| The Amur tiger(Panthera tigris altaica)is one of the most endangered species in the world and the gut microbial community is the result of the long-term co-evolution,which has an important impact on the health and survival of the Amur tiger.In order to understand the composition and potential function of the gut microbial community of the captive and wild Amur tigers,to analyzed the differences in the composition and function of the gut microbial community of the captive and wild amur tigers,and explore how the factors such as diet,drugs affect the gut microbes of captive Amur tiger,in this paper,we combined with metagenomics sequencing technology,16S rDNA sequencing technology and metabolome analysis,comprehensively studied the composition and function of gut microbiomes of captive and wild Amur tigers,in-depth analyzed the differences in composition and function of the gut microbiome between captive and wild Amur tigers,Amur tigers and other animals,and explored the effects of breastfeeding and goat milk feeding on the composition and function of gut microbiomes in Amur tiger cubs,explored the effects of Fenbendazole and Ivermectin Tablets on the gut microbe diversity and metabolic phenotype of captive Amur tiger.To lay a foundation for further research on the gut microbiome of the Amur tiger,and provide a theoretical basis for promoting the protection and management of the Amur tiger.The following results are obtained:1.We sequenced the gut metagenomes of 3 captive Amur tigers using next-generation sequencing technology to obtain a better understanding of the microbial diversity and functional capacity of captive Amur tiger gut microbiome.Taxonomic analysis of the metagenomic reads indicated that the gut microbiome of the Amur tiger were dominated by Proteobacteria,Firmicutes,Actinobacteria and Fusobacteria.Moreover,at the genus level,Escherichia,Collinsella and Fusobacterium were most abundant in the captive Amur tiger gut metagenome.At the species level,Escherichia coli,Fusobacterium ulcerans and Fusobacterium varium were the species with highest abundances in the captive Amur tiger gut microbiome.The primary functional categories were associated mainly with Carbohydrate metabolism Membrane transport and Amino acid metabolism.We sequenced the gut metagenomes of 3 wild Amur tigers using next-generation sequencing technology to obtain a better understanding of the microbial diversity and functional capacity of wild Amur tiger gut microbiome.The results showed that in the wild Amur tiger gut metagenome,Proteobacteria was the most predominant phylum,followed by Firmicutes,Bacteroidetes,Actinobacteria and Fusobacteria.Escherichia,Clostridium and Streptococcus were the most abundant genera in the wild Amur tiger gut metagenome.At the species level,Escherichia coli,Clostridium perfringens and Escherichia albertii were the species with the highest abundances in the wild Amur tiger gut microbiome.The primary functional categories were primarily associated with Carbohydrate metabolism,Amino acid metabolism and Energy metabolism.2.A comparative macrogenomic analysis of captive Amur tigers and wild Amur tigers and Amur tigers to other animal,to deeply analyz the differences in gut microbiome structure and function between captive amur tigers and wild Amur tigers and Amur tigers to other animal.The results showed that there were significant differences between the gut microbiome of the captive and wild Amur tigers in 10 phyla,177 genera and 458 species.The abundance of Proteobacteria,Chrysiogenetes and Ignavibacteriae in the gut microbiome of captive amur tigers was higher than that of wild Amur tigers,and the abundance of Microsporidia and Euglenida was lower than that of wild Amur tiger.At the genus level,the abundance of Escherichia in Amur tigers is higher than that of wild Amur tigers.At the species level,the abundance of Escherichia coli and Clostridium baratii in captive Amur tigers is higher than that of wild Amur tigers.In function,a total of 1114 KO,4477 NOG,and 36 CAZy were found to have significant differences.In the comparative analysis of the genomes of Amur tigers and other animals,it was found that Actinomycetes,Bacteroidetes,Firmicutes and Proteobacteria have high relative abundance in all animal gut microbiome,the relative abundance of Fusobacteria in the wild Amur tiger and captive Amur tiger gut microbiome is higher than that of other animals.The gut microbiome of cats,pandas and wolves are aggregated,and the gut microbiome of pigs,chickens and mice are aggregated.Functional analysis showed that Amur tigers,cats,pandas,wolves and humans gathered together,and separated from pigs,all gut microbiome metabolic subsystems were mainly carbohydrate metabolism.3.We used whole-metagenome shotgun sequencing to analyze the effects of two different feeding patterns,goat milk feeding and breast feeding,on the composition and functional structures of gut microbiota in Amur tiger cubs.The results showed that goat milk-fed cubs have fewer beneficial bacteria and more pathogenic bacteria and a higher microbial diversity in their gut than breastfed cubs.A total of 15 genera showed statistically significant differences;the relative abundances of Mesorhizobium,Spirosoma and Sorangium were significantly decreased,whereas those of Brochothrix,Syntrophococcus and Acetomicrobium were significantly increased in the goat milk-fed group compared with those in the breastfed group.At the species level,the relative abundances of Streptomyces scabiei,Streptomyces avermitilis and Streptomyces davawensis were significantly decreased,whereas those of Niabella soli,Aeromonas media and Brochothrix thermosphacta were significantly increased in the goat milk-fed group compared with those in the breastfed group.At the functional level,carbohydrate metabolism,translation and replication and repair decreased,and amino acid metabolism,membrane transport and metabolism of cofactors and vitamins increased in the gut microbiota of goat milk-fed cubs compared with breastfed cubs.Our results indicate for the first time that the different milk feeding patterns of goat milk feeding and breast feeding can change the composition and functional structures of gut microbiota in Amur tiger cubs and that breastfed tiger cubs and goat milk-fed tiger cubs have distinct microbiotas in their guts.4.We used an integrated approach of 16S rDNA sequencing combined with metabolomics to analyze the effects of Fenbendazole and Ivermectin Tablets on the gut microbiota and fecal metabolic phenotype of the captive Amur tiger.The results showed that the relative abundances of the bacterial genera Collinsella,Clostridium XI and Megamonas were decreased,whereas those of Escherichia and Clostridium sensu stricto were increased in experimental captive Amur tigers compared with those in normal controls.Meanwhile,there were significant differences in 15 metabolites in the captive Siberian tiger after feeding Fenbendazole and Ivermectin Tablets,including lower levels of 5,6-dihydrouracil,adenine hydrochloride hydrate,galactitol,etc.and catechol and higher levels of acrylic acid,acetoacetate,catechol,etc.Moreover,the differentially abundant gut microbes were substantially associated with the altered gut metabolites,especially the bacteria in the Firmicutes and Actinomycetes,which were involved in the metabolism of 5,6-dihydrouracil,6-phospho-D-gluconate and 1-methylnicotinamide.Our results indicate for the first time that Fenbendazole and Ivermectin Tablets not only disturb the gut microbiota at the abundance level but also alter the metabolic homeostasis of the captive Amur tiger. |
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