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Methanogens are diverse group of archaebacteria that produce methane as a major product of their metabolism and a basic requirement in waste water treatment and biofuel generation. There are few experiments that have focused on the study of methanogenic bacteria; this paper describes the methanogens identified in the rumen contents of cattle. Fresh rumen were collected from three different slaughter houses in Ota, Ogun State and carefully transported to the laboratory for analysis. A compounded enrichment mineral medium was employed for culturing and presumptive identification of the characteristics methanogen isolates was based on cultural and phenotypic characteristics premised on standard methods. The total plate count ranged from 5.65×1011 to 4.0×105 cfu/ml and 75% of the isolates are Gram negative bacteria while 25% are Gram positive bacteria. Also, 12 methanogens were identified in the rumen samples belonging to the genera Methanococcus, Methanobacterium and Methanosarcina. The mean difference shows that the Methanococcuswas the predominant among the other genera. The molecular analysis of some selected isolates shows the presence of DNA in the characterized sample. Methanogens isolated from remote source like cattle rumen content can serve as catalyst in biofuel production to solve the recent and future world energy problem.