摘要
【目的】为合理有效防控禽沙门菌病,从疑似沙门菌感染的病死鹅中进行鼠伤寒沙门菌的分离和鉴定。【方法】无菌采集6份病死鹅的脾脏组织样品进行病原菌分离鉴定,使用16S rRNA基因的细菌通用引物和沙门菌属的特异性基因invA引物分别进行PCR扩增及测序分析,并进行药敏试验和耐药基因检测。【结果】从6个样本中成功分离出6株沙门菌。这些菌株在麦康凯培养基中形成没有颜色的菌落,在亚硫酸铋培养基中形成的菌落为黑色。通过PCR方法分别扩增出16S rRNA与invA基因1500 bp和330 bp的条带;16S rRNA测序结果与鼠伤寒沙门菌参考菌株相似性位于99.59%~100%之间,invA基因测序结果与鼠伤寒沙门菌参考菌株相似性位于98.49%~100%之间;药敏结果显示,6株鼠伤寒沙门菌对9种药物的耐药率为100%,对多粘菌素B耐药率为0,对其他药物的耐药率介于50%~83.33%之间,多重耐药现象严重;耐药基因显示6株分离菌携带四环素类tetC基因,2株携带β-内酰胺类blaCTX-M基因。【结论】从病死鹅脾脏组织中分离鉴定了6株鼠伤寒沙门菌,分离菌株耐药严重,临床建议选用多粘菌素B等敏感药物治疗。为安徽凤阳及周边地区禽沙门菌病的防控和耐药机理研究提供了科学依据。
[Objective]To effectively and efficiently prevent and control avian salmonellosis,the isolation and identification of Salmonella typhimurium from dead geese suspected of having a Salmonella infection were carried out.[Methods]Six sterile samples of spleen tissue from dead geese were collected for pathogen isolation and identification.Universal bacterial primers targeting the 16S rRNA gene and specific primers for Salmonella invA gene were used for PCR amplification and sequencing analysis,followed by antibiotic sensitivity testing and drug resistance gene detection.[Results]Six strains of Salmonella were isolated from the samples.Colorless colonies emerged on MacConkey agar,and black colonies appeared on bismuth sulfite agar.16S rRNA and invA genes were amplified,yielding bands of 1500 bp and 330 bp,respectively.The sequencing results of the 16S rRNA gene showed similarities ranging from 99.59%to 100%compared with the reference strain of Salmonella typhimurium,while the sequencing results of the invA gene showed similarities ranging from 98.49%to 100%.Drug sensitivity testing revealed that all six strains of Salmonella typhimurium were resistant to multiple drugs,the resistance rate to nine different drugs was 100%,the resistance rate to polymyxin B was 0,and the resistance rate to other drugs was between 50%and 83.33%.The drug resistance gene analysis showed that all six isolated strains carried the tetC gene,and two strains carried the bla CTX-M gene.[Conclusion]Six strains of Salmonella typhimurium were isolated and identified from the spleens of dead geese.The results shown that the isolated strains exhibited severe drug resistance,and the sensitive drugs such as polymyxin B would be better in clinical treatment.This study provides a scientific basis for the prevention and control of avian salmonellosis and the mechanism research of drug resistance in Fengyang and surrounding areas of Anhui province.
作者
马佰贺
王紫苇
王海龙
鞠诗婷
徐洁
张航
郭伟娜
MA Baihe;WANG Ziwei;WANG Hailong;JU Shiting;XU Jie;ZHANG Hang;GUO Weina(College of Animal Science,Anhui Science and Technology University,Fengyang 233100,China)
出处
《北京农学院学报》
2024年第4期82-87,共6页
Journal of Beijing University of Agriculture
基金
安徽省教育厅自然科学研究重点项目(KJ2020A0087)
安徽科技学院稳定人才项目(DKWD201801)
安徽科技学院省级大学生创新课题项目(S202210879187)。
关键词
沙门菌
分离
鉴定
耐药性检测
Salmonella isolation
identification
drug resistance detection
