The plat should be allowed to solidify and placed immediately in the incubator in an

inverted position.
a) Incubation: - The plates should be incubated at 370C for 24 hours at 200C for 48 hours.
b) Counting: - In preparing plates, such amount of water or dilution should be planted
which will give from 30 to 300 colonies on a plate. There should be always more plates
for dilution, at least two. The results as reported shall be the average of all plates falling
within limits. It is not desirable to plant more than 30 from 1 ml sample; it is obviously
to be disregarded. In practice, we do use the counts even if the of colonies in a plate are
less than, 30, when chlorinated water samples are plated, when the number of colonies
are more the 300 in a plate, we report the count as TNC that in a plate report the count
as TNC (too numerous to count). Counting shall be done with an approved counting
aid, such as colony counter. The reported number of bacteria per ml shall not include
more than two significant figures. For example of 158 as 160 whereas a count of 35 is
recorded as 35 and counts shall be designated as Standard Plate Count at 370C or
200C.
Coliform Count by MTD Method
The coliform group includes all of the aerobic and facultative anaerobic gram negative,
no spore forming rod shaped bacteria which ferment lactose with gas formation within
48 hours at 370C. The standard test for the estimation of numbers of the coliforms group
may be carried out either by the Multiple Tube Dilution (MTD) or Membrane Filter
(MF) methods.
It is convenient to express the results of the examination of replicate and dilutions in
term of Most Probable Number (MPN). This term is actually an estimate based on
certain probability formulae. The MPN value for a given sample is obtained by the use
of MPN tables. Standard practice in water analysis is to plant five tubes for each dilution
and a minimum three different dilutions are employed. The results are to be recorded in
the proper form.
Details of procedure: The water sample is shaken thoroughly before making dilutions or
before inoculation.