1株副猪嗜血杆菌血清4型的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2022.03.032

摘要/Abstract

摘要： 【目的】阐明引起河南漯河某规模化猪场仔猪呼吸道症状的主要细菌性病原体及其生物学特性。【方法】无菌采集发病仔猪的口鼻拭子及病死猪的胸腔积液及肺脏、肝脏、脾脏等组织器官，进行病原的PCR检测，并通过细菌分离培养、形态学观察、卫星试验、16S rDNA基因PCR扩增、系统进化树构建、多重PCR鉴定分离菌的血清型、致病性试验和药敏试验等对分离菌株进行生物学特性分析。【结果】在含有V因子的BHI固体平板上生长出圆形、表面光滑湿润、无色透明的微小菌落，在不含V因子的培养基上不生长；分离菌株经革兰染色，镜下可见革兰阴性菌，多呈单个存在的球杆、长杆状；在金黄色葡萄球菌的培养物周围形成典型的"卫星菌落"；16S rDNA基因序列分析表明，分离菌株与副猪嗜血杆菌（Haemophilus parasuis，Hps）处于同一分支，相似性>97%，亚型鉴定结果判定分离菌为Hps血清4型；将分离的Hps腹腔注射感染小鼠，可导致感染小鼠多个器官不同程度出血，以肺脏最为明显；高剂量感染可导致小鼠死亡。药敏试验发现，该菌株对四环素、环丙沙星、复方新诺明具有较强抗性，合理用药后，取得了良好的治疗效果。【结论】试验成功分离出1株血清4型Hps，可致感染小鼠多个器官不同程度出血，对四环素、环丙沙星、复方新诺明耐药，研究结果为副猪嗜血杆菌病的临床诊断和治疗提供了理论依据。

关键词: 副猪嗜血杆菌(Hps); 血清4型; 分离鉴定; 耐药性

Abstract: 【Objective】 This study was aimed to clarify the main bacterial pathogens and their biological characteristics that cause respiratory symptoms of piglets in a large-scale pig farm in Luohe, Henan province.【Method】 Oral and nasal swabs of infected piglets and tissues and organs such as pleural effusion, lung, liver and spleen of dead pigs were collected aseptically for PCR detection of pathogens.The biological characteristics of the isolate was analyzed by isolation and culture of bacteria, morphological observation, satellite test, PCR amplification of 16S rDNA gene, construction of genetic evolution tree, multiple PCR identification of serotypes of isolated strains, pathogenicity test of mice and drug sensitivity test.【Result】 Round, smooth, moist, colorless and transparent microcolonies grew on the BHI solidplate containing V factor, but did not grow on the medium without V factor.The isolates were stained with Gram staining, and Gram-negative bacteria were seen under the microscope, mostly in the shape of a single ball club and long rod.Atypical 'satellite colony' was formed around the culture of Staphylococcus aureus.16S rDNA sequence analysis showed that the isolate was in the same branch as Haemophilus parasuis (Hps) with more than 97% similarity.The isolate was identified as Hps serotype 4.When the isolated Hps was injected into mice intraperitoneally, bleeding in various organs of the infected mice was induced, especially in lung.High doses of infection of Hps serotype 4 could cause death in mice.Drug sensitivity test showed that the strain had strong resistance to tetracycline, ciprofloxacin and co-trimoxazole compound sulfamethoxazole.After rational administration, a good therapeutic effect was achieved.【Conclusion】 A strain of Hps serotype 4 was successfully isolated, which could cause bleeding in different degrees in multiple organs of infected mice, and was resistant to tetracycline, ciprofloxacin and co-trimoxazole.This results provided a theoretical basis for the clinical diagnosis and treatment of Hps.

Key words: Haemophilus parasuis(Hps); serotype 4; isolation and identification; drug resistance

中图分类号:

S852.61⁺2

王同兆, 张付贤, 雷连成. 1株副猪嗜血杆菌血清4型的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2022, 49(3): 1096-1105.

WNAG Tongzhao, ZHANG Fuxian, LEI Liancheng. Isolation, Identification and Drug Resistance Analysis of a Strain of Haemophilus parasuis Serotype 4[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1096-1105.

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