Spread plate method

Enumeration of microorganisms in this experiment was done by using spread plate

method and pour plate method. In this experiment, sample that used for serial dilution
was aquarium water sample. According to the result, the colonies found through spread
plate method were more abundant than pour plate method. Based on John and Robert
(2003), spread plate method generally will get higher number of colonies formed than
pour plate method. This is because there is a possibility that the sample consisting some
of the microorganisms that indigenous to water had been killed when temperature rose
above 20˚C and the liquefy agar had a temperature of 45˚C which preventing it from
being solidify (John and Robert, 2003). Spread plate method unlikely to kill relative heat
sensitive microorganisms (Tomasiewicz et al., 1980).

Based on the Table 1, many colonies of microorganisms had been formed after 24 hours
and most of the colonies formed are creamy colour. After 48 hours, the colonies become
even bigger and most of them clumped together. Morphology of the colonies provided a
lot of information in identifying and understanding about the microorganisms. In Table 3,
there were two type of microorganisms found: Type A and Type B. Type A with irregular
shape are likely to be motility microorganisms. Type A microorganism might be Bacillus
sp. while Type B possible is Micrococcus sp..The further confirmation of the species can
be done through microscopic examination.

According to Table 4, dilution factor of 10 -2 should have higher number of colonies

formed that dilution factor of 10-4 because after a serial dilution, microorganisms
population should be decreased. For dilution factor 10-1 and 10-3, there numerous colonies
formed and listed as uncountable in the table. The possible explanation is maybe because
of the amount of aquarium sample taken already consist of compact of microorganism
before putting into the serial dilution. The phenomena for dilution factor of 10-2 occurred,
the most of the microorganisms had been killed by spreader. During the experiment, the
spreader was not fully cooled down and the temperature had eliminated many
microorganism.
References:

Buck, J.D. and Cleverdon, R.C., 2003. The spread plate as a method for the enumeration
of marine bacteria. Limnology and Oceanograhy. 5(1): 78-80.

Tomasiewicz, D.M., Hotchkiss, D.K., Reinbold, G.W. and Hartman, P.A., 1980. The
most suitable number of colonies on plates for counting. J. Food Prot. 43: 282-286.