Molecular Detection of Norwalk Virus in Carp Fish and Shrimp Ponds in Khuzestan Province, Iran by RT-PCR Method

Parisa Shojaie, Z. Mashak, Valliolah Koohdar
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Abstract

Background: Norwalk virus is one of the most common causes of viral gastroenteritis. The aquatic products are potential sources of contamination with this virus. Objectives: The main objective of the study was to investigate the presence of the Norwalk virus in different aquatic animals in Khuzestan provinces, Iran. Materials and Methods: A total of 40 pieces of fish (silver carp, common carp, big head, and grass carp species) and 10 pieces of shrimps were caught from ponds, and the samples were transferred to the laboratory in ice bags. After the separation of the intestine, the content of the intestine was extracted using two sterile filters. Then, the supernatant was used for reverse transcription polymerase chain reaction (RT-PCR) using Calicivirus-specific primers (p289/ p290). Then, Norwalk virus-specific primers (NVp36/NVp35) were detected in Calicivirus positive samples. Results: The results showed 8% (4 samples) and 6% (3 samples) of the samples were infected with Calicivirus (p289/p290 genes) and Norwalk virus (NVp36/NVp35 genes), respectively. Calicivirus positive samples included 2 common carp, 1 silver carp, and 1 shrimp. Norwalk virus-positive samples included 2 common carp and 1 shrimp. In other words, the highest prevalence of virus was observed in aquatic fish feeding from the bottom of the pool. Due to the fact that this species is bred with other species and considering that this virus lives in the gastrointestinal tract, the ingestion of feces of other infected organisms can lead to the increase of this virus in the digestive system of carp. Conclusion: Therefore, due to the importance of Norwalk as a zoonotic agent and the possibility of human infection through consumption of aquatic products, preventive measures such as not using animal manure for fertilization and preventing the growth of phytoplankton in aquaculture ponds and cooking meat properly are suggested.
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RT-PCR法检测伊朗胡齐斯坦省鲤鱼鱼虾池中诺瓦克病毒
背景:诺瓦克病毒是病毒性胃肠炎最常见的病因之一。水产品是该病毒的潜在污染源。目的:本研究的主要目的是调查诺瓦克病毒在伊朗胡齐斯坦省不同水生动物中的存在。材料与方法:从池塘中捕获鱼类(鲢鱼、鲤鱼、大头鱼、草鱼等)40条,虾10条,装在冰袋中移送实验室。分离肠后,用两个无菌过滤器提取肠内容物。然后,用杯状病毒特异性引物(p289/ p290)进行逆转录聚合酶链反应(RT-PCR)。然后在杯状病毒阳性样本中检测到诺瓦克病毒特异性引物(NVp36/NVp35)。结果:感染杯状病毒(p289/p290基因)和诺瓦克病毒(NVp36/NVp35基因)的样品分别为8%(4份)和6%(3份)。呈杯状病毒阳性的鲤鱼2例,鲢鱼1例,对虾1例。诺瓦克病毒阳性样本包括2条鲤鱼和1只虾。换句话说,从池底取食的水生鱼类中观察到的病毒流行率最高。由于该品种与其他品种杂交,再考虑到该病毒生活在胃肠道中,因此摄入其他受感染生物的粪便可导致该病毒在鲤鱼消化系统中的数量增加。结论:鉴于诺瓦克作为人畜共患病原体的重要性,以及人类通过食用水产品感染诺瓦克的可能性,建议采取不使用动物粪便施肥、防止养殖池塘浮游植物生长、适当烹调肉类等预防措施。
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