目的 了解2022年沧州市动物性水产品中副溶血性弧菌的耐药特征、毒力基因及多位点序列分型(multilocus sequence typing,MLST)情况,掌握其分子流行病学特征,为相关食品风险监测及进一步防控提供数据支持.方法 2022年随机采取沧州市市售动物性水产品样品,根据《2022年国家食品污染和有害因素风险监测工作手册》要求,对所采集的样品进行细菌培养及分离鉴定,对鉴定为副溶血性弧菌的菌株进行药敏试验、PCR毒力基因检测和全基因组序列测定,借助相关数据库获得不同菌株的耐药表型、毒力基因、耐药基因和多位点序列分型情况.结果 药敏结果提示,样品中分离的副溶血性弧菌对大多数抗生素敏感或中敏,共检出6株耐药菌株,其中包含1株多重耐药株.耐药基因共检出10种,其中β-内酰胺类blaCARB基因、四环素类的tet(35)基因和调控耐药相关基因adeF、rsmA、CRP基因全部菌株均检出.不耐热溶血素(thermolabile hemolysin gene,tlh)基因携带率 100％,耐热直接溶血素(thermostable direct hemolysin,tdh)基因和耐热相关溶血素(tdh related hemolysin,trh)基因未检出.31株副溶血性弧菌依据7个管家基因分型,得到29种序列型(sequence type,ST型).结论 沧州市动物性水产品中检出的副溶血性弧菌属于低水平耐药,未检出毒力基因,携带多个耐药基因,基因序列具有遗传多样性,应加强监测.
Study on molecular typing of Vibrio parahaemolyticus in animal aquatic products of Cangzhou City in 2022
Objective To understand the drug resistance characteristics,virulence genes and multilocus sequence typing(MLST)of Vibrio parahaemolyticus in animal aquatic products of Cangzhou City in 2022,analyze the epidemiological characteristics,and provide data support for related food risk monitoring and further prevention and control.Methods The samples of commercially available animal aquatic products were randomly collected from Cangzhou City in 2022.According to the requirements of 2022 national food pollution and harmful factors risk monitoring manual,the bacterial culture and isolation identification were performed on collected samples.The drug sensitivity testing,PCR virulence gene testing,and whole genome sequencing were conducted on strains identified as Vibrio parahaemolyticus,and the relevant databases was used to obtain the phenotypes,virulence genes,resistance genes,and multi site sequence typing of different bacterial strains.Results The susceptibility results suggested that Vibrio parahaemolyticus isolated in the samples was sensitive or moderately sensitive to most antibiotics,and six resistant strains were detected,containing a multiresistant strain.A total of 10 resistance genes were detected,among which β-lactam blaCARB gene,tet(35)gene of tetracycline,and regulatory resistance related genes adeF,rsmA and CRP genes were detected.The thermolabile hemolysin gene(tlh)was carried at 100％,while thermostable direct hemolysin(tdh)and tdh related hemolysin(trh)genes were not detected.Based on seven housekeeping genotypes,31 strains of Vibrio parahaemolyticus yielded 29 sequence types(ST).Conclusion The Vibrio parahaemolyticus detected in animal aquatic products of Cangzhou City belongs to low-level drug resistance,with no virulence genes,carrying multiple drug resistance genes,and the gene sequence has genetic diversity,so the monitoring should be strengthened.
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