C. U.

SHAH INSTITUTE OF SCIENCE

Run by C. U. Ronak Foundation
Ashram Road
Ahmedabad (380014)

Microbiology Sem III

Paper: 201

UNIT 4
MICROBIAL GROWTH

Criteria for Growth Measurement

B.Sc. course Affiliated with Gujarat University
 Determination of Cell Mass
1. Determination of cellular Nitrogen & Protein content
Cells are harvested from media washed out properly and then
used for quantitative determination.

Micro Kjeldahl method is used to measure cellular nitrogen.

The method is only applicable when population is higher in

number.
 2. Determination of other components
Cell mass can also be estimated by measuring the
concentration of some cellular substance, as long as its
concentration is constant in each cell.

For example:
i. Total protein levels, Sulfur and phosphorus level.
ii. Chlorophyll determinations can be used to measure
phototrophic protist and cyanobacterial populations.
iii. The quantity of ATP can be used to estimate the amount
of living microbial mass.
 3. Determination of dry weight of cell
It can be used when cell population is in much higher
amount and dense in the culture.

In this method:

Cells are extracted from the medium by centrifugation or

filtration and dried at 50-55 °C.

That dry mass can be use for determination.

 4. Determination of packed cell volume (PVC)

Bacterial cell belonging to particular species that have its

own specified volume.

When number increases; cell volume also get increased.

In this method it can be done by centrifugation.

The cell mass which is settled in the tube is measured as

packed cell volume.
 5. Determination by Turbidometry Method
Spectrophotometry is a more rapid and sensitive method
for measuring cell mass.

It depends on the fact that microbial cells scatter light that

strikes them.

Because microbial cells in a population are of roughly

constant size, the amount of scattering is directly
proportional to the biomass of cells present and indirectly
related to cell number.
When the concentration
of bacteria reaches about
a million cells per
milliliter, the medium
appears slightly cloudy
or turbid.
Further increases in
concentration result in
greater turbidity, and less
light is transmitted
through the medium.
The extent of light scattering (i.e., decrease in
transmitted light) can be measured by a
spectrophotometer and is called the absorbance (optical
density) of the medium.
Absorbance is almost linearly related to cell
concentration at absorbance levels less than about 0.5.
If the sample exceeds this value, it must first be diluted
and then absorbance measured.
Thus population size can be easily measured as long as
the population is high enough to give detectable
turbidity.
Pross and cons of methods for determination
of cell mass
Most convenient method is turbidometric method.

Determination of nitrogen, protein, sulphur, phosphorous

have only academic values. They have more complexity
while performing actual estimation.

Not for only living cell mass.

 Measurement of Cell Activity

When Cell number and Cell mass increases their metabolic

activity also get increased.

There are parameters selected for measurement for cell

activity is:

i. By measuring O2 consumption and CO2 evolved.

ii. By estimation of organic acids, enzymes,Alcohol, etc.
iii. By determination of carbon source.
 Continuous Culture System
It is possible to grow microorganisms in a system
with constant environmental conditions maintained
through continual provision of nutrients and
removal of wastes. Such a system is called a
continuous culture system.

There Two types of Continuous culture system:

i. Chemostats
ii. Turbidostats
 Chemostats
Chemostat is constructed so
that the rate at which sterile
medium is fed into a culture
vessel is the same as the rate at
which the medium containing
microorganism is removed.

Nutrient availability is limited

during the growth.
 Turbidostats
The Turbidostat has a photocell that measures the
turbidity (defned as the amount of light scattered) of the
culture in the growth vessel.
The flow rate of media through the vessel is automatically
regulated to maintain a predetermined turbidity; Because
turbidity is related to cell density.

The turbidostat maintains a desired cell density while in

chemostate Nutrient it is depend on the nutrient
availability.
 Ms. Aanal Patani

Assistant Professor
Dept. of Microbiology & Biotechnology
C. U. Shah Institute of Science

I shall be happy to entertain your queries

at
pataniaanal@gmail.com