Question

9 ml (a) Dilution 1:10 1:10 0.1 ml 1 ml 0.1 ml FIGURE 54.4 Heterotrophic plate count on food. a) Prepare seraldilutions of food ample (b Lab six Petri dishes with the diutions to be plated. this procedure with the 1:10 dilution and the 1:10 PROCEDURE Second Period dilution until all the dishes have been inoculated (see FIGURE 54.3a). Why can the same pipette be 1. Arrange each plate in order from lowest to hig used for each transfer? 2. Select the plate with 25 to 250 colonies. Re data for plates with fewer than 25 colonies as 6. Check the temperature of the water bath contain- few to count (TFTC) and those with more than outside of the agar. temperature of the colonies, as too numerous to count (TNTC) agar container your hand. It 3. Count the colonies on the plate selected. should be warm, but not too hot to hold. Why? 4. Calculate the number of colony forming unit the original food. For example, if 129 colonies v counted on a 1:10 dilution, calculate as follows 7. Pour the melted nutrient agar into one of the dishes (to about one-third full) (see FIGURE 54.3b). Cover 129 colonies the plate, and swirl it gently (see FIGURE 54.3c) 129,000 1 ml x 10 1.29 x 10 CFU/ml or to distribute the sample through the agar evenly. Continue until all the plates are poured. gram of milk or food 8. When each plate has solidified, seal with tape, in- vert it, and incubate all plates at 35 c for 24 to 48
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Answer #1

In the HPC or SPC i.e.abbrevated as Heterotrophic plate count or standard plate count, the same pippette is used because of same sample dilution is taken we need to obtain all the bacterial population in the dilution sample same pippette gives better removal sticked bacteria to the pippette walls also.

2) we need to check the temperature of the molten agar medium before the palting the sample innoculant as we know hetertrophic efficiency depends on the sample innoculation, type of the media used and temperature administered for the growth of the bacteria, the optimum temperature ie..35C is used for the efficient temperature for growth and withstand viability of the bacterial forms which present in the diluted samples..

After plating these are covered and incubated and colonies are developed and CFU palting efficiency is calculated after necessary incubation time.

:-)

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