Growth Of Bacteria

Bacterial Growth
With respect to humans, the term growth refers to an increase in size; for example, going from a tiny
newborn baby to a large adult.
•Although bacteria do increase in size before cell division, bacterial growth refers to an increase in the
number of organisms rather than an increase in their size.
•Bacterial growth can be defined as an orderly increase of all the chemical components of the cell.
•Cell multiplication is a consequence of growth that leads to an increase in the number of bacteria
making up a population or culture.
•Most bacteria divide by binary fission in which the bacteria undergo cell division to produce two
daughter cells identical to the parent cell.
Generation time
•The growth rate of a bacterium is measured by measuring the change in bacterial number per unit
time.
•Generation time is the time required for a bacterium to give rise to two daughter cells under
optimum conditions.
•The generation time for most of the pathogenic bacteria, such as E. coli, is about 20 minutes.
•The generation time is longer (i.e., 20 hours) for Mycobacterium tuberculosis and longest (i.e., 20
days) for M. leprae.
•A bacterium replicates and multiplies rapidly producing millions of cells within 24 hours.
Bacterial growth curve and its significance
When a broth culture is inoculated with a small bacterial inoculum, the population size of the bacteria
increases showing a classical pattern. When plotted on a graph, a distinct curve is obtained referred to as
the bacterial growth curve.

Method of Obtaining Bacterial Growth Curve

A population growth curve for any particular species of bacterium may be determined by growing a pure
culture of the organism in a liquid medium at a constant temperature.
Samples of the culture are collected at fixed intervals (e.g., every 30 minutes), and the number of viable
organisms in each sample is determined.
The data are then plotted on logarithmic graph paper.
The logarithm of the number of bacteria per milliliter of medium is plotted against time.
The bacterial growth curve shows the following four distinct phases:

1.Lag phase:
•After a liquid culture broth is inoculated, the multiplication of bacteria does not start immediately. It takes
some time to multiply.
•The time between inoculation and beginning of multiplication is known as lag phase.
•In this phase, the inoculated bacteria become adapted to the environment, switch on various enzymes, and
adjust to the environmental temperature and atmospheric conditions.
•During this phase, there is an increase in size of bacteria but no appreciable increase in number of bacterial
cells. The cells are active metabolically.
•The duration of the lag phase varies with the bacterial species, nature of culture medium, incubation
temperature, etc.
•It may vary from 1 hour to several days.
2.Log phase:
•This phase is characterized by rapid exponential cell growth (i.e., 1 to 2 to 4 to 8 and so on).
•The bacterial population doubles during every generation. They multiply at their maximum rate.
•The bacterial cells are small and uniformly stained.
•The microbes are sensitive to adverse conditions, such as antibiotics and other antimicrobial agents.
•Growth rate is the greatest during the log phase.
•The log phase is always brief, unless the rapidly dividing culture is maintained by constant addition of
nutrients and frequent removal of waste products.
•When plotted on logarithmic graph paper, the log phase appears as a steeply sloped straight line.
3.Stationary phase:
•After log phase, the bacterial growth almost stops completely due to lack of essential nutrients,
lack of water oxygen, change in pH of the medium, etc. and accumulation of their own toxic
metabolic wastes.
•It is during this phase that the culture is at its greatest population density.
•However, Death rate of bacteria exceeds the rate of replication of bacteria.
•Endospores start forming during this stage.
•Many bacteria start producing exotoxins.
4.Decline phase:
•During this phase, the bacterial population declines due to death of cells.
•The decline phase starts due to
(a) accumulation of toxic products and autolytic enzymes and
(b) exhaustion of nutrients.
•Involution forms are common in this stage. Some cells assume various shapes, becoming long,
filamentous rods or branching or globular forms that are difficult to identify.
•Some develop without a cell wall and are referred to as protoplasts, spheroplasts, or L-phase variants (L-
forms).
•When these involuted forms are inoculated into a fresh nutrient medium, they usually revert to the
original shape of the healthy bacteria.
Significance of the Bacterial Growth Curve
•The study of bacterial growth curves is important when aiming to utilize or inoculate known numbers of
the bacterial isolate, for example to enhance plant growth, increase biodegradation of toxic organics, or
produce antibiotics or other natural products at an industrial scale.
•Knowledge of bacterial growth kinetics and bacterial numbers in a culture medium is important from both
a research and commercial point of view.
•Growth kinetics is also useful for assessing whether particular strains of bacteria are adapted to
metabolize certain substrates, such as industrial waste or oil pollution.
•Bacteria that are genetically engineered to clean up oil spills, for example, can be grown in the presence of
complex hydrocarbons to ensure that their growth would not be repressed by the toxic effects of oil.
•Similarly, the slope and shape of growth curves produced from bacteria grown with mixtures of industrial
waste products can inform scientists whether the bacteria can metabolize the particular substance, and
how many potential energy sources for the bacteria can be found in the waste mixture.
BACTERIAL GENETICS
Genetics is the study of inheritance. Bacterial inherited characteristics are encoded in DNA.

Bacteria have two types of DNA that contain their genes. These are :

• Chromosome

• Extra chromosome: Plasmid

 The bacterial chromosome is circular, double stranded DNA attached to bacterial cell membrane.

DNA replication in bacteria is semi-conservative i.e. each strand of DNA is conserved intact during replication

and becomes one of the two strands of the new daughter molecules.

 Plasmids are self-replicating extra chromosomal DNA molecules. It multiplies independent of the host cell.

Multiple copies of the same plasmid may be present in each bacterial cell. Different plasmids are also

often present in the same bacterial cell.

Genetic variation in Bacteria
Mechanisms: Mutation and Gene transfer

1. Mutation: It is due to a chemical alteration in DNA. It could be spontaneous or induced by chemical and

physical means . Mutants are variants in which one or more bases in their DNA are altered; which are

heritable and irreversible

Types of mutation

1. Substitution: Change of a single base.

2. Deletion: Los of a base.

3. Insertion: Addition of a base.

2. Gene transfer

There are three types of gene transfer that alter the DNA gene

content of bacteria.

These are:

1. Transformation

2. Transduction

3. Conjugation

1. Transformation occurs when fragments of exogenous bacterial

DNA are taken up and absorbed into recipient bacterial cells.

Transformation of genes from one bacterium to another results in

• Change in pathogenicity of the bacterium.

• Change in antibiotic sensitivity pattern of bacterium

2. Transduction occurs when fragments of chromosomal DNA is

transferred or transduced into a second bacterium by phage.

During phage replication, the bacterial DNA may be accidentally

enclosed instead of the normal phage DNA, and when this

particle which enclosed the bacterial DNA infects a second

bacterial cell, the DNA from the first bacterium is released and

incorporated into The chromosome of the second bacterium.

3. Conjugation occurs when plasmid DNA is transferred from

donor to recipient bacterium by direct contact via a sex pilus.