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苏格兰设得兰群岛大西洋鲑(Salmo salar L.)后幼鱼中的传染性胰腺坏死病毒:病毒鉴定、组织病理学、免疫组织化学以及与苏格兰大陆分离株的基因比较

Infectious pancreatic necrosis virus in Atlantic salmon, Salmo salar L., post-smolts in the Shetland Isles, Scotland: virus identification, histopathology, immunohistochemistry and genetic comparison with Scottish mainland isolates.

作者信息

Smail D A, Bain N, Bruno D W, King J A, Thompson F, Pendrey D J, Morrice S, Cunningham C O

机构信息

FRS Marine Laboratory, Aberdeen, UK.

出版信息

J Fish Dis. 2006 Jan;29(1):31-41. doi: 10.1111/j.1365-2761.2005.00678.x.

Abstract

During mid-June 1999 peak mortalities of 11% of the total stock per week were seen at a sea cage site of Atlantic salmon, Salmo salar L., post-smolts in the Shetland Isles, Scotland. Virus was isolated on chinook salmon embryo (CHSE) cells in a standard diagnostic test and infectious pancreatic necrosis virus (IPNV) identified by enzyme-linked immunosorbent assay. IPNV was confirmed as serogroup A by a cell immunofluorescent antibody test using the cross-reactive monoclonal antibody AS-1. Four weeks after the main outbreak, virus titres in surviving moribund fish were assayed at >10(10) TCID50 g(-1) kidney. Histopathology of moribund fish was characterized by pancreatic acinar cell necrosis and a marked catarrhal enteritis of the intestinal mucosa. In the liver, necrosis, leucocytic infiltration and a generalized cell vacuolation were noted. IPNV-specific immunostaining was demonstrated in pancreas, liver, heart, gill and kidney tissue. The nucleotide sequence of the coding region of segment A was determined from the Shetland isolate. A 1180 bp fragment of the VP2 gene of this isolate was compared with a 1979 reference isolate from mainland Scottish Atlantic salmon, La/79 and another more recent mainland isolate, 432/00. Both A2 isolates were derived from carrier fish without signs of IPN and serotyped by a plaque neutralization test. The Shetland isolate shows a different nucleotide and amino acid sequence compared with the two isolates from carrier fish. These latter isolates showed identical amino acid sequences in the fragment examined, despite the 21 years separating the isolations. Sequence comparisons with other A2 (Sp) isolates on the database confirm all three Scottish isolates are A2 (Sp).

摘要

1999年6月中旬,在苏格兰设得兰群岛一个养殖大西洋鲑(Salmo salar L.)后幼鲑的海水网箱养殖场,每周出现的死亡高峰达总存量的11%。在标准诊断试验中,病毒在奇努克鲑胚胎(CHSE)细胞上分离出来,并通过酶联免疫吸附测定法鉴定为传染性胰腺坏死病毒(IPNV)。使用交叉反应单克隆抗体AS-1通过细胞免疫荧光抗体试验将IPNV确认为血清群A。主要疫情爆发四周后,对存活的濒死鱼的病毒滴度进行检测,肾组织病毒滴度>10(10) TCID50 g(-1)。濒死鱼的组织病理学特征为胰腺腺泡细胞坏死和肠道黏膜明显的卡他性肠炎。在肝脏中,可见坏死、白细胞浸润和普遍的细胞空泡化。在胰腺、肝脏、心脏、鳃和肾组织中发现了IPNV特异性免疫染色。从设得兰群岛分离株中确定了A节段编码区的核苷酸序列。将该分离株VP2基因的1180 bp片段与1979年来自苏格兰大陆大西洋鲑的参考分离株La/79以及另一个更近的大陆分离株432/00进行比较。这两个A2分离株均来自无IPN症状的携带病毒鱼,并通过蚀斑中和试验进行血清分型。与来自携带病毒鱼的两个分离株相比,设得兰群岛分离株显示出不同的核苷酸和氨基酸序列。尽管这两个分离株相隔21年分离,但在所检测的片段中显示出相同的氨基酸序列。与数据库中其他A2(Sp)分离株的序列比较证实,所有三个苏格兰分离株均为A2(Sp)。

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