贵州省三穗县鸭疫里默氏杆菌的分离鉴定、毒力基因检测及耐药性分析

doi:10.16431/j.cnki.1671-7236.2021.11.037

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (11): 4274-4283.doi: 10.16431/j.cnki.1671-7236.2021.11.037

贵州省三穗县鸭疫里默氏杆菌的分离鉴定、毒力基因检测及耐药性分析

吴征卓^1,4, 陈国权¹, 姚碧琼⁴, 施大军⁵, 阎朝华¹, 王娜¹, 王开功^1,2,3, 周碧君^1,2,3, 程振涛^1,2,3, 文明^1,2,3

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州大学动物疫病研究所, 贵阳 550025;
3. 贵州省动物疫病与兽医公共卫生重点实验室, 贵阳 550025;
4. 三穗县动物疫病预防控制中心, 三穗 556500;
5. 贵州千里山生态食品股份有限公司, 三穗 556500

收稿日期:2021-03-01 出版日期:2021-11-20 发布日期:2021-11-01
通讯作者: 王开功, 文明 E-mail:as.kgwang@gzu.edu.cn;as.mwen@gzu.edu.cn
作者简介:吴征卓(1988-),男,贵州三穗人,硕士,研究方向:兽医微生物与免疫学,E-mail:1037870378@qq.com
基金资助:
三穗鸭工程技术研究中心建设项目（黔科合平台人才[2019]5203号）；贵州省科技平台及人才团队计划项目（黔科合平台人才[2018]5253号）；贵州省研究生教育创新计划项目（GZZ2017002）

Isolation, Identification, Virus Gene Detection and Antibiotic Resistance Analysis of Riemerella anatipestifer in Sansui County of Guizhou Province

WU Zhengzhuo^1,4, CHEN Guoquan¹, YAO Biqiong⁴, SHI Dajun⁵, YAN Chaohua¹, WANG Na¹, WANG Kaigong^1,2,3, ZHOU Bijun^1,2,3, CHENG Zhentao^1,2,3, WEN Ming^1,2,3

1. College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Institute of Animal Disease, Guizhou University, Guiyang 550025, China;
3. Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, Guiyang 550025, China;
4. Animal Disease Prevention and Control Center of Sansui County, Sansui 556500, China;
5. Qianlishan Ecological Food Company Limited of Guizhou Province, Sansui 556500, China

Received:2021-03-01 Online:2021-11-20 Published:2021-11-01

摘要/Abstract

摘要： 为了解贵州省三穗县鸭疫里默氏杆菌（Riemerella anatipestifer，RA）的流行血清型、毒力和耐药情况，本试验从贵州省三穗县6个规模养鸭场临床疑似RA感染的病鸭体内分离出6株分离菌，对病原菌进行分离鉴定、毒力基因检测和耐药性分析。细菌分离鉴定结果显示，分离菌在巧克力琼脂培养基上长出表面光滑、圆形半透明的滴状菌落；革兰氏染色呈阴性短小杆菌，瑞氏染色呈两极浓染；分离菌均不具运动性，尿素、触酶和氧化酶试验均为阳性，符合RA生化特性，将6株分离菌分别命名为SS-RA1~SS-RA6；6株分离菌的16S rRNA基因序列与NCBI上RA参考菌株的基因序列相似性≥98%，说明6株分离菌均是RA；SS-RA1~SS-RA4为血清2型且含有8种毒力基因（OmpA、CAMP、wza、AS87_04050、Fur、SIP、TbdR1和luxE基因），SS-RA5和SS-RA6为血清11型，缺失AS87_04050基因，仅含有上述其余7种毒力基因；动物回归试验结果显示，攻毒组雏鸭均全部死亡，对照组雏鸭未表现明显临床症状，表明6株分离菌对雏鸭均有致病力；药敏试验结果显示，6株分离菌仅对羧苄西林、哌拉西林、头孢他啶、头孢氨苄、头孢曲松、头孢拉定和头孢哌酮7种抗菌药物敏感，对其他13种抗菌药物均表现不同程度的耐药，对氨基糖苷类和大环内酯类抗菌药物的耐药率为100%。本试验成功分离得到6株RA，为贵州省三穗县鸭疫里默氏杆菌病的疫苗选择和药物防治提供理论依据。

关键词: 鸭疫里默氏杆菌; 分离鉴定; 毒力基因; 耐药性分析

Abstract: In order to understand the epidemic serotype, virulence and drug resistance of Riemerella anatipestifer (RA) in Sansui county, Guizhou province, 6 isolates were isolated from clinically suspected RA infected ducks in six large-scale duck farms in Sansui county, Guizhou province. The pathogenic bacteria were isolated and identified, and virulence gene detection and drug resistance analysis were carried out. The results of bacterial isolation and identification showed that the isolates were smooth, round and translucent drop-shaped colonies on chocolate agar medium. Gram staining was negative for short bacilli, Wright staining showed bipolar staining. The isolates did not have movement. The test of urea, catalase and oxidase were positive, which was in line with the biochemical characteristics of RA. The 6 strains of isolates were named SS-RA1 to SS-RA6. The 16S rRNA gene sequence of the 6 strains of isolates was compared with that of the RA reference strain on NCBI, the gene sequence similarity was ≥ 98%, indicating that all 6 strains were RA. SS-RA1 to SS-RA4 were serotype 2 and contain 8 virulence genes (OmpA、CAMP、wza、AS87_04050、Fur、SIP、TbdR1 and luxE genes), SS-RA5 and SS-RA6 were serotype 11, lacking the AS87_04050 gene, and only containing the remaining 7 virulence genes. The results of animal regression test showed that all the ducklings in the virus attack group died, and the ducklings in the control group did not show obvious clinical symptoms, indicating that the 6 strains of bacteria were pathogenic to the ducklings. The results of drug sensitivity test showed that the 6 strains of isolates were only sensitive to 7 antibiotics:Carboxybenzyl, piperacillin, ceftazidime, cefalexin, ceftriaxone, cefradine and cefoperazone, and showed different degrees of resistance to the other 13 antibiotics. The resistance rates to aminoglycosides and macrolides were 100%. In this study, the 6 RA strains were successfully isolated, providing a theoretical basis for vaccine selection and drug control of Riemerella anatipestifer infection in Sansui county, Guizhou province.

Key words: Riemerella anatipestifer; isolation and identification; virulence genes; antibiotic resistance analysis

中图分类号:

S852.61

吴征卓, 陈国权, 姚碧琼, 施大军, 阎朝华, 王娜, 王开功, 周碧君, 程振涛, 文明. 贵州省三穗县鸭疫里默氏杆菌的分离鉴定、毒力基因检测及耐药性分析[J]. 中国畜牧兽医, 2021, 48(11): 4274-4283.

WU Zhengzhuo, CHEN Guoquan, YAO Biqiong, SHI Dajun, YAN Chaohua, WANG Na, WANG Kaigong, ZHOU Bijun, CHENG Zhentao, WEN Ming. Isolation, Identification, Virus Gene Detection and Antibiotic Resistance Analysis of Riemerella anatipestifer in Sansui County of Guizhou Province[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4274-4283.

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贵州省三穗县鸭疫里默氏杆菌的分离鉴定、毒力基因检测及耐药性分析

Isolation, Identification, Virus Gene Detection and Antibiotic Resistance Analysis of Riemerella anatipestifer in Sansui County of Guizhou Province

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