屠宰生猪副猪嗜血杆菌的分离鉴定及药物敏感性分析

doi:10.16431/j.cnki.1671-7236.2019.07.033

摘要/Abstract

摘要：

为调查屠宰生猪副猪嗜血杆菌（Hps）的隐性感染情况，本研究从陕西省某规模化生猪屠宰场采集80份病变肺脏，应用PCR方法进行Hps核酸检测，无菌取阳性肺脏，通过平板划线分离获得疑似Hps，进一步对分离菌进行形态观察、培养特性分析、卫星现象观察、生化试验和分离菌核酸片段序列分析，纯化获得Hps分离菌；用药敏纸片法分析Hps分离菌株对常用药物的敏感性。结果显示，从80份病变肺脏中检出39份Hps阳性，阳性率48.75%（39/80）；从39份Hps阳性肺脏中分离获得3株疑似Hps，分离菌在显微镜下呈革兰氏阴性、短杆状；在添加有胎牛血清和NAD的TSA平板上生长良好；在金黄色葡萄球菌周围呈现出典型的"卫星现象"生长；能发酵葡萄糖、蔗糖、果糖、半乳糖、麦芽糖和阿拉伯糖，不发酵甘露醇；PCR扩增序列与Hps同源性在98%以上。药物敏感性试验结果显示，3株分离菌对环丙沙星、恩诺沙星和氟苯尼考等抗菌药物高度敏感，对青霉素、林可霉素和甲氧苄啶等抗菌药物耐药。本研究获得的Hps分离株为疫苗研制提供了基础材料，为生猪养殖场选择敏感药物防控Hps感染提供了参考依据。

关键词: 副猪嗜血杆菌(Hps); 分离鉴定; 药物敏感性试验

Abstract:

In order to investigate the hidden infection of Haemophilus parasuis (Hps) in slaughtered pigs,this study collected 80 diseased lungs from a large-scale pig slaughterhouse in Shaanxi province.The positive lungs were aseptically sampled,and the suspected Hps was obtained by streaking through the plate.The morphology of the isolates,analysis of the culture characteristics,observation of the satellite phenomenon,biochemical test and sequence analysis of the isolates nucleic acid fragments were carried out,and the Hps isolates were purified.The sensitivity of Hps isolates to commonly used drugs was analyzed by drug sensitive paper method.The results showed that 39 Hps positive samples were detected from 80 lungs,and the positive rate was 48.75% (39/80).Three suspected Hps were isolated from 39 Hps-positive lungs.The isolate was Gram-negative,short rod under the microscope,grew well on TSA plates supplemented with fetal bovine serum and NAD,exhibited a typical "satellite phenomenon" growth around S.aureus,could ferment glucose,sucrose,fructose,galactose,maltose and arabinose without fermenting mannitol.The PCR amplification sequence was more than 98% homologous to Hps.The results of drug susceptibility test showed that the three isolates were highly sensitive to antibacterial drugs such as ciprofloxacin,enrofloxacin and florfenicol,and were not sensitive to antibacterial drugs such as penicillin,lincomycin and trimethoprim.The Hps isolates obtained in this study provided the basic materials for vaccine development,and provided a reference for the selection of sensitive drugs to prevent and control Hps infection in pig farms.

Key words: Haemophilus parasuis(Hps); isolation and identification; drug sensitivity test

中图分类号:

S852.61

田昊伦, 冯秀, 王凯, 冯全文, 李鑫鑫, 范志新, 罗丽华, 孙颖, 周宏超, 张彦明, 郭抗抗. 屠宰生猪副猪嗜血杆菌的分离鉴定及药物敏感性分析[J]. 《中国畜牧兽医》, 2019, 46(7): 2151-2158.

TIAN Haolun, FENG Xiu, WANG Kai, FENG Quanwen, LI Xinxin, FAN Zhixin, LUO Lihua, SUN Ying, ZHOU Hongchao, ZHANG Yanming, GUO Kangkang. Isolation, Identification and Drug Sensitivity Analysis of Haemophilus parasuis in Slaughter Pigs[J]. , 2019, 46(7): 2151-2158.

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