Jiraporn Srisala

National Center for Genetic Engineering and Biotechnology (BIOTEC), Bang Kadi, Pathum Thani, Thailand

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Publications (14)27.68 Total impact

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    ABSTRACT: Several shrimp diseases are new or newly emerged in Asia, including acute hepatopancreatic necrosis disease (AHPND), hepatopancreatic microsporidiosis (HPM), hepatopancreatic haplosporidiosis (HPH), aggregated transformed microvilli (ATM) and covert mortality disease (CMD). In addition to these, white spot disease (WSD), yellow head disease (YHD) and infectious myonecrosis (IMN) continue as the most serious viral threats to shrimp farmers in the region. Other diseases such as monodon slow growth syndrome (MSGS), white tail disease (WTD) and abdominal segment deformity disease (ASDD) are of less concern. In contrast, Taura syndrome virus (TSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV) have become innocuous due to the widespread use of highly tolerant specific pathogen free (SPF) stocks of Penaeus (Litopenaeus) vannamei that dominate production. Similarly, diseases caused by monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV) appear not to affect P. vannamei. Spread of diseases has been promoted by the use of live or fresh broodstock feeds such as polychaetes and clams. Also, shortages in the supply of imported SPF broodstock led some entrepreneurs to employ post larvae (PL) of imported SPF stocks to produce 2nd generation broodstock in open shrimp ponds where they became contaminated and were then used to produce PL for stocking ponds. These practices left the whole shrimp industry vulnerable to rapid spread of the new and newly emerging diseases and resulted in the current crisis in Asian shrimp culture. The situation has been exacerbated since 2009 by an almost exclusive focus on AHPND, which is only partially responsible for what has been widely called early mortality syndrome (EMS). The purpose of this review is to summarize progress of research on AHPND bacteria and also to encourage a wider focus on additional pathogens that are causing farm losses. The significance of these diseases and their implications for the future of shrimp aquaculture are discussed. Statement of relevance: This review summarizes recent information about new and newly emerging diseases of cultured shrimp in Asia and discusses the biosecurity lapses that led to the current shrimp production crisis. All industry stakeholders must be aware of this situation and of the need for regional and global collaborative efforts to stem this crisis and prevent future development of another.
    Full-text · Article · Oct 2015 · Aquaculture
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    ABSTRACT: Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7) and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV) could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral administration.
    Preview · Article · Jun 2015 · PLoS ONE
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    ABSTRACT: In our research efforts to reduce the impact of white spot syndrome virus (WSSV) disease outbreaks in shrimp aquaculture, we studied the effect of β-glucan administration to activate the prophenoloxidase (proPO) enzymatic cascade prior to WSSV challenge. Injection of a single dose of β-glucan (5 μg/g) prior to WSSV challenge resulted in activation of the proPO system and reduced shrimp mortality (25–50%) when compared to controls (100%). By contrast, no significant reduction was observed using yellow head virus (YHV) in a similar protocol. We subsequently hypothesized that administration of a second dose of β-glucan after WSSV challenge might reduce shrimp mortality further. Surprisingly, the opposite occurred, and mortality of the WSSV-infected shrimp increased to 100% after the second β-glucan dose. Both immunofluorescence and RT-PCR assays revealed low WSSV levels in hemocytes of shrimp collected after the second dose of β-glucan administration, suggesting that the cause of increased mortality was unlikely to be increased WSSV replication. We found from measured phenoloxidase acitivity (PO) and H2O2 production that the higher mortality may have resulted from a combination of WSSV infection plus over-production of reactive oxygen species (ROS) stimulated by two doses of β-glucan. Thus, caution may be prudent in continuous or prolonged activation of the shrimp immune system by β-glucan administration lest it exacerbate shrimp mortality in the event of WSSV infection.
    Full-text · Article · Oct 2014 · Fish & Shellfish Immunology
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    ABSTRACT: The Thai Department of Fisheries (DOF), 2013 estimated that outbreaks of acute early mortality (often called early mortality syndrome or EMS) in cultivated shrimp were responsible for a 33% drop in shrimp production during the first quarter of 2013. Similar early mortality in Vietnam was ascribed to specific isolates of Vibrio parahaemolyticus that caused acute hepatopancreatic necrosis disease (AHPND) but the status of EMS/AHPND in Thailand was unclear. Here we describe the isolation and characterization of bacteria isolated from the hepatopancreas (HP) of shrimp collected from an early mortality outbreak farm in Thailand. Four independent bacterial isolates were identified as V. parahaemolyticus by BLAST analysis and by gene-specific marker detection of a lecithin dependent hemolysin (LDH) considered to be specific for the species. Immersion challenges with 3 of these and a reference isolate, obtained from China in 2010, using a previously published laboratory infection model caused very high mortality accompanied by characteristic AHPND histopathology in the shrimp HP. Tests with one of these isolates (5HP) revealed that rate of mortality was dose dependent. Using the same challenge protocol, the 4th isolate (2HP) also caused high mortality, but it was not accompanied by AHPND histopathology. Instead, it caused a different histopathology of the HP including collapsed epithelia and unique vacuolization of embryonic cells (E-cells). These results revealed the possibility of diversity in isolates of V. parahaemolyticus that may cause early mortality in shrimp cultivation ponds. Genomic and episomic DNA of these isolates and isolates of V. parahaemolyticus that cause no disease need to be compared to better understand the molecular basis of bacterial virulence in AHPND.
    No preview · Article · May 2014 · Aquaculture
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    ABSTRACT: Self-assembled monolayers (SAM) of 3-mercaptopropionic acid (MPA) were applied on tin-doped indium oxide (ITO) surfaces and used as a working electrode for sensing DNA hybridization. The concentration of probe single stranded DNA (ssDNA), complemented with target DNA, was optimized for the highest yield immobilization on MPA/ITO platform. The ssDNA/MPA/ITO was allowed to hybridize to target DNA prepared from PCR amplification that first tested by the synthesized complementary sequences. Both cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were employed for investigating probe ssDNA immobilization and target DNA hybridization. For fast and low concentration detecting purposes, methylene blue (MB) coupled with differential pulse voltammetry (DPV) was used for detecting the target DNA hybridization events.
    Full-text · Article · Sep 2013
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    ABSTRACT: The microsporidian Enterocytozoon hepatopenaei was first described from Thailand in 2009 in farmed, indigenous giant tiger shrimp Penaeus (Penaeus) monodon. The natural reservoir for the parasite is still unknown. More recently, a microsporidian closely resembling it in morphology and tissue preference was found in Thai-farmed, exotic, whiteleg shrimp Penaeus (Litopenaeus) vannamei exhibiting white feces syndrome (WFS). Our objective was to compare the newly found pathogen with E. hepatopenaei and to determine its causal relationship with WFS. Generic primers used to amplify a fragment of the small subunit ribosomal RNA (ssu rRNA) gene for cloning and sequencing revealed that the new parasite from WFS ponds had 99% sequence identity to that of E. hepatopenaei, suggesting it was conspecific. Normal histological analysis using tissue sections stained with hematoxylin and eosin (H&E) revealed that relatively few tubule epithelial cells exhibited spores, suggesting that the infections were light. However, the H&E results were deceptive since nested PCR and in situ hybridization analysis based on the cloned ssu rRNA gene fragment revealed very heavy infections in tubule epithelial cells in the central region of the hepatopancreas in the absence of spores. Despite these results, high prevalence of E. hepatopenaei in shrimp from ponds not exhibiting WFS and a pond that had recovered from WFS indicated no direct causal association between these infections and WFS. This was supported by laboratory oral challenge trials that revealed direct horizontal transmission to uninfected shrimp but no signs of WFS. The microsporidian newly found in P. vannamei is conspecific with previously described E. hepatopenaei and it is not causally associated with WFS. However, the deceptive severity of infections (much greater than previously reported in P. monodon) would undoubtedly have a negative effect on whiteleg shrimp growth and production efficiency and this could be exacerbated by the possibility of horizontal transmission revealed by laboratory challenge tests. Thus, it is recommended that the PCR and in situ hybridization methods developed herein be used to identify the natural reservoir species so they can be eliminated from the shrimp rearing system.
    Full-text · Article · Jul 2013 · BMC Veterinary Research
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    ABSTRACT: Disease outbreaks caused by viral pathogens constitute a major limitation to development of the shrimp aquaculture industry. Many research have been conducted to better understand how host shrimp respond to viral infections with the aim of using the gained knowledge to develop better strategies for disease management and control. One approach has been to study the interactions between host and viral proteins, and particularly host virus-binding proteins that might play an important role in the viral infection process. Within the past five years, increasing numbers of virus-binding proteins (VBPs) have been reported in shrimp. Characterization of these molecules has emphasized on their potential therapeutic applications by demonstrating their activities in inhibition of viral replication via in vivo neutralization assay. However, signaling to induce innate antiviral immune responses as a consequence of binding between viral proteins and VBPs remain to be fully elucidated.
    No preview · Article · Feb 2013 · Fish & Shellfish Immunology
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    ABSTRACT: Enterocytozoon hepatopenaei is an emerging microsporidian parasite that has been linked to recent losses caused by white faeces syndrome (WFS) in cultivated giant or black tiger shrimp Penaeus (Penaeus) monodon and whiteleg shrimp Penaeus (Litopenaeus) vannamei in Asia. To more accurately assess its impact on shrimp production and to determine reservoir carriers for control measures, our objective was to establish a loop-mediated isothermal amplification (LAMP) assay combined with colorimetric nanogold (AuNP) for rapid, sensitive and inexpensive detection of this parasite. A set of six specific primers was designed to successfully detect the SSU rRNA gene of E. hepatopenaei by a LAMP reaction of 45 min at 65°C combined with visual detection of the amplification product via hybridization at 65°C for 5 min with a ssDNA-labelled nanogold probe, followed by salt-induced AuNP aggregation (total assay time, approximately 50 min). This method gave similar results to LAMP followed by electrophoresis or spectrophotometric detection, and it was more sensitive (0·02 fg total DNA) than a conventional nested PCR (0·2 fg total DNA). The new method gave negative results with shrimp DNA templates extracted from diseased shrimp containing other pathogens, indicating that the LAMP-AuNP assay was specific for E. hepatopenaei. Without sacrificing sensitivity or specificity, the new LAMP-AuNP assay significantly reduced the time, ease and cost for molecular detection of E. hepatopenaei in shrimp. The new method employs simple, inexpensive equipment and involves simple steps making it applicable for small field laboratories. Wider application of the method to screen broodstock before use in a hatchery, to screen postlarvae before stocking shrimp ponds, to test for natural carriers and to monitor shrimp in rearing ponds would help to assess and reduce the negative impact of this parasite in shrimp farming.
    Full-text · Article · Feb 2013 · Journal of Applied Microbiology
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    ABSTRACT: Disease outbreaks caused by viral pathogens constitute a major limitation to development of the shrimp aquaculture industry. Many research have been conducted to better understand how host shrimp respond to viral infections with the aim of using the gained knowledge to develop better strategies for disease management and control. One approach has been to study the interactions between host and viral proteins, and particularly host virus-binding proteins that might play an important role in the viral infection process. Within the past five years, increasing numbers of virus-binding proteins (VBPs) have been reported in shrimp. Characterization of these molecules has emphasized on their potential therapeutic applications by demonstrating their activities in inhibition of viral replication via in vivo neutralization assay. However, signaling to induce innate antiviral immune responses as a consequence of binding between viral proteins and VBPs remain to be fully elucidated.
    No preview · Article · Sep 2012 · Fish & Shellfish Immunology
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    Full-text · Article · Sep 2012
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    ABSTRACT: White spot syndrome virus is currently the leading cause of production losses in the shrimp industry. Penaeus monodon Rab7 protein has been recognized as a viral-binding protein with an efficient protective effect against white spot syndrome infection. Plant-derived recombinant PmRab7 might serve as an alternative source for in-feed vaccination, considering the remarkable abilities of plant expression systems. PmRab7 was introduced into the Arabidopsis thaliana T87 genome. Arabidopsis-derived recombinant PmRab7 showed high binding activity against white spot syndrome virus and a viral envelope, VP28. The growth profile of Arabidopsis suspension culture expressing PmRab7 (ECR21# 35) resembled that of its counterpart. PmRab7 expression in ECR21# 35 reached its maximum level at 5 mg g(-1) dry weight in 12 days, which was higher than those previously reported in Escherichia coli and in Pichia. Co-injection of white spot syndrome virus and Arabidopsis crude extract containing PmRab7 in Litopenaeus vannamei showed an 87% increase in shrimp survival rate at 5 day after injection. In this study, we propose an alternative PmRab7 source with higher production yield, and cheaper culture media costs, that might serve the industry's need for an in-feed supplement against white spot syndrome infection.
    Full-text · Article · May 2012 · Journal of Biotechnology
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    ABSTRACT: The aim of this study was to examine whether shrimp yellow head virus (YHV) from processed shrimp tissue infected at the pre-patent disease level could be transmitted to naïve shrimp in a laboratory setting. In a preliminary test, 120 YHV-free shrimp were injected intramuscularly with a virulent YHV stock to yield 5 × 105 (pre-patent disease level) and 2500 (carrier level) viral copies/g shrimp tissue (60 shrimp each dose). These are possible infection levels for grossly normal shrimp from normal harvest ponds (i.e., not shrimp from disease outbreak ponds). These yielded 1-step and 2-step positive (nested) RT-PCR reactions, respectively, in pleopods at 6 h post-injection of the viral stock. After being subjected to standard industrial processing conditions, only fresh frozen whole or peeled shrimp injected with pre-patent dose gave positive RT-PCR test results for YHV. None of the naïve shrimp exposed to the chopped processed products for 24 h and then reared on a standard diet for 14 days showed any significant mortality or gave any positive test results for YHV using nested RT-PCR assays. Based on these preliminary test results, a large-scale test was carried out using only the high, pre-patent injection dose with 1000 fresh frozen whole shrimp. The negative control consisted of 1000 buffer-injected shrimp. A random sample of 60 shrimp from the YHV-injected group after processing, revealed 57 positive for YHV by 1-step RT-PCR assay. Of the 3 remaining, 2 were positive and 1 negative by nested RT-PCR assay. All 60 shrimp from the buffer-injected, control group were negative for YHV by nested RT-PCR assay. Exposure of these whole shrimp to naïve shrimp resulted in no significant mortality and no positive RT-PCR test results for YHV by nested RT-PCR assay in the exposed naïve shrimp. Our results showed that grossly normal whole shrimp processed by chilling and freezing would present negligible YHV disease transmission risks, even if they were 1-step RT-PCR positive for YHV. Thus, shrimp subjected to any additional processing steps such as peeling or cooking should present even lower transmission risks.
    No preview · Article · Feb 2010 · Aquaculture
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    ABSTRACT: White spot syndrome virus (WSSV) PCR-detection methods that used electrophoresis or lateral flow chromatographic strips (LFCS) were to compare and visualize PCR amplicons. Real-time PCR was used to prepare a stock template solution containing 2.85 x 10 (6) copies WSSV/microl from WSSV-infected shrimp. Serial stock dilutions were used as templates for PCR amplification of a WSSV-specific DNA fragment that was detected either in ethidium bromide stained agarose electrophoresis gels or on a chromatographic strip where it interacted with antibody to markers labeled on hybridization complex. PCR amplification employed both 1-step PCR and semi-nested PCR methods. By using 1-step PCR, the LFCS method (100 copies) gave 10 times higher sensitivity than gel electrophoresis (10(3) copies). A combination of a semi-nested PCR with LFCS gave a comparable sensitivity to those with commercial kits for nested PCR (20 copies). In addition, LFCS confirmed amplicon identity, avoided handling of carcinogenic ethidium bromide and could be completed in approximately 20-30 min post-PCR compared with 1h for gel electrophoresis. The costs for the two methods were comparable. In conclusion, semi-nested PCR followed by LFCS is a safe and rapid alternative method for detection of WSSV that provides sensitivity similar to that obtained by standard nested PCR methods.
    No preview · Article · Sep 2008 · Journal of Virological Methods
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    ABSTRACT: In Thailand, several PCR-based methods are used by private and public service laboratories for the detection of white spot syndrome virus (WSSV) infection in penaeid shrimp post larvae (PL) before they are stocked in rearing ponds. Conflicting test results for similar samples sent to two service laboratories has decreased confidence in PCR testing. Thus, we compared the sensitivity of several PCR methods commonly employed in Thailand using Taqman real-time PCR as the gold standard with a purified WSSV template stock. Using this stock for assays, we found no significant inhibitory effect by WSSV-free host shrimp DNA over the range 0 to 300 ng per reaction or by added DNA from WSSV-infected shrimp. Real-time PCR could detect WSSV with certainty at dilutions of approximately 5 copies per reaction while 1000 copies were needed for a common one-step PCR method and 50 for a common single-tube nested PCR (1N-PCR) method. Of 2 two-tube nested PCR protocols tested, one required 100 and the other 1000 copies. In addition to these sensitivity tests, a triple-blind ring test was carried out employing sets of 10 WSSV-infected DNA extracts sent to 12 commercial and public laboratories in Thailand, without specifying the PCR method to be used. Returned results included no false positives and two false negatives, the latter both from light infection vials. This translated into a test sensitivity of 97.3% and a specificity of 100%. Overall, the results confirmed the validity of PCR-based methods in Thailand for detection of WSSV in shrimp DNA extracts.
    No preview · Article · May 2006 · Aquaculture

Publication Stats

130 Citations
27.68 Total Impact Points

Institutions

  • 2012-2015
    • National Center for Genetic Engineering and Biotechnology (BIOTEC)
      Bang Kadi, Pathum Thani, Thailand
  • 2006-2015
    • Mahidol University
      • • Faculty of Science
      • • Department of Biotechnology
      • • Center for Shrimp Molecular Biology and Biotechnology (Centex Shrimp)
      Siayuthia, Bangkok, Thailand