Study On The Antibiotic Resistance Of Giant Panda Gut Microbiome,Habitat Soil Microorganisms And Giant Pandas Staple Bamboo

Giant panda is an endemic protected animal in China.It is known as "living fossil" and "Chinese national treasure".The gut microbiome of giant pandas plays a great important role in the growth and development,physiological metabolism,digestion and absorption,as well as immunity to diseases.As a star species,the society attach a great importance to the protection of giant pandas and have carried out in-depth research.As a new type of environmental pollutants,antibiotic resistance genes(ARGs)pose a serious threat to human health and ecosystem security.Based on the current research,ARGs are detected in different environments,and can spread in a wider range through a variety of transmission modes such as physical modes,horizontal and vertical transmission of genes.The continuous increase of bacterial tolerance has caused varying degrees of harm to the ecological environment.As a flagship species of biodiversity protection,it is of great significance to carry out research on new environmental pollutants in giant panda habitat.Based on the summary of the ecological risks of ARGs,85 wild giant panda feces from Wolong Nature Reserve,Mabian Dafengding Nature Reserve,Guanyinshan Nature Reserve,Wanglang Nature Reserve,and Fushuiyuan State-owned Forest Farm,14 fecal samples of Wolong captive giant pandas,30 giant panda habitat soil samples and giant panda staple food bamboo samples have been selected in this study.Highthroughput quantitative PCR(HT-QPCR)based on 16 s r RNA gene was used to detect321 pairs of ARGs and 56 pairs of Mobile Genetic Elements(MGEs)to explore the different of ARGs distribution and its differences in the gut microbes of giant pandas.The gut microbes of giant pandas and soil microorganism were sequenced by metagenomic sequencing technology to explore the diversity and composition of giant panda gut microbes and soil microorganism between different regions.The potential effects of ARGs and MGEs in soil and staple food bamboo on ARGs in gut microbes of giant pandas were analyzed using Venn diagram visualization results.The main results are as follows:(1)Shannon index showed that there is no significant difference in the diversity of gut microbiome of giant pandas in various regions,including captive and wild giant pandas.The results of PCo A analysis showed that there was no significant difference in the composition of gut microbiome between Mabian Dafengding Nature Reserve and Wanglang Nature Reserve,but there were significant differences among other regions.At the phylum level,Firmicutes and Proteobacteria are the main dominant flora in the gut microbiome of giant pandas.(2)In the gut microbiome of giant pandas,232,179,8,119 and 90 ARGs were detected in the gut of wild giant pandas in Wolong Nature Reserve,Mabian Dafengding Nature Reserve,Guanyinshan Nature Reserve,Wanglang Nature Reserve and Fushuiyuan State-owned Forest Farm respectively;The number of MGEs detected were45,30,1,25 and 21 respectively.118 ARGs and 30 MGEs were detected in the gut of Wolong captive giant panda.PCo A analysis showed that there is no significant difference in the composition of ARGs in the gut of giant pandas between Wolong Nature Reserve and Fushuiyuan State-owned Forest Farm,Guanyinshan Nature Reserve and Mabian Dafengding Nature Reserve,Wanglang Nature Reserve and Guanyinshan Nature Reserve.The relative abundance of ARGs in Mabian Dafengding Nature Reserve was significantly higher than that in other areas;The relative abundance of multidrug ARGs in all regions was significantly higher than that of other categories,it was the main ARGs category in the gut of giant pandas in the study area.(3)The diversity of soil microorganisms in Wolong Nature Reserve is significantly higher than that in Fushuiyuan State-owned Forest Farm,and there is no significant difference in the diversity of soil microorganisms among other regions.PCo A analysis showed that there were significant differences in soil microbial composition between Wanglang Nature Reserve and Guanyinshan Nature Reserve(P < 0.05),and there were extremely significant differences in soil microbial composition among other regions(P< 0.01).At the phylum level,the dominant flora of soil microorganisms were concentrated in proteobacteria,actinobacteriota,acidobacteriota and Chloroflexi.(4)Among soil microorganisms,49,72,47,48 and 41 ARGs were detected in Wolong Nature Reserve,Mabian Dafengding Nature Reserve,Guanyinshan Nature Reserve,Wanglang Nature Reserve and Fushuiyuan State-owned Forest Farm respectively;The number of MGEs detected were 11,11,12,8 and 16 respectively.PCo A analysis showed that there was no significant difference in the composition of ARGs in soil microorganisms in each study area.Like the relative abundance of ARGs in the gut microbiome of giant pandas,the abundance of ARGs in Mabian Dafengding Nature Reserve is higher than that in other areas;Multidrug ARGs have a high relative abundance in soil microorganisms and are the main ARGs in the soil of the study area.(5)In the phyllospheric microorganisms of bamboo,the number of ARGs detected in Wolong Nature Reserve,Mabian Dafengding Nature Reserve,Guanyinshan Nature Reserve,Wanglang Nature Reserve and Fushuiyuan State-owned Forest Farm were 129,143,92,81 and 141 respectively;The number of MGEs detected were 26,29,22,20 and 30 respectively.The results of PCo A analysis showed that the clustering of ARGs in bamboo microorganisms,the staple food of giant pandas in each region,was obvious,and its composition was significantly different.Aminoglycoside ARGs have high abundance in Wolong Nature Reserve,Mabian Dafengding Nature Reserve and Fushuiyuan State-owned Forest Farm,and were the main ARGs category in the staple food bamboo of giant pandas;The relative abundance of multidrug ARGs in Mabian Dafengding Nature Reserve,Guanyinshan Nature Reserve and Wanglang Nature Reserve is high,which was the main ARGs category in the staple food bamboo of giant pandas.