一株血清9型副猪嗜血杆菌的分离鉴定与药敏试验

Isolation,Identification and Drug Sensitivity Test of Haemophilus parasuis in One Strain of Serovars 9

【目的】确定河南某猪场送检病猪疑似副猪嗜血杆菌的血清型,为其提临床用药提供参考.【方法】经实验室分离培养,对分离到的病原菌进行生化鉴定、药敏试验和血清型分型.[结果]该分离菌革兰氏染色为阴性短杆菌,具有多形性;发酵葡萄糖、果糖、蔗糖、麦芽糖,甘露醇、木糖、尿素、氧化酶阴性,硝酸还原酶、接触酶阳性,MR和VP试验阴性,有卫星现象;16Sr RNA扩增出了821 bp目的片段,测序结果与GenBank公布的副猪嗜血杆菌NR_118757.1同源性达98%以上;分离到的细菌对头孢噻呋、阿米卡星、硫酸黏菌素、甲砜霉素、卡那霉素、庆大霉素、多西环素、氟苯尼考高敏,对阿莫西林、恩诺沙星耐药;采用14对分型引物对分离菌进行血清型分型,多重PCR扩增到700 bp左右大小的目的基因片段,测序结果与GenBank公布的KC795429.1副猪嗜血杆菌funV同源性达99.85%,确定该分离菌株为9型副猪嗜血杆菌.【结论】本研究结果将为副猪嗜血杆菌的防治提供参考.

【Objective】The purpose of this study was to determine the serotypes of suspected Haemophilus parapigs in diseased sample from a pig farm in Henan Province,and to provide a drug regimen.【Method】The strain was isolated and cultured in the laboratory,identified by biochemistry,drug sensitivity test and serotyping.【Result】The results showed that Gram staining was negative short bacillus,which was polymorphic.It fermented glucose,fructose,sucrose,maltose,mannitol,xylose,urea,oxidase negative,nitrate reductase,contact enzyme positive,MR and VP test negative,and therewas satellite phenomenon;The target fragment of 821 bp was amplified by 16Sr RNA,and the homology between the sequencing results and the NR_118757.1 of Haemophilus parasuis published by GenBank was more than 98%.It was sensitive to 8 antibiotics including Ceftiofur,Amikacin,Colistin Sulfate,Thiamphenicol,Kanamycin,Gentamicin,Doxycycline,Florfenicol,and was resistant to Amoxicillin and Enrofloxacin.The serotyping of the isolated bacteria was carried out with 14 pairs of typing primers,and the target gene fragment about the size of 700 bp was amplified by multiplex PCR.The homology between the sequencing results and the funV published by GenBank of KC795429.1 was 99.85%,which was confirmed that this isolated strain was serovars 9 Hemophilus paracinis.【Conclusion】The results provided reference for the prevention and treatment of Haemophilus parapigs.