3.3 Microbiological Analysis Of The Bottomer
3.3 Microbiological Analysis
3.3.1 Bacteria count using Heamocytometer
This method was used to determine the of three types of dessert but does not differentiate the type of bacteria. 25 g of food samples will be homoginesed in a sterile stomacher bag and shaken for two minutes with 225 ml of peptone water to obtain the food mixture. Using separate sterile pipets, decimal dilutions of 10-2, 10-3, 10-4, 10-5 will be prepared and others as appropriate, of food homogenate by transferring 10 ml of previous dilution to 90 ml of diluent. Using a micropipette, add 10 μl of dilution quickly and smoothly without interruption, to the v-shaped groove on each side of the hemacytometer. Hemocytometer was cover using coverslip counted under a microscope.(FDA, …show more content…
The tubes were incubated at 35°C for 24 hours, following which, the cloudiness and bubble trapped in Durham tube was continued for further analysis. Next, 1 ml of each tube of LST were transferred into tube of 9 ml of Brillian Green Lactose Bile (BGLB) for confirmation of total coliform. BGLB tubes were incubated at 35°C and gas production was examined after 48 hour. The confirmations were determined by the cloudiness and trapped bubble in the Durham tube (FDA, …show more content…
Citrate Test
The citrate test screens a bacterial isolate for the ability to utilize citrate as its carbon and energy source. Simmon’s citrate agar will be prepared by using Oxoid well-prepared Simmon’s citrate agar powder. The Simmon’s citrate agar was prepared by using Oxoid well-prepared Simmon’s citrate agar powder. The preparation was began by suspending 5.75g of the powder in 250 ml of distilled water and mixed thoroughly. The following shows the amount of each composition in 1 litre of Simmon’s citrate agar solution prepared using Simmon’s citrate agar powder.
This Simmon’s citrate agar solution was then heated gently. Next, the mixture was autoclaved at 121°C under 15 psi pressure for 15 minutes. After autoclaved, 4 ml of medium was dispensed into the new falcon tubes using micropipette and this step was performed in laminar flow. The agar was allowed to cool down in slanted position (long slant, shallow butt) before the tubes were closed with caps. The tubes were labelled and stored in refrigerator at
3.3.1 Bacteria count using Heamocytometer
This method was used to determine the of three types of dessert but does not differentiate the type of bacteria. 25 g of food samples will be homoginesed in a sterile stomacher bag and shaken for two minutes with 225 ml of peptone water to obtain the food mixture. Using separate sterile pipets, decimal dilutions of 10-2, 10-3, 10-4, 10-5 will be prepared and others as appropriate, of food homogenate by transferring 10 ml of previous dilution to 90 ml of diluent. Using a micropipette, add 10 μl of dilution quickly and smoothly without interruption, to the v-shaped groove on each side of the hemacytometer. Hemocytometer was cover using coverslip counted under a microscope.(FDA, …show more content…
The tubes were incubated at 35°C for 24 hours, following which, the cloudiness and bubble trapped in Durham tube was continued for further analysis. Next, 1 ml of each tube of LST were transferred into tube of 9 ml of Brillian Green Lactose Bile (BGLB) for confirmation of total coliform. BGLB tubes were incubated at 35°C and gas production was examined after 48 hour. The confirmations were determined by the cloudiness and trapped bubble in the Durham tube (FDA, …show more content…
Citrate Test
The citrate test screens a bacterial isolate for the ability to utilize citrate as its carbon and energy source. Simmon’s citrate agar will be prepared by using Oxoid well-prepared Simmon’s citrate agar powder. The Simmon’s citrate agar was prepared by using Oxoid well-prepared Simmon’s citrate agar powder. The preparation was began by suspending 5.75g of the powder in 250 ml of distilled water and mixed thoroughly. The following shows the amount of each composition in 1 litre of Simmon’s citrate agar solution prepared using Simmon’s citrate agar powder.
This Simmon’s citrate agar solution was then heated gently. Next, the mixture was autoclaved at 121°C under 15 psi pressure for 15 minutes. After autoclaved, 4 ml of medium was dispensed into the new falcon tubes using micropipette and this step was performed in laminar flow. The agar was allowed to cool down in slanted position (long slant, shallow butt) before the tubes were closed with caps. The tubes were labelled and stored in refrigerator at