Detection of white spot syndrome virus (WSSV) in the Pacific oyster Crassostrea gigas

Vázquez-Boucard, Celia; Álvarez-Ruíz, Píndaro; Escobedo‐Fregoso, Cristina; Anguiano-Vega, Gerardo; Durán‐Avelar, Ma. de Jesús; Pinto, Vania Serrano; Escobedo-Bonilla, César Marcial

doi:10.1016/j.jip.2010.04.004

Cited by 30 publications

(17 citation statements)

References 26 publications

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“…Shellfish are filter‐feeding animals and may concentrate pathogens in the surrounding water areas (Metcalf, ; Rippey, ). Therefore, it seems reasonable they might be associated with the transmission of various pathogenic viruses (Kim et al., ; Vazquez‐Boucard et al., ). It is unfortunate that, our knowledge of the interactions between NNV and shellfish as carriers that could accumulate viruses in the marine environment is very limited.…”

Section: Introductionmentioning

confidence: 99%

Isolation and initial characterization of new betanodaviruses in shellfish

Kim

Kwon

Min

et al. 2018

Transbound Emerg Dis

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Betanodaviruses cause the disease viral nervous necrosis (VNN) in finfish. Using a novel approach with two consecutive PCRs, detection semi-nested two-step RT-PCR (DSN-2 RT-PCR) and discriminative multiplex two-step RT-PCR (DMT-2 RT-PCR), we have identified the presence of a new type of betanodavirus in shellfish and called it Korean shellfish nervous necrosis virus (KSNNV). Partial nucleotide sequences of the T4 region in RNA2 fragment of KSNNVs were 73%-75% homologous to those of other reported genotypes and formed a new cluster of betanodavirus in phylogenetic tree analysis. Successful isolation of KSNNV was achieved in two of six shellfish samples containing high concentrations of virus using the blind passage method, and the typical shapes of betanodavirus were confirmed in KSNNV-KOR1 by electron microscopy. In the experimental infection test, seven of 14 fish species showed susceptibility to KSNNV-KOR1 isolate but without clinical signs or death. Although the range of susceptible host species was not significantly different from the RGNNV type, the concentration of KSNNV in the brain of infected fish (10 -10 copies/mg brain) was much lower compared to that found in sevenband grouper (Epinephelus septemfasciatus Thunberg) sampled in the moribund stage with RGNNV infection (10 -10 copies/mg brain). However, histopathological analyses showed the presence of multiple vacuoles in brains of all KSNNV-infected fish at 14 days postinjection. In detection test, as a single or multiple type with the other genotype(s) (RGNNV or BFNNV), the prevalence of KSNNV was 8.4% and 8.7% in domestic (62 of 741 samples) and Chinese samples (12 of 138 samples), respectively, but not in finfish. We propose that KSNNVs obtained from shellfish be classified into a separate and new genotype of betanodavirus.

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Section: Introductionmentioning

confidence: 99%

Isolation and initial characterization of new betanodaviruses in shellfish

Kim

Kwon

Min

et al. 2018

Transbound Emerg Dis

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“…Because of this, they have long been used as environmental biomonitors to detect waterborne chemicals and metals (Claisse 1989, Kwan et al 2003, as well as bacteria, parasites and viruses impacting humans (Miller et al 2005). Previously, we reported the presence of WSSV in the gills and digestive glands of Crassostrea gigas oys-ters placed in water supply canals at a farm where a WSD outbreak had occurred (Vazquez-Boucard et al 2010). Here, oysters were investigated for their potential use as an early warning bioindicator of WSSV in shrimp farm water inlets.…”

Section: Introductionmentioning

confidence: 93%

“…Resale or republication not permitted without written consent of the publisher Dis Aquat Org 98: 201-207, 2012 ters placed in water supply canals at a farm where a WSD outbreak had occurred (Vazquez-Boucard et al 2010). Here, oysters were investigated for their potential use as an early warning bioindicator of WSSV in shrimp farm water inlets.…”

Section: Abstract: Wssv · Disease Management · Bivalve · Sentinel Spmentioning

confidence: 99%

Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus

et al. 2012

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This study explored whether Crassostrea gigas oysters can be used as a bioindicator of white spot syndrome virus (WSSV) in shrimp farm water canals. Bioassays showed that C. gigas can accumulate WSSV in their gills and digestive glands but do not become infected, either by exposure to seawater containing WSSV or by cohabitation with infected shrimp. The use of a WSSV nested PCR to screen oysters placed in water canals at the entry of a shrimp farm allowed WSSV to be detected 16 d prior to the disease occurring. The finding that C. gigas can concentrate small amounts of WSSV present in seawater without being harmed makes it an ideal sentinel species at shrimp farms. KEY WORDS: WSSV · Disease management · Bivalve · Sentinel speciesResale or republication not permitted without written consent of the publisher Dis Aquat Org 98: 201-207, 2012 ters placed in water supply canals at a farm where a WSD outbreak had occurred (Vazquez-Boucard et al. 2010). Here, oysters were investigated for their potential use as an early warning bioindicator of WSSV in shrimp farm water inlets. Before examining the use of C. gigas oysters at a local shrimp farm, laboratory bioassays were performed to confirm their ability to accumulate WSSV and insusceptibility to infection and disease. MATERIALS AND METHODS Virus inoculumAbdominal tissue of WSSV-infected Litopenaeus vannamei shrimp was homogenized with phosphatebuffered saline (PBS, pH 7.4, 1:10 v/v) and clarified at 3500 × g for 20 min at 4°C. The supernatant was transferred to a clean tube and clarified further at 13 000 × g for 20 min at 4°C. This highly clarified supernatant was then filtered through a 0.2 µm membrane filter and serial 10-fold dilutions (10 ) were prepared in PBS. Aliquots (50 µl) of each dilution were injected into tail muscle of 5 L. vannamei weighing 40 ± 4 g (mean ± SE). Prior to inoculation, haemolymph collected from several shrimp was tested by PCR to confirm the absence of WSSV infection. Individual shrimp were held in 10 l tanks containing aerated sea water at 27 ± 1°C and 35 g l −1 salinity and ob served every 12 h; any dead or moribund shrimp were removed and frozen at −80°C. At Day 5 post-injection (p.i.), muscle tissue from all stored moribund and dead shrimp was tested for WSSV using a nested PCR. The virus infection dose 50% endpoint (ID 50 ) was calculated using the method of Reed & Muench (1938), as adapted by EscobedoBonilla et al. (2005) for WSSV in Litopenaeus vannamei, and was expressed per ml of filtrate. Briefly, the percent mortality (M ) was determined at each inoculum dilution and the proportional distance (50% − M b )/(M a − M b ) was calculated using the mortalities at the dilutions directly above (a) and below (b) the 50% mortality point. The proportional distance was then added to the log 10 of Dilution b in order to derive ID 50 ml −1 for the inoculum following correction for the injection of a 50 µl volume. Infection bioassaysTo determine whether Crassostrea gigas is susceptible to WSSV infection, (1) WSSV-free oysters an...

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“…(filter-feeding) norovirus human enteric virus marine birnavirus (MABV), white spot syndrome virus (WSSV) (digestive gland) (Atmar et al, 1995;Gerba et al, 1978;Goyal et al, 1979;Suzuki and Nojima, 1999;Vazquez-Boucard et al, 2010).…”

Section: (Paralichthys Olivaceus) (Scophthalmus Maximus) Flounder Irimentioning

confidence: 99%

“…HRM 분석법을 이용한 패류 내 Megalocytiviruses의 검출과 유전적 분석 고 찰 norovirus human enteric virus , MABV, WSSV (Atmar et al, 1995;Gerba et al, 1978;Goyal et al, 1979;Suzuki and Nojima, 1999;Vazquez-Boucard et al, 2010).…”

Section: Megalocytivirusunclassified

Detection and Genetic Differentiation of Megalocytiviruses in Shellfish, via High-Resolution Melting (HRM) Analysis

Kim¹,

Jin²,

Kim³

et al. 2014

Korean Journal of Fisheries and Aquatic Sciences

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Viruses in the genus Megalocytivirus have been subdivided into four subgroups. Among these subgroups 2 and 4, represented by the red sea bream iridovirus (RBIV) and the olive flounder iridovirus (FLIV), respectively, are nonexotic. subgroups 1 and 3, represented by the red sea bream iridovirus (RSIV) and the infectious spleen and kidney necrosis virus (ISKNV), respectively, have not been detected in Korea and are known as exotic. Shellfish are filterfeeders, and can thus filter and accumulate Megalocytivirus in their digestive glands, allowing us to track viral contamination in surrounding aquatic environment. In this study, we developed a high-resolution melting (HRM) analysis to differentiate among subgroups of Megalocytivirus accumulated in shellfish, and confirmed the convenience and efficiency of this method. More than two subgroups of Megalocytivirus were found in the digestive gland of a single shellfish. We classified all Megalocytivirus viruses from shellfish in Korea into subgroups 2 and 4, although proportions of subgroups were different among regions. Compared to nucleotide sequencing analysis, HRM analysis is a simple and rapid method for differentiating of Megalocytivirus subgroups.

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Detection of white spot syndrome virus (WSSV) in the Pacific oyster Crassostrea gigas

Cited by 30 publications

References 26 publications

Isolation and initial characterization of new betanodaviruses in shellfish

Isolation and initial characterization of new betanodaviruses in shellfish

Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus

Detection and Genetic Differentiation of Megalocytiviruses in Shellfish, via High-Resolution Melting (HRM) Analysis

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