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利用 Bacteroidales 和 F 特异性 RNA 噬菌体标记物鉴定河口牡蛎中的粪便污染来源。

Identification of the origin of faecal contamination in estuarine oysters using Bacteroidales and F-specific RNA bacteriophage markers.

机构信息

Laboratoire Santé Environnement et Microbiologie, Unité SG2M, Département RBE, IFREMER, Plouzané, France.

出版信息

J Appl Microbiol. 2013 Sep;115(3):897-907. doi: 10.1111/jam.12260. Epub 2013 Jun 14.

Abstract

AIMS

The aim of this study was to identify the origin of faecal pollution impacting the Elorn estuary (Brittany, France) by applying microbial source tracking (MST) markers in both oysters and estuarine waters.

METHODS AND RESULTS

The MST markers used were as follows: (i) human-, ruminant- and pig-associated Bacteroidales markers by real-time PCR and (ii) human genogroup II and animal genogroup I of F-specific RNA bacteriophages (FRNAPH) by culture/genotyping and by direct real-time reverse-transcriptase PCR. The higher occurrence of the human genogroup II of F-specific RNA bacteriophages using a culture/genotyping method, and human-associated Bacteroidales marker by real-time PCR, allowed the identification of human faecal contamination as the predominant source of contamination in oysters (total of 18 oyster batches tested) and waters (total of 24 water samples tested). The importance of using the intravalvular liquids instead of digestive tissues, when applying host-associated Bacteroidales markers in oysters, was also revealed.

CONCLUSIONS

This study has shown that the application of a MST toolbox of diverse bacterial and viral methods can provide multiple lines of evidence to identify the predominant source of faecal contamination in shellfish from an estuarine environment.

SIGNIFICANCE AND IMPACT OF THE STUDY

Application of this MST toolbox is a useful approach to understand the origin of faecal contamination in shellfish harvesting areas in an estuarine setting.

摘要

目的

本研究旨在通过在牡蛎和河口水中应用微生物源追踪(MST)标记物,确定影响埃洛恩河口(法国布列塔尼)的粪便污染来源。

方法和结果

使用的 MST 标记物如下:(i)通过实时 PCR 检测人、反刍动物和猪相关的拟杆菌标记物,(ii)通过培养/基因分型和直接实时逆转录 PCR 检测人基因 II 组和动物基因 I 组的 F 特异性 RNA 噬菌体(FRNAPH)。使用培养/基因分型方法检测到人基因 II 组的 F 特异性 RNA 噬菌体的更高发生率,以及实时 PCR 检测到的人相关拟杆菌标记物,可确定人类粪便污染是牡蛎(总共测试了 18 批牡蛎)和水体(总共测试了 24 个水样)中污染的主要来源。当在牡蛎中应用宿主相关拟杆菌标记物时,还揭示了使用瓣膜内液体而不是消化组织的重要性。

结论

本研究表明,应用多种细菌和病毒方法的 MST 工具包可以提供多条证据来确定河口环境中贝类粪便污染的主要来源。

意义和影响

该 MST 工具包的应用是了解河口贝类养殖区粪便污染来源的一种有用方法。

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