Comparative Analysis of Hindgut Microbiota Variation in Protaetia brevitarsis Larvae across Diverse Farms

Abstract

Protaetia brevitarsis larvae are farm-raised for food, are used in traditional East Asian medicine, and convert organic waste into biofertilizers. Here, the comparative analysis of the gut microbiota of third-instar larvae obtained from five different farms was investigated using 16S rRNA microbial profiling. Species richness, evenness, and diversity results using alpha-diversity analysis (observed species, Chao1, Shannon, Simpson) were similar between farms, except for those between the TO and KO farms. beta-diversity was significantly different in distribution and relative abundance between farms (PERMANOVA, pseudo-F = 13.20, p = 0.001). At the phylum level, Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota were the most dominant, accounting for 73-88% of the hindgut microbial community. At the genus level, Tuberibacillus, Proteiniphilum, Desulfovibrio, Luoshenia, and Thermoactinomyces were the most abundant. Although oak sawdust was the main feed component, there were large variations in distribution and relative abundance across farms at the phylum and genus levels. Venn diagram and linear discriminant analysis effect size analyses revealed large variations in the hindgut microbial communities of P. brevitarsis larvae between farms. These results suggest environmental factors were more important than feed ingredients or genetic predisposition for the establishment of the intestinal microbiota of P. brevitarsis larvae. These findings serve as reference data to understand the intestinal microbiota of P. brevitarsis larvae.