Abstract

A study aiming to investigate the rumen methanogens diversity using PCR- DGGE in Indian buffaloes (N= 10) was undertaken. Rumen fluid samples were collected from the male fistulated buffaloes. DNA from the rumen fluid samples was isolated using RBB+C method. DGGE gel band pattern was analysed and bands with differential expression were excised, cloned and sequenced for exploring the methanogens diversity in buffaloes. Sequences were aligned using Codon Code Aligner. The phylogeny was constructed based on neighbour joining method with Jukes Cantor nucleotide substitution model. Based on the differential patterns, total 31 bands were analyzed for the methanogen diversity through phylogenetic approach. Results from the study established that Methanobrevibacter is the most prominent genus of methanogens in Indian buffaloes. However, a considerable diversity exists within this genus at species level. Methanobrevibacter woesi represented the largest cluster of methanogens; while Methanobrevibacter millerae constituted the second largest fraction of archaeal community. Methanogens affiliated to Methanomicrobiales and Thermoplasmatales were not detected through DGGE in present study. To identify the methanogens from other minor groups such as Methanomicrobiales, Thermoplasmatales, Methanosarcinales further studies using high throughput techniques are warranted in buffaloes.

Keywords: Archaea; Buffalo; DGGE; Methanogens; Rumen