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Donald V. Lightner - One of the best experts on this subject based on the ideXlab platform.
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RESEARCH Nonsusceptibility of Primate Cells to Taura Syndrome Virus
2013Co-Authors: Carlos R. Pantoja, Solangel A. Navarro, Jaime Naranjo, Donald V. Lightner, Charles P. GerbaAbstract:Taura Syndrome Virus (TSV), a pathogen of penaeid shrimp and member of the family Dicistroviridae, was recently reported to have the ability to infect primate cells. We independently retested this hypothesis. Three lines of primate cells FRhK-4, MA-104, and BGMK, which are highly susceptible to infection by human picornaViruses, were challenged with TSV. Viral replication was assayed by realtime reverse transcription–polymerase chain reaction using cell media samples collected on days 0, 4, and 7 postchallenge. By day 7, genome copy numbers had decreased 25%–99%. No cytopathic effect was observed after 7 days. An in situ hybridization assay, with gene probes specific for detection of TSV, was negative for TSV in challenged cells. The infectivity of residual Virus in the cell culture media at day 7 was confirmed by bioassay using TSV-free indicator shrimp (Litopenaeus vannamei). TSV did not infect the primate cells tested, and no evidence of zoonotic potential was found. The general assumption is that the viral agents that cause disease in penaeid shrimp do not infect vertebrates. Supporting this assumption is the absence of documented cases of any shrimp Virus causing disease in any animal species other than crustaceans. In a recent article, Taura Syndrome Virus (TSV), exclusively a pathogen of penaeid shrimp, was attributed a zoonotic potential because of its reported ability to infect cultured human and monkey cells (1). Aside from the food safety issues raised by this report, we were very interested in confirming those results because of the practical value of this new option for growing TSV in vitro. To date, TSV (or any other of the known Viruses of the penaeid shrimp) has not yet been successfully cultured in any invertebrate or vertebrate cellculture system. Hence, if viable, the use of primate cells for propagation of TSV would prove to be an important advancement in the study of TSV, and perhaps of other crustacean Viruses. While the experiment reported in this study did not include all of the cell lines used by Audelodel-Valle et al. (1), namely human rhabdomyosarcom
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New genotypes of white spot Syndrome Virus (WSSV) and Taura Syndrome Virus (TSV) from the Kingdom of Saudi Arabia.
Diseases of Aquatic Organisms, 2012Co-Authors: Kathy F.j. Tang, Carlos R. Pantoja, Solangel A. Navarro, Fernando Aranguren, Donald V. LightnerAbstract:White spot Syndrome Virus (WSSV) and Taura Syndrome Virus (TSV) are highly patho genic to penaeid shrimp and have caused significant economic losses in the shrimp culture industry around the world. During 2010 and 2011, both WSSV and TSV were found in Saudi Ara- bia, where they caused severe mortalities in cultured Indian white shrimp Penaeus indicus. Most outbreaks of shrimp Viruses in production facilities can be traced to the importation of infected stocks or commodity shrimp. In an attempt to determine the origins of these viral outbreaks in Saudi Arabia, we performed variable number of tandem repeat (VNTR) analyses for WSSV iso- lates and a phylogenetic analysis for TSV isolates. From the WSSV genome, the VNTR in open reading frames (ORFs) 125 and 94 were investigated with PCR followed by DNA sequence analy- sis. The genotypes were categorized as {N125, N94} where N is the number of repeat units in a spe- cific ORF, and the subscript indicates the ORF (i.e. ORFs 125 and 94 in this case). From 15 Saudi Arabia WSSV isolates, we detected 3 genotypes: {6125, 794}, {7125, del94}, and {8125, 1394}. The WSSV genotype of {7125, del94} appears to be a new variant with a 1522 bp deletion encompassing com- plete coding regions of ORF 94 and ORF 95 and the first 82 bp of ORF 93. For TSV genotyping, we used a phylogenetic analysis based on the amino acid sequence of TSV capsid protein 2 (CP2). We analyzed 8 Saudi Arabian isolates in addition to 36 isolates from other areas: SE Asia, Mexico, Venezuela and Belize. The Saudi Arabian TSV clustered into a new, distinct group. Based on these genotyping analyses, new WSSV and TSV genotypes were found in Saudi Arabia. The data suggest that they have come from wild shrimp Penaeus indicus from the Red Sea that are used for broodstock.
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Protection from yellow head Virus (YHV) infection in Penaeus vannamei pre-infected with Taura Syndrome Virus (TSV).
Diseases of Aquatic Organisms, 2012Co-Authors: Luis Fernando Aranguren, Kathy F.j. Tang, Donald V. LightnerAbstract:Pacific white shrimp Penaeus vannamei that were pre-exposed to Taura Syndrome Virus (TSV) and then challenged with yellow head Virus (YHV) acquired partial protection from yellow head disease (YHD). Experimental infections were carried out using specific-pathogen-free (SPF) shrimp which were first exposed per os to TSV; at 27, 37 and 47 d post infection they were then challenged by injection with 1 × 104 copies of YHV per shrimp (designated the TSV-YHV group). Shrimp not infected with TSV were injected with YHV as a positive control. Survival analyses comparing the TSV-YHV and YHV (positive control) groups were conducted, and significant survival rates were found for all the time groups (p < 0.001). A higher final survival was found in the TSV-YHV group (mean 55%) than in the positive control (0%) (p < 0.05). Duplex reverse transcription quantitative PCR was used to quantify both TSV and YHV. Lower YHV copy numbers were found in the TSV-YHV group than in the positive control in pleopods (3.52 × 109 vs. 1.88 × 1010 copies µg RNA-1) (p < 0.001) and lymphoid organ (LO) samples (3.52 × 109 vs. 1.88 × 1010 copies µg RNA-1) (p < 0.01). In situ hybridization assays were conducted, and differences in the distribution of the 2 Viruses in the target tissues were found. The foci of LO were infected with TSV but were not infected with YHV. This study suggests that a viral interference effect exists between TSV and YHV, which could, in part, explain the absence of YHD in the Americas, where P. vannamei are often raised in farms where TSV is present.
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Hyperthermia does not protect Kona stock Penaeus vannamei against infection by a Taura Syndrome Virus isolate from Belize.
Diseases of Aquatic Organisms, 2010Co-Authors: Isabelle Côté, Donald V. LightnerAbstract:This study evaluated the susceptibility of Penaeus vannamei, Kona stock-line, to infection by an isolate of Taura Syndrome Virus from Belize (TSV-BZ) under hyperthermic conditions (32 degrees C). Shrimp exposed to the reference Hawaii-94 isolate of TSV (TSV-HI) showed resistance to infection at 32 degrees C as demonstrated by the absence of mortality, histopathological lesions and decreased viral load by qPCR. However, at 32 degrees C, shrimp were fully susceptible to the disease caused by TSV-BZ, exhibiting high mortality, severe histopathological lesions and increased viral load. This susceptibility of shrimp to TSV-BZ infection under hyperthermic conditions was independent of the route of infection (injection vs. per os) and the salinity of the water (11 vs. 28). TSV-BZ might be a temperature-permissible mutant of TSV.
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Development and characterization of a monoclonal antibody against Taura Syndrome Virus.
Journal of Fish Diseases, 2009Co-Authors: I. Côté, Bonnie T. Poulos, Rita M. Redman, Donald V. LightnerAbstract:We produced a panel of monoclonal antibodies (MAbs) from the fusion of Taura Syndrome Virus variants from Belize (TSV-BZ) immunized BALB/cJ mouse spleen cells and non-immunoglobulin secreting SP2/0 mouse myeloma cells. One antibody, 2C4, showed strong specificity and sensitivity for TSV in dot-blot immunoassay and immunohistochemistry (IHC) analysis. The MAb reacted against native TSV-BZ, TSV variants from Sinaloa, Mexico (TSV-SI) and TSV variants from Hawaii (TSV-HI) in dot-blot immunoassay. By IHC, the antibody identified the Virus in a pattern similar to the digoxigenin-labelled TSV-cDNA probe for the TSV-BZ, TSV-HI and TSV-SI variants, but not for the TSV variants from Venezuela (TSV-VE) and the TSV variants from Thailand (TSV-TH). MAb 2C4 did not react against other shrimp pathogens or with normal shrimp tissue. Western blot analysis showed a strong reaction against CP2, a region of high antigenic variability amongst TSV variants. This antibody has potential diagnostic application in detection and differentiation of certain TSV biotypes.
J. M. Lotz - One of the best experts on this subject based on the ideXlab platform.
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Identification of Genes Involved in Taura Syndrome Virus Resistance in Litopenaeus Vannamei
Journal of Aquatic Animal Health, 2014Co-Authors: Idrissa Boube, J. M. Lotz, Alex E. Pozhitkov, Robert J. GriffittAbstract:AbstractThe goal of the present research was to identify the genes that are differentially expressed between two lineages of Pacific white shrimp Litopenaeus vannamei displaying different susceptibilities to Taura Syndrome Virus (TSV) and to understand the molecular pathways involved in resistance to the disease. An oligonucleotide microarray was constructed and used to identify several genes that were differentially expressed in the two L. vannamei lineages following infection with TSV. Individual L. vannamei from either resistant or susceptible lineages were exposed via injection to TSV. Individuals were removed at 6 and 24 h postinfection, and gene expression was assessed with the in-house microarray. The microarray data resulted in the selection of a set of 397 genes that were altered by TSV exposure between the different lineages. Significantly differentially expressed genes were subjected to hierarchical clustering and revealed a lineage-dependent clustering at 24 h postinoculation, but not at 6 h p...
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Estimation of genetic parameters for survival to multiple isolates of Taura Syndrome Virus in a selected population of Pacific white shrimp Penaeus (Litopenaeus) vannamei
Aquaculture, 2013Co-Authors: Dustin R. Moss, Shaun M. Moss, J. M. LotzAbstract:Abstract Taura Syndrome Virus (TSV) is an economically important pathogen of the Pacific white shrimp, Penaeus ( Litopenaeus ) vannamei . To date, > 40 unique TSV isolates have been identified and phylogenetic analyses of these isolates have revealed four distinct genetic groups named according to their geographic origin: Americas, Belize, South East Asia, and Venezuela. Although there is evidence that virulence varies among different TSV isolates, little is known about how shrimp survival is correlated among isolates (i.e. genetic correlations). In addition, estimates of genetic correlation between TSV survival and other commercially important traits are limited. The objectives of this study were to (1) estimate genetic correlations for shrimp survival to a genetically diverse suite of TSV isolates and (2) estimate genetic correlations between isolate-specific TSV survival and growout performance traits (i.e. growth and growout survival). A total of 180 full-sib families were challenged with TSV: 130 families challenged with Americas and Belize group isolates and 50 families challenged with isolates from all four genetic groups. In addition, 100 of these families were tested for growout performance in a recirculating aquaculture system (RAS) at intensive stocking densities (> 230 shrimp/m 2 ). All families were from a shrimp line selected for TSV resistance and growth over multiple generations. Genetic correlations for survival among TSV isolates were positive and of moderate to high magnitude ( r G = 0.35–0.99). Genetic correlations for TSV survival and RAS growth were all negative, but of low magnitude ( r G = − 0.07 to − 0.29). Correlations between TSV survival and RAS survival varied from slightly negative to moderately positive. These results indicate that breeding for survival to any one of the four TSV isolates evaluated in this study should, in general, improve survival to the other isolates. Results also suggest that there are no significant costs associated with selection for TSV resistance relative to growout performance.
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Taura Syndrome Virus loads in Litopenaeus vannamei hemolymph following infection are related to differential mortality
Diseases of Aquatic Organisms, 2010Co-Authors: Zhiming Cao, Shiao Y. Wang, Verlee Breeland, Anne-marie Moore, J. M. LotzAbstract:Taura Syndrome is an economically important disease that can cause catastrophic losses of farmed shrimp. Without effective treatments for Taura Syndrome Virus (TSV), one approach to managing the problem is to selectively breed shrimp populations with increased disease resistance. To better understand why some shrimp can survive exposure to TSV, information is needed on how viral loads progress and persist following infection. Data reported here show that mortalities occurring mostly within 1 wk of infection are associated with high viral titers, and titers as high as 10(8.7) genome copies per microl hemolymph can persist for up to 3 wk in survivors. Thereafter, and up to approximately 9 wk post-exposure, most surviving shrimp remain chronically infected with TSV loads ranging from 10(4) to 10(8) genome copies per microl hemolymph. Challenging shrimp from families with varying TSV resistance showed that in shrimp from less resistant families, the TSV load in hemolymph increased earlier and reached higher peaks than in shrimp from more resistant families. Although TSV loads in moribund shrimp from families differing in resistance did not differ significantly, infection loads among survivors were lower in shrimp from more resistant families. Taken together, the data suggest that lethal infection loads can occur in both more and less susceptible shrimp and that survivors represent shrimp in which viral expansion is better contained.
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Effect of Chronic Taura Syndrome Virus Infection On Salinity Tolerance of Litopenaeus vannamei
Diseases of Aquatic Organisms, 2005Co-Authors: J. M. Lotz, Lesber Salazar Anton, M. Andres SotoAbstract:Taura Syndrome Virus (TSV) is one of the most important shrimp Viruses affecting farmed shrimp worldwide. After an acute phase during which the likelihood of mortality is elevated, infected shrimp enter a chronic phase during which shrimp appear to resume normal behavior and display no gross signs of infection. This study was designed to determine if chronically TSV-infected shrimp Litopenaeus vannamei are compromised by the infection. Specifically we investigated whether chronically infected shrimp could tolerate a drop in salinity as strongly as uninfected shrimp. The study consisted of 3 trials that compared survival of uninfected and chronically TSV-infected L. vannamei after drops in salinity from 24 ppt to salinities varying from 18 to 0 ppt. Logistic regression detected a significant effect of TSV infection on survival of chronically infected shrimp (p < 0.05). Salinity drops from 24 ppt to 3 and 6 ppt resulted in statistically different survivals (p < 0.05). Survival rates were similar among groups for salinity drops to greater than 6 ppt or less than 3 ppt. Salinities at which 50% of the shrimp died (LC 50 ) were 3.06 ppt for the uninfected and 6.65 ppt for the chronically infected groups. Moreover, histopathological analysis of chronically infected shrimp that were moribund or recently dead showed no signs of having reverted to the acute stage of the disease. These results suggest that chronically infected shrimp are not able to tolerate a salinity drop as strongly as uninfected shrimp.
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The use of differential display to isolate viral genomic sequence for rapid development of PCR-based detection methods. A test case using Taura Syndrome Virus.
Journal of Virological Methods, 2004Co-Authors: Shiao Y. Wang, J. M. LotzAbstract:The purpose of this study was to explore the efficacy of using differential display (DD) to isolate viral genomic sequence using tissues from infected organisms so that a PCR procedure to detect the pathogen may be developed rapidly. The model Virus used was the Taura Syndrome Virus (TSV), a ssRNA Virus that cause high rates of mortality at shrimp farms. Two random primers in combination with four anchored primers were used to isolate five cDNAs, ranging in size from 241 to 822 bp, that were differentially expressed in TSV-infected shrimp (Litopenaeus vannamei). PCR experiments revealed that four of the five encoded shrimp genes while the fifth was likely to be a TSV gene. Evidence that the putative TSV sequence is part of the TSV genome was obtained by the 97% sequence identity it shared with the published TSV genome. PCR primers were designed successfully using the differential display sequence to develop a RT-PCR-based method to detect TSV. Because differential display does not require physical isolation of the Virus and only a small amount of infected sample is needed, the technique may be useful as a method to isolate nucleic acid sequences from emerging pathogens so that PCR primers for their detection may be developed rapidly.
Kathy F.j. Tang - One of the best experts on this subject based on the ideXlab platform.
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Detection of the microsporidian Enterocytozoon hepatopenaei (EHP) and Taura Syndrome Virus in Penaeus vannamei cultured in Venezuela
Aquaculture, 2017Co-Authors: Kathy F.j. Tang, Luis Fernando Aranguren, Patharapol Piamsomboon, Jee Eun Han, Irina Y. Maskaykina, Margeaux M. SchmidtAbstract:Abstract The microsporidian Enterocytozoon hepatopenaei (EHP) is an intracellular parasite that has become a critical threat to the shrimp farming industry in SE Asia. This parasite replicates in the hepatopancreas and midgut, and infected shrimp exhibit reduced feeding and severely retarded growth. In this study, we describe the first case of EHP-infected Penaeus vannamei cultured in Venezuela. Its histopathology is very similar to that of SE Asia EHP, with infected shrimp showing basophilic inclusions in hepatopancreas. Upon in situ hybridization of an EHP 18S rRNA gene fragment labeled with digoxigenin, the probe reacted intensely to the basophilic inclusions within the cytoplasm of the infected cells. We also compared the nucleotide sequence similarities in 18S rRNA gene (1095-bp), in β-tubulin (583-bp) and spore wall protein (431-bp) genes between the Venezuela EHP and SE Asia isolates; and the results showed 99%, 93% and 91% identities, respectively. This suggests that the Venezuela EHP was not recently introduced from SE Asia. In a shrimp farm that was affected by EHP, Taura Syndrome Virus (TSV) was also detected in the P. vannamei by RT-qPCR and histological examinations. TSV is a major pathogen in penaeid shrimp aquaculture and has caused substantial economic losses during the last 2 decades, however, TSV has not been reported since 2011. This TSV was determined to be a Venezuela genotype based on the phylogenetic analysis in the capsid protein 2 gene and it showed to have sequence identities of 97% (nucleotide) and 98% (amino acid) to a Venezuela isolate found in 2005. The emergence of EHP and occurrence of TSV in the Venezuela will have a significant impact on the shrimp production if they spread to other farms. Therefore, the shrimp cultured within the country should be monitored closely for the presence of EHP and TSV.
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New genotypes of white spot Syndrome Virus (WSSV) and Taura Syndrome Virus (TSV) from the Kingdom of Saudi Arabia.
Diseases of Aquatic Organisms, 2012Co-Authors: Kathy F.j. Tang, Carlos R. Pantoja, Solangel A. Navarro, Fernando Aranguren, Donald V. LightnerAbstract:White spot Syndrome Virus (WSSV) and Taura Syndrome Virus (TSV) are highly patho genic to penaeid shrimp and have caused significant economic losses in the shrimp culture industry around the world. During 2010 and 2011, both WSSV and TSV were found in Saudi Ara- bia, where they caused severe mortalities in cultured Indian white shrimp Penaeus indicus. Most outbreaks of shrimp Viruses in production facilities can be traced to the importation of infected stocks or commodity shrimp. In an attempt to determine the origins of these viral outbreaks in Saudi Arabia, we performed variable number of tandem repeat (VNTR) analyses for WSSV iso- lates and a phylogenetic analysis for TSV isolates. From the WSSV genome, the VNTR in open reading frames (ORFs) 125 and 94 were investigated with PCR followed by DNA sequence analy- sis. The genotypes were categorized as {N125, N94} where N is the number of repeat units in a spe- cific ORF, and the subscript indicates the ORF (i.e. ORFs 125 and 94 in this case). From 15 Saudi Arabia WSSV isolates, we detected 3 genotypes: {6125, 794}, {7125, del94}, and {8125, 1394}. The WSSV genotype of {7125, del94} appears to be a new variant with a 1522 bp deletion encompassing com- plete coding regions of ORF 94 and ORF 95 and the first 82 bp of ORF 93. For TSV genotyping, we used a phylogenetic analysis based on the amino acid sequence of TSV capsid protein 2 (CP2). We analyzed 8 Saudi Arabian isolates in addition to 36 isolates from other areas: SE Asia, Mexico, Venezuela and Belize. The Saudi Arabian TSV clustered into a new, distinct group. Based on these genotyping analyses, new WSSV and TSV genotypes were found in Saudi Arabia. The data suggest that they have come from wild shrimp Penaeus indicus from the Red Sea that are used for broodstock.
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Protection from yellow head Virus (YHV) infection in Penaeus vannamei pre-infected with Taura Syndrome Virus (TSV).
Diseases of Aquatic Organisms, 2012Co-Authors: Luis Fernando Aranguren, Kathy F.j. Tang, Donald V. LightnerAbstract:Pacific white shrimp Penaeus vannamei that were pre-exposed to Taura Syndrome Virus (TSV) and then challenged with yellow head Virus (YHV) acquired partial protection from yellow head disease (YHD). Experimental infections were carried out using specific-pathogen-free (SPF) shrimp which were first exposed per os to TSV; at 27, 37 and 47 d post infection they were then challenged by injection with 1 × 104 copies of YHV per shrimp (designated the TSV-YHV group). Shrimp not infected with TSV were injected with YHV as a positive control. Survival analyses comparing the TSV-YHV and YHV (positive control) groups were conducted, and significant survival rates were found for all the time groups (p < 0.001). A higher final survival was found in the TSV-YHV group (mean 55%) than in the positive control (0%) (p < 0.05). Duplex reverse transcription quantitative PCR was used to quantify both TSV and YHV. Lower YHV copy numbers were found in the TSV-YHV group than in the positive control in pleopods (3.52 × 109 vs. 1.88 × 1010 copies µg RNA-1) (p < 0.001) and lymphoid organ (LO) samples (3.52 × 109 vs. 1.88 × 1010 copies µg RNA-1) (p < 0.01). In situ hybridization assays were conducted, and differences in the distribution of the 2 Viruses in the target tissues were found. The foci of LO were infected with TSV but were not infected with YHV. This study suggests that a viral interference effect exists between TSV and YHV, which could, in part, explain the absence of YHD in the Americas, where P. vannamei are often raised in farms where TSV is present.
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An improved Taura Syndrome Virus (TSV) RT-PCR using newly designed primers.
Aquaculture, 2009Co-Authors: Solangel A. Navarro, Kathy F.j. Tang, Donald V. LightnerAbstract:Taura Syndrome Virus (TSV)-specific primers, designated as 7171F/7511R, were designed to improve the sensitivity of RT-PCR detection. This pair of primers was shown to detect TSV isolates representative of four phylogenetic lineages: Belize, Americas, SE Asia, and Venezuela. Its detection limit was determined to be 20 copies of the TSV genome, 100 times more sensitive than the TSV primers 9992/9195 currently being used by many laboratories and recommended for surveillance and diagnostic applications by the Office International des Epizooties. Primers 7171F/7511R were found to be specific to TSV and did not to react to either Infectious hypodermal and hematopoietic necrosis Virus (IHHNV), White spot Syndrome Virus (WSSV), Yellow head Virus (YHV), or Infectious myonecrosis Virus (IMNV). This new RT-PCR method was shown to detect TSV in chronically infected Penaeus vannamei that had survived up to 236 days after exposure to TSV.
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A quick fuse and the emergence of Taura Syndrome Virus.
Virology, 2009Co-Authors: Joel O. Wertheim, Solangel A. Navarro, Kathy F.j. Tang, Donald V. LightnerAbstract:Abstract Over the last two decades, Taura Syndrome Virus (TSV) has emerged as a major pathogen in penaeid shrimp aquaculture and has caused substantial economic loss. The disease was first discovered in Ecuador in 1991, and the Virus is now globally distributed with the greatest concentration of infections in the Americas and Southeast Asia. To determine the evolutionary history of this Virus, we constructed a phylogeny containing 83 TSV isolates from 16 countries sampled over a 16-year period. This phylogeny was inferred using a relaxed molecular clock in a Bayesian Markov chain Monte Carlo framework. We found phylogenetic evidence that the TSV epidemic did indeed originate in the Americas sometime around 1991 (1988–1993). We estimated the TSV nucleotide substitution rate at 2.37 × 10 − 3 (1.98 × 10 − 3 to 2.82 × 10 − 3 ) substitutions/site/year within capsid gene 2. In addition, the phylogeny was able to independently corroborate many of the suspected routes of TSV transmission around the world. Finally, we asked whether TSV emergence in new geographic locations operates under a quick fuse (i.e. rapid appearance of widespread disease). Using a relaxed molecular clock, we determined that TSV is almost always discovered within a year of entering a new region. This suggests that current monitoring programs are effective at detecting novel TSV outbreaks.
Wansika Kiatpathomchai - One of the best experts on this subject based on the ideXlab platform.
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RT-LAMP detection of shrimp Taura Syndrome Virus (TSV) by combination with a nanogold-oligo probe
Aquaculture Research, 2013Co-Authors: Jurairat Phromjai, Adisorn Tuantranont, Thitima Mathuros, D. Phokharatkul, Potchanathorn Prombun, Rungkarn Suebsing, Wansika KiatpathomchaiAbstract:This study describes the application and optimization of a newly developed specific gold nanoparticle (AuNP) visual detection method to detect a reverse transcription loop-mediated isothermal amplification (RT-LAMP) product of Taura Syndrome Virus (TSV) in shrimp with simple and cheaper in comparison to conventional gel electrophoresis. Briefly, the visual detection method is based on prevention of a salt induced DNA-labelled AuNP colour change from red to blue–purple due to hybridization with complementary DNA amplicons that arise from a positive TSV RT-LAMP reaction. RT-LAMP combined with visual amplicon detection using the DNA-labelled AuNP-probe had high sensitivity and the probe did not cross react with amplicons obtained using specific LAMP methods with other shrimp Viruses. The advantages of this assay include simplicity, short analysis time and low-cost, suitable for field laboratory applications.
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in vitro cultivation of shrimp Taura Syndrome Virus tsv in a c6 36 mosquito cell line
Journal of Fish Diseases, 2011Co-Authors: Narong Arunrut, Jurairat Phromjai, Warachin Gangnonngiw, Nipaporn Kanthong, S Sriurairatana, Wansika KiatpathomchaiAbstract:Journal of Fish Diseases Volume 34, Issue 10, pages 805–810, October 2011 Additional Information How to Cite Arunrut, N., Phromjai, J., Gangnonngiw, W., Kanthong, N., Sriurairatana, S. and Kiatpathomchai, W. (2011), In vitro cultivation of shrimp Taura Syndrome Virus (TSV) in a C6/36 mosquito cell line. Journal of Fish Diseases, 34: 805– 810. doi: 10.1111/j.1365-2761.2011.01286.x Author Information 1 Page 1 of 4 In vitro cultivation of shrimp Taura Syndrome Virus (TSV) in a C6/36 mosquito cell line Arunrut 20...
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In vitro cultivation of shrimp Taura Syndrome Virus (TSV) in a C6/36 mosquito cell line.
Journal of Fish Diseases, 2011Co-Authors: Narong Arunrut, Jurairat Phromjai, Warachin Gangnonngiw, Nipaporn Kanthong, S Sriurairatana, Wansika KiatpathomchaiAbstract:Journal of Fish Diseases Volume 34, Issue 10, pages 805–810, October 2011 Additional Information How to Cite Arunrut, N., Phromjai, J., Gangnonngiw, W., Kanthong, N., Sriurairatana, S. and Kiatpathomchai, W. (2011), In vitro cultivation of shrimp Taura Syndrome Virus (TSV) in a C6/36 mosquito cell line. Journal of Fish Diseases, 34: 805– 810. doi: 10.1111/j.1365-2761.2011.01286.x Author Information 1 Page 1 of 4 In vitro cultivation of shrimp Taura Syndrome Virus (TSV) in a C6/36 mosquito cell line Arunrut 20...
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Detection of shrimp Taura Syndrome Virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter.
Journal of Virological Methods, 2011Co-Authors: Assawapong Sappat, Wansadaj Jaroenram, Tanom Lomas, Adisorn Tuantranont, Teeranart Puthawibool, Wansika KiatpathomchaiAbstract:Abstract In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura Syndrome Virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg2P2O7). The device incorporated a heating block that maintained an optimal temperature of 63 °C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp Viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10 min for rapid RNA preparation with 30 min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1 h compared to 4–8 h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field.
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Turbidity detection of shrimp Taura Syndrome Virus by loop-mediated isothermal amplification reaction
2010 3rd International Nanoelectronics Conference (INEC), 2010Co-Authors: Assawapong Sappat, Wansika Kiatpathomchai, Wansadaj Jaroenram, Tanom Lomas, Adisorn TuantranontAbstract:Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a novel method of gene amplification that amplifies nucleic acid, which can be applied for disease diagnosis in shrimp aquaculture. During the LAMP reaction, the white precipitate of magnesium pyrophosphate (Mg2P2O7) is formed correlates with the amount of synthesized DNA. So, the turbidity can be measured. In this study, a portable turbidimeter has been developed for field to detection of Taura Syndrome Virus (TSV) that causes large economic losses to most major shrimp-producing countries including Thailand. The device could maintain an optimal temperature (63 °C) for 25 ¿l of LAMP sample solution contained in a 0.2 ml commercial PCR tube. We also applied the spectroscopic measurement technique to monitor a by-product of LAMP reaction, light emitting diode (LED) was used as a light source. Light dependent resistance (LDR) was used as detector. The results obtained from turbidity measurement revealed the same detection limit to those from agarose gel electrophoresis method.
Nastiti Wijayanti - One of the best experts on this subject based on the ideXlab platform.
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Molecular Detection of Taura Syndrome Virus Infections in White Shrimp (Litopenaeus vannamei) and Giant Prawn (Macrobrachium rosenbergii)
2014Co-Authors: Fariha Willisiani, Nur Rohmah, Irma Nur Rahmawati, Nastiti WijayantiAbstract:Abstract Giant prawn ( Macrobrachium rosenbergii ) and vaname shrimp ( Litopenaeus vannamei ) are types of shrimp that became excellent commodities in the fisheries sector. However, one of the obstacles in the vaname shrimp aquaculture is a disease caused by the infection of Taura Syndrome Virus (TSV). One of the consequence of raising the vaname shrimp in Indonesia is the possibility of spreading TSV infection in another shrimp species. TSV infection in giant prawns in Indonesia has not been reported. The aims of this study were: 1. To determine the resistance of giant prawns toward TSV infection and 2. To detect molecularly using RT-PCR technique the presence of Viruses TSV on vaname shrimp or giant prawns infected with 3 different doses (0.05 ml; 0.10 ml and 0.15 ml) of TSV inoculum using a pair of specific primers for TSV 9992F (5'-AAG CTT GCG TAG ACA GCC-3') and 9195R (5'-TCA AGA ATG GCT TCC TGG-3'). The research results showed that vaname shrimp mortality infected by 0.05 ml; 0.10 ml and 0.15 ml TSV inoculums were 14.28%, 42.86%, and 57.14%, respectively. Whereas the giant prawns mortality that were infected using the same dose of TSV inoculums were 0%, 8.33%, and 8.33%, respectively. Positive result was detected molecularly only from haemolymph of vaname shrimp infected using 0.15 ml of TSV inoculum. On the other hand, positive results were detected in pleopod and the gill of vaname shrimp infected using 0.05 ml; 0.10 ml or 0.15 ml of TSV inoculums. In giant prawns, infection using 3 different doses of TSV inoculums causes negative result molecularly. Based on all of the facts, it can be concluded that, giant prawns has the higher resistance to TSV infection than that of vaname shrimp.
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Deteksi Molekuler Infeksi Taura Syndrome Virus Pada Udang Vaname (Litopenaeus vannamei) dan Udang Galah (Macrobrachium rosenbergii) Molecular Detection of Taura Syndrome Virus Infections in White Shrimp (Litopenaeus vannamei) and Giant Prawn (Macrobr
2013Co-Authors: Fariha Wilisiani, Nur Rohmah, Irma Nur Rahmawati, Nastiti WijayantiAbstract:Giant prawn (Macrobrachium rosenbergii) and vaname shrimp (Litopenaeus vannamei) are types of shrimp that became excellent commodities in the fisheries sector. However, one of the obstacles in the vaname shrimp aquaculture is a disease caused by the infection of Taura Syndrome Virus (TSV). One of the consequence of raising the vaname shrimp in Indonesia is the possibility of spreading TSV infection in another shrimp species. TSV infection in giant prawns in Indonesia has not been reported. The aims of this study were: 1. To determine the resistance of giant prawns toward TSV infection and 2. To detect molecularly using RT-PCR technique the presence of Viruses TSV on vaname shrimp or giant prawns infected with 3 different doses (0.05 ml; 0.10 ml and 0.15 ml) of TSV inoculum using a pair of specific primers for TSV 9992F (5'-AAG CTT GCG TAG ACA GCC-3 ') and 9195R (5'-TCA AGA ATG GCT TCC TGG-3'). The research results showed that vaname shrimp mortality infected by 0.05 ml; 0.10 ml and 0.15 ml TSV inoculums were 14.28%, 42.86%, and 57.14%, respectively. Whereas the giant prawns mortality that were infected using the same dose of TSV inoculums were 0%, 8.33%, and 8.33%, respectively. Positive result was detected molecularly only from haemolymph of vaname shrimp infected using 0.15 ml of TSV inoculum. On the other hand, positive results were detected in pleopod and the gill of vaname shrimp infected using 0.05 ml; 0.10 ml or 0.15 ml of TSV inoculums. In giant prawns, infection using 3 different doses of TSV inoculums causes negative result molecularly. Based on all of the facts, it can be concluded that, giant prawns has the higher resistance to TSV infection than that of vaname shrimp.
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Deteksi Molekuler Infeksi Taura Syndrome Virus Pada Udang Vaname (Litopenaeus vannamel) dan Udang Galah (Macrobrachium rosenbergil) = Molecular Detection of Taura Syndrome Virus Infections in White Shrimp (Litopenaeus va
[Yogyakarta] : Universitas Gadjah Mada, 2013Co-Authors: Fariha Wilisiani, Nur Rohmah, Irma Nur Rahmawati, Nastiti WijayantiAbstract:Giant prawn (Macrobrachium rosenbergii) and vaname shrimp (Litopenaeus vannamei) are types of shrimp that became excellent commodities in the fisheries sector. However, one of the obstacles in the vaname shrimp aquaculture is a disease caused by the infection ofTaura Syndrome Virus (TSV). One ofthe consequence of raising the vaname shrimp in Indonesia is the possibility of spreading TSV infection in another shrimp species. TSV infection in giant prawns in Indonesia has not been reported. The aims of this study were: 1. To determine the resistance of giant prawns toward TSV infection and 2. To detect molecularly using RT-PCR technique the presence of Viruses TSV on vaname shrimp or giant prawns infected with 3 different doses (0.05 ml0.10 ml and 0.15 ml) ofTSV inoculum using a pair of specific primers for TSV 9992P (5\u27-AAG CTT GCG TAG ACA GCC-3 \u27) and 9195R (5\u27- TCAAGAATG GCT TCC TGG-3\u27). The research results showed that vaname shrimp mortality infected by 0.05 ml0.10 ml and 0.15 ml TSV inoculums were 14.28%,42.86%, and 57.14%, respectively. Whereas the giant prawns mortality that were infected using the same dose ofTSV inoculums were 0%, 8.33%, and 8.33%, respectively. Positive result was detected molecularly only from haemolymph of vaname shrimp infected using 0.15 ml ofTSV inoculum. On the other hand, positive results were detected in pleopod and the gill of vaname shrimp infected using 0.05 ml0.10 ml orO.15 ml ofTSV inoculums. In giant prawns, infection using 3 different doses of TSV inoculums causes negative result molecularly. Based on all of the facts, it can be concluded that, giant prawns has the higher resistance to TSV infection than that of vaname shrimp. Udang galah (Macrobrachium rosenbergii) dan udang vaname (Litopenaeus vannamei) merupakanjenis udang yang menjadi komoditas unggulan dalam sektor perikanan. Salah satu kendala dalam budidaya udang vaname adalah penyakit yang disebabkan oleh infeksi Taura Syndrome Virus (TSV). Dengan dibudidayakannya udang vaname di Indonesia, memungkinkan terjadinya penyebaran infeksi TSV pada udangjenis lain. Hingga saat ini, infeksi TSV pada udang galah belum pernah dilaporkan. Tujuan dari penelitian ini adalah: 1. Mempelajari ketahanan udang galah terhadap infeksi TSV dan 2. Mendeteksi secara molekuler Virus TSV dengan metode RT-PCR pada udang galah dan udang vaname yang diinfeksi dengan dosis inokulum TSV yang berbeda (0,05 ml0,1 ml dan 0,15 m!) menggunakan pasanganprimer spesifik untuk TSVyaitu 9992P (5\u27-AAG TAG ACA ACA GCC GCG CTT-3\u27) dan 9195R (5\u27TCA ATG AGA GCT TGG TCC-3\u27). Hasil penelitian menunjukkan bahwa mortalitas udang vaname akibat infeksi Virus TSV dengan dosis 0,05 ml0,1 ml dan 0, 15ml masing-masing adalah 14,28%,42,86% dan 57,14%. Sedangkan mortalitas pada udang galah yang diinfeksi Virus TSV dengan dosis yang sarna masing-masing adalah 0%,8,33% dan 8,33%. Hasil positif secara molekuler terdeteksi dari sampel hemolimfa udang vaname yang diinfeksi 0,15 ml inokulum TSV. Sebaliknya, hasil positif terdeteksi dari sampel pleopod dan insang udang vaname yang diinfeksi 0,05 ml, 0,1 ml dan 0,15 ml inokulum TSV. Pada udang galah, infeksi menggunakan inokulum TSV dengan 3 dosis yang berbeda secara molekuler hasilnya negatif. Berdasarkan semua fakta dapat disimpulkan bahwa udang galah memiliki ketahanan terhadap infeksi TSV yang lebih tinggi dibandingkan udang vaname
