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Role of cyclophilin A as an immunostimulator in responses to Vibrio parahaemolyticus AHPND infection in the black tiger shrimp Penaeus monodon
(2025-11-01)
Towittayapanchai, Pornsawan
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Ponprateep, Sirikwan
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Supungul, Premruethai
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Tang, Sureerat
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Jaree, Phattarunda
Cyclophilin A (CypA) is a highly conserved peptidyl-prolyl isomerase that plays a crucial role in protein folding, immune regulation, and cellular stress responses. In shrimp, CypA has been implicated in innate immunity, particularly in response to viral and bacterial infections. This study investigates the role of the Penaeus monodon cyclophilin A (PmCypA) in shrimp infected with V. parahaemolyticus AHPND (VP<inf>AHPND</inf>). During VP<inf>AHPND</inf> infection, the expression levels of PmCypA gene and PmCypA protein were significantly increased at 24 h post-infection (h.p.i.) in shrimp hemocyte. To investigate the role of PmCypA in shrimp immune response, shrimp were administrated with the recombinant PmCypA protein (rPmCypA) and submerged in VP<inf>AHPND</inf> suspension. The results showed that the rPmCypA could rapidly stimulate the expression of immune-related genes involved in several immune pathways, such as immune recognition receptors, clotting system, phagocytosis and homeostasis. Moreover, the injected rPmCypA led to an increase in the proportion of hyaline cells and the greater quantity of granulocytes that contained PmCypA. Furthermore, the rPmCypA could prolong shrimp survival during VP<inf>AHPND</inf> infection. These results suggest that the PmCypA is a bacterial-responsive gene in shrimp immune defense response and potentially an immunostimulator in shrimp.
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Antiviral properties of Penaeus monodon cyclophilin A in response to white spot syndrome virus infection in the black tiger shrimp
(2024-01-01)
Vatanavicharn, Tipachai
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Matjank, Watchalaya
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Masrinoul, Promsin
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Supungul, Premruethai
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Tassanakajon, Anchalee
Cyclophilin A (CypA) or peptidylprolyl isomerase A, plays an important role in protein folding, trafficking, environmental stress, cell signaling and apoptosis etc. In shrimp, the mRNA expression level of PmCypA was stimulated by LPS. In this study, all three types of shrimp hemocytes: hyaline cell, granulocyte and semi-granulocyte expressed the PmCypA protein. The mRNA expression level of PmCypA was found to be up-regulate to four-fold in white spot syndrome virus (WSSV) infected hemocytes at 48 h. Interestingly, PmCypA protein was only detected extracellularly in shrimp plasma at 24 h post WSSV infection. To find out the function of extracellular PmCypA, the recombinant PmCypA (rPmCypA) was produced and administrated in shrimp primary hemocyte cell culture to observe the antiviral properties. In rPmCypA-administrated hemocyte cell culture, the mRNA transcripts of WSSV intermediate early gene, ie1 and early gene, wsv477 were significantly decreased but not that of late gene, vp28. To explore the antiviral mechanism of PmCypA, the expression of PmCypA in shrimp hemocytes was silenced and the expression of immune-related genes were investigated. Surprisingly, the suppression of PmCypA affected other gene expression, decreasing of penaeidin, PmHHAP and PmCaspase and increasing of C-type lectin. Our results suggested that the PmCypA might plays important role in anti-WSSV via apoptosis pathway. Further studies of PmCypA underlying antiviral mechanism are underway to show its biological function in shrimp immunity.
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Production of Herbal Vinegar Using Isolated Microorganisms from Traditional Herbal Vinegar Fermentation
(2023-01-01)
Thongluedee, Rujira
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Vatanavicharn, Tipachai
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Sutthiphatkul, Tanyarat
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Ochaikul, Duangjai
Schizosaccharomyces pombe YM-19 and Acetobacter pasteurianus EM2-03 were isolated and identified from the traditional herbal vinegar fermentation process using cultural and molecular techniques. The two microorganisms were prepared to inoculum for an experimental herbal vinegar fermentation, which is called “Loog Plaag Mea.” The first starter culture was 10% v/v S. pombe YM1-19. Fermentation was carried out under anaerobic conditions at 30°C for 4 days. The alcohol content was 6.18±0.13% v/v and the pH value was 3.52±0.02 on day 4 of the fermentation period. Subsequently, 10% v/v A. pasteurianus EM2-03 was added to the fermentation process under aerobic conditions at 30°C for 22 days. The final herbal vinegar product contained 4.91±0.15% v/v acetic acid, and a pH value of 3.04±0.04. Its total phenolic content and IC<inf>50</inf> value of DPPH radical scavenging were 1,908.38±38.75 µgGAE/mL and 0.017±0.001 µL/mL, respectively. The experimental fermented herbal vinegar had physicochemical properties that very similar to traditional vinegar. As a result, the isolated microorganisms can be used to improve product consistency and quality control in the mass production of vinegar.
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Production of Polyhydroxybutyrate (PHB) by Novel Bacillus megaterium SWU01 Isolated from Activated Sludge
(2022-11-01)
Chuavong, Wisan
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Ponprateep, Sirikwan
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Ajawatanawong, Pravech
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Vatanavicharn, Tipachai
Polyhydroxybutyrate (PHB) is a natural polymer accumulated in granules as intracellular energy storage in stress condition. The aim of this research was to isolate potential PHB producing bacteria from canned tuna wastewater activated sludge and cow dung. Five positive bacterial isolates were confirmed the accumulation of PHB using lipophilic staining and TEM. To identify the bacterial species, the phylogenetic analyses inferred from 16S rRNA gene sequence demonstrated that the bacterial strains were closely related to Bacillus genera. The comparison of PHB production from bacterial isolates showed that SWU01 strain isolated from the activated sludge exhibited the highest yield of PHB production. The cultivation of SWU01 stain in optimization condition (modified M9 medium supplemented with 2% sodium acetate and 0.5% yeast extract at pH 7 for 48 h) exhibited the yield of PHB up to 0.95 g/L or 7.5 compared with M9 medium supplemented with 2% glucose. Transmission electron micrograph showed high among of PHB in the granule of the bacterial cell. From all results, the bacterial strain SWU01, efficient PHB producers, exhibited the potential for their utilization in commercial PHB production.
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Isolation and identification of fungi with glucoamylase activity from loog-pang-khao-mak (A Thai Traditional Fermentation Starter)
(2020-03-01)
Roongrojmongkhon, Nawaphorn
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Rungjindamai, Nattawut
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Vatanavicharn, Tipachai
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Ochaikul, Duangjai
Loog-pang-khao-mak is a Thai traditional fermentation starter that has been used for production of Thai fermented foods for decades. This research aimed to isolate and identify the fungi that produce effective glucoamylase but low ethanol content from the starter. A total of 166 isolates were screened from twelve samples of Loog-pang-khao-mak accumulated from 12 provinces in Thailand using dichloran rose bengal chloramphenicol agar (DRBC Agar). Seventy-nine isolates that effectively hydrolyze starch were selected for glucoamylase activity and alcohol production assay. Three yeast isolates exhibited high glucoamylase activity ranging from 139.14 to 140.94 unit/ml and lowest alcohol yield of 0.41% (v/v) were Saccharomycopsis fibuligera using morphological and molecular identification. The five isolates of mold exhibited high glucoamylase activity (149.20 to 152.60 unit/ml) were identified as Aspergillus Niger, Aspergillus oryzae and Amylomyces rouxii. These findings provide further knowledge on the fungi and their potential use as traditional inocula for fermentation of food products.
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Plasmolipin, PmPLP1, from Penaeus monodon is a potential receptor for yellow head virus infection
(2018-11-01)
Matjank, Watchalaya
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Ponprateep, Sirikwan
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Rimphanitchayakit, Vichien
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Tassanakajon, Anchalee
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Somboonwiwat, Kunlaya
Plasmolipin has been characterized as a cell entry receptor for mouse endogenous retrovirus. In black tiger shrimp, two isoforms of plasmolipin genes, PmPLP1 and PmPLP2, have been identified from the Penaeus monodon EST database. The PmPLP1 is highly up-regulated in yellow head virus (YHV)-infected shrimp. Herein, the function of PmPLP1 is shown to be involved in YHV infection. The immunoblotting and immunolocalization showed that the PmPLP1 protein was highly expressed and located at the plasma membrane of gills from YHV-infected shrimp. Moreover, the PmPLP1 expressed in the Sf9 insect cells resided at the cell membrane rendering the cells more susceptible to YHV infection. Using the ELISA binding and mortality assays, the synthetic external loop of PmPLP1 was shown to bind the purified YHV and neutralize the virus resulting in the decrease in YHV infection. Our results suggested that the PmPLP1 was likely a receptor of YHV in shrimp.
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Alpha-2-macroglobulin is a modulator of prophenoloxidase system in pacific white shrimp Litopenaeus vannamai
(2017-03-01)
Ponprateep, Sirikwan
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Vatanavicharn, Tipachai
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Lo, Chu Fang
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Tassanakajon, Anchalee
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Rimphanitchayakit, Vichien
The shrimp multifunctional protein alpha-2-macroglobulin (A2M) is abundantly expressed in plasma, highly up-regulated upon microbial infection and involved in several immune pathways such as blood clotting system, phagocytosis and melanization. Herein, the function of LvA2M from Litopenaeus vannamei on the prophenoloxidase (proPO) system is reported. The recombinant (r)LvA2M produced strongly and specifically inhibited trypsin and the PO activity in shrimp plasma in a dose-dependent manner. Silencing of LvA2M led to an increase in the PO activity in shrimp plasma although the expression of proPO-associated genes, proPO-activating enzyme (PPAE) and prophenoloxidase (proPO) but not the proPO-activating factor (PPAF) was down-regulated. In Vibrio parahaemolyticus AHPND-infected shrimp, the LvA2M activity was suppressed in an early phase of infection while the PO activity was increased. Thus, the proPO-activating system was regulated by the LvA2M.
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PmVRP15, a novel viral responsive protein from the black tiger shrimp, Penaeus monodon, promoted white spot syndrome virus replication
(2014-03-17)
Vatanavicharn, Tipachai
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Prapavorarat, Adisak
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Jaree, Phattarunda
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Somboonwiwat, Kunlaya
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Tassanakajon, Anchalee
Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410-fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript was obtained and showed no significant similarity to any known gene in the GenBank database. The predicted open reading frame of PmVRP15 encodes for a deduced 137 amino acid protein containing a putative transmembrane helix. Immunofluorescent localization of the PmVRP15 protein revealed it accumulated around the nuclear membrane in all three types of shrimp hemocytes and that the protein was highly up-regulated in WSSV-infected shrimps. Double-stranded RNA interference-mediated gene silencing of PmVRP15 in P. monodon significantly decreased WSSV propagation compared to the control shrimps (injected with GFP dsRNA). The significant decrease in cumulative mortality rate of WSSV-infected shrimp following PmVRP15 knockdown was observed. These results suggest that PmVRP15 is likely to be a nuclear membrane protein and that it acts as a part of WSSV propagation pathway. Copyright: © 2014 Vatanavicharn et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Two plasmolipins from the black tiger shrimp, Penaeus monodon and their response to virus pathogens
(2012-01-01)
Vatanavicharn, Tipachai
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Pongsomboon, Siriporn
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Tassanakajon, Anchalee
Two isoforms of plasmolipin were initially identified from the black tiger shrimp (Penaeus monodon) EST database and completed using 5' RACE to reveal complete cDNAs of 558. bp (PmPLP1) and 537. bp (PmPLP2) with 87% nucleotide sequence identity. The deduced amino acid sequences contained four-transmembrane domains and showed the highest amino acid identity (49% and 51%, respectively) to the honey bee (Apis mellifera) chemokine-like factor (CKLF), with a very similar hydrophobic pattern to other plasmolipins. Transcripts of PmPLP1 and PmPLP2 were observed in all tested shrimp tissues with the highest expression levels in the gill and epipodite for PmPLP1 and in the hemocytes and antennal gland for PmPLP2. PmPLP1 transcript levels were significantly upregulated in hemocytes at 24 and 72. h post infection (hpi) with yellow head virus (YHV) (7.4- and 14.7- fold, respectively), but only after 72. hpi by white spot syndrome virus (WSSV). In contrast, PmPLP2 was only slightly (but statistically significant) up-regulated with YHV and WSSV. Thus, PmPLPs have the potential to be a part of viral infection mechanism or defense response. This is the first characterization of a plasmolipin gene in crustaceans. © 2012 Elsevier Ltd.

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