16s rRNA Amplicon Sequencing Approach for Community and Predictive Functional Diversity of Therapeutically Valuable Formulation of Cow-derivatives

Abstract

Panchagavya is the blend of five ingredients obtained from cows and used in traditional Hindu rituals. Panchagavya preparation is a microbial mediated process that possibly involves microbial succession. The present study was conducted to decode the microbial community that exists in the preparation of three hours of old Panchagavya. DNA was isolated from Panchagavya using HiPurA™ Stool DNA Purification Kit followed by quality checking by Agarose electrophoresis and Qubit fluorometer. The V3 and V4 region of 16s rRNA based amplicon sequencing was performed using the Illumine MiSeq platform. Taxonomic profile encoded by using One Codex, kraken and MG-RAST. Functional traits detected through the abundance of specific genes using Tax4Fun. Taxonomic result suggests the total 2000 species were identified. The most abundant was Streptomyces griseocarneus (2.65%) followed by Clostridiales bacterium (2.26%), Bacteroidales bacterium (1.38%), and Verrucomicrobia bacterium (1.13%). Community based analysis revealed the microbial diversity and presence of anaerobic, unclassified, and uncultivable microbes in metagenomes, which may be associated with the pharmacological properties of Panchagavya. Functional analysis predicts around 351 metabolic pathways for metabolism of carbohydrates, synthesis of secondary metabolites and degradation of xenobiotic compounds. The detection of various secondary metabolites genes associated with pharmacological molecules correlated with its traditional clinical applications. The present study revealed the advantages of cultivation approach for exploring untapped and unique bacterial diversity, and also utilities for various biotechnological and environmental applications.