Agusan del Sur State College of Agriculture and

Technology
San Teodoro, Bunawan, Agusan del Sur College of
Teacher Education

WRITTEN REPORT
in
MEASUREMENT OF
MICROBIAL GROWTH
(Chapter 3)
S112 – Microbiology and Parasitology

Submitted by: Jeremy Nest A. Manulat

BSEd – Science 3A
 Submitted to: Dr. Vivian C. Peligro
Instructor

Objectives:
At the end of the lesson, the students will be able to;
a) Define the three types of measurement
b) Determine the different procedures in measuring the microbial growth
c) Identify the direct and indirect using a table

Measurement of Microbial Growth

• According to Adarsh Pandey (2020), there are various ways to measures
microbial growth for the determination of growth rates and generation times.
• For the measurement of growth either mass or population number is followed
because growth leads to increase in both.
• Growth can be measured by one of the following types of measurements:
1. Cell count this method involves the measurement of growth either by
microscopy or by using an electronic particle counter or indirectly by a
colony count.
2. Cell mass in this growth can be measured directly by weighing or by a
measurement of nitrogen concentration in cells or indirectly by the
determination of turbidity using spectrophotometer.
3. Cell activity in this growth can be measured indirectly by analysis of the
degree of biochemical activity to the size of population.

GROWTH CURVE
 Since bacteria are easy to grow in the lab, their growth has been studied
extensively.
 It has been determined that in a closed system or batch culture (no food
added, no wastes removed) bacteria will grow in a predictable pattern,
resulting in a growth curve composed of four distinct phases of growth: the
lag phase, the exponential or log phase, the stationary phase, and the death or
decline phase.
 Additionally, this growth curve can yield generation time for a particular
organism – the amount of time it takes for the population to double.
LAG PHASE
 The lag phase is an adaptation period, where the bacteria are adjusting to
their new conditions.
 The length of the lag phase can vary considerably, based on how different
the conditions are from the conditions that the bacteria came from, as well as
the condition of the bacterial cells themselves.
 Actively growing cells transferred from one type of media into the same type
of media, with the same environmental conditions, will have the shortest lag
period. Damaged cells will have a long lag period, since they must repair
themselves before they can engage in reproduction.
EXPONENTIAL OR LOG PHASE

 The exponential or log phase of growth is marked by predictable doublings

of the population, where 1 cell become 2 cells, becomes 4, becomes 8 etc.
 The slope of the line is equal to 0.301/g. Alternatively one can rely on the
fixed relationship between the initial number of cells at the start of the
exponential phase and the number of cells after some period of time, which
can be expressed by:
 N=N02𝑛
 Where N is the final cell concentration, N0 is the initial cell concentration,
and n is the number of generations that occurred between the specified
period of time
 The lag phase is an adaptation period, where the bacteria are adjusting to
their new conditions.
 The length of the lag phase can vary considerably, based on how different
the conditions are from the conditions that the bacteria came from, as well as
the condition of the bacterial cells themselves.
 Actively growing cells transferred from one type of media into the same type
of media, with the same environmental conditions, will have the shortest lag
period. Damaged cells will have a long lag period, since they must repair
themselves before they can engage in reproduction.
 STATIONARY PHASE
 At this point the number of new cells being produced is equal to the number
of cells dying off or growth has entirely ceased, resulting in a flattening out
of growth on the growth curve

DEATH OR DECLINE PHASE

 In the last phase of the growth curve, the death or decline phase, the number
of viable cells decreases in a predictable (or exponential) fashion.
 The steepness of the slope corresponds to how fast cells are losing viability.
It is thought that the culture conditions have deteriorated to a point where the
cells are irreparably harmed, since cells collected from this phase fail to
show growth when transferred to fresh medium.

SOME SPECIFIC PROCEDURE WILL ILLUSTRATE THE APPLICATION

OF EACH TYPE OF MEASUREMENT

1. DIRECT MICROSCOPIC COUNT

• The most obvious way to count microbial numbers is through direct

counting.
• Petroff-hausser counting is one of the easiest and accurate way to count
bacteria.

 Concentration of the cells can be calculated by using the average no. of

bacteria the avg. number of bacteria in these squares.
  There are 25 squares covering a part of area of 1 mm2, then the entire number
of bacteria in 1 mm2 of the chamber is (number/square) (25 squares). The
chamber is 0.02 mm deep and thus, bacteria/mm3 = (bacteria/square) (25
squares) (50).
 The amount of bacteria per cm3 is 103 times this value. For example, imagine
the average count per square is 28 bacteria: bacteria/cm3 = (28 bacteria) (25
squares) (50) (103) = 3.5X 107.

2. ELECTRONIC ENUMERATION OF CELL NUMBERS

• In this method of microbial growth measurement, bacterial suspension is

kept inside an electronic particle counter, within which the bacteria are
passed through tiny orifice 10 to 30 μm in diameter.
• This orifice is then connected to the two compartments of the counter which
contains an electrically conductive solution.
• The electrical resistance between two compartments will increases
momentarily, when bacterium passes through the orifice. This generates an
electrical signal which is automatically counted.
• The main disadvantage of this method is that there is no way to determine
whether the cell counted is viable or not.

3. THE PLATE COUNT METHOD

Methods of preparing plates for plate counts. (a) The pour plate method. (b) The spread
plate method.

• Plate count method can be done in two ways either by spread plate method or
by pour plate method.
• This technique has some drawbacks because some relatively heat-sensitive
microorganisms may be damaged by the melted agar and will therefore be
unable to form colonies

4. TURBIDITY ESTIMATION OF BACTERIAL NUMBERS

• Turbidity is the Cloudiness or haziness of a media or fluid caused by large

no. of individual particles.
• The instrument used to measure turbidity is a spectrophotometer (or
colorimeter).
• Microbial mass can be determined by determination of absorption of light.
• In the spectrophotometer, a beam of light is transmitted through a bacterial
suspension to a light-sensitive detector, as the bacterial numbers increase,
less light will reach the detector.
• As the population increases, absorbance of the light increases by the cells, so
the turbidity also increases. Turbidity can be measured by using an
instrument spectrophotometer.
• The absorbance is used to plot bacterial growth.
5. DETERMINATION OF NITROGEN CONTENT

• The major constituents of cell material are protein, and since nitrogen is
characteristics part of proteins.
• Bacterial population or cell crop can measure in terms of bacterial nitrogen.
• In this growth can be measured by first harvesting the cells and wash them
free of medium and then perform a quantitative chemical analysis of
nitrogen.

6. DETERMINATION OF DRY WEIGHT OF CELLS

• For filamentous bacteria and molds, the usual measuring methods are less
satisfactory. A plate count would not measure this increase in filamentous
mass.
• One of the better ways to measure the growth of filamentous organisms is by
dry weight.
• In this procedure, the fungus is removed from the growth medium, filtered to
remove extraneous material, and dried in a desiccator, it is then weighed.
• Growth measurement by measuring cell mass is one of the easiest ways, a
known volume of culture sample from the ferment or withdrawn and
centrifuged.
• It is the most direct approach for quantitative measurement of a mass of
cells.
7. COUNTING BACTERIA BY FILTRATION METHOD

• When the number of bacteria is extremely few, as in lakes or relatively pure

streams, bacteria are often counted by filtration methods.
• During this technique, a minimum of 100 ml of water are passed through a
thin membrane filter whose pores are too tiny to permit bacteria to pass.
• After filtration bacteria are filtered out and present on the surface of the
filter. Then filter is transferred to a Petri plate containing a in liquid nutrient
medium, where colonies grow from the bacteria on the filter’s surface.
• This method is applied frequently to detection and enumeration of coliform
bacteria, which are indicators of fecal contamination of food or water.
8. MOST PROBABLE NUMBER (MPN) METHOD

• This statistical estimating technique is based on the fact that the greater the
number of bacteria in a sample, the more dilution is needed to reduce the
density to the point at which no bacteria are left to grow in the tubes in a
dilution series.
• The MPN method is most useful when the microbes being counted will not
grow on solid media (such as the chemoautotrophic nitrifying bacteria).
• It is also useful when the growth of bacteria in a liquid differential medium is
used to identify the microbes (such as coliform bacteria, which selectively
ferment lactose to acid, in water testing).
• The MPN is only a statement that there is a 95% chance that the bacterial
population falls within a certain range and that the MPN is statistically the
most probable number.
 Activity 1:
Direction: Identify the measurement procedure if it is a direct or indirect
measurement.

M Microscopic count Determination of nitrogen content

Electronic enumeration Determination of dry weight
Plate count method Filtration method
Turbidity estimation MPN method

Direct Measurement Indirect Measurement

Answer key:
Direct measurement
1. Microscopic count
2. Plate count method
3. Filtration method
4. MPN method
Indirect measurement
1. Turbidity estimation
2. Determination of dry weight
3. Determination of nitrogen content
4. Electronic enumeration
Quiz
Direction: Choose the letter of the correct answer.

1. It has been determined that in a closed system or batch culture bacteria will
grow in a predictable pattern.
a) Growth Curve
b) Growth Population
c) Measurement of growth
d) Microbial Growth
2. In this phase the number of cell decreases in the predictable or exponential
fashion.
a) Stationary Phase
b) Decline Phase
c) Lag Phase
d) Log Phase
3. This method involves the measurement of growth either by microscopy or by
using an electronic particle counter or indirectly by a colony count.
a) Cell mass
b) Cell activity
c) Cell count
d) Cell Viability
4. In this bacteria are adjusting to their new conditions.
a) Log Phase
b) Stationary Phase
c) Decline Phase
d) Lag Phase
5. In this growth can be measured directly by weighing or by measuring nitrogen
concentration.
a) Cell count
b) Cell viability
c) Cell mass
d) Cell activity
6. Cells in this phase of growth are the healthiest and most uniform.
a) Exponential Phase
b) Lag Phase
c) Stationary Phase
d) Death Phase
7. At this point the number of new cells being produced is equal to the number of
cells dying off or growth has entirely ceased
a) Log Phase
b) Stationary Phase
c) Decline Phase
d) Lag Phase
8. Growth can be measured by one of the following types of measurements
EXCEPT:
a) Cell viability
b) Cell mass
c) Cell count
d) Cell activity
9. In this growth can be measured indirectly by analysis of the degree of
biochemical activity to the size of population.
a) Cell activity
b) Cell viability
c) Cell count
d) Cell mass
10. It is a specialized glass slide and one of the easiest and accurate way to count
bacteria.
a) Plate count method
b) Turbidity estimation
c) Petroff-hausser counting
d) MPN method
11. How many squares covering a 1𝑚𝑚2 grid in a slide?
a) 28
b) 25
c) 23
d) 50
12. Microbial growth measurement is bacterial suspension is kept inside an
electronic particle counter.
a) Plate count method
b) Electron counter method
c) Electronic enumeration
d) Turbidity estimation
13. The most obvious way to count microbial numbers is through direct counting.
a) Direct microscopic count
 b) Turbidity estimation
c) Petroff-hausser counting
d) MPN method
14. It is measured by first harvesting the cells and wash them free of medium and
then perform a quantitative chemical analysis of nitrogen.
a) Chemical analysis of nitrogen
b) Plate count method
c) MPN method
d) Determination of nitrogen content
15. This method allows the determination of the number of cells that will multiply
under certain defined conditions.
a) MPN method
b) Plate count method
c) Electronic enumeration
d) Turbidity estimation
16. In this method it uses light and spectrophotometer in monitoring a bacterial
growth.
a) MPN method
b) Plate count method
c) Electronic enumeration
d) Turbidity estimation
17. It is the most direct approach for quantitative measurement of a mass of cells.
a) Direct microscopic count
b) Determination of dry weight of cell
c) MPN method
d) Electronic enumeration
18. Calculate the average count per square is 28 bacteria: bacteria/cm3 = (28
bacteria) (25 squares) (50) (103) =?
a) 3.5X 107

b) 2. 5X 107
c) 3.5X 106
d) 2. 5X 106
19. Growth curve composed of four distinct phases of growth:
a) Anaphase, prophase, metaphase, telophase
b) lag phase, log phase, anaphase, telophase
c) Lag phase, exponential phase, stationary phase, death phase,
 d) lag phase, log phase, stationary phase, decease phase
20. This method is applied frequently to detection and enumeration of coliform
bacteria.
a) Filtration method
b) MPN method
c) Plate count method
d) Electronic enumeration

Reference: Adarsh Pandey. (December 2020).Measurement of Microbial Growth.

Retrieved from https://microbiologynotes.org/measurements-of-microbial-growth/