline phase. This is seen in a liquid medium.
z In the lag phase, the bacteria adapt to the environment. There is no
appreciable increase in cell number.
z In the log phase, there is exponential increase in the number of bacterial
cells.
z In the stationary phase, there is no increase or decrease in the number of
bacterial cells.
z In the decline phase, there is a decrease in the bacterial population due to
cell death.

TERMINAL QUESTIONS
1. Describe the classification of bacteria based on their nutritional requirements.
2. Explain the temperature requirements of bacteria with suitable examples.
3. Describe the gaseous requirements of bacteria.
4. Define fermentation and give an example of fermentation.
4. Describe the bacterial growth curve with suitable diagrams.

ANSWERS TO INTEXT QUESTIONS

1. C 2. B 3. A 4. C 5. B

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