Molecular Genetic Diversity and Quantitation of Methanogen in Ruminal Fluid of Buffalo (Bubalus bubalis) Fed Ration (Wheat Straw and Concentrate Mixture Diet)
High roughage diet causes more methane emissions; however, the total methanogen abundance is not influenced by roughage proportion. Technologies to reduce methane emissions are lacking, and development of inhibitors and vaccines that mitigate rumen-derived methane by targeting methanogens relies on present knowledge of the methanogens. In this work, we have investigated molecular diversity of rumen methanogens of Surti buffalo. DNA from rumen fluid was extracted, and 16S rRNA encoding genes were amplified using methanogen specific primer to generate 16S rDNA clone libraries. Seventy-six clones were randomly selected and analysed by RFLP resulting in 21 operational taxonomic units (OTUs). BLAST analysis with available sequences in database revealed sequences of 13 OTUs (55 clones) showing similarity with Methanomicrobium sp, 3 OTUs (15 clones) with Methanobrevibacter sp. The remaining 5 OTUs (6 clones) belonged to uncultured archaea. The phylogenetic analysis indicated that methanogenic communities found in the library were clustered in the order of Methanomicrobiales (18 OTUs) and Methanobacteriales (3 OTUs). The population of Methanomicrobiales, Methanobacteriales, and Methanococcales were also observed, accounting for 1.94%, 0.72%, and 0.47% of total archaea, respectively. 1. Introduction Methanogens, members of the domain Archaea, fall within the kingdom euryarchaeota [1]. They are obligate anaerobes and can be unmistakably differentiated from other organisms since they all produce methane as a major catabolic end-product [2]. The most common species of methanogens isolated from the rumen are Methanobrevibacter ruminantium, Methanomicrobium mobile, Methanobacterium formicicum, Methanobacterium smithii, Methanobacterium olleyae, Methanobacterium bryantii, Methanosarcina barkeri, and Methanoculleus olentangyi [3–9]. The diversity of archaea found in the rumen has been recently reviewed [10–12]. Interest in methanogens from ruminants has resulted from the role of methane in global warming and from the fact that enteric methane emission is a major source of greenhouse gas in agriculture sector. Currently, India possesses the world’s largest livestock population of 485 million, which accounts for 13% of the global livestock population (Intergovernmental Panel on Climate Change, 2001). It has 57% of the world’s buffalo and 16% of the cattle population. Buffalo contributes to 42% of the total methane emission by livestock in India [13]. Reducing enteric methane emissions has been identified as one way of lowering global methane emissions. However, the
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