Question

1) A patient has a high fever and is sweating profusely from a new and persistent bacterial infection. The clinician wants to isolate and identify this unique bacterium, and knows that this bacterium will only grow at high temperatures and can survive only in sweat/high salt conditions. Describe the steps for this isolation.

Answer 1

As the detail given about bacteria it says that this bacteria is highly thermostable and Holophile that is salt loving bacteria to isolate such bacteria we can follow the below step:

1. Take a sample from the patient which can be of any type,saliva or any typle.
2. Dilute this salvia in autoclave water and do a serial dilution upto 10 times.
3. Make a growing media for this bacteria which will contain high salt concentration(NaCl),make this media upto 50ml.
4. Disslove this serial dilluted sample from the 10^TH number test tube about 50microlitre into this gowing media and close the flask with cotton plug tightly.
5. Now the next condition is highly thermostable so we have to put this media at above 50 degree temperature as we know optimal growth for most bacteria is 37 degree so other bacteria will not grow at this temperature and this isolated bacteria from patient will only grow.
6. Keep this flask for 18-24 hr at 220 RPM in incubator.

Next morning take out this media and spread on petri plate and leave for next 18-24 hr at same temperature with new meadia transefer and you will see single colong of desire bacteria.