Publications by authors named "Eakapol Wangkahart"

11 Publications

  • Page 1 of 1

Psychobiotic Effects of Multi-Strain Probiotics Originated from Thai Fermented Foods in a Rat Model.

Food Sci Anim Resour 2020 Nov 1;40(6):1014-1032. Epub 2020 Nov 1.

Faculty of Science and Marine Environment, Universiti Malaysia Terengganu, Kuala Nerus, Terengganu 21030, Malaysia.

This work aimed to investigate the psychobiotic effects of six bacterial strains on the mind and behavior of male Wistar rats. The probiotic (PRO) group (n=7) were rats pre-treated with antibiotics for 7 days followed by 14-day probiotic administration, antibiotics (ANT) group (n=7) were rats treated with antibiotics for 21 days without probiotics. The control (CON) group (n=7) were rats that received sham treatment for 21 days. The six bacterial strains with probiotic properties were mostly isolated from Thai fermented foods; WS11, SK321, SK324, TRBC 3003, TBRC 7154 and subsp. TBRC 375. The probiotics were freeze-dried into powder (6×10 CFU/5 g) and administered to the PRO group via oral gavage. Behavioral tests were performed. The PRO group displayed significantly reduced anxiety level and increased locomotor function using a marble burying test and open field test, respectively and significantly improved short-term memory performance using a novel object recognition test. Antibiotics significantly reduced microbial counts in rat feces in the ANT group by 100 fold compared to the PRO group. Probiotics significantly enhanced antioxidant enzymatic and non-enzymatic defenses in rat brains as assessed using catalase activity and ferric reducing antioxidant power assay, respectively. Probiotics also showed neuroprotective effects with less pyknotic cells and lower frequency of vacuolization in cerebral cortex. This multi-strain probiotic formulation from Thai fermented foods may offer a potential to develop psychobiotic-rich functional foods to modulate human mind and behaviors.
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http://dx.doi.org/10.5851/kosfa.2020.e72DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7713776PMC
November 2020

Ancient Cytokine Interleukin 15-Like (IL-15L) Induces a Type 2 Immune Response.

Front Immunol 2020 29;11:549319. Epub 2020 Oct 29.

Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan.

Related interleukin-2, -15, and -15-like (IL-2, -15, and -15L) are ancient cytokines, with all three genes surviving in extant fish and some mammals. The present study is the first to identify IL-15L functions, namely in rainbow trout. In isolated trout splenocytes, and , purified recombinant IL-15L+IL-15Rα molecules induced expression of and homologs, which are markers of type 2 immunity. In contrast, trout IL-15 stimulated type 1 immunity markers, thus IL-15 and IL-15L can have opposing functions. Trout IL-15L was more dependent on "in " presentation by the receptor chain IL-15Rα than IL-15, and stimulated CD4CD8(IgM) lymphocytes from thymus and spleen. We propose an important role for IL-15L early in the type 2 immunity cytokine cascade. Trout IL-2 and IL-15 exhibited features reminiscent of their mechanistic and functional dichotomy observed in mammals; for example, IL-15 but not IL-2 required a receptor alpha chain (only IL-15Rα in the case of fish) for its stability, and only IL-15 was efficient in stimulating lymphocytes from mucosal tissues. Data suggest that IL-15L and IL-15 may be particularly effective in stimulating innate lymphocyte type 2 cells (ILC2) and natural killer (NK) cells, respectively, but further identification of the cell types is needed. An interesting finding different from in mammals was the efficient stimulation of CD4CD8 thymocytes by IL-2. In short, this study presents fundamental information on the evolution of the IL-2/15/15L cytokine family.
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http://dx.doi.org/10.3389/fimmu.2020.549319DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7658486PMC
May 2021

Effect of dietary selenium from selenium-enriched kale sprout, selenomethionine, and sodium selenite on performance and selenium concentrations in the tissues of growing quails.

Anim Biosci 2021 Apr 12;34(4):692-700. Epub 2020 May 12.

Department of Chemistry and Center of Excellence for Innovation in Chemistry (PERCH-CIC), Faculty of Sciences, Mahasarakham University, Kantharawichai, Mahasarakham, 44150, Thailand.

Objective: This study aimed to determine the effect of dietary selenium (Se) from Se-enriched kale sprout (SeKS), selenomethionine (SeMet), and sodium selenite (SS) on performance, carcass characteristics and Se concentrations in the tissues, and to study the relationship between Se concentrations in muscle and feather in growing quails.

Methods: The 320 quails (7 d of age) were divided into four treatments, according to a completely randomized design. The treatments were T1: control diet; T2, T3, and T4: control diets plus 0.2 mg Se/kg from SS, SeMet, and SeKS, respectively. The performance, carcass characteristics, and Se concentrations in the tissues of quails were determined.

Results: The results indicated no effect (p>0.05) of Se supplementation on performance, carcass characteristics and glutathione peroxidase (GSH-Px) activity in breast muscle of quails. Supplemental Se from SS, SeMet, and SeKS increased greater (p<0.05) Se concentrations in breast muscle, liver, kidney, heart, and feather, compared to those of quails fed the control diet. Quails fed Se from SeMet had greater (p<0.05) Se concentrations in the tissues than quails fed Se from SeKS and SS. In addition, Se concentrations in breast muscle and feather of quails at 21 and 42-d-old were highly correlated (R2 0.714 to 0.756) (p<0.05).

Conclusion: Performance, carcass characteristics and GSH-Px activity in breast muscle of quails were not affected (p>0.05) by dietary Se supplementation. The Se from SeMet was more effective in increasing Se concentrations in the tissues of quails than Se from SeKS and SS. Feather Se concentrations of 21 and 42-d-old quails can be used for assessment of Se bioavailability of Se sources.
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http://dx.doi.org/10.5713/ajas.20.0111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961293PMC
April 2021

Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture.

Front Immunol 2018 9;9:3054. Epub 2019 Jan 9.

Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.

Immunostimulants and vaccines are important for controlling infectious diseases in fish aquaculture. In this study we assess the potential of flagellin to be used for such purposes in rainbow trout (). A recombinant flagellin from the salmonid pathogen (YRF) has been produced previously by us and shown to be a potent activator of inflammatory cytokines, acute phase proteins and antimicrobial peptides . Here we show that YRF is the most potent inflammatory activator of three bacterial PAMPs (LPS, peptidoglycan and flagellin) tested. The host response to flagellin was next studied . The YRF modulated gene expression was examined in two systemic (spleen and liver) and two mucosa-associated (gills and skin) tissues. YRF injection initiated a transient systemic inflammatory response with key pro-inflammatory cytokines (IL-1β, TNFα, IL-6, and IL-11 etc.) and chemokines (CXCL_F4 and CXCL-8) induced rapidly (by 6 h) but subsiding quickly (by 24 h) in multiple tissues. Consequently, a variety of anti-microbial pathways were activated systemically with heightened expression of acute phase proteins, antimicrobial peptides and complement genes in multiple tissues, which was sustained to 24 h in the liver and mucosal tissues. The Th17 cytokine IL-17A/F1 was also induced in the spleen and liver, and Th2 cytokine IL-4/13 was induced in the liver. However, the anti-inflammatory IL-10 and the Th1 cytokine IFNγ were refractory. A secreted form of TLR5 (TLR5s) was induced by flagellin in all tissues examined whilst the membrane form was refractory, suggesting that TLR5s may function as a negative feedback regulator. Trout liver appeared to be an important organ responding to flagellin stimulation, with marked induction of IL-11, IL-23P19, IL-17C1, SAA, and cathelicidin-2. YRF induced a strong antibody response. These antibodies reacted against the middle domain of YRF and were able to decrease YRF bioactivity. Intact YRF was necessary for its bioactivity, as deletion of the N-terminal, C terminal or middle domain of YRF led to functional loss. This study suggests that flagellin could be a potent immunostimulant and vaccine adjuvant for fish aquaculture.
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http://dx.doi.org/10.3389/fimmu.2018.03054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6333709PMC
October 2019

Insights into the Evolution of the Suppressors of Cytokine Signaling (SOCS) Gene Family in Vertebrates.

Mol Biol Evol 2019 02;36(2):393-411

Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen, United Kingdom.

The SOCS family are key negative regulators of cytokine and growth factor signaling. Typically, 8-17 SOCS genes are present in vertebrate species with eight known in mammals, classified as type I (SOCS4-7) and type II (CISH and SOCS1-3) SOCS. It was believed that the type II SOCS were expanded through the two rounds of whole genome duplication (1R and 2R WGDs) from a single CISH/SOCS1-3 precursor. Previously, 12 genes were identified in rainbow trout but here we report 15 additional loci are present, and confirm 26 of the genes are expressed, giving rainbow trout the largest SOCS gene repertoire identified to date. The discovery of the additional SOCS genes in trout has led to a novel model of SOCS family evolution, whereby the vertebrate SOCS gene family was derived from CISH/SOCS2, SOCS1/SOCS3, SOCS4/5, SOCS6, and SOCS7 ancestors likely present before the two WGD events. It is also apparent that teleost SOCS2b, SOCS4, and SOCS5b molecules are not true orthologues of mammalian SOCS2, SOCS4, and SOCS5, respectively. The rate of SOCS gene structural changes increased from 2R vertebrates, to 4R rainbow trout, and the genes with structural changes show large differences and low correlation coefficient of expression levels relative to their paralogues, suggesting a role of structural changes in expression and functional diversification. This study has important impacts in the functional prediction and understanding of the SOCS gene family in different vertebrates, and provides a framework for determining how many SOCS genes could be expected in a particular vertebrate species/lineage.
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http://dx.doi.org/10.1093/molbev/msy230DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368001PMC
February 2019

Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent Paralogs in Salmonids.

Front Immunol 2018 26;9:1683. Epub 2018 Jul 26.

Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.

Mammalian interleukin (IL)-2 is a cytokine centrally involved in the differentiation and survival of CD4+ T helper subsets and CD4+ T regulatory cells and in activation of cytotoxic effector lymphocytes. In bony fish, orthologs have been identified with an additional divergent gene on the same locus present in several fish species. We report here two divergent paralogs, and , in salmonids that originated from the whole genome duplication event in this fish lineage. The salmonid paralogs differ not only in sequence but also in exon sizes. The IL-2 isoforms that are encoded have disparate pI values and may have evolved to preferentially bind specific IL-2 receptors. Rainbow trout paralogs are highly expressed in thymus, spleen, gills, kidney and intestine, important tissues/organs in fish T cell development and function. Their expression in peripheral blood leukocytes (PBL) is low constitutively but can be upregulated by the mixed leukocyte reaction, by the T cell mitogen phytohemagglutinin and by signal mimics of T cell activation (phorbol 12-myristate 13-acetate and calcium ionophore). Both trout IL-2 isoforms promoted PBL proliferation and sustained high-level expression of and , suggesting that trout IL-2 isoforms are T cell growth/survival factors mainly expressed by activated T cells. The recombinant proteins for these two trout paralogs have been produced in and possess shared but also distinct bioactivities. IL-2A, but not IL-2B, induced and expression in PBL. IL-2B had a stronger effect on upregulation of the T helper 1 (Th1) cytokine () and could sustain and expression levels. Nevertheless, both cytokines upregulated key Th1 ( and ) and T helper 2 (Th2) cytokines ( and ), cytokine and chemokine receptors and the antimicrobial peptide but had limited effects on T helper 17 cytokines and in PBL. They could also enhance PBL phagocytosis. These results suggest, for the first time in fish, that IL-2 isoforms may have an important role in regulating Th1 and Th2 cell development, and innate and adaptive host defenses in fish, and shed light on lineage-specific expansion, evolution, and functional diversification of in vertebrates.
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http://dx.doi.org/10.3389/fimmu.2018.01683DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6070626PMC
September 2019

Rainbow trout (Oncorhynchus mykiss) adipose tissue undergoes major changes in immune gene expression following bacterial infection or stimulation with pro-inflammatory molecules.

Dev Comp Immunol 2018 04 7;81:83-94. Epub 2017 Nov 7.

Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Tillydrone Avenue, Aberdeen, AB24 2TZ, UK. Electronic address:

In mammals, visceral adipose is increasingly seen as playing an important role in immune function with numerous pro-inflammatory, anti-inflammatory and immune-modulating proteins and peptides being identified in adipocytes. Adipose is also now known as a tissue that has an important role in the regulation of peritoneal immune responses. Despite this, only lately has consideration been given to visceral adipose as an important immune tissue in fish, especially in the context of intraperitoneal vaccination. The present study demonstrates that fish visceral adipose is capable of expressing a large range of immune molecules in response to stimulation with a live bacterium (A. salmonicida), a bacterial PAMP (Y. ruckeri flagellin), and the pro-inflammatory cytokines IL-1β, TNF-α3 and IFN-γ. Following infection and stimulation with flagellin and IL-1β a large upregulation of pro-inflammatory and antimicrobial molecules was seen, with a high degree of overlap. TNF-α treatment affected relatively few genes and the effects were more modest. IFN-γ had the smallest impact on adipose but IFN-γ inducible genes showed some of the largest effects. Overall, it is clear that adipose tissue should be considered an active immune site in fish, capable of participating in and influencing immune responses.
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http://dx.doi.org/10.1016/j.dci.2017.11.001DOI Listing
April 2018

Dissecting the immune pathways stimulated following injection vaccination of rainbow trout (Oncorhynchus mykiss) against enteric redmouth disease (ERM).

Fish Shellfish Immunol 2019 Feb 27;85:18-30. Epub 2017 Jul 27.

Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, UK. Electronic address:

Enteric redmouth disease (ERM or yersiniosis) is one of the most important diseases of salmonids and leads to significant economic losses. It is caused by the Gram-negative bacterium Yersinia ruckeri but can be controlled by bacterin vaccination. The first commercial ERM vaccine was licenced in 1976 and is one of the most significant and successful health practices within the aquaculture industry. Although ERM vaccination provides complete protection, knowledge of the host immune response to the vaccine and the molecular mechanisms that underpin the protection elicited is limited. In this report, we analysed the expression in spleen and gills of a large set of genes encoding for cytokines, acute phase proteins (APPs) and antimicrobial peptides (AMPs) in response to ERM vaccination in rainbow trout, Oncorhynchus mykiss. Many immune genes in teleost fish are known to have multiple paralogues that can show differential responses to ERM vaccination, highlighting the necessity to determine whether all of the genes present react in a similar manner. ERM vaccination immediately activated a balanced inflammatory response with correlated expression of both pro- and anti-inflammatory cytokines (eg IL-1β1-2, TNF-α1-3, IL-6, IL-8 and IL-10A etc.) in the spleen. The increase of pro-inflammatory cytokines may explain the systemic upregulation of APPs (eg serum amyloid A protein and serum amyloid protein P) and AMPs (eg cathelicidins and hepcidin) seen in both spleen and gills. We also observed an upregulation of all the α-chains but only one β-chain (p40B2) of the IL-12 family cytokines, that suggests specific IL-12 and IL-23 isoforms with distinct functions might be produced in the spleen of vaccinated fish. Notably the expression of Th1 cytokines (IFN-γ1-2) and a Th17 cytokine (IL-17A/F1a) was also up-regulated and correlated with enhanced expression of the IL-12 family α-chains, and the majority of pro- and anti-inflammatory cytokines, APPs and AMPs. These expression profiles may suggest that ERM vaccination activates host innate immunity and expression of specific IL-12 and IL-23 isoforms leading to a Th1 and Th17 biased immune response. A late induction of Th2 cytokines (IL-4/13B1-2) was also observed, that may have a homeostatic role and/or involvement in antibody production. This study has increased our understanding of the host immune response to ERM vaccination and the adaptive pathways involved. The early responses of a set of genes established in this study may provide essential information and function as biomarkers in future vaccine development in aquaculture.
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http://dx.doi.org/10.1016/j.fsi.2017.07.056DOI Listing
February 2019

Characterisation of arginase paralogues in salmonids and their modulation by immune stimulation/ infection.

Fish Shellfish Immunol 2017 Feb 23;61:138-151. Epub 2016 Dec 23.

Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, UK. Electronic address:

In this study we show that four arginase isoforms (arg1a, arg1b, arg2a, arg2b) exist in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). We have characterised these molecules in terms of a) sequence analysis, b) constitutive expression in different tissues, and modulated expression following c) stimulation of head kidney macrophages in vitro, or d) vaccination/infection with Yersinia ruckeri and e) parasite infection (AGD caused by Paramoeba perurans and PKD caused by Tetracapsuloides bryosalmonae). Synteny analysis suggested that these arginase genes are paralogues likely from the Ss4R duplication event, and amino acid identity/similarity analyses showed that the proteins are relatively well conserved across species. In rainbow trout constitutive expression of one or both paralogues was seen in most tissues but different constitutive expression patterns were observed for the different isoforms. Stimulation of rainbow trout head kidney macrophages with PAMPs and cytokines also revealed isoform specific responses and kinetics, with arg1a being particularly highly modulated by the PAMPs and pro-inflammatory cytokines. In contrast the type II arginase paralogues were induced by rIl-4/13, albeit to a lesser degree. Vaccination and infection with Y. ruckeri also revealed isoform specific responses, with variation in tissue expression level and kinetics. Lastly, the impact of parasite infection was studied, where down regulation of arg1a and arg1b was seen in two different models (AGD in salmon and PKD in trout) and of arg2a in AGD. The differential responses seen are discussed in the context of markers of type II responses in fish and paralogue subfunctionalization.
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http://dx.doi.org/10.1016/j.fsi.2016.12.024DOI Listing
February 2017

Sequence and Expression Analysis of Interferon Regulatory Factor 10 (IRF10) in Three Diverse Teleost Fish Reveals Its Role in Antiviral Defense.

PLoS One 2016 19;11(1):e0147181. Epub 2016 Jan 19.

Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen AB24 2TZ, United Kingdom.

Background: Interferon regulatory factor (IRF) 10 was first found in birds and is present in the genome of other tetrapods (but not humans and mice), as well as in teleost fish. The functional role of IRF10 in vertebrate immunity is relatively unknown compared to IRF1-9. The target of this research was to clone and characterize the IRF10 genes in three economically important fish species that will facilitate future evaluation of this molecule in fish innate and adaptive immunity.

Molecular Characterization Of Irf10 In Three Fish Species: In the present study, a single IRF10 gene was cloned in grass carp Ctenopharyngodon idella and Asian swamp eel Monopterus albus, and two, named IRF10a and IRF10b, in rainbow trout Oncorhynchus mykiss. The fish IRF10 molecules share highest identities to other vertebrate IRF10s, and have a well conserved DNA binding domain, IRF-associated domain, and an 8 exon/7 intron structure with conserved intron phase. The presence of an upstream ATG or open reading frame (ORF) in the 5'-untranslated region of different fish IRF10 cDNA sequences suggests potential regulation at the translational level, and this has been verified by in vitro transcription/translation experiments of the trout IRF10a cDNA, but would still need to be validated in fish cells.

Expression Analysis Of Irf10 In Vivo And In Vitro: Both trout IRF10 paralogues are highly expressed in thymus, blood and spleen but are relatively low in head kidney and caudal kidney. Trout IRF10b expression is significantly higher than IRF10a in integumentary tissues i.e. gills, scales, skin, intestine, adipose fin and tail fins, suggesting that IRF10b may be more important in mucosal immunity. The expression of both trout IRF10 paralogues is up-regulated by recombinant IFN-γ. The expression of the IRF10 genes is highly induced by Poly I:C in vitro and in vivo, and by viral infection, but is less responsive to peptidoglycan and bacterial infection, suggesting an important role of fish IRF10 in antiviral defense.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147181PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4718558PMC
July 2016

Re-examination of the rainbow trout (Oncorhynchus mykiss) immune response to flagellin: Yersinia ruckeri flagellin is a potent activator of acute phase proteins, anti-microbial peptides and pro-inflammatory cytokines in vitro.

Dev Comp Immunol 2016 Apr 21;57:75-87. Epub 2015 Dec 21.

Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, AB24 2TZ, UK. Electronic address:

Flagellin is the principal component of bacterial flagellum and a major target of the host immune system. To provide new insights into the role of flagellin in fish immune responses to flagellated microorganisms, a recombinant flagellin from Yersinia ruckeri (rYRF) was produced and its bioactivity investigated in the trout macrophage cell line RTS-11 and head kidney cells. rYRF is a potent activator of pro-inflammatory cytokines, acute phase proteins, antimicrobial peptides and subunits of the IL-12 cytokine family. This and the synergy seen with IFN-γ to enhance further expression of specific IL-12 and TNF-α isoforms may suggest that flagellin could be a useful immune stimulant or adjuvant for use in aquaculture. Gene paralogues were often differentially modulated, highlighting the need to study all of the paralogues of immune genes in fish to gain a full understanding of the effects of PAMPs or other stimulants, and the potential immune responses elicited.
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http://dx.doi.org/10.1016/j.dci.2015.12.017DOI Listing
April 2016
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