广西某猪场猪链球菌分离鉴定及其生物学特性研究

doi:10.16431/j.cnki.1671-7236.2021.02.036

摘要/Abstract

摘要： 2019年12月广西玉林某猪场猪连续发生多起保育猪突然死亡病例，为确诊该猪场猪发病死亡原因并调查分离菌株的致病性与耐药情况，对发病猪场进行采样和细菌分离培养，并对分离菌株进行形态学观察、生化鉴定、gdh基因PCR扩增、血清型和7个毒力基因的鉴定、耐药性检测以及耐药基因的鉴定。结果显示，从两头病死猪中各分离出1株细菌，在TSB固体培养基表面培养为灰白色、表面光滑、边缘整齐且大小一致的圆形菌落，初步鉴定为革兰阳性链球菌，分别命名为GXYL-F1、GXYL-N2。经猪链球菌特异性基因gdh及血清型引物鉴定，两分离株均为9型猪链球菌。生化鉴定结果显示，两分离菌株均可发酵水杨素、麦芽糖、M.R.和七叶苷。PCR检测菌株的毒力基因结果显示，gdh、fbps、sao、sbp2'和orf2均为阳性，mrp、epf、sly基因均为阴性。药敏试验结果显示，两分离株均对新霉素、链霉素、替米考星、泰乐菌素、红霉素、四环素、左氧氟沙星、青霉素和磺胺嘧啶这9种药物呈耐药性，此外分离菌株GXYL-F1还对恩诺沙星耐药。两分离菌株均扩增出耐药基因aadA1、qnrB、qnrS。表明该猪场保育猪突然死亡是由9型猪链球菌引起。上述研究结果为该猪场制定9型猪链球菌防治措施提供参考依据，并为了解猪链球菌流行血清型多样性与防控技术研究提供了新的数据。

关键词: 猪; 猪链球菌; PCR鉴定; 耐药性; 毒力基因; 药敏试验

Abstract: In December 2019,a series of sudden deaths of protected pigs occurred in a swine farm in Yulin,Guangxi.In order to confirm the cause of death and investigate the pathogenicity and drug resistance of isolated strains,samples and bacterial isolation and culture were conducted in the sick swine farm.Morphological observation,biochemical identification,PCR amplification of gdh gene,identification of serum type and some virulence genes,detection of drug resistance and identification of drug resistance genes were performed on the isolates.One bacterial strain was isolated from two dead pigs separately.On TSB solid medium,the colonies were gray white,smooth,and uniform in size.They were preliminarily identified as Gram-positive Streptococcus and named GXYL-F1 and GXYL-N2,respectively.Both isolates were identified as Streptococcus suis type 9 by specific gene gdh and serum primer.Biochemical identification results showed that the two isolated strains could ferment salicylline,maltose,M.R.and aescinate.The virulence gene of the strain detected by PCR showed that genes gdh,fbps,sao,sbp2' and orf2 were positive,while mrp,epf and sly genes were negative.Drug sensitivity test results showed that GXYL-F1 and GXYL-N2 were all resistant to neomycin,streptomycin,timicoxacin,tylosin,erythromycin,tetracycline,levofloxacin,penicillin and sulfadiazine.In addition,the isolated strain GXYl-F1 was also resistant to enrofloxacin.The drug-resistant gene aadA1,qnrB and qnrS were amplified in both isolates.The study showed that the disease was caused by Streptococcus suis type 9.The above research results provided a reference for this swine farm to develop the control measures of Streptococcus suis type 9 and a new data for the serotype diversity and prevention of technology reaserch.

Key words: swine; Streptococcus suis; PCR identification; drug resistance; virulence gene; drug sensitive test

中图分类号:

S852.61

郭长明, 陈怀君, 朱善元, 袁敬知, 葛强, 李珣, 王晓晔. 广西某猪场猪链球菌分离鉴定及其生物学特性研究[J]. 中国畜牧兽医, 2021, 48(2): 736-746.

GUO Changming, CHEN Huaijun, ZHU Shanyuan, YUAN Jingzhi, GE Qiang, LI Xun, WANG Xiaoye. Isolation and Identification of Streptococcus suis and Biological Characteristics Research on a Swine Farm in Guangxi[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 736-746.

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