BACTERIAL GROWTH

In microbiology, growth is defined as an increase in the number of cells. Most

bacterial organisms reproduce by an asexual process called binary fission,

Binary Fission includes the stages:

 Cell Elongation - Biosynthesis of new cell wall and membrane and intracellular
proteins occurs. Cells elongate to approximately twice their original length.
 DNA replication. A new copy of the cell's chromosome is made.
 Septum formation- then form a partition that eventually separates the cell
into two daughter cells. This partition is called a septum and is a result of the
inward growth of the cytoplasmic membrane and cell wall from opposing
directions.

This process divides the cell into two daughter cells. Division planes can be of one
(leads to chains), two (leads to sheets), three (cuboidal packets) or many (grape-like
formation).

By definition, when one cell divides to form two, one generation has occurred.
During the growth cycle all cellular constituents increase proportionally. Each
daughter cell receives a chromosome and sufficient copies of ribosomes and all other
macromolecular complexes, monomers, and inorganic ions to exist as an
independent cell.

Cell division apparatus

A series of proteins present in all Bacteria, called Fts proteins (filamentous

temperature sensitive), are essential for cell division. Many Fts proteins such as
FtsZ, FtsA, ZipA, FtsK interact to form a division apparatus in the cell called the
divisome, responsible for causing cell division.

There is great practical interest in understanding bacterial cell division in great detail
because such knowledge could lead to the development of new drugs that target
specific steps in the growth of pathogenic bacteria.

Generation time

The interval for the formation of two cells from one is called a generation, the time
required for this to occur is called the generation time.
The time required for a cell to divide (and its population to double) is called the
generation time.

In bacteria, each new division cycle or generation increases the population by a

factor of 2, or doubles it. Thus, the initial parent stage consists of 1 cell, the first
generation consists of 2 cells, the second 4, the third 8, then 16, 32, 64, and so on.

As long as the environment remains favorable, this doubling effect can continue at a
constant rate. With the passing of each generation, the population will double, over
and over again.

The average generation time of bacteria is 30 to 60 minutes under optimum

conditions. The shortest generation times can be 10 to 12 minutes, and longer
generation times require days. For example, Mycobacterium leprae, has a generation
time of 20 days—as long as that of some animals. Most pathogens have relatively
short doubling times. Salmonella and Staphylococcus bacteria that cause food-borne
illness, double in 20 to 30 minutes, which is why leaving food at room temperature
even for a short period has caused many cases of food-borne disease. In a few
hours, a population of these bacteria can easily grow from a small number of cells to
several million.
(The generation time is useful to determine as faster division times often mean a shorter incubation
period for a disease. For example, suppose you eat an undercooked hamburger contaminated with
the pathogen E. coli O157:H7, which has one of the shortest generation times—just 20 minutes under
optimal conditions. If you ingested one cell (more likely several hundred at least) at 8:00 PM this
evening, two would be present by 8:20, four by 8:40, and eight by 9:00. You would have over 4,000
by midnight. By 3:00 AM, there would be over 2 million).

The cell population size can be represented by the number 2 with an exponent (2 1,
22, 23, 24); the exponent increases by one in each generation; and
the number of the exponent is also the number of the generation. This growth
pattern is termed exponential.
Because these populations often contain very large numbers of cells, it is useful to
express them by means of exponents or logarithms. The data from a growing
bacterial population are graphed by plotting the number of cells as a function of
time. The cell number can be represented logarithmically or arithmetically.
Plotting the logarithm number over time provides a straight line indicative of
exponential growth. Plotting the data arithmetically gives a constantly curved slope.
In general, logarithmic graphs are preferred because an accurate cell number is
easier to read, especially during early growth phases.

The Mathematics of Growth

During the exponential phase each microorganism is dividing at constant intervals.
Thus the population will double in number during a specific length of time called the
generation time or doubling time. This situation can be illustrated with a simple
example.
Suppose that a culture tube is inoculated with one cell that divides every 20 minutes.
The population will be 2 cells after 20 minutes, 4 cells after 40 minutes, and so
forth. Because the population is doubling every generation, the increase in
population is always 2n where n is the number of generations. The resulting
population increase is exponential or logarithmic.
These observations can be expressed as equations for the generation time.
Once we know there are n generations in t hours, we can find the growth rate
constant (The length of the generation time is a measure of the growth rate of an
organism), i.e no. of generations per hour.

Thus, we can determine the generation time which is equal to 1/K hour.
The Microbial Growth Cycle or Bacterial Growth Curve

Microorganisms growing in an enclosed vessel, such as a tube or a

flask (a growth condition called a batch culture) in a liquid media,
show a characteristic growth pattern. The growth of microorganisms
reproducing by binary fission, can be plotted as the logarithm of the
number of viable cells versus the incubation time. The resulting
growth curve describes an entire growth cycle consisting of four
distinct phases- the lag phase, exponential phase,stationary phase, and
death phase.

Lag Phase
When a microbial culture is inoculated into a fresh medium,growth
usually begins only after a period of time called the lag phase. No
immediate increase in cell number occurs in this phase. The lag phase
may be brief or extended, can be observed because of many reasons-
 The cells may be old and depleted of ATP, essential cofactors,
and ribosomes; these must be synthesized before growth can
begin.
 The medium may be different from the one the microorganism
was growing in previously. Here new enzymes would be needed
to use different nutrients.
  Possibly the microorganisms have been injured and require time
to recover.
Hence, this is a period of adaptation to new environmental conditions.
It is only when an exponentially growing culture is transferred into
the same medium under the same conditions of growth (temperature,
aeration,and the like), there is no lag phase and growth begins
immediately.

Log or exponential phase

At the late lag phase, the cells have adjusted to the new environment
and begin to grow. During the exponential phase of growth each cell
divides to form two cells, each of which also divides to form two
more cells, and so on.
The cell number increases exponentially.
The cells grow at their maximum but constant growth rate that is, they
are completing the cell cycle and doubling in number at regular
intervals.
The cells in exponential growth are usually in their healthiest state.
Hence, cells in "mid-exponential" phase are often desirable for studies
of their enzymes or other cell components.
The log phase is time when cells are most active metabolically and is
preferred for industrial purposes.
Cells are most susceptible to adverse environmental factors at this
stage such as Radiations, Antibiotics etc.
This phase will continue as long as cells have adequate nutrients and
the environment is favorable.
Stationary Phase
In a closed system such as a batch culture, exponential growth is
limited. The growing bacterial culture eventually reaches a phase
during which there is no net increase in the number of cells and the
growth curve becomes horizontal.
This stationary phase usually is attained by bacteria at a population
level of around 10 9 cells per ml.
In the stationary phase, the total number of viable microorganisms
remains constant as a result from a balance between cell division and
cell death. No. of cells grow=no.of cells die. In some organisms, slow
growth may actually occur during the stationary phase, but no net
increase in cell number occurs.
Microbial populations enter the stationary phase for several
reasons. One obvious factor is nutrient limitation;
if one or more essential nutrient is severely depleted, population
growth will slow.
Aerobic organisms often are limited by O 2 availability.
Population growth also may cease due to the accumulation of toxic
waste products. For example, streptococci can produce so much lactic
acid and other organic acids from sugar fermentation that their
medium becomes acidic and growth is inhibited. This probably occurs
often in nature because many environments have low nutrient levels.
Procaryotes have evolved a number of strategies to survive starvation.
Organisms survive starvation by:
Some bacteria respond with obvious morphological changes such as
endospore formation,
Survive on nutrients leaking from dead siblings,
Starving bacteria frequently produce a variety of starvation proteins,
which make the cell much more resistant to damage.
They increase peptidoglycan crosslinking and cell wall strength. The
Dps (DNA binding protein) protein protects DNA.
Chaperone proteins prevent protein denaturation and renature
damaged proteins.
Because of these and many other mechanisms, starved cells become
harder to kill and more resistant to starvation, damaging temperature
changes, oxidative and osmotic damage, and toxic chemicals such as
chlorine.

Death Phase:
The number of deaths eventually exceeds the number of new cells
formed, and the population enters the death phase, or logarithmic
decline phase. This phase continues until the population is diminished
to a tiny fraction of the number of cells in the previous phase or until
the population dies out entirely.