洛阳某猪场猪传染性胸膜肺炎放线杆菌的分离鉴定

doi:10.16431/j.cnki.1671-7236.2020.04.031

摘要/Abstract

摘要： 为确定洛阳某猪场疑似猪传染性胸膜肺炎的病原种类及其血清型，为其临床科学防疫和合理用药提供参考方案，本试验无菌采集了病猪肺脏等病料，采用病菌培养后形态观察、革兰氏染色、生理生化特性试验、PCR诊断等技术对病原菌进行实验室分离鉴定，通过基因测序和同源性分析方法对其进行确诊，通过动物攻毒试验和药敏试验分析分离菌株的致病性和耐药性，采用5对分型引物对分离菌进行基因分型以确定病原菌血清型。结果显示，分离菌2007菌落形态为光滑、透明、边缘整齐、圆形、隆起的小菌落，革兰氏染色特征为阴性杆菌且呈多型性，具有猪传染性胸膜肺炎放线杆菌（APP）的典型培养特征，且生化结果符合猪传染性胸膜肺炎放线杆菌的生化特性。同源性分析结果显示，分离菌株2007与猪传染性胸膜肺炎放线杆菌菌株FJ848573.1同源性为99.4%，说明分离株2007是猪传染性胸膜肺炎放线杆菌。动物攻毒试验结果显示，分离菌株2007对小鼠有强致病性。药敏试验结果显示，分离株2007对泰妙菌素、氨苄西林和头孢噻呋钠等高敏，对替米考星、红霉素、氟苯尼考等20种抗菌药完全耐药，对34种抗生素的耐药率高达76.5%，具有较强的多重耐药性。分离菌2007基因分型扩增到目的基因片段与血清型5型结果相符。本试验结果为发病猪场提出了解决该病的具体防治措施，并最终获得了良好的效果，为猪传染性胸膜肺炎放线杆菌临床耐药折点的制定提供了一些参考。

关键词: 猪传染性胸膜肺炎; 药敏试验; 基因分型; 分离鉴定

Abstract: In order to determine the pathogenic species and serotypes of suspected infectious pleuropneumonia in a pig farm in Luoyang,and to provide a reference plan for its clinical scientific epidemic prevention and rational drug use,in this experiment,the diseased pig lungs and other diseased materials were collected aseptically,and the pathogenic bacteria were isolated and identified in laboratory by morphological observation,Gram staining,physiological and biochemical characteristics test,PCR diagnosis and other technologies.It was confirmed by gene sequencing and homology analysis.The pathogenicity and drug resistance of the isolated strains were analyzed by animal attack test and drug sensitivity test.Five pairs of primers were used for genotyping to determine the serotypes.The results showed that the colony morphology of 2007 was smooth,transparent,orderly,round and protuberant,Gram staining was negative and polytypic.It had the typical culture characteristics of Actinobacillus pleuropneumoniae (APP),and the biochemical results were consistent with the biochemical characteristics of Actinobacillus pleuropneumoniae.The results of homology analysis showed that the homology between the isolate 2007 and Actinobacillus pleuropneumoniae strain FJ848573.1 was 99.4%,indicating that the isolate 2007 was Actinobacillus pleuropneumoniae.The results showed that the isolate 2007 had strong pathogenicity to mice.The results of drug sensitivity test showed that the isolate 2007 was highly sensitive to temicillin,ampicillin and ceftiofur sodium,completely resistant to 20 kinds of antimicrobial agents,such as tilmicosin,erythromycin and florfenicol,and the resistance rate to 34 kinds of antibiotics was 76.5%,with strong multiple drug resistance.The results of the isolate 2007 genotyping showed that the target gene fragment was consistent with serotype 5.The results of this study provided specific prevention and treatment measures for the pig farms,and finally achieved good results,which provided some references for the establishment of clinical resistant fold points of Actinobacillus pleuropneumoniae.

Key words: porcine contagious pleuropneumonia; isolation and identification; genotyping; drug sensitivity

中图分类号:

S852.61⁺9

李凯明, 董发明, 刘守川, 刘杰, 梁立钦, 李凯强. 洛阳某猪场猪传染性胸膜肺炎放线杆菌的分离鉴定[J]. 中国畜牧兽医, 2020, 47(4): 1241-1249.

LI Kaiming, DONG Faming, LIU Shouchuan, LIU Jie, LIANG Liqin, LI Kaiqiang. Isolation and Identification of Porcine Contagious Actinobacillus pleuropneumoniae in Luoyang[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1241-1249.

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