番鸭源鸭疫里默氏杆菌的分离鉴定及其耐药性分析

doi:10.16431/j.cnki.1671-7236.2020.12.033

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (12): 4085-4092.doi: 10.16431/j.cnki.1671-7236.2020.12.033

番鸭源鸭疫里默氏杆菌的分离鉴定及其耐药性分析

肖琨¹, 陈广³, 张军², 温贵兰³, 文明³, 程振涛³

1. 六盘水市动物疫病预防控制中心, 六盘水 553000;
2. 六盘水市动物卫生监督所, 六盘水 553000;
3. 贵州大学动物科学学院, 动物疫病研究所, 贵阳 550016

收稿日期:2020-06-08 出版日期:2020-12-20 发布日期:2020-12-18
通讯作者: 温贵兰 E-mail:524340732@qq.com
作者简介:肖琨(1974-),男,贵州六盘水人,本科,高级兽医师,研究方向:动物疫病预防控制,E-mail:891556831@qq.com
基金资助:
2019年度贵州省农业科技支撑计划项目（黔科合支撑[2019]2286号）；贵州大学博士基金（贵大人基合字[2013]12号）；贵州省高层次创新型人才培养（黔财教[2017]号）；贵州省科技平台及人才团队计划（黔科合平台人才[2018]5253）

Isolation,Identification and Drug Resistance Analysis of Riemerella anatipestifer from Muscovy Duck

XIAO Kun¹, CHEN Guang³, ZHANG Jun², WEN Guilan³, WEN Ming³, CHENG Zhentao³

1. Liupanshui Animal Disease Prevention and Control Center, Liupanshui 553000, China;
2. Liupanshui Animal Health Supervision Institute, Liupanshui 553000, China;
3. Institute of Animal Diseases, School of Animal Science, Guizhou University, Guiyang 550016, China

Received:2020-06-08 Online:2020-12-20 Published:2020-12-18

摘要/Abstract

摘要： 为了解番鸭源鸭疫里默氏杆菌的耐药性，本研究对疑似鸭疫里默氏杆菌感染的发病番鸭进行细菌分离培养、革兰氏染色镜检、病鸭病原检测、生化试验、16S rRNA序列分析、PCR鉴定、药敏试验和耐药基因检测。细菌分离结果显示，分离菌在鲜血琼脂培养基上长出表面光滑、边缘整齐、有光泽、半透明的奶油状针尖大小菌落；革兰氏染色镜检呈革兰氏阴性短小杆菌，命名为GZQN201907。GZQN201907生化试验中尿素反应阳性，葡萄糖、麦芽糖、乳糖等生化反应呈阴性。其16S rRNA系统进化树与鸭疫里默氏杆菌处于同一分支；并且分离菌鸭疫里默氏杆菌OmpA基因PCR鉴定结果为阳性。其对头孢呋辛、红霉素、头孢他啶等18种抗菌药耐药，对羧苄西林和环丙沙星中度敏感，对新霉素和复方新诺明敏感。而耐药基因能检测出β-内酰胺类耐药基因VIM、TEM，四环素类耐药基因tetB，大环内酯类耐药基因ermB和ermF。药敏试验与耐药基因检测结果说明，GZQN201907对β-内酰胺类、四环素类、大环内酯类3类药物的耐药表型和耐药基因检测结果一致。动物回归试验中接种GZQN201907的雏鸭在72 h内全部死亡，而对照组雏鸭未出现任何症状，说明GZQN201907对雏鸭有致病力。试验成功分离到1株番鸭源鸭疫里默氏杆菌，为鸭疫里默氏杆菌病的防治奠定基础。

关键词: 番鸭; 鸭疫里默氏杆菌; 耐药基因

Abstract: In order to understand the drug resistance of Riemerella anatipestifer from Muscovy duck,this study carried out bacterial isolation and culture,Gram staining microscopy,pathogen detection,biochemical test,16S rRNA sequence analysis,PCR identification,drug sensitivity test and drug resistance gene detection for Muscovy duck suspected of Riemerella anatipestifer infection.The results of bacterial isolation showed that on the blood agar medium,the isolated bacteria grew creamy needle tip size colonies with smooth surface,neat edge,luster and translucency.The Gram-negative bacillus brevis was detected by Gram-negative staining microscopy,and it was named GZQN201907.In the biochemical test of GZQN201907,urea reaction was positive,but glucose,maltose,lactose and other biochemical reactions were negative.The 16S rRNA phylogenetic tree was in the same branch as Riemerella anatipestifer.And the OmpA gene of PCR identification results of the isolated strain were positive.The drug sensitivity test was sensitive to 18 antibiotics including cefuroxime,erythromycin and ceftazidime,moderately sensitive to carboxypicillin and ciprofloxacin,and sensitive to neomycin and cotrimoxazole.And the resistance genes could detect the β-lactam resistance genes VIM and TEM,tetracycline resistance genes tetB,macrolide resistance genes ermB and ermF.The results of drug sensitivity test and drug resistance gene detection indicated that GZQN201907 showed the same resistance phenotype and gene detection results for β-lactam,tetracycline and macrolide.In the animal regression test,all the ducklings inoculated with GZQN201907 died within 72 h,while the control group showed no symptoms,indicating that GZQN201907 was virulent to the ducklings.One strain of Riemerella anatipestifer from Muscovy duck was successfully isolated,which laid a foundation for the prevention and treatment of Riemerella anatipestifer from muscovich.

Key words: Muscovy duck; Riemerella anatipestifer; drug resistance gene

中图分类号:

S855.1⁺2

肖琨, 陈广, 张军, 温贵兰, 文明, 程振涛. 番鸭源鸭疫里默氏杆菌的分离鉴定及其耐药性分析[J]. 中国畜牧兽医, 2020, 47(12): 4085-4092.

XIAO Kun, CHEN Guang, ZHANG Jun, WEN Guilan, WEN Ming, CHENG Zhentao. Isolation,Identification and Drug Resistance Analysis of Riemerella anatipestifer from Muscovy Duck[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 4085-4092.

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番鸭源鸭疫里默氏杆菌的分离鉴定及其耐药性分析

Isolation,Identification and Drug Resistance Analysis of Riemerella anatipestifer from Muscovy Duck

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