ENUMERATION OF MICROORGANISMS

Mansoura university, Faculty of pharmacy,

Microbiology and Immunology department
Methods:

1. Plate count methods

A. pour plate method

B. Surface drop method

2. Most-Probable-Number (MPN) Method

Mansoura university, Faculty of pharmacy,

Microbiology and Immunology department
B. Surface drop method
B. Surface drop method:
● The sample solution is serially diluted by adding 1ml of suspension to 9ml of
saline.

● One agar plate is poured and allowed to solidify, the surface of the plate need
to be sufficiently dry to allow a 10μl drop to be absorbed in 15–20 minutes.

● Plate is divided into equal 4 sectors. The sectors are labelled with the
dilutions.

Mansoura university, Faculty of pharmacy,

Microbiology and Immunology department
● In each sector, 3 x 10 μl of the appropriate dilution
is dropped onto the surface of the agar and the drop
allowed to spread naturally.

● The plates are left upright on the bench to dry

before inversion and incubation at 37 °C for 18 – 24
hours.
Mansoura university, Faculty of pharmacy,
Microbiology and Immunology department
• Each sector is observed for growth, high concentrations will give a
confluent growth over the area of the drop, or a large number of
small/merged colonies. Colonies are counted in the sector where the highest
number of full-size discrete colonies can be seen (usually drops containing
between 2-20 colonies are counted).
• The following equation is used to calculate the number of colony forming
units (CFU) per ml from the original aliquot / sample:
CFU per ml = Average number of colonies for a dilution x 100 x dilution
factor
Mansoura university, Faculty of pharmacy,
Microbiology and Immunology department
Mansoura university, Faculty of pharmacy,
Microbiology and Immunology department
Mansoura university, Faculty of pharmacy,
Microbiology and Immunology department