河南省安阳市副猪嗜血杆菌流行菌株调查与耐药性分析  

Investigation on the Prevalent Strains of Haemophilus parasuis in Anyang City,Henan Province and Analysis on Its Drug Resistance

作　　者：杜娟 刘亚辉 苏玉贤 Du Juan;Liu Yahui;Su Yuxian(Kaifeng Livestock Workstation,Kaifeng,Henan 475000,China;Pingdingshan Animal Husbandry Development Center,Pingdingshan,Henan 467000,China)

机构地区：[1]开封市畜牧工作站,河南开封475000 [2]平顶山市畜牧发展中心,河南平顶山467000

出　　处：《中国动物检疫》2023年第8期20-27,共8页China Animal Health Inspection

基　　金：农业农村部2022年兽药质量监督抽检和风险监测计划项目(农办牧〔2022〕1号)。

摘　　要：为了解河南省安阳市副猪嗜血杆菌流行菌株的血清型和耐药性情况,2022年从该地区25家养猪场无菌采集临床疑似感染副猪嗜血杆菌的病死猪肝脏、肺脏、关节渗出液等组织病料175份,采用细菌分离纯化培养、形态学观察、“卫星生长”培养、生化反应和PCR鉴定等方法进行副猪嗜血杆菌分离鉴定,应用琼脂免疫扩散法进一步进行血清分型,采用Kirby-Bauer纸片法检测分离菌株对16种抗生素的耐药性。结果显示:从175份组织病料中检测出34株副猪嗜血杆菌,检出率为19.43%;分离菌在含有NAD和犊牛血清的TSA培养基上产生针尖大小、无色透明、光滑湿润的菌落,镜检可见革兰氏阴性、长杆状、细丝状等多形态菌体,在金黄色葡萄球菌周围菌落出现“卫星生长”现象,且无溶血,分离培养结果符合副猪嗜血杆菌特征。经PCR扩增测序,分离菌株与GenBank中公布的副猪嗜血杆菌16S rRNA序列同源性在97%以上;共鉴定出4型14株、5型4株、6型3株、13型8株和未定型5株,分别占41.18%、11.76%、8.82%、23.53%和14.71%;34株副猪嗜血杆菌对对阿莫西林、青霉素G、链霉素、土霉素4种抗生素耐药性最高,耐药率为88.24%~94.12%,多重耐药较严重,至少对5种以上抗生素产生耐药性,以9重耐药最多。结果表明,河南省安阳市猪场存在一定程度的副猪嗜血杆菌流行,流行菌株血清型复杂,耐药严重。本调查为河南省安阳市副猪嗜血杆菌病防控提供了数据支撑。In order to investigate the serotype and drug resistance of Haemophilus parasuis(HPS)prevalent in Anyang City,Henan Province,in 2022,175 tissue samples including livers,lungs,joint exudates and other tissues of dead pigs clinically suspected of infection with HPS were aseptically collected from 25 swine farms for isolation and identification of HPS using bacterial isolation and purification culture,morphological observation,satellite growth culture,biochemical reaction and PCR assay,as well as further serotyping by agar immunodiffusion method,then the resistance of the isolates to 16 antibiotics was tested by Kirby-Bauer method.The results revealed that 34 strains of HPS were detected with a detection rate of 19.43%;bacterial colonies that were of pinhead size,colorless,transparent,smooth and humid were produced by the isolates on TSA medium containing NAD and calf serum;Gram-negative bacteria with various forms such as long rod-shaped and filamentous,were observed through microscopic examination;the phenomenon of“satellite growth”appeared in the bacterial colonies around Staphylococcus aureus,without hemolysis,and the results of isolation and culture were consistent with the characteristics of HPS.PCR amplification and sequencing results showed that the isolates were 97%identity to 16S rRNA sequence of HPS published in GenBank;14 strains of serotype 4,4 strains of serotype 5,3 strains of serotype 6,8 strains of serotype 13 and 5 unidentified strains were identified,accounting for 41.18%,11.76%,8.82%,23.53%and 14.71%respectively;34 strains of HPS were most resistant to amoxicillin,penicillin G,streptomycin and oxytetracycline,with the drug resistance rate ranged from 88.24%to 94.12%,especially resistant to multiple drugs,to 5 antibiotics at least and to 9 at most.In conclusion,HPS was prevalent in swine farms in Anyang City to a certain extent,and the prevalent strains contained complex serotypes and were highly resistant to drugs.Data support was thus provided for the prevention and control of HPS in Anyang

关 键 词：副猪嗜血杆菌 分离鉴定 血清分型 耐药试验 

分 类 号：S858.28[农业科学—临床兽医学]