Gut analysis using 16S rRNA for bacterial identification in the Pure Mysore and FC1xFC2 breeds of silkworm Bombyx mori

Abstract

Silkworm Bombyx mori, a typical lepidoptera insect, is extremely important in agriculture with economic importance. Its short life cycle, clear genetic background, rich genetic resources, and a significant number of genes similar to humans made it famous for many scientific investigations. Two silkworm strains with one Multivoltine (Pure Mysore) and one Bivoltine (FC1xFC2) were considered for gut microbial profiling. The two strains were grouped into two groups namely Healthy and Unhealthy (starved) and were reared. The stage-specific gut tissues were collected for Gut bacteria culturing from two groups of both strains. Gut bacteria were isolated and cultured from all healthy and unhealthy considered groups. Genomic DNA was isolated for further 16S rRNA gene amplification. 16S rRNA gene sequence was determined by Sanger sequencing and the results were queried against the NCBI database. The Blast analysis revealed the predominant bacterial species in healthy and unhealthy groups of Pure Mysore and FC1xFC2. The Proteus sps, Proteus mirabilis, Flavobacterium sps, Pseudomonas, Bacillus licheniformis were disclosed through the 16S rRNA sequencing and few more gut bacteria belonging to Klebsiella and Enterobacter were revealed through the morphological and a biochemical characterization. The present study unveils the predominant existence of above bacterial species in the intestinal tract of multivoltine and bivoltine breeds of Bombyx mori. In recent times, clear knowledge of intestinal microbial diversity and their role in the host metabolism has gained interest for improvement in commercial sericulture. The focus on microbial profiling would pave insights on the insect-gut microbiome interaction and their role in a beneficial way