Bacterial Growth
Curve
Nandhana Satheesan
I MSc Microbiology
DBT, CUSAT
Growth
It is a complex process involving
1. Entrance of basic nutrients into the cell
2. Conversion of these compounds into energy and vital cell constituents
3. Replication of the chromosome
4. Increase in size and mass of the cell
5. Division of the cell into two daughter cells, each containing a copy of the genome
and other vital components
The Growth Curve
• The most common means of bacterial reproduction is binary fission; one cell
divides, producing two cells.
• Population growth is studied by analyzing the growth curve of a microbial
culture.
• Evident in batch culture or closed system where no nutrient medium is provided
and there is depletion of waste materials
.
• 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
curve has four distinct phases.
Lag Phase
• When microorganisms are introduced into fresh culture medium, usually no
immediate increase in cell number occurs, so this period is called the lag phase.
• No net increase in mass, the cell is synthesizing new components.
Necessity of lag phase
[Link] cells may be old and depleted of ATP, essential cofactors, and ribosomes; these
must be synthesized before growth can begin.
2. The medium may be different from the one the microorganism was growing in
previously.
3. Possibly the microorganisms have been injured and require time to recover.
The lag phase varies considerably in length with the condition
of the microorganisms and the nature of the medium.
• This phase may be quite long if the inoculum is from an old culture or one that has
been refrigerated.
• Inoculation of a culture into a chemically different medium also results in a longer
lag phase.
• When a young, vigorously growing exponential phase culture is transferred to fresh
medium of the same composition, the lag phase will be short or absent.
Exponential Phase
• microorganisms are growing and dividing at the maximal rate possible.
• Their rate of growth is constant that is, the microorganisms are dividing and doubling
in number at regular intervals ; the curve rises smoothly.
• The population is most uniform in terms of chemical and physiological properties,
exponential phase cultures are usually used in biochemical and physiological studies.
• Exponential growth is balanced growth - all cellular constituents are manufactured at
constant rates relative to each other.
• If nutrient levels or other environmental conditions change, unbalanced growth
results where the synthesis of cell components vary relative to one another until a
new balanced state is reached.
• Unbalanced growth is readily observed in two types of experiments- shift up and
shift down.
shift-up experiment
* culture is transferred from a nutritionally poor medium to a richer one
• There is a lag while the cells first construct new ribosomes to enhance their capacity
for protein synthesis. This is followed by increases in protein and DNA synthesis.
* Finally, the expected rise in reproductive rate takes place.
shift-down experiment
• A culture is transferred from a rich medium to a poor one.
• there is a lag in growth because cells need time to make the enzymes required for the
biosynthesis of unavailable nutrients.
• Consequently cell division and DNA replication continue after the shift-down, but net
protein and RNA synthesis slow.
• The cells become smaller and reorganize themselves metabolically until they are able
to grow again
These shift-up and shift-down experiments demonstrate that microbial growth is under
precise, coordinated control and responds quickly to changes in environmental
conditions.
Stationary phase
• Population growth ceases and the growth curve becomes horizontal.
• This stationary phase usually is attained by bacteria at a population level of around
109 cells per ml.
• In the stationary phase the total number of viable microorganisms remains constant.
This may result from a balance between cell division and cell death, or the
population may simply cease to divide but remain metabolically active.
Microbial populations enter the stationary phase for several reasons
• nutrient limitation
• 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.
Bacteria in a batch culture may enter stationary phase in response to starvation.
Prokaryotes have evolved a number of strategies to survive starvation.
• Many do not respond with obvious morphological changes such as endospore formation,
but only decrease in overall size
• They increase peptidoglycan crosslinking and cell wall strength
• .
• The Dps (DNA binding protein from starved cells) protein protects DNA.
• Chaperone proteins prevent protein denaturation and renature damaged proteins.
• As a result of these and many other mechanisms, the starved cells become harder to
kill and more resistant to starvation itself, damaging temperature changes, oxidative
and osmotic damage, and toxic chemicals such as chlorine.
There is even evidence that Salmonella enterica serovar Typhimurium (S. typhimurium),
and some other bacterial pathogens become more virulent when starved.
Senescence and Death
• It was assumed that detrimental environmental changes like nutrient deprivation and
the buildup of toxic wastes caused irreparable harm resulting in loss of viability.
• Loss of viability was often not accompanied by a loss
in total cell number, it was assumed that cells
died but did not lyse.
• This view is currently under debate. There are
two alternative hypotheses for this.
Hypothesis 1 : Programmed Cell Death
Programmed cell death predicts that a fraction of
the microbial population is genetically
programmed to commit suicide.
• In this case, nonculturable cells are dead and the
nutrients that they leak enable the eventual
growth of those cells in the population that did
not initiate suicide.
• The dying cells are thus altruistic—that is to
say, they sacrifice themselves for the benefit of
the larger population.
Hypothesis 2 : Viable But Non Culturable (VBNC)
• Some microbiologists believe starving cells that show an
exponential decline in density have not irreversibly lost their
ability to reproduce. Rather, they suggest that microbes are
temporarily unable to grow, at least under the laboratory
conditions used.
• This phenomenon, in which the cells are called viable but
nonculturable (VBNC), is thought to be the result of a genetic
response triggered in starving, stationary phase cells.
• Once the appropriate conditions are available VBNC microbes resume growth.
• VBNC microorganisms could pose a public health threat.
Phase of Prolonged Decline
Exponential decline in viability is sometimes replaced by a gradual decline in the
number of culturable cells.
The bacterial population continually evolves so that actively reproducing cells are those
best able to use the nutrients released by their dying siblings and best able to tolerate the
accumulated toxins.
• This dynamic process is marked by successive waves of genetically distinct variants.
Thus natural selection can be witnessed within a single culture vessel.
Mathematics of Growth
• If we start with a single bacterium, the increase in population is by geometric
progression:
n = the number of generations
The total population N at the end of a given time period would be
expressed as
However, under practical conditions, the number of bacteria N0 inoculated at
time zero is not I but more likely several thousand, so the formula now becomes
Solving for n,
The rate of growth during the exponential phase in a batch culture can be expressed in terms
of the mean growth rate constant (k).
This is the number of generations per unit time, often expressed as the generations per
hour.
the mean generation time or mean doubling time (g)—can now be calculated.
The mean generation time is the reciprocal of the mean growth rate constant.
For example, suppose that a bacterial population increases from 10 3 cells to 109 cells
in 10 hours.
Thank You
Reference
• Presscott’s Microbiology