1株血清2型鸭疫里默氏杆菌的分离鉴定、致病性研究及灭活疫苗制备  

Isolation,Identification,Pathogenicity Analysis and Inactivated Vaccine Preparation of a Strain of Riemerella anatipestifer Serotype 2

作　　者：戴婷婷 刘荣昌[3] 陈海玉 庄丽云 韦玉帆 谭慧敏 刘洋 唐福川 黄瑜[3] 程艳青 包银莉 郑新添[2] DAI Tingting;LIU Rongchang;CHEN Haiyu;ZHUANG Liyun;WEI Yufan;TAN Huimin;LIU Yang;TANG Fuchuan;HUANG Yu;CHENG Yanqing;BAO Yinli;ZHENG Xintian(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;School of Life Sciences,Longyan University,Longyan 364012,China;Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention,Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350003,China)

机构地区：[1]福建农林大学动物科学学院,福州350002 [2]龙岩学院生命科学学院,龙岩364012 [3]福建省农业科学院畜牧兽医研究所,福建省禽病防治重点实验室,福州350003

出　　处：《中国畜牧兽医》2024年第12期5499-5506,共8页China Animal Husbandry & Veterinary Medicine

基　　金：财政部和农业农村部:国家现代农业产业技术体系(CARS-42);福建省自然科学基金项目(2023J01363);龙岩学院预防兽医学与生物技术福建省高等学校重点实验室开放基金(2022KF03)。

摘　　要：【目的】本研究拟对从1例死亡雏鸭体内分离获得的鸭疫里默氏杆菌进行生物学特性和疫苗效果分析,旨在为鸭传染性浆膜炎的防治提供指导。【方法】采集送检雏鸭的脑、心脏和肝脏组织进行细菌分离培养,通过菌落形态观察、革兰染色镜检、16S rDNA PCR扩增测序鉴定菌株并分析其遗传进化关系;采用玻片凝集法鉴定菌株血清型,检测菌株对常见抗菌药物的敏感性及致病性;利用分离株制备灭活疫苗,测定其免疫保护效果。【结果】分离株在血平板上呈半透明、露滴状圆形小菌落,革兰染色可见多个丝状排列的阴性短杆菌,瑞氏染色可见两级浓染的短小杆菌。16S rDNA测序结果显示,分离株与鸭疫里默氏杆菌参考株CZ-RA03处于同一分支,相似性>99.9%。血清型鉴定结果显示,分离株为鸭疫里默氏杆菌血清2型。药敏试验结果显示,分离株对氧氟沙星、链霉素、卡那霉素等药物耐药,对氟苯尼考、四环素、多西环素等药物敏感。致病性试验结果显示,分离株对7日龄雏鸭的半数致死量(LD 50)为1.51×10^(7)CFU,雏鸭感染后肾脏载菌量最高,为2.15×10^(6)CFU/g。利用分离株制备油乳剂灭活疫苗免疫7日龄雏鸭,其免疫攻毒后存活率为86.67%。【结论】本研究从发病雏鸭中分离鉴定出1株具有较高致病性的血清2型鸭疫里默氏杆菌,以该菌制备的油乳剂灭活疫苗具有良好的免疫保护效果。研究结果为鸭传染性浆膜炎的防治提供了理论基础。【Objective】The objective of this study was to analyze the biological characteristics and vaccine effectiveness of Riemerella anatipestifer isolated from a dead duckling,in order to provide guidance for the prevention and treatment of infectious serositis of duck.【Method】The brain,heart and liver tissues of the duckling were collected for bacterial isolation and culture.The strain was identified by colony morphology observation,Gram staining microscopy,16S rDNA PCR amplification and sequencing,and the genetic evolution relationship was analyzed.The serotype of the strain was identified by slide agglutination method,and the susceptibility to common antibiotics and pathogenicity of the strain were detected.The inactivated vaccine was prepared by the isolate and its immunoprotective effect was measured.【Result】The isolate showed semi-transparent,dewdrop-like round colonies on the blood plate.Gram staining showed multiple filamentous arrangement of negative short bacilli,and Wright’s staining showed two stages of intensely stained short bacilli.16S rDNA sequencing showed that the isolate was in the same branch as the reference strain of Riemerella anatipestifer CZ-RA03,and the similarity was over 99.9%.Serotype identification showed that the isolate was Riemerella anatipestifer serotype 2.The results of drug sensitivity test showed that the isolate was resistant to ofloxacin,streptomycin and kanamycin,and sensitive to flufenicol,tetracycline and doxycycline.The results of pathogenicity test showed that the median lethal dose(LD 50)of the isolate to 7-day-old duckling was 1.51×10^(7)CFU,and the kidney bacterial load of infected duckling was the highest(2.15×10^(6)CFU/g).The oil emulsion inactivated vaccine was used to immunize 7-day-old duckling,and the surviral rate was 86.67%.【Conclusion】In this study,a high pathogenicity strain of Riemerella anatipestifer serotype 2 was isolated and identified from infected ducklings,and the oil emulsion inactivated vaccine prepared by the strain had good immune

关 键 词：鸭疫里默氏杆菌 生物学特性 免疫原性 

分 类 号：S852.61[农业科学—基础兽医学]