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Molecular Surveillance of Enterovirus and Norwalk-Like Virus in Oysters Relocated to a Municipal-Sewage-Impacted Gulf Estuary

机译:肠道病毒和挪沃样病毒在牡蛎中的分子监测

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摘要

An 18-month survey was conducted to examine the prevalence of enteric viruses and their relationship to indicators in environmentally polluted shellfish. Groups of oysters, one group per 4 weeks, were relocated to a coastal water area in the Gulf of Mexico that is impacted by municipal sewage and were analyzed for enteroviruses, Norwalk-like viruses (NLV), and indicator microorganisms (fecal coliform, Escherichia coli, and male-specific coliphages). The levels of indicator microorganisms were consistent with the expected continuous pollution of the area. Fourteen of the 18 oyster samples were found by reverse transcription (RT)-PCR to harbor NLV and/or enterovirus sequences. Of the four virus-negative oysters, three had exposure to water temperatures of >29°C. Concomitant with these findings, two of these four oysters also accumulated the lowest levels of coliphages. PCR primers targeting pan-enteroviruses and the NLV 95/96-US common subset were utilized; NLV sequences were detected more frequently than those of enteroviruses. Within the 12-month sampling period, NLV and enterovirus sequences were detected in 58 and 42%, respectively, of the oysters (67% of the oysters tested were positive for at least one virus) from a prohibited shellfish-growing area approximately 30 m away from a sewage discharge site. Eight (4.6%) of the 175 NLV capsid nucleotide sequences were heterogeneous among the clones derived from naturally polluted oysters. Overall, enteric viral sequences were found in the contaminated oysters throughout all seasons except hot summer, with a higher prevalence of NLV than enterovirus. Although a high percentage of the oysters harbored enteric viruses, the virus levels were usually less than or equal to 2 logs of RT-PCR-detectable units per gram of oyster meat.
机译:进行了为期18个月的调查,以检查肠病毒的流行率及其与环境污染贝类指标之间的关系。每四周一组牡蛎,被转移到墨西哥湾的沿海水域,受到市政污水的影响,并进行了肠病毒,诺沃克样病毒(NLV)和指示微生物(粪大肠菌,大肠埃希氏菌)的分析。大肠杆菌和雄性特异的噬菌体)。指示微生物的水平与该地区预期的持续污染相一致。通过逆转录(RT)-PCR发现18个牡蛎样品中有14个带有NLV和/或肠病毒序列。在四只病毒阴性牡蛎中,有三只暴露于> 29°C的水温。与这些发现相伴的是,这四只牡蛎中有两只的噬菌体含量也最低。利用针对全肠病毒和NLV 95 / 96-US常见亚组的PCR引物; NLV序列比肠病毒更频繁地被检测到。在12个月的采样期内,从大约30 m的禁止贝类养殖区域中分别检测到58和42%的牡蛎中的NLV和肠病毒序列(测试的牡蛎中67%的至少一种病毒呈阳性)。远离污水排放地点。 175个NLV衣壳核苷酸序列中的八个(4.6%)在源自自然污染牡蛎的克隆中异质。总体而言,除炎热的夏季外,所有季节都在受污染的牡蛎中发现了肠道病毒序列,与肠道病毒相比,NLV的发病率更高。尽管牡蛎中有很大比例的牡蛎都带有肠道病毒,但该病毒水平通常小于或等于每克牡蛎肉2个对数的RT-PCR可检测单位。

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