Bacterial growth

Mohd Nazmul Hasan

Bacterial growth

Bacterial growth is considered at two levels, increase in the

size of the individual cell and increase in the number of
cells. When a bacterial cell reaches a certain size, it divides
to form two daughter cells by means of binary fission.

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 known as the generation
time or population doubling time.

Two types of bacterial count 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. The
viable count measures the number of living cells. i.e., cells
capable of multiplication.

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 growth curve shows 4 different phases

1) Lag phase
2) Log phase
3) Stationary phase
4) Phase of decline

Lag phase

Immediately after inoculation of the cells into fresh

medium, the population remains temporarily
unchanged. Although there is no apparent cell
division occurring, the cells may be growing in
volume or mass, synthesizing enzymes, proteins,
RNA, etc., and increasing in metabolic activity.

Exponential (log) Phase

The exponential phase of growth is a pattern of

balanced growth wherein all the cells are dividing
regularly by binary fission, and are growing by
geometric progression with time. The cells divide at
a constant rate depending upon the composition of
the growth medium and the conditions of
incubation.

Stationary Phase

Exponential growth cannot be continued forever in a

batch culture (e.g. a closed system such as a test
tube or flask).

Population growth is limited by one of three factors:

exhaustion of available nutrients; accumulation of
inhibitory metabolites or end products; exhaustion
of space, in this case called a lack of "biological
space".

The number of progeny cells formed is just enough

to replace the number of cells that die. The viable
count remains stationary as an equilibrium exists
between the dying cells and the new formed cells.

Phase of decline / Death Phase

If incubation continues after the population

reaches stationary phase, a death phase follows,
in which the viable cell population declines.

During the death phase, the number of viable

cells decreases geometrically (exponentially),
essentially the reverse of growth during the log
phase.

Besides nutritional exhaustion and toxic

accumulation, cell death may be caused by
autolytic enzymes also.

Factors Influencing Bacterial

Growth
Physical requirements
a. Temperature
Bacteria vary in their requirements 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
the range.
The temperature at which growth occurs best is known as
optimum temperature which is in the case of most
pathogenic bacteria is 370C.

Bacteria can be divided into few groups based on their

optimum growth temperature:

1. Psychrophiles are cold-loving bacteria. Their optimum growth

temperature is between -50C and 150C. They are usually found in
the Arctic and Antarctic regions and in streams fed by glaciers.

2. Mesophiles are bacteria that grow best at moderate

temperatures. Their optimum growth temperature is between
250C and 450C. Most bacteria are mesophilic and include common
soil bacteria and bacteria that live in and on the body.

3. Thermophiles are heat-loving bacteria. Their optimum growth

temperature is between 450C and 700C and are commonly found
in hot springs and in compost heaps.

4. Hyperthermophiles are bacteria that grow at very high

temperatures. Their optimum growth temperature is between
700C and 1100C. They are usually members of the Archae and are
found growing near hydrothermal vents at great depths in the
ocean.

b. Oxygen requirements
Microorganisms show a great deal of variation in
their requirements for gaseous oxygen. Most can
be placed in one of the following groups:

1. Obligate aerobes are organisms that grow

only in the presence of oxygen. They obtain their
energy through aerobic respiration.

2. Microaerophiles are organisms that require a

low concentration of oxygen (2% to 10%) for
growth, but higher concentrations are inhibitory.
They obtain their energy through aerobic
respiration.

3. Obligate anaerobes are organisms that grow

only in the absence of oxygen and, in fact, are often
inhibited or killed by its presence. They obtain their
energy through anaerobic respiration or fermentation.

4. Aerotolerant anaerobes like obligate anaerobes

cannot use oxygen to transform energy but can grow
in its presence. They obtain energy only by
fermentation and are known as obligate fermenters.

5. Facultative anaerobes are organisms that grow

with or without oxygen, but generally better with
oxygen. They obtain their energy through aerobic
respiration if oxygen is present, but use fermentation
or anaerobic respiration if it is absent. Most bacteria
are facultative anaerobes.

c. pH

Microorganisms can be placed in one of the

following groups based on their optimum pH
requirements:
1. Neutrophiles grow best at a pH range of 5
to 8.
2. Acidophiles grow best at a pH below 5.5.
3. Allaliphiles grow best at a pH above 8.5.

d. Osmotic pressure

Osmosis is the diffusion of water across a

membrane from an area of higher water
concentration (lower solute concentration) to
lower water concentration (higher solute
concentration). A cell can find itself in one of
three environments: isotonic, hypertonic, or
hypotonic.

Most bacteria require an isotonic environment

or a hypotonic environment for optimum
growth. Organisms that can grow at relatively
high salt concentration (up to 10%) are said to be
osmotolerant.

Nutritional requirements
In addition to a proper physical environment,
microorganisms also depend on an available source of
chemical nutrients. Microorganisms are often grouped
according to their energy source and their source of
carbon.

a. Energy source
1.

Phototrophs use radiant energy (light) as their

primary energy source.

2.

Chemotrophs use the oxidation and reduction of

chemical compounds as their primary energy
source.

b. Carbon source
Carbon is the structural backbone of the organic compounds that
make up a living cell. Based on their source of carbon bacteria
can be classified as autotrophs or heterotrophs.
1.

An

Autotrophs: require only carbon dioxide as a carbon source.

autotroph can synthesize organic molecules from inorganic
nutrients.

2.

Heterotrophs: require organic forms of carbon. A

heterotroph
cannot synthesize organic molecules from inorganic nutrients.

c. Nitrogen source
Nitrogen is needed for the synthesis of such molecules as amino
acids, DNA, RNA and ATP. Depending on the organism, nitrogen,
nitrates, ammonia, or organic nitrogen compounds may be used
as a nitrogen source.

d. Minerals
1.

Sulfur: Sulfur is needed to synthesize sulfur-containing amino

acids and certain vitamins. Depending on the organism,
sulfates, hydrogen sulfide, or sulfur-containing amino acids
may be used as a sulfur source.

2.

Phosphorus: Phosphorus is needed to synthesize

phospholipids, DNA, RNA, and ATP Phosphate ions are the
primary source of phosphorus.

3.

Potassium, magnesium, and calcium: These are required

for certain enzymes to function as well as additional functions.

4.

Iron: Iron is a part of certain enzymes.

5. Trace elements: Trace elements are elements required in

very minute amounts, and like potassium, magnesium,
calcium, and iron, they usually function as cofactors in
enzyme reactions. They include sodium, zinc, copper,
molybdenum, manganese, and cobalt ions. Cofactors usually
function as electron donors or electron acceptors during
enzyme reactions.

e. Water
f. Growth factors
Growth factors are organic compounds such
as amino acids, purines, pyrimidines, and
vitamins that a cell must have for growth but
cannot synthesize itself.
Organisms having complex nutritional
requirements and needing many growth
factors are said to be fastidious.

Culture media
Bacteria can be grown in laboratory using different
media. Because different pathogenic bacteria have
different nutritional needs, various types of culture
media have been developed for use in diagnostic
microbiology. Growth media are used in either of two
phases:
1. Solid media: e.g. Blood agar, Nutrient agar etc.
(Agar
1.5-2%)
2. Liquid media: e.g. Nutrient broth, Peptone water
etc.
(Agar 0%)

In diagnostic microbiology there are few

categories of media used for different types of
microorganisms. They are:

Basic media

Enriched and enrichment media

Selective media

Differential media

Transport media

Basic media

These are simple media that will support the growth

of microorganisms that do not have special
nutritional requirements. E.g. Nutrient agar.

Enriched and enrichment media

These are media that are enriched with blood, extra

peptones, serum, egg or vitamins to support the
growth of pathogens that require additional
nutrients or growth stimulants. E.g. Blood agar,
chocolate agar.

The term enrichment is used to describe a fluid

medium that increases the numbers of a pathogen
by containing enrichments and or substances that
inhibit the multiplication of unwanted bacteria. E.g.
Selenite F broth.

Selective media

Contains one or more agents that are inhibitory to all

organisms except those being sought. E.g. Deoxycholate
citrate medium.

Differential media

These are media that employs some factors like indicators or

dyes that allow colonies of one bacterial species or type to
exhibit certain cultural characteristics that can be used to
distinguish them from other bacteria growing on the same agar
plate. E.g. Thiosulphate citrate bile-salt sucrose (TCBS) agar,
MacConkey agar.

Transport media

These are mostly semisolid media that contain ingredients to

prevent the overgrowth of commensals and ensure the survival
of pathogens when specimen cannot be cultured soon after
collection. E.g. Amies transport medium, Stuarts media.

Thank you
for not snoring