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首页> 外文期刊>Applied and Environmental Microbiology >Protocol for Specific Isolation of Virulent Strains of Vibrio vulnificus Serovar E (Biotype 2) from Environmental Samples
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Protocol for Specific Isolation of Virulent Strains of Vibrio vulnificus Serovar E (Biotype 2) from Environmental Samples

机译：从环境样品中特异性分离Vulioificus Serovar E（生物型2）毒力毒株的协议

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摘要

The eel pathogen Vibrio vulnificus biotype 2 comprises at least three serovars, with serovar E being the only one involved in both epizootics of eel vibriosis and sporadic cases of human infections. The virulent strains of this serovar (VSE) have only been recovered from clinical (mainly eel tissue) sources. The main objective of this work was to design and validate a new protocol for VSE-specific isolation from environmental samples. The key element of the new protocol is the broth used for the first step (saline eel serum broth [SEB]), which contains eel serum as a nutritive and selective component. This approach takes advantage of the ability of VSE cells to grow in eel serum and thus to separate themselves from the pool of competitors. The growth yield in SEB after 8 h of incubation was 1,000 times higher for VSE strains than for their putative competitors (including biotype 1 strains of the species). The selective and differential agar Vibrio vulnificus medium (VVM) was selected from five selective media for the second step because it gave the highest plating efficiency not only for the VSE group but also for other V. vulnificus groups, including biotype 3. The entire protocol was validated by field studies, with alkaline peptone water plus VVM as a control. V. vulnificus was isolated by both protocols, but serovar E was only recovered by the new method described here. All selected serovar E isolates were identified as VSE since they were virulent for both eels and iron-overloaded mice and resisted the bactericidal action of eel and iron-overloaded human sera. In conclusion, this new protocol is a suitable method for the isolation of VSE strains from environmental samples and is recommended for epidemiological studies of the pathogenic serovar E.
机译：鳗病病原体弧菌生物型2至少包含三个血清型，其中血清型E是唯一参与鳗弧菌病流行和人间偶发感染的病例。仅从临床（主要是鳗鱼组织）来源中回收了该血清型（VSE）的强毒株。这项工作的主要目的是设计和验证一种用于从环境样品中分离VSE的新协议。新协议的关键要素是第一步所用的肉汤（盐水鳗鱼血清肉汤[SEB]），其中含有鳗鱼血清作为营养和选择性成分。这种方法利用了VSE细胞在鳗鱼血清中生长并因此使其自身与竞争者分离的能力。 VSE菌株孵育8小时后SEB的生长产量比其假定的竞争者（包括该物种的生物型1菌株）高1,000倍。从第二步的五个选择性培养基中选择了选择性和差异琼脂弧菌培养基（VVM），因为它不仅对VSE组而且对于其他V. vulnificus组（包括生物型3）都具有最高的铺板效率。整个方案通过碱性蛋白ept水加VVM作为对照，通过现场研究进行了验证。两种方法均分离出了V. vulnificus，但仅通过此处描述的新方法回收了血清型E。所有选定的血清型E分离株均被鉴定为VSE，因为它们对鳗鱼和铁超负荷的小鼠均具有毒性，并能抵抗鳗鱼和铁超负荷的人血清的杀菌作用。总之，该新方案是从环境样品中分离VSE菌株的合适方法，建议用于病原性血清型E的流行病学研究。

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