人工养殖林麝脓肿病病原菌的分离与鉴定  

Isolation and Identification of Pathogenic Bacteria for Abscess in Captive Forest Musk Deer(Moschus berezovskii)

作　　者：吴世洲 杨林富 钱丽敏 杨明华[2] 李琦华[2] 陈培富 WU Shizhou;YANG Linfu;QIAN Limin;YANG Minghua;LI Qihua;CHEN Peifu(College of Veterinary Medicine,Yunnan Agricultural University,Kunming,650201,China;College of Animal Science and Technology,Yunnan Agricultural University,Kunming,650201,China)

机构地区：[1]云南农业大学动物医学院,昆明650201 [2]云南农业大学动物科学技术学院,昆明650201

出　　处：《野生动物学报》2021年第3期872-878,共7页CHINESE JOURNAL OF WILDLIFE

基　　金：云南省重大科技专项计划(生物医药)项目(2018ZF012)。

摘　　要：滇西某林麝养殖场频发林麝(Moschus berezovskii)脓肿病,死亡率高,经济损失大。因此,有必要尽快明确林麝脓肿病的具体病原,以有效防治该病。对该养殖场健康和患病林麝各4只(即2组),采集抗凝血液测定其生理生化指标,并对组间差异做配对t检验;采集皮下及肝脏脓肿内容物接种血琼脂,对分离菌株做形态观察、生化试验、药敏试验和重要基因的聚合酶链式反应(PCR)扩增;取其中1病死个体肝脏病变组织制作石蜡切片,做病理显微观察。结果表明:皮下和肝脏脓肿血琼脂培养均获得乳白色菌落,16S rRNA基因序列分析显示,其病原菌分别为黏质沙雷菌(Serratia marcescens,3株)和大肠杆菌(Escherichia coli,1株)。PCR扩增发现,前者携带phlA、swr、shlA、shlB毒力基因,后者具有肠外致病性大肠杆菌特征基因ompA。所有分离菌株均对青霉素和氨苄西林耐药,对头孢曲松、卡那霉素和恩诺沙星敏感;镜检见肝组织出现大面积干酪样坏死,相关结缔组织有大量炎性细胞浸润。结合已有相关研究报道,提示黏质沙雷菌和肠外致病性大肠杆菌是引起该场林麝脓肿病的重要机会致病菌。Captive forest musk deer(Moschus berezovskii)frequently contracted abscess disease at a deer farm in West Yunnan,which caused high mortality and important economic loss.Specific pathogens causing abscess disease need to be identified as soon as possible for achieving effective prevention and treatment.Anticoagulated blood samples were collected from 2 groups of musk deer(4 healthy and 4 sick deers each)at the farm.Physiological and biochemical indexes were then measured and the difference between the 2 groups was subjected to t-paired test.The samples of content in the subcutaneous and hepatic abscesses from the sick deer were inoculated to blood agar.The isolates were subjected to morphological observation,biochemical tests,drug susceptibility tests,and amplification of important genes using polymerase chain reaction(PCR).The paraffin sections of lesion tissues from the liver of a dead deer were made for pathological microscopy.Milky white colonies formed on the blood agar for both the subcutaneous and hepatic abscess tissue samples,and 16S rRNA gene sequencing showed that they were Serratia marcescens(3 isolates)and Escherichia coli(1 isolate),respectively.Using PCR amplification,we detected four virulence genes including phlA,swr,shlA,and shlB in the former,and extraintestinal pathogenic E.coli characteristic gene ompA in the latter.All the isolates were resistant to penicillin and ampicillin,and were susceptible to ceftriaxone,kanamycin and enrofloxacin.Massive caseous necrosis in the effected hepatic tissue and infiltration of numerous inflammatory cells into the related connective tissue were observed under microscope.Combined with the reported findings,the results of this study indicated that S.marcescens and extraintestinal pathogenic E.coli were important opportunistic bacteria that caused abscess disease at the forest musk deer farm.

关 键 词：林麝 脓肿 黏质沙雷菌 肠外致病性大肠杆菌 人工养殖 

分 类 号：S858.9[农业科学—临床兽医学]