Cultivation, Isolation And Identification Of Sulfate Reducing Bacteria Employing Hungate Technique

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Vipul Kashyap and Chand Ram Grover

Synbiotic Functional Foods & Bioremediation Research Laboratory, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India

 

 

Obligate anaerobes are the most complicated establishment in the nature, which survive only in absence of molecular oxygen. The major challenge to cultivate anaerobes is lack of well-equipped anaerobic facility in the laboratory. Hungate roll tube technique is generally used for anaerobic isolation, wherein roll tubes are employed for isolation of bacterial colonies. In the present study the roll tubes were replaced with serum bottles to cultivate and isolate sulfate reducing bacteria as these are more convenient and reliable. A total 5 isolates were recovered from open drains of Tung Dhab region of Amritsar district (Punjab), India based on colony morphology which were further screened on the basis of gram staining, and 16S rRNA analysis. Sulfate reducing bacteria (BN-15) was finally identified as Clostridium indolis BN-15. Substrate i.e. short chain volatile fatty acids and NaCl utilization patterns were also observed for finally screened isolate.

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