Don't complicate matters too much if you are a beginner to thermophile research. Dilute Nutrient broth (0.1%) or dilute Trypticase Soy Broth (0.1%) is a good complex medium to start off with. TSB can be pHed to alkaline pH if required.

If you need to isolate colonies than 2.5% agar can be added.However make sure that you autoclave the medium for at least 30 mins. Agar is loaded with spore from thermophilic bacilli and they grow as contaminants if you fail to autoclave the medium.

To avoid water condensation on agar surface, you should try to dry the plates though this may not be entirely necessary.

Culturing anaerobes is another matter ................

PMID:10653739

Culture-dependent and culture-independent characterization of microbial assemblages associated with high-temperature petroleum reservoirs.

Orphan VJ, Taylor LT, Hafenbradl D, Delong EF.

SourceMarine Science Institute, University of California, Santa Barbara, California 93106, USA.

Abstract

Recent investigations of oil reservoirs in a variety of locales have indicated that these habitats may harbor active thermophilic prokaryotic assemblages. In this study, we used both molecular and culture-based methods to characterize prokaryotic consortia associated with high-temperature, sulfur-rich oil reservoirs in California. Enrichment cultures designed for anaerobic thermophiles, both autotrophic and heterotrophic, were successful at temperatures ranging from 60 to 90 degrees C. Heterotrophic enrichments from all sites yielded sheathed rods (Thermotogales), pleomorphic rods resembling Thermoanaerobacter, and Thermococcus-like isolates. The predominant autotrophic microorganisms recovered from inorganic enrichments using H(2), acetate, and CO(2) as energy and carbon sources were methanogens, including isolates closely related to Methanobacterium, Methanococcus, and Methanoculleus species. Two 16S rRNA gene (rDNA) libraries were generated from total community DNA collected from production wellheads, using either archaeal or universal oligonucleotide primer sets. Sequence analysis of the universal library indicated that a large percentage of clones were highly similar to known bacterial and archaeal isolates recovered from similar habitats. Represented genera in rDNA clone libraries included Thermoanaerobacter, Thermococcus, Desulfothiovibrio, Aminobacterium, Acidaminococcus, Pseudomonas, Halomonas, Acinetobacter, Sphingomonas, Methylobacterium, and Desulfomicrobium. The archaeal library was dominated by methanogen-like rDNAs, with a lower percentage of clones belonging to the Thermococcales. Our results strongly support the hypothesis that sulfur-utilizing and methane-producing thermophilic microorganisms have a widespread distribution in oil reservoirs and the potential to actively participate in the biogeochemical transformation of carbon, hydrogen, and sulfur in situ.

GELRITE is a media which is in use nwadays for culturing Thermophiles.

It is a low-acetyl,clarified gellan gum.

It is a gelling agent with good thermal stability and significantly better clarity than agar.GELRITE forms a clear gel( with the aid of a cation such as Mg or Ca) after heating and cooling.

Don't complicate matters too much if you are a beginner to thermophile research. Dilute Nutrient broth (0.1%) or dilute Trypticase Soy Broth (0.1%) is a good complex medium to start off with. TSB can be pHed to alkaline pH if required.

If you need to isolate colonies than 2.5% agar can be added.However make sure that you autoclave the medium for at least 30 mins. Agar is loaded with spore from thermophilic bacilli and they grow as contaminants if you fail to autoclave the medium.

To avoid water condensation on agar surface, you should try to dry the plates though this may not be entirely necessary.

Culturing anaerobes is another matter ................

Neha,

As B.patel said don't complicate it.You can follow above statement or Please go through APHA Book May be chapter 23 or 24. That will use full for you to isolate and identify Thermophilic acidophilic bactera.Since you have to identify the Thermophlic you must give a heat shock in the respective diluent then go for the Plating.

APHA will guide you clearly.

If gelification is the problem, making gels with potasium silicate neutralize with HCl is the solution. Silica Gel as a Microbiological Medium :Potentialities and a New Method of Preparation. John M. Kingsbury and Elso S. Barghoorn Appl. Microbiol. 1954, 2(1):5. Applied and inveronmental microbiology.

You can make the gel, esterilizae, dry if you want and then add the oslution you need to isolate what ever you want

Bahram, if you want to isolate and set up media for that, I highly recommand you to look for references from B.K.C Patel (from Griffith University, the same person whon commented your post) and Bernard Ollivier (IRD or ORSTOM depending of the date but it's the same person, Marseille). I know perfectly you will find some excellent answers. They worked on methanogens, thermophiles bacteria and especially on SRB.

If you need more advice.... post again!!

for Halophile bacteria, see also Bernard Ollivier or Mr. Oren (forget his first name)

It is also possible to grow thermophillic heterotrophs by serial transfers in nutrient broth and incubation at high temperatures for selective enrichment and then go for isolation on 4% agar containing nutrient medium provided , a beaker containing water is placed in the incubator to maintain humidity .

Priya, you do definitelly wrong, after that an enrichment come up, you have to do direct serial dilution in agar tube otherwise you will lost some bacteria and for sure favorized some other one. a direct serail dilution after agear tube is the best.

2.5% agar till 65 degree Celcius then after yes gelrite is mostly used but hard to work with.

Thanks Patrick . Is agar tube suitable for aerobic thermophiles or would there be any chance of loosing them . Also for some thermophiles nutrient broth with 0.2% peptone is suggested for seriel transfers . Does it carry an advantage .

for an aerobic thermophiles bacteria, you can use plates but you have to put them in a bag/comtainer closed (and you have to change each 2 days the air inside) to preserve the humidity and avoid the dehydration of your agar. Or, you can definitelly use some aerobic media in Hungate tubes but you have to renew each day the air inside the tubes using for sure a 0.22um filter for the inlet .

when you start an isolation process, just use all the time the same media composition otherwise you will change your community . So if you added AT THE BEGINNING 0.2% of peptone, keep it, if not, don't add it.

I am also working on thermophiles.. so I suggest that these microbes can be grown on NA media, LB media, and also basal liquid media as I have done this. so let you try this... all d best !!!

Mr. Pankaj will you tell me how much percentage is used for the agar. And also tell me the temperature used for thermophiles

Mr Jagdish Parihar....

Sorry for being late in replying... but I want to tell you that I took 2% agar in media at 50 degree temp.