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Damiles, Johmar, De Vera, Alyssa Gail, Elio, Charlene Lorraine, Fernandez, Gino
Miguel*
2BIOLOGY6, Department of Biology, College of Science, UST, Manila
*fginomiguel@yahoo.com
Abstract
Microorganisms are within nature, they occur and are studied within the living environment or inside a laboratory, they
are not isolated then studied as different and separate organisms, but as a whole, as populations within nature, either
in colonies or as dispersed units suspended in a medium or in a suspended animation. Pour plate and Spread Plate
method are the two most common methods used in the isolation of microorganisms from their natural habitat.
Additionally, these two processes give the most approximate or estimate number of organisms present in a
population of microorganisms.
Keywords: Spread Plate Method, Pour Plate Method, Isolation, Suspended in a Liquid Medium
Introduction
Microorganisms are within nature, they occur
and are studied within the living environment or
inside a laboratory, they are not isolated then
studied as different and separate organisms, but
as a whole, as populations within nature, either in
colonies or as dispersed units suspended in a
medium or in a suspended animation. Pour plate
and Spread Plate method are the two most
common methods used in the isolation of
microorganisms from their natural habitat.
Additionally, these two processes give the most
approximate or estimate number of organisms
present in a population of microorganisms. The
pour plate technique can be used to determine the
number of microbes/mL or microbes/gram in a
specimen. It has the advantage of not requiring
previously prepared plates, and is often used to
assay bacterial contamination of foodstuffs. The
principle steps are to 1) prepare/dilute the sample
2) place an aliquot of the diluted sample in an
empty sterile plate 3) pour in 15 mL of melted agar
which has been cooled to 45 o C, swirl to mix well
4) let cool undisturbed to solidify on a flat table top
5) invert and incubate to develop colonies. Each
colony represents a "colony forming unit" (CFU).
For optimum accuracy of a count, the
Methodology
organisms.
Conclusion
From the experiment, the researchers have
inferred that the best way of quantitative analysis
in microbial populations would be the spread plate
technique. Spread plate technique is a better way
of determining the amount of colonies in the
sample because it does not damage the colonies
in any way, and the colonies would be almost
evenly distributed through the plate thus making it
easier to count. Pour plate could kill some of the
microorganisms because the researchers would
have to pour molten agar into their diluted sample,
and the heat might have killed off some of the
References
[1] http://www2.hendrix.edu/biology/CellWeb/Techniques/microspread.htm
[2] http://biology.clc.uc.edu/fankhauser/Labs/Microbiology/Meat_Milk/Pour_Plate.html
[3] Laboratory Manual in General Microbiology. (2014). Manila: University of Santo Tomas.