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THE WHITE SPOT SYNDROME VIRUS (WSSV) LOAD IN Dendronereis spp.  [cached]
Desrina, Sarjito, Alfabetian Harjuno Condro Haditomo, Diana Chilmawati
Journal of Coastal Development , 2012,
Abstract: The white spot syndrome virus (WSSV), the causative agent of White Spot Syndrome Disease (WSSD), is a major shrimp pathogen in Indonesia. Dendronereis spp. is a ubiquitous Polychaetes and natural food of shrimp raised in brackishwater pond in Indonesia. The objective of this research is to determine the occurrence of WSSV and the viral load in Dendronereis spp. obtained from the shrimp pond. Dendronereis spp. was obtained with PVC (10 cm in diameter) from a traditional shrimp pond in Semarang vicinity. As acomparison, healthy looking Penaeus monodon was also obtained from the same pond. The occurrence of WSSV in Dendronereis spp. was determined with 1-step and nested PCR using primer for WSSV major envelope protein, VP 28. The viral load was counted with 1-step Real Time PCR. The WSSV was detected in Dendronereis spp. with 1-step and nested PCR. The point prevalence of WSSV infection in Dendronereis spp. is 90 %. The viral load ranged from 0 to 1.9 x 104 copy of DNA/μg total DNA. The viral load in Dendronereis is comparable with that of naturally infected and at carrier state P.monodon from the same pond. This is the first report of WSSV load in naturally infected Dendronereis spp.
The Shrimp NF-κB Pathway Is Activated by White Spot Syndrome Virus (WSSV) 449 to Facilitate the Expression of WSSV069 (ie1), WSSV303 and WSSV371  [PDF]
Pei-Hui Wang, Zhi-Hua Gu, Ding-Hui Wan, Ming-Yan Zhang, Shao-Ping Weng, Xiao-Qiang Yu, Jian-Guo He
PLOS ONE , 2011, DOI: 10.1371/journal.pone.0024773
Abstract: The Toll-like receptor (TLR)-mediated NF-κB pathway is essential for defending against viruses in insects and mammals. Viruses also develop strategies to utilize this pathway to benefit their infection and replication in mammal hosts. In invertebrates, the TLR-mediated NF-κB pathway has only been well-studied in insects and has been demonstrated to be important in antiviral responses. However, there are few reports of interactions between viruses and the TLR-mediated NF-κB pathway in invertebrate hosts. Here, we studied Litopenaeus vannamei Pelle, which is the central regulator of the Toll pathway, and proposed that a similar TLR/MyD88/Tube/Pelle/TRAF6/NF-κB cascade may exist in shrimp for immune gene regulation. After performing genome-wild analysis of white spot syndrome virus (WSSV) encoded proteins, we found that WSSV449 shows 15.7-19.4% identity to Tube, which is an important component of the insect Toll pathway. We further found that WSSV449 activated promoters of Toll pathway-controlled antimicrobial peptide genes, indicating WSSV449 has a similar function to host Tube in activating the NF-κB pathway. We suspected that WSSV449 activated the Toll-mediated NF-κB pathway for regulating viral gene expression. To test this hypothesis, we analyzed the promoters of viral genes and found 40 promoters that possess NF-κB binding sites. A promoter screen showed that the promoter activities of WSSV069 (ie1), WSSV303 and WSSV371 can be highly induced by the shrimp NF-κB family protein LvDorsal. WSSV449 also induced these three viral promoter activities by activating the NF-κB pathway. To our knowledge, this is the first report of a virus that encodes a protein similar to the Toll pathway component Tube to upregulate gene expression in the invertebrate host.
Early Detection of White Spot Syndrome Virus (WSSV) in Isolated Hemocytes of Litopenaeus vannamei  [PDF]
Ariadne Hernández-Pérez, Rossanna Rodríguez-Canul, Edgar Torres-Irineo, Fernando Mendoza-Cano, Daniel Eduardo Coronado-Molina, Jesús Alejandro Zamora-Brise?o, Jorge Hernández-López
CellBio (CellBio) , 2017, DOI: 10.4236/cellbio.2017.61001
Abstract: To date, White Spot Syndrome (WSS) produced by the White Spot Syndrome Virus (WSSV) causes one of the most severe diseases infecting penaeid shrimps worldwide. Although a vast amount of studies has elucidated pathogenesis in live infection models, there is still little information about the interaction of WSSV infections using in vitro models in the whiteleg shrimp Litopenaeus vannamei (L. vannamei) hemocytes. In this study, a WSSV infection kinetics was performed using total hemocytes isolated from healthy L. vannamei organisms and maintained in in vitro conditions using isotonic solution for shrimp (ISS). The infected experimental cells received ≈ 30,000 viral copies of WSSV. The viability of the hemocytes (control and infected group) was measured during the kinetics with trypan blue exclusion method and cells were maintained up to 6 hpi (post-infection) with non-significant differences of viability between both groups. WSSV replication was assessed using RT- PCR at the RNA expression level of the early viral gene Ie1 and transcripts were detected as early as 30 min pi. Hemocytes from WSSV group showed disrupted integrity, degranulation and irregular shape. This study provides evidence of the capability of WSSV to infect and replicates in L. vannamei hemocytes using in vitro assays in short times as 30 min.
Immune response of black tiger shrimp (Penaeus monodon Fabricius) to yellow head virus (YHV) and white spot syndrome virus (WSSV)
Ruangsri, J.,Phongdara, A.,Prasertsan, P.,Supamattaya, K.
Songklanakarin Journal of Science and Technology , 2005,
Abstract: Immunological responses of black tiger shrimp (Penaeus monodon) were induced under laboratory conditions by injecting white spot syndrome virus (WSSV) and yellow head virus (YHV). The survivors from WSSV and YHV infection showed improvement in their immunity against the re-challenging by both viruses. The WSSV survivors showed higher capability than that of YHV survivors in developing the immunity. The highest relative percent survival (RPS) against WSSV and YHV noted after a 43-day period of WSSV injection was 67 and 37.5%, respectively. While the RPSs against both viruses after periods of 46 and 60 days of the YHV injection were 37.5 and 12.1%. Similarly, an in vitro neutralization activity of plasma separated from the survivors with WSSV injection showed preference of virus being eliminated over the plasma from YHV injection.Blood parameters for survivors with WSSV injection for 43 days showed an increase in phenoloxidase activity, while the YHV injected survivors exhibited higher level of total hemocytes and phenoloxidase activity. Histopathological examinations in survivors revealed changes of lymphatic tubes into spheroids higher than those in normal individual.

WEI Xiu-Mei,SHENG Xiu-Zhen,TANG Xiao-Qian,XING Jing,ZHAN Wen-Bin,

海洋与湖沼 , 2010,
Abstract: During the past more than ten years since a white spot syndrome virus (WSSV) disease outbreak in 1993, many research papers have been published on the pathogen, pathology, detection and diagnostic methods of the disease. Recent works focused mainly on the infection mechanism and searching the strategies of prophylaxis and control of WSSV infection. The purpose of this work was to produce rabbit anti-idiotypic antibody of WSSV (Ab2), analyze its characteriza-tion, and find a new way to study WSSV infection m...
Screening White Spot Syndrome Virus(WSSV)-Resistant Molecular Markers from Fenneropenaeus chinensis Screening White Spot Syndrome Virus(WSSV)-Resistant Molecular Markers from Fenneropenaeus chinensis  [PDF]
WU Yingying,MENG Xianhong,KONG Jie,LUAN Sheng,LUO Kun,WANG Qingyin,ZHENG Yongyun
- , 2017,
Abstract: White spot syndrome virus(WSSV)-resistant molecular markers were screened from the selectively bred new variety ‘Huanghai No. 2' of Fenneropenaeus chinensis using unlabeled-probe high-resolution melting(HRM) technique. After the artificial infection with WSSV, the first 96 dead shrimps and the last 96 surviving shrimps were collected, representing WSSV-susceptible and-resistant populations, respectively. The genotypes at well-developed 39 single nucleotide polymorphisms(SNPs) loci were obtained. As revealed in the Chi-square test, 3 SNPs, genotype A/A of contig C364-89 AT, genotype A/A of C2635-527 CA and genotype C/T of contig C12355-592 CT, were positively correlated with disease-resistance traits. Other 2 SNPs, genotype G/G of contig C283-145 AG and genotype C/C of contig C12355-592 CT, were negatively correlated. Moreover, analysis with Blast X program for disease-resistant SNPs indicated that 3 contigs, Contig283, Contig364 and Contig12355, matched to the functional genes of effector caspase of Penaeus monodon, peptide transporter family 1-like protein, and 40 S ribosomal protein S2 of Perca flavescens with high sequence similarity. The results will be helpful to provide theoretical and technical supports for molecular marker-assisted selective breeding of F. chinensis
PCR detection of white spot syndrome virus (WSSV) from farmed Pacific white shrimp (Litopenaeus vannamei) in selected sites of the Philippines  [PDF]
Benedict A. Maralit,Christopher M. A. Caipang,Mudjekeewis D. Santos,Mary Beth B. Maningas
Aquaculture, Aquarium, Conservation & Legislation , 2011,
Abstract: Great losses caused by white spot syndrome virus (WSSV) in shrimp culture have beenattributed to poor screening procedures in farms and the lack of sufficient access to specific pathogenfree brood stock. Thus, early detection of the virus is considered the best option for shrimp farmers. Thestudy, thus, assessed viral incidence in the Philippines and partially sequenced and characterized thePhilippine WSSV isolate with regards to other isolates in GenBank. Developed primers for PCR can detecttarget genes from 0.4 pg of DNA extract from shrimp samples. PCR detection revealed that 6.67 %(1/15) of market samples from Zambales are infected with WSSV. Shrimp samples from a local shop anda public market in General Santos City showed 46.67% (7/15) and 20% (3/15) WSSV-positive samplesrespectively. Shrimp sources from Capiz and Batangas, however, showed negative detection for WSV. Nosignificant difference in the number of infected samples from the sampling sites was found. Combineddetections reveal that the Philippines has a low infection rate of 14.67%. The study has partiallysequenced and characterized Philippine isolate. During the sampling period, most shrimps in GeneralSantos City were WSSV-positive by PCR detection.
WSSV ie1 promoter is more efficient than CMV promoter to express H5 hemagglutinin from influenza virus in baculovirus as a chicken vaccine
Fang He, YuenFern Ho, Li Yu, Jimmy Kwang
BMC Microbiology , 2008, DOI: 10.1186/1471-2180-8-238
Abstract: White spot syndrome virus (WSSV) immediate-early promoter one (ie1) was shown to be a stronger promoter for gene expression in insect cells compared with Cytomegalovirus immediate-early (CMV) promoter in luciferase assays. In an attempt to improve expression efficiency, a recombinant baculovirus was constructed expressing hemagglutinin (HA) of H5N1 influenza virus under the control of WSSV ie1 promoter. HA expression in SF9 cells increased significantly with baculovirus under WSSV ie1 promoter, compared with CMV promoter based on HA contents and hemagglutination activity. Further, immunization with baculovirus under WSSV ie1 promoter in chickens elicited higher level anti-HA antibodies compared to CMV promoter, as indicated in hemagglutination inhibition, virus neutralization and enzyme-linked immunosorbent assays. By immunohistochemistry, strong HA antigen expression was observed in different chicken organs with vaccination of WSSV ie1 promoter controlled baculovirus, confirming higher efficiency in HA expression by WSSV ie1 promoter.The production of H5 HA by baculovirus was enhanced with WSSV ie1 promoter, especially compared with CMV promoter. This contributed to effective elicitation of HA-specific antibody in vaccinated chickens. This study provides an alternative choice for baculovirus based vaccine production.The spread of highly pathogenic avian influenza A (H5N1) viruses from Asia to the Middle East, Europe, and Africa poses the threat of an influenza pandemic. Vaccination of poultry is an effective measure to control virus spread [1]. Current production of inactivated influenza vaccine requires high-level biocontainment facilities and large numbers of embryonated chicken eggs, while baculovirus surface displayed recombinant hemagglutinin may be an attractive alternative to the effective influenza vaccine [2-5].White spot syndrome virus (WSSV), a major pathogen in shrimp, can infect a wide range of invertebrate tissues and cells. WSSV genome has 9 repeated
First report on White Spot Syndrome Virus (WSSV) infection in white leg shrimp Litopenaeus vannamei (Crustacea, Penaeidae) under semi intensive culture condition in India
Gunalan Balakrishnan,Soundarapandian Peyail,Ramachandran Kumaran,Anand Theivasigamani
Aquaculture, Aquarium, Conservation & Legislation , 2011,
Abstract: Scientific shrimp culture began in India in the late eighties along the east coast particularly inAndrapradesh and Tamilnadu. Continuous success of shrimp culture was affected by mass mortalities ofcultured shrimp in 1994. Thereafter disease infection on survival and production of shrimps get itsimportance in culture. The present study is the first report on WSSV (white spot syndrome virus)infection in cultured Litopenaeus vannamei (Boone, 1931) in India. WSSV infection was observed on 70thdays of culture due to cross contamination of white spot infected shrimp from the neighboring farmbecause of birds. Due to this infection within two days the mortality ratio has gone up to 25% in pond 1and 12% in pond 2. So this present study strongly recommends to every shrimp farmers to go for birdfencing & crab fencing to avoid horizontal contamination, before stocking the good quality seed, thenthey will have the risk free WSSV culture.
Selection of a Novel Single-chain Variable Fragment Antibody Specifically Against a Linear Epitope of White Spot Syndrome Virus

Yuzhen Wang,Xiaohua Zhang,Nan Xiao,Min Zhang,Heping Dai,

生物工程学报 , 2008,
Abstract: White spot syndrome virus (WSSV) is one of the most important pathogens in shrimp farm throughout the world. Many researches on WSSV have been done, but no efficient approach has been gained to protect and cure the disease. In this study, we constructed a single-chain fragment variable (scFv) antibody library displayed on phage using spleen cells from mice immunized with denatured WSSV. After several rounds of panning respectively against purified intact WSSV virions and purified VP28 expressed in Escherichia coli, five novel scFv antibodies specifically against WSSV were selected, one of which, clone P75E8, recognized a linear epitope. The location in virions of the epitopes recognized by the five scFv clones was determined by immunoelectron microscopy. This study provides a new way to obtain more different antibodies specifically binding to WSSV, and especially provides a new strategy to obtain scFvs against linear epitopes.
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