Occurrence of pathogenic vibrios in coastal areas of France
Occurrence of pathogenic vibrios in coastal areas of France
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DOI：
10.1046/j.1365-2672.2002.01663.x
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发表时间：
2002-01-01
影响因子：
4
通讯作者：
Pommepuy, M
中科院分区：
文献类型：
--
作者：
Hervio-Heath, D;Colwell, RR;Pommepuy, M
目的：调查法国海洋和河口环境中潜在致病性弧菌的发生情况。方法与结果：采用硫代硫酸盐-柠檬酸盐-胆汁盐-蔗糖和改性纤维素-多粘菌素-琼脂等选择性培养基，对1999年7 - 9月采集的沿海水域和贻贝样品进行了培养分析。假定的弧菌菌落分离和鉴定使用选定的生化试验。采用聚合酶链式反应（PCR）技术，根据细胞溶解素、vvhA基因、pR72H DNA片段和16S-23S rRNA基因间间隔区（ISR）特异性引物，分别鉴定出创伤弧菌、副溶血性弧菌和霍乱弧菌。189株中以溶藻弧菌（99株）为优势菌种，其次为副溶血性弧菌（41株）、创伤弧菌（20株）和非o1 /非o139型霍乱弧菌（3株）。20株创伤弧菌均PCR扩增出vvhA基因，其中16株分离自河口水体。对41株副溶血性弧菌的pR72H DNA片段进行PCR扩增，得到两个独特的扩增片段，分别为387和320 bp。后者出现在24.4%的分离株中，以前在迄今检查的副溶血性弧菌菌株中未发现。其中两个分离株的trh基因扩增表明它们是毒性菌株。经16S-23S rRNA ISR扩增鉴定为霍乱弧菌的3株菌株均为非霍乱（非o1 /非o139）菌株。结论：本研究结果表明法国沿海水域存在致病性弧菌。此外，PCR方法对物种鉴定的快速和可靠的确认是有用的。研究的意义和影响：这些发现表明，在养殖贻贝和水生环境中存在致病性弧菌可能存在卫生风险。该方法可用于环境样品中病原弧菌的直接检测。
Aims: This study was carried out to investigate the occurrence of potentially pathogenic species of Vibrio in French marine and estuarine environments.Methods and Results: Samples of coastal waters and mussels collected between July and September 1999 were analysed by culture, using selective media including thiosulphate-citrate-bile salts-sucrose and modified cellobiose-polymixin B-colistin agar. Presumptive Vibrio colonies were isolated and identified using selected biochemical tests. Specific primers based on flanking sequences of the cytolysin, vvhA gene, pR72H DNA fragment and 16S-23S rRNA intergenic spacer region (ISR) were used in a polymerase chain reaction (PCR) to confirm the identification of Vibrio vulnificus , V. parahaemolyticus and V. cholerae , respectively. In this study, V. alginolyticus (99 of 189) was the predominant species, followed by V. parahaemolyticus (41 of 189), V. vulnificus (20 of 189) and non-O1/non-O139 V. cholerae (three of 189). All 20 V. vulnificus isolates showed PCR amplification of the vvhA gene, 16 of which had been isolated from estuarine water. The PCR amplification of the pR72H DNA fragment in 41 V. parahaemolyticus isolates generated two unique amplicons of 387 and 320 bp. The latter, present in 24.4% of these isolates, had not previously been found in V. parahaemolyticus strains examined to date. Amplification of the trh gene in two of the isolates suggested these to be virulent strains. Three strains identified as V. cholerae by amplification of the 16S-23S rRNA ISR were confirmed to be non-cholera (non-O1/non-O139) strains.Conclusions: The results of this study demonstrated the presence of pathogenic Vibrio species in French coastal waters. Furthermore, the PCR approach proved useful for the rapid and reliable confirmation of species identification.Significance and Impact of the Study: These findings indicate the potential sanitary risk associated with the presence of pathogenic Vibrio spp. in cultivated mussels and in the aquatic environment. The PCR can be used to detect pathogenic vibrios directly in environmental samples.