Salmonella Essay

In order to isolate bacteriophage specific for Salmonella, we will following procedures below. Firstly, we will inoculate 5 mL tryptic soy broth with Salmonella. Then we touch a colony of Salmonella by using a sterile needle and transfer it to the tube consisting LB broth. After that, we incubate the culture overnight at 37oC. The next day, we will inoculate another tube with 4.5 mL of water sample that we take from poultry farm nearby that we suspect as source of Salmonella contamination on the squid water farm. We also add 0.5 mL of overnight Salmonella culture and 0.5 mL of 10X tryptic soy broth. The mixture will be incubate for about 24 to 48 hours at 37oC. During this incubation period, we expect phage in water sample will be able to bind to Salmonella. The phage also will replicate and lyse the bacteria. Therefore, the significance of this step is to amplify the phage that can infect the Salmonella.

After the incubation period, the tube will be centrifuge using table top centrifuge machine for 10 minutes at 2500 rpm. Centrifuging process should causing the bacteria and other cell debris to form a pellet at the bottom of the tube. The supernatant produce will be taken up about 3 mL using a syringe. A 0.22 micron filter will be attach to the base of the syringe and the supernatant will be filter into a sterile tube. Any remaining bacteria are prevented from passing through the filter except for phage. The filtrate will be label as ‘Enrich Phage’ and store it in refrigerator. By the way, one more overnight culture of Salmonella will be set up.

The next day, 100 µL of an overnight culture of Salmonella growth will be transfer onto the center of a Petri plate containing tryptic soy agar where taken out from refrigerator. Sprea...

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... squid farm where and the waste water from poultry farm nearby for laboratory testing and isolation of Salmonella with respective bacteriophage. After we get the positive result, we continue our activity by characterization of bacteriophage. This activity is expected to take about 2 month since we need to do several repeating processes including culturing the bacteriophage, staining and observing under microscope. For the next step, we will take about 3 months for studying the biocontrol effect of bacteriophage toward Salmonella. This period is the longest among the activities that we will do because this activity involving try and error method where we may get negative result due to error during the past procedure. Lastly, we will analyze our result for about 2 months involving the data taken throughout our research so that it will compatible with our objectives.