Difference Between CFU and MPN
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Key Difference - CFU vs MPN
Colony forming unit (CFU) and Most probable number (MPN) are two methods used to
enumerate microorganisms in samples. Both parameters are used to detect water quality and
fecal indicator bacteria in water samples. Colony forming unit is a measure used to enumerate
the number of viable bacterial cells or fungal cells in specific volume or weight of a given
sample. The standard unit for this parameter is CFU/ml or CFU/g. Most probable number is
another unit used to measure the number of viable bacterial cells in a liquid sample. The key
difference between CFU and MPN is that CFU is calculated from the bacterial and fungal
colonies growing on a solid agar plate while MPN is calculated from viable bacteria
growing in a liquid medium.
What is CFU?
Colony forming unit (CFU) is a parameter which measures the number of viable bacterial or
fungal cells in a given sample. The method which counts colony forming units is referred as
standard plate count. The viable colonies that appear on the agar plates are expressed as CFU
per 1 ml (colony forming unit per milliliter) of the sample for liquids or CFU per 1 g (colony
forming unit per one gram) of the sample for solids.
There are two common methods used to measure CFU in a sample. They are spread plate
method and pour plate method. These two methods are supported by a technique called serial
dilution. Serially diluted samples enable obtaining a countable number of colonies on the agar
surface. A known volume of sample can be spread onto the surface of an agar plate, or mixed
with agar and poured onto a plate. The plate is then incubated and the arising colonies are
counted. The number of colonies relates to the number of microorganisms within the original
sample. The plates which show too many colonies or too few colonies are excluded from
counting because the results may not statistically accurate on those plates. Statistically, the best
range is 30 – 300 colonies on an agar plate. Hence, the correct plates should be selected for
accurate enumeration. Serial dilution is performed for the above function.
Once you count the number of viable colonies on plates, CFU/ml can be calculated using the
following equation.
CFU per ml of original sample = number of colonies on a plate X dilution factor
Dilution factor = (1/ Dilution of the plate)
For example, if you get 149 colonies on the plate of the 10-4 dilution, then the number of
bacteria in 1 ml of the original sample can be calculated as follows:
CFU/ml = (149) x (1/10-4)
= 149 × 104 or 1490000
= 1. 49 x 106
Figure 01: Colony Forming Unit
What is MPN?
The most probable number is an alternative measure to CFU/ml. MPN also estimates the
viable cells in a liquid sample. It counts organisms growing in a liquid culture and is a
predominantly bacteriological technique. This method is particularly useful for samples which
contain low concentrations of bacterial cells; for example, milk, potable water etc. MPN value
is expressed for 100 ml of volume. MPN relies on a statistical method based on probability
theory. There are statistical tables designed to find MPN values per 100 ml of the sample. These
tables show the results at 95% confidence limits.
MPN value is calculated after performing a technique called multiple tube fermentation method.
Three sets of tubes containing a suitable culture medium are inoculated with three different
volumes of the sample such as 10 ml, 1 ml, and 0.1 ml and incubated for the growth. After the
incubation period, tubes are scored + (positive) or – (negative) for the presence or absence of
growth. The pattern of positive and negative results are then compared with an MPN table of
statistical probabilities to estimate the number of microorganisms. Then MPN value is given for
100 ml of the sample. MPN is widely used to detect coliform bacteria present in water samples.
Figure 02: MPN table
What is the difference between CFU and MPN?
CFU vs MPN
CFU is a measure used to express the MPN is an alternative measure to CFU and
number of viable bacterial or fungal measures the number of viable bacterial cells
colonies in a given sample. in a liquid sample.
Unit
CFU/ml or CFU/g MPN/100 ml
Calculation
CFU is calculated by counting the number MPN is calculated by comparing positive
of colonies grown on agar plates. and negative patterns of the tubes with MPN
statistical table.
Serial Dilution Technique
Serial dilution is performed before Serial dilution is not normally performed
placing the samples on agar plates. when MPN is calculated
Methods
Spread plate method and pour plate Multiple tube fermentation is the method
method are two types of methods perform performs to obtain MPN value.
to obtain CFU.
Summary - CFU vs MPN
Measuring the microbial growth is required for many reasons. In food processing plants, it is
necessary to measure the level & type of microorganisms in food. In food industry and
medicine, it is necessary to ensure that sterilization treatments are effectively applied. In sewage
treatment plants, it is required to take microbial count routinely. When optimizing procedure in
molecular biology, it is required to measure the number of colonies on plates. Hence, there are
different enumeration and growth measurement methods available. CFU and MPN are two such
methods widely adopting in various fields. CFU is a measure of the number of viable bacterial
and fungal colonies present in a given sample. It is calculated using standard plate count method
or viable plate count method. MPN is another measure which expresses the number of bacterial
cells present in a given volume of the liquid sample. It is calculated using multiple tube
fermentation method and MPN table. This is the difference between CFU and MPN.
References:
1. "Colony-forming unit." Wikipedia. Wikimedia Foundation, 28 May 2017. Web. Available here. 01 June
2017.
2. "Microbiology - 014 - Most Probable Number." Microbiology - 014 - Most Probable Number | Microbiology
Undergraduate Program. N.p., n.d. Web. Available here. 01 June 2017.
Image Courtesy:
1." Manual CFU counting" By Quentin Geissmann - Own work (CC BY-SA 3.0) via Commons Wikimedia
2. "OSC Microbio 00 EE MPNTable" By CNX OpenStax - (CC BY 4.0) via Commons Wikimedia
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