Metagenomic Analysis of Mangshan Pit Viper (Protobothrops mangshanensis) Gut Microbiota Reveals Differences among Wild and Captive Individuals Linked to Hibernating Behaviors

Gut microbiota play important roles in the immunity,digestion,and energy meta bolism of their reptile hosts.Mangshan pit viper(Protobothrops mangshanensis)is a critically endangered snake species that is a Class I national protected species in China.Little is known regarding the relationship between P.mangshanensis and their gut microbial communities.In this study,the gut microbiota of wild P.mangshanensis individuals,artificially hiberna ting captive individuals,and non-hibernating captive individuals were compared across nine samples.Comparative shotgun metagenomic analysis was used to investigate the taxonomic composition,diversity,and function of P.mangshanensis gut microbial communities and assess whether their gut microbiomes were affected by their living environments and captivity conditions.The dominant phyla within P.mangshanensis gut microbial communities were Proteobacteria(65.55%),Bacteroidetes(15.97%),and Firmicutes(8.11%).Enriched functional pathways within the gut microbiota included meta bolism(54.9%),environmental information processing(9.67%),and genetic information processing(9.37%).Wild snake gut communities exhibited higher microbial diversity than the other two groups.The gut microbiomes of wild and hibernating captive snakes may be more reflective of healthy intestinal homeostasis than that in nonhibernating snakes.Specifically,non-hibernating snakes exhibited increased levelsof potentially pathogenic populations and functional specialization within gut microbial communities.Thus,different livingenvironments and captivitymethodsmay correspond to major shifts in microbiota composition,diversity,and function within P.mangshanensis.This study provides important insights to help guide the conservation of P.mangshanensis,while also carrying broad implications for our understanding of the effects of living environments and non-hibernating captivity conditions on the gut microbiota of snakes.