Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water
Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water
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DOI：
10.1016/j.jviromet.2015.03.001
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发表时间：
2015-06-15
影响因子：
3.1
通讯作者：
Rajan, J. Joseph Sahaya
中科院分区：
文献类型：
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作者：
Alavandi, S. V.;Bharathi, R. Ananda;Rajan, J. Joseph Sahaya
水是水产养殖中白斑综合征病毒（WSSV）传播途径中最重要的组成部分，但相关信息很少。在水中检测病毒是一项挑战，因为它们的数量往往太低，无法通过聚合酶链反应（PCR）等现有方法检测到。为了克服这一困难，必须在检测之前从大量水中浓缩水中的病毒。本研究采用切向流过滤（TFF）超滤（UF）法对3条小溪、10个对虾养殖池、3个对虾养殖池和2个幼虾养殖池的水产养殖生态系统水样和1个孵化场出水处理池水样进行了病毒浓度检测。从这些来源中提取20至1001份水，浓缩至最终体积为100毫升（200-1000倍）。TFF对WSSV的回收率为7.5 ~ 89.61%。从3个对虾养殖池、对虾养殖池、幼虾养殖池和对虾孵化出水处理池各1个水样中成功检测到WSSV病毒浓缩液，实时荧光定量PCR检测到WSSV拷贝数为6 ~ 157 mL（-1）。超滤病毒浓缩技术可以有效地检测水产养殖设施水中的虾类病毒病原体。它可以作为了解水产养殖设施采用的生物安全方案的有效性和开展水生病毒病原体流行病学调查的重要工具。(C) 2015 Elsevier B.V.版权所有
Water represents the most important component in the white spot syndrome virus (WSSV) transmission pathway in aquaculture, yet there is very little information. Detection of viruses in water is a challenge, since their counts will often be too low to be detected by available methods such as polymerase chain reaction (PCR). In order to overcome this difficulty, viruses in water have to be concentrated from large volumes of water prior to detection. In this study, a total of 19 water samples from aquaculture ecosystem comprising 3 creeks, 10 shrimp culture ponds, 3 shrimp broodstock tanks and 2 larval rearing tanks of shrimp hatcheries and a sample from a hatchery effluent treatment tank were subjected to concentration of viruses by ultrafiltration (UF) using tangential flow filtration (TFF). Twenty to 1001 of water from these sources was concentrated to a final volume of 100 mL (200-1000 fold). The efficiency of recovery of WSSV by TFF ranged from 7.5 to 89.61%. WSSV could be successfully detected by PCR in the viral concentrates obtained from water samples of three shrimp culture ponds, one each of the shrimp broodstock tank, larval rearing tank, and the shrimp hatchery effluent treatment tank with WSSV copy numbers ranging from 6 to 157 mL(-1) by quantitative real time PCR. The ultrafiltration virus concentration technique enables efficient detection of shrimp viral pathogens in water from aquaculture facilities. It could be used as an important tool to understand the efficacy of biosecurity protocols adopted in the aquaculture facility and to carry out epidemiological investigations of aquatic viral pathogens. (C) 2015 Elsevier B.V. All rights reserved.