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Vanvimon Saksmerprome - One of the best experts on this subject based on the ideXlab platform.
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Use of microalgae Chlamydomonas reinhardtii for production of double-stranded RNA against Shrimp Virus
Elsevier, 2016Co-Authors: Parinyachat Somchai, Sarocha Jitrakorn, Siripong Thitamadee, Metha Meetam, Vanvimon SaksmerpromeAbstract:RNA interference has been proposed to be a promising tool for combating Shrimp Viruses. Antiviral double-stranded (ds)RNA has been mostly produced in Escherichia coli-expression system because of its high efficiency and inexpensive operations. However, overusing the bacteria may raise concerns regarding public health and environmental contamination, and seeking for a new dsRNA production platform would be alternative for future molecular farming. In this study, we exploited the green microalgae Chlamydomonas reinhardtii to produce dsRNA targeting the lethal Shrimp yellow head Virus (YHV). The expression plasmid pSL18 for C. reinhardtii was constructed to contain YHV-specific hairpin RNA expression cassette, and the successful assembly of pSL18-YHV was confirmed by PCR and enzymatic digestions. Glass bead method was employed for transformation of C. reinhardtii nuclear genome with pSL18-YHV. Microalgal expression of dsRNA-YHV, approximately 45 ng from 100-mL culture, was detected by qRT-PCR. Oral feeding experiment on postlarval Shrimp revealed that the formulated feed with C. reinhardtii expressing dsRNA-YHV, at the ratio of 1 × 108 transformants per gram feed, improved 22% survival rate after YHV challenge. The present study suggests that C. reinhardtii can be bioengineered to produce viral-specific dsRNA for Shrimp viral disease control, and the developed qRT-PCR could detect microalgal dsRNA with detection limit of subpicogram. Keywords: Chlamydomonas reinhardtii, Double-stranded RNA, Shrimp, Yellow head Virus, Nuclear transformation, qRT-PC
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biology genome organization and evolution of parvoViruses in marine Shrimp
Advances in Virus Research, 2014Co-Authors: Arun K Dhar, Vanvimon Saksmerprome, Refugio Roblessikisaka, Dilip K LakshmanAbstract:Abstract As Shrimp aquaculture has evolved from a subsistent farming activity to an economically important global industry, viral diseases have also become a serious threat to the sustainable growth and productivity of this industry. ParvoViruses represent an economically important group of Viruses that has greatly affected Shrimp aquaculture. In the early 1980s, an outbreak of a Shrimp parvoVirus, infectious hypodermal and hematopoietic necrosis Virus (IHHNV), led to the collapse of penaeid Shrimp farming in theAmericas. Since then, considerable progress has been made in characterizing the parvoViruses of Shrimp and developing diagnostic methods aimed to preventing the spread of diseases caused by these Viruses. To date, four parvoViruses are known that infect Shrimp; these include IHHNV, hepatopancreatic parvoVirus (HPV), spawner-isolated mortality Virus (SMV), and lymphoid organ parvo-like Virus. Due to the economic repercussions that IHHNV and HPV outbreaks have caused to Shrimp farming over the years, studies have been focused mostly on these two pathogens, while information on SMV and LPV remains limited. IHHNV was the first Shrimp Virus to be sequenced and the first for which highly sensitive diagnostic methods were developed. IHHNV-resistant lines of Shrimp were also developed to mitigate the losses caused by this Virus. While the losses due to IHHNV have been largely contained in recent years, reports of HPV-induced mortalities in larval stages in hatchery and losses due to reduced growth have increased. This review presents a comprehensive account of the history and current knowledge on the biology, diagnostics methods, genomic features, mechanisms of evolution, and management strategies of Shrimp parvoViruses. We also highlighted areas where research efforts should be focused in order to gain further insight on the mechanisms of parvoviral pathogenicity in Shrimp that will help to prevent future losses caused by these Viruses.
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additional random single to multiple genome fragments of penaeus stylirostris densoVirus in the giant tiger Shrimp genome have implications for viral disease diagnosis
Virus Research, 2011Co-Authors: Vanvimon Saksmerprome, Timothy W. Flegel, Sarocha Jitrakorn, Kanokporn Chayaburakul, Seansook Laiphrom, Khanittha BoonsuaAbstract:Scattered reports of viral inserts in Shrimp and insect genomes led to the hypothesis that random, autonomous insertion of such sequences occurs in these organisms and leads to specific, heritable immunity. To test the prediction regarding random insertion of viral sequences into the Shrimp genome, we examined the giant tiger Shrimp for random genomic insertions of Penaeus stylirostris densoVirus (also called IHHNV). By PCR analysis using a set of 7 overlapping primer pairs to cover the whole IHHNV genome (4 kb), PCR failure with some pairs indicated sequence gaps that revealed a random pattern of putative viral inserts in the genomes of individual Shrimp. Targeting a putative insert from one arbitrarily selected specimen, we used genome walking to reveal a viral insert linked to a host microsattelite-like fragment. This differed from 2 previously reported inserted fragments of IHHNV in P. monodon. In one specimen, 2 slightly different inserts were revealed, probably on paired chromosomes. By design and use of chimeric Shrimp/Virus primer pairs we proved that similar insertions occurred in several Shrimp specimens, including those infected with IHHNV but showing no signs of disease. For the infected specimens, the inserts gave false positive PCR test results using 309F/R primers and a new IQ2000 test protocol currently recommended for detection of infectious IHHNV. This is the first experimental support for the hypothesis-based prediction that a random number and length of sequence fragments from a single Virus genome may occur in the Shrimp genome. Since some inserts can give false positive results for infectious IHHNV with the recommended methods above, they may have a negative effect on international seafood trade. In addition, discard of domesticated Shrimp breeding stocks based on such false positive results might have negative consequences, if such inserts are related to Shrimp viral disease tolerance, as also hypothesized.
Kayhooi Khoo - One of the best experts on this subject based on the ideXlab platform.
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to complete its replication cycle a Shrimp Virus changes the population of long chain fatty acids during infection via the pi3k akt mtor hif1α pathway
Developmental and Comparative Immunology, 2015Co-Authors: Yun-chieh Hsieh, Suh-yuen Liang, Shu-yu Lin, Yi Min Chen, Yu Han Chang, Chang Yi Lin, Sheng Hsiung Chang, Yi Jan Wang, Kayhooi KhooAbstract:White spot syndrome Virus (WSSV), the causative agent of white spot disease (WSD), is a serious and aggressive Shrimp viral pathogen with a worldwide distribution. At the genome replication stage (12 hpi), WSSV induces a metabolic rerouting known as the invertebrate Warburg effect, which boosts the availability of energy and biosynthetic building blocks in the host cell. Here we show that unlike the lipogenesis that is seen in cancer cells that are undergoing the Warburg effect, at 12 hpi, all of the long chain fatty acids (LCFAs) were significantly decreased in the stomach cells of WSSV-infected Shrimp. By means of this non-selective WSSV-induced lipolysis, the LCFAs were apparently diverted into β-oxidation and used to replenish the TCA cycle. Conversely, at 24 hpi, when the Warburg effect had ceased, most of the LCFAs were significantly up-regulated and the composition was also significantly altered. In crayfish these changes were in a direction that appeared to favor the formation of WSSV virion particles. We also found that, at 24 hpi, but not at 12 hpi, the PI3K-Akt-mTOR-HIF1α pathway induced the expression of fatty acid synthase (FAS), an enzyme which catalyzes the conversion of acetyl-CoA into LCFAs. WSSV virion formation was impaired in the presence of the FAS inhibitor C75, although viral gene and viral DNA levels were unaffected. WSSV therefore appears to use the PI3K-Akt-mTOR pathway to induce lipid biosynthesis at 24 hpi in order to support viral morphogenesis.
Timothy W. Flegel - One of the best experts on this subject based on the ideXlab platform.
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additional random single to multiple genome fragments of penaeus stylirostris densoVirus in the giant tiger Shrimp genome have implications for viral disease diagnosis
Virus Research, 2011Co-Authors: Vanvimon Saksmerprome, Timothy W. Flegel, Sarocha Jitrakorn, Kanokporn Chayaburakul, Seansook Laiphrom, Khanittha BoonsuaAbstract:Scattered reports of viral inserts in Shrimp and insect genomes led to the hypothesis that random, autonomous insertion of such sequences occurs in these organisms and leads to specific, heritable immunity. To test the prediction regarding random insertion of viral sequences into the Shrimp genome, we examined the giant tiger Shrimp for random genomic insertions of Penaeus stylirostris densoVirus (also called IHHNV). By PCR analysis using a set of 7 overlapping primer pairs to cover the whole IHHNV genome (4 kb), PCR failure with some pairs indicated sequence gaps that revealed a random pattern of putative viral inserts in the genomes of individual Shrimp. Targeting a putative insert from one arbitrarily selected specimen, we used genome walking to reveal a viral insert linked to a host microsattelite-like fragment. This differed from 2 previously reported inserted fragments of IHHNV in P. monodon. In one specimen, 2 slightly different inserts were revealed, probably on paired chromosomes. By design and use of chimeric Shrimp/Virus primer pairs we proved that similar insertions occurred in several Shrimp specimens, including those infected with IHHNV but showing no signs of disease. For the infected specimens, the inserts gave false positive PCR test results using 309F/R primers and a new IQ2000 test protocol currently recommended for detection of infectious IHHNV. This is the first experimental support for the hypothesis-based prediction that a random number and length of sequence fragments from a single Virus genome may occur in the Shrimp genome. Since some inserts can give false positive results for infectious IHHNV with the recommended methods above, they may have a negative effect on international seafood trade. In addition, discard of domesticated Shrimp breeding stocks based on such false positive results might have negative consequences, if such inserts are related to Shrimp viral disease tolerance, as also hypothesized.
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persistent expression of Shrimp Virus antigens in two insect cell lines challenged with two Shrimp Viruses
Fish Pathology, 2009Co-Authors: Anuwat Sriton, Warachin Gangnonngiw, Nipaporn Kanthong, Timothy W. Flegel, Siriporn Sriurairatana, Sukathida UbolAbstract:Experiments with crustacean Viruses are hampered by lack of susceptible continuous cell lines. To overcome this problem, immortal mosquito and lepidopteran cell lines were both separately challenged with a Shrimp DNA Virus (white spot syndrome Virus: WSSV, = PRDV) and RNA Virus (yellow head Virus: YHV) followed by serial, split-passage with immunohistochemical monitoring by confocal laser microscopy using labeled monoclonal antibodies to Shrimp viral antigens. Stable, immortal cultures with 100% of the cells expressing Shrimp-Virus antigens were obtained, although the infected cells appeared grossly normal by phase contrast microscopy. Nor did they show any ultrastructural modifications characteristic of the challenge Viruses. These persistently-expressing insect cell cultures were stable and could be continuously passaged, stored and revived as required. Since disparate Viruses and insect cells were used, this appears to be a generic process that may be applicable to other Shrimp Viruses as well.
Yun-chieh Hsieh - One of the best experts on this subject based on the ideXlab platform.
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to complete its replication cycle a Shrimp Virus changes the population of long chain fatty acids during infection via the pi3k akt mtor hif1α pathway
Developmental and Comparative Immunology, 2015Co-Authors: Yun-chieh Hsieh, Suh-yuen Liang, Shu-yu Lin, Yi Min Chen, Yu Han Chang, Chang Yi Lin, Sheng Hsiung Chang, Yi Jan Wang, Kayhooi KhooAbstract:White spot syndrome Virus (WSSV), the causative agent of white spot disease (WSD), is a serious and aggressive Shrimp viral pathogen with a worldwide distribution. At the genome replication stage (12 hpi), WSSV induces a metabolic rerouting known as the invertebrate Warburg effect, which boosts the availability of energy and biosynthetic building blocks in the host cell. Here we show that unlike the lipogenesis that is seen in cancer cells that are undergoing the Warburg effect, at 12 hpi, all of the long chain fatty acids (LCFAs) were significantly decreased in the stomach cells of WSSV-infected Shrimp. By means of this non-selective WSSV-induced lipolysis, the LCFAs were apparently diverted into β-oxidation and used to replenish the TCA cycle. Conversely, at 24 hpi, when the Warburg effect had ceased, most of the LCFAs were significantly up-regulated and the composition was also significantly altered. In crayfish these changes were in a direction that appeared to favor the formation of WSSV virion particles. We also found that, at 24 hpi, but not at 12 hpi, the PI3K-Akt-mTOR-HIF1α pathway induced the expression of fatty acid synthase (FAS), an enzyme which catalyzes the conversion of acetyl-CoA into LCFAs. WSSV virion formation was impaired in the presence of the FAS inhibitor C75, although viral gene and viral DNA levels were unaffected. WSSV therefore appears to use the PI3K-Akt-mTOR pathway to induce lipid biosynthesis at 24 hpi in order to support viral morphogenesis.
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An invertebrate Warburg effect: a Shrimp Virus achieves successful replication by altering the host metabolome via the PI3K-Akt-mTOR pathway.
PLoS pathogens, 2014Co-Authors: Yun-tzu Huang, I-tung Chen, Der-yen Lee, Yun-chieh Hsieh, Suh-yuen Liang, Shu-yu Lin, Shiao Wei HuangAbstract:In this study, we used a systems biology approach to investigate changes in the proteome and metabolome of Shrimp hemocytes infected by the invertebrate Virus WSSV (white spot syndrome Virus) at the viral genome replication stage (12 hpi) and the late stage (24 hpi). At 12 hpi, but not at 24 hpi, there was significant up-regulation of the markers of several metabolic pathways associated with the vertebrate Warburg effect (or aerobic glycolysis), including glycolysis, the pentose phosphate pathway, nucleotide biosynthesis, glutaminolysis and amino acid biosynthesis. We show that the PI3K-Akt-mTOR pathway was of central importance in triggering this WSSV-induced Warburg effect. Although dsRNA silencing of the mTORC1 activator Rheb had only a relatively minor impact on WSSV replication, in vivo chemical inhibition of Akt, mTORC1 and mTORC2 suppressed the WSSV-induced Warburg effect and reduced both WSSV gene expression and viral genome replication. When the Warburg effect was suppressed by pretreatment with the mTOR inhibitor Torin 1, even the subsequent up-regulation of the TCA cycle was insufficient to satisfy the Virus's requirements for energy and macromolecular precursors. The WSSV-induced Warburg effect therefore appears to be essential for successful viral replication.
Shu-yu Lin - One of the best experts on this subject based on the ideXlab platform.
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to complete its replication cycle a Shrimp Virus changes the population of long chain fatty acids during infection via the pi3k akt mtor hif1α pathway
Developmental and Comparative Immunology, 2015Co-Authors: Yun-chieh Hsieh, Suh-yuen Liang, Shu-yu Lin, Yi Min Chen, Yu Han Chang, Chang Yi Lin, Sheng Hsiung Chang, Yi Jan Wang, Kayhooi KhooAbstract:White spot syndrome Virus (WSSV), the causative agent of white spot disease (WSD), is a serious and aggressive Shrimp viral pathogen with a worldwide distribution. At the genome replication stage (12 hpi), WSSV induces a metabolic rerouting known as the invertebrate Warburg effect, which boosts the availability of energy and biosynthetic building blocks in the host cell. Here we show that unlike the lipogenesis that is seen in cancer cells that are undergoing the Warburg effect, at 12 hpi, all of the long chain fatty acids (LCFAs) were significantly decreased in the stomach cells of WSSV-infected Shrimp. By means of this non-selective WSSV-induced lipolysis, the LCFAs were apparently diverted into β-oxidation and used to replenish the TCA cycle. Conversely, at 24 hpi, when the Warburg effect had ceased, most of the LCFAs were significantly up-regulated and the composition was also significantly altered. In crayfish these changes were in a direction that appeared to favor the formation of WSSV virion particles. We also found that, at 24 hpi, but not at 12 hpi, the PI3K-Akt-mTOR-HIF1α pathway induced the expression of fatty acid synthase (FAS), an enzyme which catalyzes the conversion of acetyl-CoA into LCFAs. WSSV virion formation was impaired in the presence of the FAS inhibitor C75, although viral gene and viral DNA levels were unaffected. WSSV therefore appears to use the PI3K-Akt-mTOR pathway to induce lipid biosynthesis at 24 hpi in order to support viral morphogenesis.
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An invertebrate Warburg effect: a Shrimp Virus achieves successful replication by altering the host metabolome via the PI3K-Akt-mTOR pathway.
PLoS pathogens, 2014Co-Authors: Yun-tzu Huang, I-tung Chen, Der-yen Lee, Yun-chieh Hsieh, Suh-yuen Liang, Shu-yu Lin, Shiao Wei HuangAbstract:In this study, we used a systems biology approach to investigate changes in the proteome and metabolome of Shrimp hemocytes infected by the invertebrate Virus WSSV (white spot syndrome Virus) at the viral genome replication stage (12 hpi) and the late stage (24 hpi). At 12 hpi, but not at 24 hpi, there was significant up-regulation of the markers of several metabolic pathways associated with the vertebrate Warburg effect (or aerobic glycolysis), including glycolysis, the pentose phosphate pathway, nucleotide biosynthesis, glutaminolysis and amino acid biosynthesis. We show that the PI3K-Akt-mTOR pathway was of central importance in triggering this WSSV-induced Warburg effect. Although dsRNA silencing of the mTORC1 activator Rheb had only a relatively minor impact on WSSV replication, in vivo chemical inhibition of Akt, mTORC1 and mTORC2 suppressed the WSSV-induced Warburg effect and reduced both WSSV gene expression and viral genome replication. When the Warburg effect was suppressed by pretreatment with the mTOR inhibitor Torin 1, even the subsequent up-regulation of the TCA cycle was insufficient to satisfy the Virus's requirements for energy and macromolecular precursors. The WSSV-induced Warburg effect therefore appears to be essential for successful viral replication.