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在鲑鱼(大西洋鲑)幼鲑后同居挑战期间海水中鲑鱼α病毒的过滤、浓缩和检测

Filtration, concentration and detection of salmonid alphavirus in seawater during a post-smolt salmon (Salmo salar) cohabitant challenge.

作者信息

Bernhardt Lisa-Victoria, Myrmel Mette, Lillehaug Atle, Qviller Lars, Chioma Weli Simon

机构信息

Norwegian Veterinary Institute, PO Box 750 Sentrum, 0106 Oslo, Norway.

出版信息

Dis Aquat Organ. 2021 Mar 25;144:61-73. doi: 10.3354/dao03572.

DOI:10.3354/dao03572
PMID:33764314
Abstract

Currently, the prevalence of salmonid alphavirus (SAV) in Norwegian Atlantic salmon farms is largely surveyed via sacrificing fish and sampling of organ tissue on a monthly basis. However, a more cost-efficient, straightforward, rapid, reliable, reproducible and animal welfare friendly method based on the detection of SAV in water could be considered as an alternative method. In the present study, such a method was developed and optimized through a 6 wk cohabitant challenge trial, using post-smolt Atlantic salmon Salmo salar L challenged with high or low doses of SAV subtype 3 (SAV3). Tank water and tissue samples from cohabitant fish were collected at 16 time points. SAV3 was concentrated from the water by filtration, using either electronegative or electropositive membrane filters, which were subsequently rinsed with one of 4 different buffer solutions. SAV3 was detected first in tank water (7 d post-challenge, DPC), and later in cohabitant fish organ tissue samples (12 DPC). The electronegative filter (MF-Millipore™) and rinsing with NucliSENS® easyMAG® Lysis Buffer presented the best SAV3 recovery. A significant positive correlation was found between SAV3 in the tank water concentrates and the mid-kidney samples. Based on these results, detection of SAV3 in filtrated seawater is believed to have the potential to serve as an alternative method for surveillance of SAV in Atlantic salmon farms.

摘要

目前,挪威大西洋鲑鱼养殖场中鲑鱼α病毒(SAV)的流行情况主要通过每月宰杀鱼类并采集器官组织样本进行调查。然而,一种基于检测水中SAV的更具成本效益、直接、快速、可靠、可重复且对动物福利友好的方法可被视为替代方法。在本研究中,通过为期6周的同居挑战试验开发并优化了这样一种方法,使用高剂量或低剂量的SAV 3型(SAV3)对后洄游大西洋鲑鱼(Salmo salar L)进行挑战。在16个时间点采集同居鱼类的养殖池水和组织样本。使用带负电或带正电的膜过滤器通过过滤从水中浓缩SAV3,随后用4种不同缓冲溶液之一冲洗。在养殖池水中首先检测到SAV3(攻毒后7天,DPC),随后在同居鱼类器官组织样本中检测到(攻毒后12天,DPC)。带负电的过滤器(MF - Millipore™)并用NucliSENS® easyMAG®裂解缓冲液冲洗时,SAV3回收率最高。在养殖池水浓缩物中的SAV3与中肾样本之间发现了显著的正相关。基于这些结果,认为在过滤后的海水中检测SAV3有潜力作为大西洋鲑鱼养殖场中SAV监测的替代方法。

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