It is always happened that while working on any extremophiles we got extremotolerants first. Here in this case I want to isolate obligate alkalophiles from sea sediments and water. I went through many literatures but can anyone help me for the same? What is the initial steps I should take for obligates only. Any experiences regarding this please share.
Thank you in advance.
Hi Abhas
What you say is true, when searching for strains resistant to a phenomena, we always get also some that are tolerant to the same. For your "problem" i'd suggest you to use
This is a very common problem in isolation of different kind of microorganism. Depending upon type of microbes, the distinction between obligate and tolerant is also crude.
In order to isolate obligate alkalophiles you can do a two step process. First isolate all the microbes growing at the condition desired by you, in your case, i guess it would a alkaline pH value (e.g. pH 9). The tolerant microbes should also grow at pH value lower (pH6-8) /more(pH 10-11) than the pH value required by you. So grow all the isolated microbes at different lower and higher pH value. The microbes which will grow at other pH should be tolerant ones, while the isolated microbes which does not grow at other pH values should be idealy an obligate one.
Perform this growth experiment in a broth rather than plates.
Set condition (required pH range for your obligate microbe) as per your requirement of an obligate microbe. In your future work, if the alkanity of the medium is going to vary or change during course of experiment, then isolate a tolerant microbe rather and obligate one.
Hope it helps. Best of luck for your experiments
Use Horikoshi medium I or II to get alkaphilic organisms growth or collect water frm lonar lake and prepare nut agar by this lon lake water
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