Section 2

• Bacterial culture characteristics

• Establishing pure culture
• Isolation of discrete colonies
• Enumeration of bacteria
Bacterial culture characteristics

1- Size
2- Form (Whole colony)
3- Elevation
4- Margin (Edge)
5- Pigmentation
1- Size
2- Form
3- Elevation
4- Margin
5- Pigmentation
 Pure culture
Pure culture, in microbiology, a laboratory
culture containing a single species of organism.

A pure culture is usually derived from a mixed

culture (one containing many species) by
transferring a small sample into new, sterile
growth medium
 Isolation of discrete colonies
from mixed culture

1 2
Streak plate method Spread plate method
1- Streak plate method
Zigzag shape
provide large surface with better separation
Advantage of streak method
• Simple
• Rapid

Disadvantage
• Cloudiness of colonies
2- Spread method
Advantage of spread method
• Spread the cell allover the plate
• It has accountable number of isolated
bacteria
Determination of bacterial growth

Microbial enumeration is routinely used

in the areas of public health. Food or water
microbiologists test food, milk or water for
the numbers of microbial pathogens to
determine if these products are safe for
human consumption.
Determination of bacterial growth

1 2
Spectrophotometer Serial dilution agar plate
(Turbidity method) (Colony forming unit)
(CFU)

Liquid media Solid media

1- Spectrophotometer
Spectrophotometry is a method to
measure how much a substance absorbs
light by measuring the intensity of light
as a beam of light passes through sample
solution. The basic principle is that each
compound absorbs or transmits light over
a certain range of wavelength.
With increasing the growth, the turbidity increase
Advantage
• Rapid
• Easy

Disadvantage
• Total count include living and dead cell
• Used only for large number (million
cell)
 2- Serial dilution agar plate
The first step in making a serial dilution is to
take a known volume (usually 1ml) of stock and
place it into a known volume of distilled water
(usually 9ml). This produces 10ml of the dilute
solution. This dilute solution has 1ml of
extract /10ml, producing a 10-fold dilution.
(i.e. the amount of stock in each ml of the
diluted solution is 0.1ml.)
Advantage
• Accurate (count only living cell)
• Cheaper
• Simpler
• Repeatable results

Disadvantage
• Time consuming (Slow)
• Count colony not individual cells
• Media must be support the growth of all microorganism