| 高山兀鹫肠道微生物的分离鉴定与耐药性分析 |
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| 作者姓名： | 候建平 郑思思 龙鑫 海佳怡 王稳 朱丽琳 |
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| 作者单位： | 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;青海大学生态环境工程学院;西宁市野生动物疫源疫病监测站 |
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| 基金项目： | 国家自然科学基金地区科学基金项目(31960277);青海省自然科学基金青年项目(2018-ZJ-932Q);生态环境部生物多样性保护专项(2018QHU63)。 |
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| 摘 要： | 在青海高原,国家二级保护鸟种,高山兀鹫(Gyps himalayensis)以动物和人类的尸体为食,对维持生态系统平衡至关重要。然而,腐尸中存在诸多有害因素(如兽药残留,病原菌或病毒感染等),对高山兀鹫种群的生存和保护构成威胁。肠道微生物调控宿主的免疫、营养物质代谢及发育等诸多生理过程,因此,也成为野生动物保护领域的研究热点。为了解高山兀鹫肠道微生物的多样性及其耐药情况,本研究采集野生高山兀鹫的新鲜粪便在实验室进行细菌分离培养、16S rRNA基因PCR扩增和序列分析、生化鉴定以及细菌耐药性试验。结果显示:从高山兀鹫新鲜粪便中共分离到119株细菌,隶属于6个属、8个种,分别是大肠埃希氏菌(Escherichia coli)、藤黄微球菌(Micrococcus luteus)、奥斯陆莫拉氏菌(Moraxella osloensis)、蜡样芽孢杆菌(Bacillus cereus)、索氏类梭菌(Paeniclostridium sordellii)、蒙氏肠球菌(Enterococcus mundtii)、海氏肠球菌(Enterococcus hirae)以及耐久肠球菌(Enterococcus durans)。选取含有34种不同抗生素的药敏纸片对分离的8种代表性菌株进行耐药性分析,发现这些菌株对8到26种不等的抗生素表现出耐药性。进一步分析发现,氧氟沙星、哌拉西林、诺氟沙星、环丙氟哌酸、左氧氟沙星以及强力霉素能够抑制本研究中分离到的所有8种代表性菌株。而制霉菌素以及林可霉素对所有分离到的8种代表性菌株均无抑制作用。本研究利用培养的手段分析了高山兀鹫肠道微生物中潜在的致病菌、益生菌及其耐药性,增添了对高山兀鹫肠道微生物的新认知,对其保护具有重要意义。
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| 关 键 词： | 高山兀鹫 肠道微生物 分离鉴定 耐药性 |
Isolation,Identification and Antimicrobial Susceptibility of Gut Microbiota from Himalayan Vulture(Gyps himalayensis) |
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| Authors: | HOU Jianping ZHENG Sisi LONG Xin HAI Jiayi WANG Wen ZHU Lilin |
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| Institution: | (State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining,810016,China;College of Eco-Environmental Engineering,Qinghai University,Xining,810016,China;Xining Wild Animal Epidemic Disease Monitoring Station,Xining,810001,China) |
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| Abstract: | Himalayan vulture(Gyps himalayensis)is a Class II national protected bird species in China.On the Qinghai Plateau,these are important scavengers(feeding on the carcasses of animals and humans).However,there are many potentially toxic components of carcasses(e.g.,veterinary drug residues,pathogens,viruses),which pose a serious threat to the survival and protection of Himalayan vultures populations.The gut microbiome regulates a range of host physiological functions including immunity,nutrient metabolism and development.This has become a research hotspot in the field of wildlife protection.To investigate the diversity and antibiotic susceptibility of gut microbiota from Himalayan vultures,we collected their fresh feces for bacterial isolation,16S rRNA gene PCR amplification and sequence analysis,biochemical identification and antibiotic resistance tests.The results showed that 119 strains of bacteria of 6 genera and 8 species,were isolated from the fresh feces of Himalayan vultures.They were Escherichia coli,Micrococcus luteus,Moraxella osloensis,Bacillus cereus,Paeniclostridium sordellii,Enterococcus mundtii,Enterococcus hirae,and Enterococcus durans.Drug resistance analysis was conducted on the eight representative,isolated strains or species with 34 different antibiotic susceptibility slips.The eight strains showed resistance to 8 to 26 different antibiotics.Further analysis showed that ofloxacin,piperacillin,norfloxacin,ciproxan,levofloxacin and doxycycline inhibited all eight representative strain/species.Nystatin and lincomycin had no inhibitory effect on any of the eight strains.Our study objective was to enhance understanding of the intestinal microorganisms of Himalayan vultures as a basis for their protection and conservation. |
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| Keywords: | Himalayan vulture Gut microbiota Isolation and identification Antibiotic resistance |
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