摘要
为明确贵阳市某公园动物园1只亚洲黑熊死亡的原因,本试验采集死亡黑熊内脏器官,进行细菌分离培养、菌体形态观察、生理生化鉴定、16SrRNA基因序列分析和小鼠感染试验。结果显示,从死亡黑熊肺脏样本分离出一种圆形、凸起、表面光滑、边缘整齐、中等大小的乳白色菌落,经染色镜检为呈单在或双排列的革兰氏阴性小杆菌。该分离菌可利用葡萄糖、蔗糖、甘露醇、山梨醇和枸橼酸盐,不发酵乳糖,不产生硫化氢,精氨酸脱羧酶阳性,赖氨酸脱羧酶阴性,符合阴沟肠杆菌生化特征。经细菌16SrRNA通用引物扩增,可得到大小为1 419bp的片段,与NCBI上相应序列进行BLAST比对,结果显示与阴沟肠杆菌序列相似度达99.9%,确认该分离菌为阴沟肠杆菌。用该分离菌腹腔接种小鼠,可致死小鼠,半数致死量(LD50)为7.94×108 CFU/mL。药敏试验发现,该分离菌对丁胺卡那霉素、氧氟沙星、诺氟沙星和环丙沙星等常用抗菌药均保持较高的敏感性。本试验结果表明,阴沟肠杆菌是导致该亚洲黑熊死亡的病原。
To determine the cause of the death of an Asian Black bear（Ursus thibetanus）at a park zoo in Guiyang,the death Black bear internal organs were collected for bacterial isolation and culture,cell morphology observation,physiological and biochemical identification,16 SrRNA gene sequence analysis and mouse infection tests.The results showed that a round,convex,smooth surface,well-defined,medium-sized milky white colony was isolated from the dead Black bear lung specimen as gram-negative bacillus by microscopy.It could utilize the glucose,sucrose,mannitol,sorbitol,and citrate,non-lactose fermentation,non-generated sulfuretted hydrogen,positive arginine decarboxylation test and negative lysine decarboxylase,consistent with the biochemical characteristics of Enterobacter cloacae.1 419 bp fragments were generated by 16 SrRNA primer amplification,and the similarity reached to 99.9% with the sequences of Enterobacter cloacae by BLAST comparison with the sequences on NCBI.Mice were inoculated intraperitoneally with theisolated bacteria,and the mice were lethal.The median lethal dose（LD50）was 7.94×108 CFU/mL.Drug susceptibility testing found that the isolates maintained high sensitivity to commonly used antimicrobial agents such as amikacin,ofloxacin,norfloxacin and ciprofloxacin.The results of this test showed that Enterobacter cloacae caused the death of the Asian Black bear.
作者
粟海军
王野影
贺欣薇
万润
文明
SU Haijun1 , WANG Yeying2 , HE Xinwei3 , WAN Run3 , WEN Ming3.(1. Research Center for Bio diversity and Nature Conservation , Guizhou University , Ouiyang 550025, China; 2. College of Life Science, Ouizhou Normal University, Guiyang 550025, China ; 3. College of Animal Science, Guizhou University, Guiyang 550025, Chin)
出处
《中国畜牧兽医》
CAS
北大核心
2018年第8期2341-2346,共6页
China Animal Husbandry & Veterinary Medicine
基金
贵州省科技联合基金项目(黔科合LH号[2015]7665号)
贵州省留学人员科技创新项目(黔人项目资助合同(2016)18号)
关键词
亚洲黑熊
阴沟肠杆菌
分离
鉴定
Ursus thibetanus
Hnterobacter cloacae
isolation
identification
