Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Italy.
Departamento de Microbiologia y Parasitologia, Centro de Investigationes Biologicas (CIBUS), Faculty of Biology, Universidad de Santiago de Compostela, Spain.
Food Microbiol. 2019 Dec;84:103258. doi: 10.1016/j.fm.2019.103258. Epub 2019 Jul 6.
The aim of the present study was to investigate the genetic variability of Vibrio parahaemolyticus strains isolated from naturally contaminated Mediterranean mussels (Mytilus galloprovincialis) and Grooved carpet shells (Ruditapes decussatus) from three harvesting areas of Sardinia (Italy) using a combination of different typing methods: traditional phenotypic systems and molecular techniques. Ninety-nine putative V. parahaemolyticus strains isolated from shellfish collected before and after purification were included in the study. Seventy-two isolates were confirmed as V. parahaemolyticus and were submitted to REP, ERIC and BOX PCRs. The combined dendrogram showed the similarity of the data set of the three typing methods and demonstrates how the different techniques grouped the strains in two clusters in accordance with each singular dendrogram. Several strains rendered a unique pattern regardless of the typing method, which indicates the high discriminatory power of the methods. Moreover, the use of multiple typing methods allowed a more accurate characterization of the genetic profiles of isolates and the identification of clones hardly revealed through the common techniques. The intraspecific typing of environmental V. parahaemolyticus can be of great interest in order to recognize clonal relationships between environmental contamination, foodborne disease, and geographical/temporal distribution of this pathogen. The comparative analysis focusing on the obtained genetic profiles supports the possibility for typing methods to discriminate strains with similar phenotypic profile, identifying the level of genetic correlation among the strains and the presence of genetic clones.
本研究旨在使用多种不同的分型方法(传统表型系统和分子技术),研究从意大利撒丁岛三个捕捞区天然污染的贻贝(Mytilus galloprovincialis)和糙扇贝(Ruditapes decussatus)中分离的副溶血性弧菌菌株的遗传变异性。本研究纳入了 99 株从贝类采集前后分离的疑似副溶血性弧菌。72 株分离株被确认为副溶血性弧菌,并进行了 REP、ERIC 和 BOX PCRs 分析。联合聚类图显示了三种分型方法数据集的相似性,并证明了不同技术如何根据每个单独的聚类图将菌株分为两个聚类。一些菌株无论采用哪种分型方法,都呈现出独特的模式,这表明这些方法具有很高的分辨能力。此外,使用多种分型方法可以更准确地描述分离株的遗传特征,并识别出常见技术难以揭示的克隆。对环境副溶血性弧菌进行种内分型对于识别环境污染、食源性疾病以及该病原体在地理和时间上的分布之间的克隆关系具有重要意义。对获得的遗传图谱进行的比较分析支持了分型方法区分具有相似表型图谱的菌株的可能性,确定了菌株之间遗传相关性的水平以及遗传克隆的存在。