龙鑫;候建平;郑思思;李佳妮;王稳;朱丽琳;
LONG Xin;HOU Jian-ping;ZHENG Si-si;LI Jia-ni;WANG Wen;ZHU Li-lin;State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University;Xi'ning Wild Animal Epidemic Disease Monitoring Station;

• 1:青海大学省部共建三江源生态与高原农牧业国家重点实验室
• 2:西宁市野生动物疫源疫病监测站

摘要(Abstract)：

为探究高原兔(Lepus oiostolus)肠道菌群多样性，本试验利用16S rDNA高通量测序方法分析高原兔小肠和大肠部位菌群种类及相对丰度，同时利用不同培养基对高原兔新鲜粪便中的微生物进行分离培养，并进一步对获得的菌株进行耐药性分析。16S rDNA高通量测序结果显示，高原兔小肠和大肠部位菌群在菌门水平主要由拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、互养菌门(Synergistetes)、蓝细菌门(Cyanobacteria)组成。优势菌属主要包括肠杆菌属(Enterobacter)、理研菌科RC9肠道群(Rikenellaceae RC9 gut group)、拟杆菌属(Bacteroides)、克里斯滕森菌科R-7群(Christensenellaceae R-7 group)。基于细菌培养分离方法共分离到25株细菌，隶属于6属、9种。耐药性分析发现，分离菌株对3～27种抗菌药物表现出耐药性。其中2种潜在有益菌株对5种常见肠道致病菌表现出一定的抑制能力。本试验结果为深入了解高原兔肠道微生物组提供了基础数据。
To explore the diversity of gut microbiota of Tibetan hare(Lepus oiostolus), this study applied 16S rDNA high-throughput sequencing method to determine the species and relative abundance of microbiota in its small and large intestines, and utilized different culture media to isolate microorganisms from its fresh feces followed by drug resistance testing on the obtained strains. The results of 16S rDNA high-throughput sequencing showed that the microbiota of small and large intestines of Lepus oiostolus comprised mainly Bacteroidetes, Firmicutes, Actinobacteria, Synergistetes and Cyanobacteria, with the genera Enterobacter, Rikenellaceae RC9 gut group, Bacteroides, and Christensenellaceae R-7 group being dominant. A total of 25 bacterial strains were isolated and assigned to 6 genera and 9 species. Drug resistance testing showed that the isolated strains were resistant to 3-27 kinds of antibacterial drugs. Two potential beneficial strains demonstrated certain inhibitory ability against 5 common intestinal pathogens. These preliminary data on the gut microbiome of Lepus oiostolus open the way for further investigation on the microbial ecology as well as microbe-host relationship during health and disease states in Lepus oiostolus.

关键词(KeyWords)： 高原兔;肠道菌群;16S rDNA高通量测序技术;细菌分离培养;耐药性
Lepus oiostolus;gut microbiota;16S rDNA high-throughput sequencing;bacterial isolation and culture;drug resistance

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区科学基金项目(31960277);国家自然科学基金国际(地区)合作与交流项目(32111530018,21-54-53031)

作者(Authors): 龙鑫;候建平;郑思思;李佳妮;王稳;朱丽琳;
LONG Xin;HOU Jian-ping;ZHENG Si-si;LI Jia-ni;WANG Wen;ZHU Li-lin;State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University;Xi'ning Wild Animal Epidemic Disease Monitoring Station;

参考文献(References)：

• [1] 王振宇，李叶，张翔，等.高原兔夏季卧栖地生境利用关键因子分析[J].四川动物，2015,34(1):47- 52.
• [2] Wei F,Wu Q,Hu Y,et al.Conservation metagenomics:a new branch of conservation biology[J].Sci China Life Sci,2019,62(2):168- 178.
• [3] 赵宇卓，吴尽，杨天雄，等.基于宏基因组学的野生动物肠道微生物研究进展[J].野生动物学报，2021,42(2):568- 574.
• [4] 国家卫生健康委员会.WS/T 639—2018 抗菌药物敏感性试验的技术要求[S].北京：中国标准出版社，2018-12-11.
• [5] 宋冰，任战军，宫瑞光，等.工厂兔、散养兔和野兔盲肠菌群多样性及菌群结构分析[J].家畜生态学报，2017,38(3):25- 30.
• [6] 祝辉.五种植食性动物肠道菌群结构及多样性的比较研究[D].雅安：四川农业大学，2018.
• [7] Valdes A M,Walter J,Segal E,et al.Role of the gut microbiota in nutrition and health[J].BMJ,2018,361:k2179.
• [8] Zafar H,Saier M H Jr.Gut Bacteroides species in health and disease[J].Gut Microbes,2021,13(1):e1848158.
• [9] Tavella T,Rampelli S,Guidarelli G,et al.Elevated gut microbiome abundance of Christensenellaceae,Porphyromonadaceae and Rikenellaceae is associated with reduced visceral adipose tissue and healthier metabolic profile in Italian elderly[J].Gut Microbes,2021,13(1):e1880221.
• [10] 王恩召，范分良，李艳玲，等.水稻根系细菌挥发性有机物对小孢根霉的非接触性抑制作用[J].中国农业科学，2020,53(10):1986- 1996.
• [11] 王雅君，刘长建，周玉荣，等.蒙氏肠球菌Sna的抗氧化及降胆固醇特性[J].食品安全质量检测学报，2017,8(9):3445- 3449.
• [12] 董晓飞，李伟，张晓晗.托尔豪特链球菌感染引起脓胸1例[J].检验医学与临床，2021,18(8):1183- 1184.
• [13] 李俊琴.藏羚羊粪便和高原鼠兔肠内容物可培养菌群结构与新种发现[D].太原：山西医科大学，2020.
• [14] 黄坚，童京京，岳华，等.牦牛发酵酸奶中耐久肠球菌的筛选鉴定和益生特性[J].食品科学，2017,38(12):43- 49.
• [15] 孙康泰，张建民，蒋大伟，等.我国动物源细菌耐药性的研究进展及防控策略[J].中国农业科技导报，2020,22(5):1- 5.
• [16] 张红娜，董梦洁，周玉法，等.动物粪便施肥措施促进耐药基因在粪便—土壤—蔬菜之间的散播[J].环境科学，2021,42(4):2080- 2088.