Comparative DNA sequence analysis of 16S/18S rRNA genes (rDNA) were undertaken to further our understanding of the make-up of micro-organisms communities in the feces of Gayals. Total DNA were extracted from the feces of 5 Gayals. Two rDNA libraries (16S/18S rDNA) were constructed. In the 16S rDNA library, phylogenetic and sequence similarity analyses of the resultant 71 clone sequences revealed the presence of 67 operational taxonomic units (OTUs) or phylotypes and defined as having more than 97% of sequence similarity. The sequences were affiliated with the following phyla: Firmicutes (34.3%), Bacteroidetes (6.0%), Proteobacteria (4.5%), and uncultured bacteria (55.2%). A set of 58 sequences were analyzed in the 18S rDNA library, which were classified into 27 OTUs. They were mainly affiliated with the following phyla: Protozoa (25.9%), Basidiomycota (3.7%), Ascomycota (11.1%), and uncultured eukaryotes (59.3%). The sequence analysis indicated that more than half of the species, harbored in Gayals fecal belonged to the not-yet-cultured groups at 90% 16S/18S similarity levels with cultured species. In addition, micro-organisms of Chytridiomycetes was one of the most significant cellulose producing species obtained from the Gayal feces as well.
Key words: Gayal, culture-independent, fecal, phylogenetic analysis, 16S/18S rDNA.
VFA, Volatile fatty acids; OTUs, operational taxonomic units; PCR,polymerase chain reaction; IPTG, isopropyl–β–D-thiogalactopyranoside; X-Gal, 5–bromo–4–chloro–3–indolylb–D-galactopyranoside; RFLP, restriction fragment length polymorphism; LB, Luria-Bertani; BLAST, basic local alignment search tool.
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