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Detection of Early Stages of Myxobolus Cerebralis in Fin Clips from Rainbow Trout (Onchorynchus mykiss)

机译:虹鳟鱼鳍夹中脑粘液虫早期的检测

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摘要

A nested polymerase chain reaction (PCR) assay was used to detect early stages of Myxobolus cerebralis in caudal and adipose fin samples from rainbow trout (RT). To determine sensitivity, groups of 10 RT were exposed to 2,000 M. cerebralis triactinomyxons/fish for 1 hour at 15 degrees C and subsequently moved to clean recirculating water. Fish were held for 2 and 6 hours and 1, 2, 3, 5, 7, 10, 30, and 60 days before sampling by nonlethal fin biopsy. Nested PCR performed on fin clips showed that M. cerebralis DNA was detected in caudal fin tissue in 100% of fish up to 5 days postexposure. At days 7 and 10 postexposure, 80% of fish were positive, and at 60 days postexposure, 60% of fish were positive using this technique. Conversely, testing on adipose fin clips proved less sensitive, as positive fish dropped from 80% at day 7 to below 20% at day 10 postinfection. Since detection of M. cerebralis infection using caudal fin samples coupled with nested PCR is an effective method for detection of early parasite stages, use of this technique provides for accurate, nonlethal testing.
机译:巢式聚合酶链反应(PCR)分析用于检测虹鳟鱼(RT)尾鳍和脂肪鳍样本中脑粘液的早期阶段。为了确定灵敏度,将10个RT组在15摄氏度下暴露于2,000 M. Brainis triactinomyxons /鱼中1小时,然后移至干净的循环水中。在通过非致死性鳍活检取样之前,将鱼饲养2和6小时,以及1、2、3、5、7、10、30和60天。在鳍片上进行的巢式PCR显示,暴露后5天之内,在100%的鱼的尾鳍组织中检测到了M. brainis DNA。使用该技术,在暴露后第7天和第10天,有80%的鱼呈阳性,在暴露后60天,有60%的鱼呈阳性。相反,对脂肪翅片的测试证明敏感性较低,因为阳性鱼从感染后第7天的80%下降到第10天的20%以下。由于使用尾鳍样品和巢式PCR结合检测脑支原体感染是检测早期寄生虫阶段的有效方法,因此使用该技术可进行准确的非致死性检测。

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