摘要
为探究漳州市养殖环境中副溶血弧菌流行状况和基因多态性,无菌采集2019年1~7月份本地区7个养殖场中罗非鱼、石斑鱼和虾的新鲜样品。用硫代硫酸盐柠檬酸盐胆盐蔗糖琼脂培养基(thiosulfate citrate bile salts sucrose agar culture medium,TCBS)对样品的疑似副溶血弧菌进行分离纯化,用聚合酶链式反应(polymerase chain reaction,PCR)扩增tlh基因和16S rDNA基因片段并测序,鉴定疑似菌株。用随机扩增多态性法(random amplified polymorphic DNA,RAPD)对分离株进行分型。实验共采集228份样品,微生物学初筛得到111株疑似菌株,PCR检测并测序鉴定69株为副溶血弧菌,阳性率为30.26%。RAPD技术对分离株分型,得到了较清晰的电泳条带,分析指纹图谱发现69株副溶血弧菌可分为9个主要类型,遗传相似性在83%~100%范围内。养殖场中的水产品一定程度上受到副溶血弧菌的污染,且不同水产品、不同养殖场、不同时间副溶血弧菌的污染程度都不同。
In order to investigate the prevalence and gene polymorphism of Vibrio parahaemolyticus in the farming environment of Zhangzhou city,fresh samples of Oreochroms mossambcus,Epinephelus spp.and Penaeus vannamei were collected from 7 farms in this region from January to July in 2019.The suspected isolates were cultured and purified by thiosulfate citrate bile salts sucrose agar culture medium(TCBS).The fragments of tlh gene and 16S rDNA gene were amplified by PCR and sequenced to identify the suspected strains.The genotyping of Vibrio parahaemolytic isolates was performed by random amplified polymorphic DNA(RAPD)method.A total of 228 samples were collected in the experiment,and 111 suspected strains were obtained through microbiological screening.A total of 69 strains were identified as Vibrio parahaemolyticus by PCR and sequencing with a positive rate of 30.26%.RAPD technique was used to type isolates of Vibrio parahaemolyticus,and clearer DNA bands were obtained.Analysis of the fingerprints showed that 69 Vibrio parahaemolytic can be divided into 9 main types,with genetic similarity in the range of 83%to 100%.To some extent,the aquatic products in the farm will be contaminated by Vibrio parahaemolyticus,and the degree of contamination of Vibrio parahaemolyticus is variable in different aquatic products,different farms and at different times.
作者
胡元庆
陈锦芳
肖芸
朱秋强
张丹凤
HU Yuanqing;CHEN Jinfang;XIAO Yun;ZHU Qiuqiang;ZHANG Danfeng(School of Biological Science and Biotechnology,Minnan Normal University,Zhangzhou 363000,China)
出处
《食品工业科技》
CAS
北大核心
2021年第8期117-124,共8页
Science and Technology of Food Industry
基金
福建省自然科学基金面上项目(2017J01453)
江苏人兽共患病学重点实验室开放课题(R1402)
福建省高校青年自然基金重点项目(JZ160454)
闽南师范大学新世纪优秀人才支持计划(MX13004)。
关键词
副溶血弧菌
养殖环境
RAPD
分子分型
Vibrio parahaemolyticus
farming environment
random amplified polymorphic DNA(RAPD)
molecular typing
