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首页> 外文期刊>Journal of Fish Diseases >Negligible risk associated with the movement of processed rainbow trout,Oncorhynchus mykiss (Walbaum), from an infectious haematopoietic necrosisvirus (IHNV) endemic area
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Negligible risk associated with the movement of processed rainbow trout,Oncorhynchus mykiss (Walbaum), from an infectious haematopoietic necrosisvirus (IHNV) endemic area

机译：与来自传染性造血坏死病毒（IHNV）流行地区的虹鳟（Oncorhynchus mykiss（Walbaum））移动相关的风险可忽略不计

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摘要

To assess the risk of transmission of infectious haematopoietic necrosis virus (IHNV) associated with the movement of processed rainbow trout, Oncorhynchus mykiss, from an area where the virus is endemic, 240 freshly eviscerated fish (225-500 g) exhibiting spinal curvature or spinal compression types of deformities were tested for IHNV by virus isolation and polymerase chain reaction (PCR) techniques. Commercially produced rainbow trout, approximately 1-year-old, that exhibited spinal deformities were considered to have had a high likelihood of having survived an outbreak of IHN. Serological analysis of fish exhibiting spinal curvature or spinal compression types of deformities for anti-IHNV antibodies resulted in 71 and 50% of the serum samples, respectively, with detectable neutralization activity suggesting previous infection with IHNV. A portion of the skin and muscle in the area of the deformity was collected, as well as brain tissue from each commercially processed fish. Tissue homogenates were tested for IHNV using the epithelioma papulosum cyprini (EPC) cell line pretreated with polyethylene glycol and the chinook salmon embryo (CHSE-214) cell line using standard methods. Nested, reverse transcriptase (RT)-PCR for the detection of IHNV used the central 1231 by portion of the glycoprotein (G) gene. All brain and skin-muscle homogenates were negative for IHNV by virus isolation and by nested RT-PCR. To assess virus clearance experimentally, groups of 100 specific-pathogen-free (SPF) rainbow trout (mean weight, 100 g) were either intraperitoneally injected with IHNV or sham infected and held separately in 385 L aquaria in SPF water at 15 degreesC. In the first study, a total of 33% of the infected fish died. Ten days after the last mortality or 34 days after post-infection, six fish from each treatment were killed at weekly intervals for 6 weeks. Kidney and brain homogenates from individual fish were tested by virus isolation and nested RT-PCR using the nucleoprotein (N) gene. All tissue homogenates were negative for IHNV. When this study was repeated, 54% of the infected fish died and beginning 34 days post-infection, six fish from each treatment were killed at biweekly intervals for 8 weeks, and kidney and brain homogenates from individual fish were tested by virus isolation and quantitative real-time PCR utilizing primers and fluorescent labelled probes generated for the specific identification of the N and G genes of IHNV. One brain specimen from an IHN survivor obtained 34 days post-infection was positive by both virus isolation and quantitative PCR; however, all remaining tissue specimens were negative over the 8-week period. Intraperitoneal injection of supernatants from pooled kidney or brain tissue obtained from control or IHN survivor fish into SPF rainbow trout (mean weight, 10 g) also failed to establish virus infection. A strong humoral response was detected in the IHN survivors in both challenge studies and is suggested as a mechanism responsible for virus clearance. These results provide scientific information that can be used to assess the risk associated with the movement of processed rainbow trout from an IHNV endemic area.
机译：为了评估与加工的虹鳟鱼Oncorhynchus mykiss的传播相关的传染性造血坏死病毒（IHNV）从该病毒的流行地区传播的风险，有240具新鲜内脏鱼（225-500 g）表现出脊柱弯曲或脊柱通过病毒分离和聚合酶链反应（PCR）技术测试了IHNV压缩类型的畸形。商业生产的虹鳟鱼（约1岁）表现出脊椎畸形，被认为极有可能在IHN爆发后幸存下来。对抗IHNV抗体表现出脊柱弯曲或脊柱压缩类型畸形的鱼进行血清学分析，分别产生了71％和50％的血清样品，可检测到的中和活性表明先前感染了IHNV。收集畸形区域的部分皮肤和肌肉，以及每条商业加工鱼的脑组织。使用标准方法，使用经聚乙二醇预处理的乳头丘疹上皮（EPC）细胞系和奇努克鲑鱼胚胎（CHSE-214）细胞系，测试组织匀浆的IHNV。巢式逆转录酶（RT）-PCR检测IHNV使用了部分糖蛋白（G）基因的中央1231。通过病毒分离和巢式RT-PCR，所有脑和皮肤肌肉匀浆均对IHNV阴性。为了通过实验评估病毒清除率，将100只无特定病原体（SPF）虹鳟鱼（平均体重100克）的组腹膜内注射IHNV或进行假感染，并分别保存在15摄氏度SPF水中的385 L水族馆中。在第一项研究中，共有33％的受感染鱼死亡。最后一次死亡后十天或感染后34天，每种处理方法将六只鱼每周杀死六只。通过病毒分离和使用核蛋白（N）基因的嵌套式RT-PCR测试了单个鱼类的肾脏和脑匀浆。所有组织匀浆均对IHNV阴性。重复这项研究后，感染的鱼中有54％的鱼死亡，并在感染后第34天开始，每两周一次杀死6条鱼，每8周一次，并通过病毒分离和定量检测了每条鱼的肾脏和脑匀浆利用引物和荧光标记探针进行实时PCR，以特异性鉴定IHNV的N和G基因。通过病毒分离和定量PCR，感染后34天获得的一名来自IHN幸存者的脑标本是阳性的。但是，所有剩余的组织标本在8周内均为阴性。将对照或IHN存活鱼的合并肾脏或脑组织的上清液腹膜内注射到SPF虹鳟鱼（平均体重10 g）中也未能建立病毒感染。在两项挑战研究中，IHN幸存者均检测到强烈的体液反应，并被认为是导致病毒清除的机制。这些结果提供了科学信息，可用于评估与IHNV流行地区加工虹鳟的活动有关的风险。

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