I have 500 ml stock culture which consist of 10^8 bacteria cells. I need to inoculate 10^5, 10^6 and 10^7 cfu/ml to my fish samples. Which way should I follow from there? Should I harvest the cells with centrifuge or directly dilute from the stock culture? Could someone explain a way of obtaining this cell culture?
Hi,
I would say both the way you can use to obtain your desired cell density. either you go with dilution or you harvest your cells according to your need. then inject it in the fish flesh through intradermal inoculation.
Thanks
Hi Surjit,
You mean it depends on my decision? If I harvest the cells via centrifuge, which amount should I use from bacteria culture and how much psyhlogical saline should I add after harvesting?
I dont know the clear way. Do you mind if you explain the method more clear or describe with numbers?
Thanks a lot,
Ilknur
Use isotonic solution for inoculation that is 0.89% saline and try to use cultures in log phase that will help you and regarding the dose consult papers.
if you will harvest the cells via centrifuge, you must not use high speed for a long time to avoid cell destruction. i suggest you use 5000 rpm / 5 min then inoculate the cells (pellet) in measured volume of the isotonic solution 0.85% saline
I m beginner in Microbiology and I have enough time to learn something untill my experiment Mr. Burzyski. thats why I ask and try to learn. after I m sure from my knowledge I will start to my experiment
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