A non-lethal method for detection of Bonamia ostreae in flat oyster (Ostrea edulis) using environmental DNA.
A non-lethal method for detection of Bonamia ostreae in flat oyster (Ostrea edulis) using environmental DNA.
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DOI：
10.1038/s41598-020-72715-y
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发表时间：
2020-09-30
影响因子：
4.6
通讯作者：
Buchmann K
中科院分区：
文献类型：
--
作者：
Jørgensen LVG;Nielsen JW;Villadsen MK;Vismann B;Dalvin S;Mathiessen H;Madsen L;Kania PW;Buchmann K
牡蛎寄生蜂感染的监测和诊断是保护和管理野生牡蛎种群的前提。此外，在水产养殖生产中，需要可靠的和非致死的检测方法来选择健康的育牡蛎。在这里，我们提出了一个非致命的诊断技术的基础上，环境DNA（eDNA）从水样和实验室试验中的应用程序。将来自丹麦Limfjorden的40只牡蛎保存在30 ppt海水中的单独罐中。6天后对水取样，之后对所有牡蛎实施安乐死并应用PCR检查感染。四只牡蛎（10%）被发现感染了B。在鳃和外套膜组织中有牡蛎。从这些牡蛎周围的水中纯化的eDNA含有寄生虫DNA。随后从现场取样，包括来自5个不同地点的20个牡蛎和15个水样。只有一只牡蛎呈阳性反应，而所有水样本对B呈阴性反应。牡蛎eDNA用这种新方法B.可以通过仅从隔离牡蛎或隔离牡蛎种群的环境中取样水来检测牡蛎。这种非致命的eDNA诊断方法可能有潜力为未来的调查和牡蛎育种计划，旨在生产无病牡蛎。
Surveillance and diagnosis of parasitic Bonamia ostreae infections in flat oysters (Ostrea edulis) are prerequisites for protection and management of wild populations. In addition, reliable and non-lethal detection methods are required for selection of healthy brood oysters in aquaculture productions. Here we present a non-lethal diagnostic technique based on environmental DNA (eDNA) from water samples and demonstrate applications in laboratory trials. Forty oysters originating from Limfjorden, Denmark were kept in 30 ppt sea water in individual tanks. Water was sampled 6 days later, after which all oysters were euthanized and examined for infection, applying PCR. Four oysters (10%) were found to be infected with B. ostreae in gill and mantle tissue. eDNA purified from the water surrounding these oysters contained parasite DNA. A subsequent sampling from the field encompassed 20 oysters and 15 water samples from 5 different locations. Only one oyster turned out positive and all water samples proved negative for B. ostreae eDNA. With this new method B. ostreae may be detected by only sampling water from the environment of isolated oysters or isolated oyster populations. This non-lethal diagnostic eDNA method could have potential for future surveys and oyster breeding programs aiming at producing disease-free oysters.
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