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expression profiling of immune-related genes from japanese flounder paralichthys olivaceus kidney cells using cdna microarrays.a japanese flounder paralichthys olivaceus cdna microarray containing 871 unique cdnas including 91 putative immune-related genes from our est studies was constructed and used to characterize of gene expression of in vitro grown kidney cells stimulated with mitogens such as cona, pma, lps or infected with hirame rhabdovirus (hrv). the numbers of genes whose expressions were increased or decreased by these factors were: 17 by con a, 139 by pma, 76 by lps and 182 by hrv infection. the treatment of ...200515752548
isolation and characterization of ornithine decarboxylase gene from flounder (paralichthys olivaceus).ornithine decarboxylase (odc) is a homodimeric enzyme dependent on pyridoxal 5'-phosphate. we identified a complementary dna clone corresponding to odc from the brain of adult flounder (paralichthys olivaceus). the flounder odc cdna consisted of 2939 bp encoding 272 amino acid residues. the flounder odc showed 80.3% sequence identity to zebrafish and 70.8% to rat at the amino acid level. comparison of the structure and nucleotide sequence of the odc genes revealed that the gene is highly conserv ...200415791490
identification of intestinal bacteria from japanese flounder (paralichthys olivaceus) and their ability to digest chitin.the aim of the present study was to clarify the taxonomic status of intestinal bacteria isolated from japanese flounder (paralichthys olivaceus) and describe their ability to digest chitin.200616910942
molecular cloning and mrna profile of insulin-like growth factor type 1 receptor in orange-spotted grouper, epinephelus coioides.the insulin-like growth factor type 1 receptor (igf-ir) belongs to the tyrosine kinase (tk) receptor family. besides being mitogenic, igf-ir plays a crucial role in cell survival, transformation and maintenance of the malignant phenotype. in this study, we cloned the cdna from the hypothalamus of the orange-spotted grouper (epinephelus coioides) using reverse transcription pcr (rt-pcr) and the rapid amplification of cdna ends (race) method. the deduced amino acid sequence showed that the recepto ...200515880261
phylogenetic analysis of the major capsid protein gene of iridovirus isolates from cultured flounders paralichthys olivaceus in korea.in 2003, 13 isolates of iridovirus were obtained from cultured flounders paralichthys olivaceus during epizootics in korea. the full open reading frames (orfs) encoding the major capsid protein (mcp) (1362 bp) from the 13 flounder iridoviruses (flivs) were sequenced and the deduced amino acid sequences were phylogenetically analyzed. phylogenetic analysis of the mcp revealed that all 13 flivs were the same species as rock bream iridovirus (rbiv), red sea bream iridovirus (rsiv), and infectious s ...200515997817
characterization of japanese flounder (paralichthys olivaceus) nk-lysin, an antimicrobial peptide.the nk-lysin cdna of japanese flounder, paralichthys olivaceus, consists of 657bp, containing an open reading frame (orf) of 444bp, which encodes 147 amino acid residues. the amino acid sequence of japanese flounder nk-lysin has 21% identity to porcine nk-lysin and bovine nk-lysin, 23% to equine nk-lysin, and 46% to zebrafish nk-lysin-like protein. multiple alignments of japanese flounder nk-lysin and other known saposin-like proteins revealed that the six cysteine residues important for structu ...200717046282
molecular cloning and characterization of cathepsin b from a scuticociliate, uronema marinum.a cdna encoding cathepsin b was cloned from the scuticociliate, uronema marinum, which invades the olive flounder, paralichthys olivaceus, leading to high mortalities in culturing fish. the full-length scuticociliate cathepsin b (scctb) gene contains an open reading frame of 1053 base pairs encoding 350 amino acids. a homology search revealed that scctb shares sequence identity with several piscine cathepsin bs (48%-45%). the protein of scctb from u. marinum extracts was purified 12.8-fold by a ...200516172011
molecular identification and expression analysis of natural resistance associated macrophage protein (nramp) cdna from japanese flounder (paralichthys olivaceus).natural resistance associated macrophage protein (nramp) controls partially innate resistance to intracellular parasites. its function is to enhance the ability of macrophages to kill pathogens. however, little is known about the structure and function of nramp in lower vertebrates such as teleosts. we have recently isolated a cdna encoding nramp from japanese flounder (paralichthys olivaceus). the full-length cdna of the nramp is 3066 bp in length, including 224 bp 5' terminal utr, 1662 bp enco ...200615998592
characterization of monoclonal antibodies against heavy and light chains of flounder (paralichthys olivaceus) immunoglobulin.flounder (paralichthys olivaceus) immunoglobulins (igs) were purified from the serum of mouse igg-immunized flounder by using affinity chromatography. under denaturing conditions in sds-page, the flounder igs appeared to be composed of 2 heavy (h) chains (72 and 77 kda) and two light (l) chains (26 and 28 kda). monoclonal antibodies (mabs) were produced by the fusion of myeloma cells (sp2/0) with balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder igs. in ...200415469712
a new genotype of lymphocystivirus, lcdv-rf, from lymphocystis diseased rockfish.lymphocystis disease virus (lcdv) is the causative agent of lymphocystis disease. in this study, nucleotide sequences of the major capsid protein (mcp) gene were analyzed among lcdv isolates from japanese flounder and rockfish. a phylogenetic tree revealed three clusters for lymphocystiviruses. the first cluster included japanese flounder isolates; the second cluster consisted of rockfish isolates; and the remaining one consisted of lcdv-1. nucleotide sequence identities were > or =99.6% among j ...200616328139
susceptibility of marine fish species to a megalocytivirus, turbot iridovirus, isolated from turbot, psetta maximus (l.).turbot iridovirus (tbiv), a member of the genus megalocytivirus in the family iridoviridae, was isolated from diseased turbot, psetta maximus (l.), in korea in 2003. in this study, experimental infection of turbot, japanese flounder, paralichthys olivaceus (temminck & schlegel), and rock bream, oplegnathus fasciatus (temminck & schlegel), with tbiv was performed to evaluate the viral susceptibility of these fish species. after virus exposure, the mortalities of turbot reared at 22 and 25 degrees ...200616866925
major histocompatibility complex class iib allele polymorphism and its association with resistance/susceptibility to vibrio anguillarum in japanese flounder (paralichthys olivaceus).the full length of major histocompatibility complex (mhc) class iib cdna was cloned from a chinese population of paralichthys olivaceus by homology cloning and rapid amplification of cdna ends-polymerase chain reaction (race-pcr). the mhc iib genomic sequence is 1,864 bp long and consists of 34-bp 5'utr, 741-bp open reading frame, 407-bp 3'utr, 96-bp intron1, 392-bp intron2, 85-bp intron3, and 109-bp intron4. phylogenetic analysis showed that the putative mhc class iib amino acid of the chinese ...200616874444
application of immunoproteomics in developing a streptococcus iniae vaccine for olive flounder (paralichthys olivaceus).streptococcus iniae is the major etiological agent of streptococcosis, which is responsible for hemorrhagic septicemia in fish, particularly olive flounder (paralichthys olivaceus). in the present study, we sought to understand the pathogenicity and immunogenicity of s. iniae in order to develop a vaccine for streptococcosis. immunoproteomics, a technique involving two-dimensional gel electrophoresis (2-de) followed by immunoblotting, was employed to investigate the pathogenicity and immunogenic ...200717140864
[extraction, identification and primary investigation of adhesion specificity of lactobacillus sp].lactobacillus l15, a strain of lactobacillus sp. isolated from the intestinal tract of healthy paralichthys olivaceus, and lactobacillus acidlophilus atcc4356 were used to investigate their specific adhesions. it was shown that cell surface proteins of two strains could be extracted by 5 mol/l lithium chloride( licl). two surface proteins with molecular weights of 61.8 kda and 54.6 kda were identified to mediate the adhesion to intestinal mucus of paralichthys olivaceus and common carp respectiv ...200616933608
pathogenicity of edwardsiella tarda to olive flounder, paralichthys olivaceus (temminck & schlegel).the ld50 and cytotoxic and enzymatic activities of both cells and extracellular products (ecps) of eight edwardsiella tarda strains were determined and their bacterial superoxide dismutase gene (sodb) and catalase gene (katb) were sequenced. strains were also examined for their ability to resist the immune responses of olive flounder, paralichthys olivaceus. ld50 values of strains (fsw910410, ke1, 2, 3, 4, 5 and 6) in olive flounder ranged between 10(2.5) and 10(5.3) cfu (colony forming units) p ...200617026669
detection of dnas homologous to betanodavirus genome rnas in barfin flounder verasper moseri and japanese flounder paralichthys olivaceus.i investigated the presence of dna homologous to genome rna1 and rna2 (rna1 dna and rna2 dna) of betanodaviruses - the causative agent of viral nervous necrosis (vnn) --in eggs, sperm, ovarian cavity fluid, larvae, and juveniles of barfin flounder verasper moseri and larvae and juveniles of japanese flounder paralichthys olivaceus collected at 6 sites in hokkaido, japan, from 1994 to 2001. rna1 dna and rna2 dna were detected by pcr in 13 and 33 % of barfin flounder samples and 0 and 69% of japan ...200617190201
role of cellular response in elimination of the monogenean neoheterobothrium hirame in japanese flounder paralichthys olivaceus.adult worms of the blood-feeding monogenean parasite neoheterobothrium hirame, which cause anemia in the japanese flounder paralichthys olivaceus, attach to the host fish by embedding their posterior part deeply into the host tissue. to investigate the possibility that cellular responses of the host fish can eliminate n. hirame, flounder were experimentally infected with n. hirame larvae and reared in either fed or starved conditions. mature parasites were identified on the buccal cavity wall of ...200515918475
complete small subunit rrna gene sequence of the scuticociliate miamiensis avidus pathogenic to olive flounder paralichthys olivaceus.eight isolates of miamiensis avidus (scuticociliates) were collected from olive flounder paralichthys olivaceus with symptoms of severe ulcers and haemorrhages at several culture farms in 1999 and 2003. cloned strains were produced and the complete small subunit ribosomal rna gene (ssu rrna) of each strain was sequenced for classification and phylogenic study. the ssu rrna is 1759 bp in length and the sequence was deposited in the genbank under accession number ay550080. all 8 strains exhibited ...200515918479
the interferon system of teleost fish.interferons (ifns) are secreted proteins, which induce vertebrate cells into an antiviral state. in mammals, three families of ifns (type i ifn, type ii ifn and ifn-lambda) can be distinguished on the basis of gene structure, protein structure and functional properties. type i ifns, which include ifn-alpha and ifn-beta, are encoded by intron lacking genes and have a major role in the first line of defense against viruses. the human ifn-lambdas have similar biological properties as type i ifns, b ...200615939626
ultrastructure and phylogeny of philasterides dicentrarchi (ciliophora, scuticociliatia) from farmed turbot in nw spain.several species of opportunistic histophagous scuticociliates have been implicated in systemic infections of farmed fish. in turbot, scuticociliatosis is an emerging disease, and the identification of the parasite species involved is controversial. we have previously isolated philasterides dicentrarchi from farmed turbot scuticociliatosis outbreaks in northwest spain. in the present study, we report detailed ultrastructural studies of this parasite, and investigate phylogenetic relations with ot ...200616388691
identification and characterization of a myeloid differentiation factor 88 (myd88) cdna and gene in japanese flounder, paralichthys olivaceus.the interleukin-1 receptor/toll-like receptor (il-1r/tlr) superfamily signaling involves myeloid differentiation factor 88 (myd88) that acts as an important adapter protein. a japanese flounder (paralichthys olivaceus) myd88 (jfmyd88) cdna and gene were cloned, and found to have lengths of 1.5 and 3.01 kb, respectively. the orf encodes 285 amino acids that contain a death domain and a toll/il-1 receptor domain. the gene is composed of 5 exons and 4 introns. the jfmyd88 gene is highly expressed i ...200616412507
functional characterisation of the japanese flounder, paralichthys olivaceus, mx promoter.the japanese flounder, paralichthys olivaceus, genome appears to encode a single mx gene based on southern blotting and previous cdna studies. the 5' flanking region of the japanese flounder mx gene was cloned and analysed for its regulatory regions. a tata box (-24 to -30), two interferon-stimulated response elements (isres) (-69 to -80 and -508 to -521) and two sp1 sites (-563 to -572 and -994 to -1003) were identified relative to the transcription start site. the effects of various stimuli, a ...200616551503
identification of two isoforms of atlantic halibut insulin-like growth factor-i receptor genes and quantitative gene expression during metamorphosis.insulin-like growth factor-i (igf-i) is an important regulator of growth and development in vertebrates. both the endocrine and paracrine actions of igf-i are mediated through ligand-binding to a membrane-bound igf-i receptor (igf-ir). the characterization of this receptor and subsequent expression studies thus help elucidate the endocrine regulation of developmental processes. as other flatfish species, the atlantic halibut (hippoglossus hippoglossus) undergoes a dramatic larval metamorphosis. ...200717395512
protection of flounder against hirame rhabdovirus (hirrv) with a dna vaccine containing the glycoprotein gene.hirame rhabdovirus (hirrv) is an important virus of cultured flounder (paralichthys olivaceus). we tested the protective immunogenicity of dna-based vaccines against this virus. genes encoding the nucleocapsid protein (n) and the c-terminal half of the glycoprotein (g) were amplified by rt-pcr and separately cloned into the eukaryotic expression vector pcdna 3.1. the g protein expressed by transfected cells was detected by western blot analysis. pcr analyses demonstrated the presence of injected ...200616176849
comparative immune responses in japanese flounder, paralichthys olivaceus after vaccination with viral hemorrhagic septicemia virus (vhsv) recombinant glycoprotein and dna vaccine using a microarray analysis.a viral hemorrhagic septicemia virus (vhsv), recombinant glycoprotein vaccine (vhsvg) and a dna vaccine (pcmv-vhsg) were injected in japanese flounder (paralichthys olivaceus). each fish was injected with 10 microg vaccine dissolved in 50 microl phosphate buffer saline (pbs). one month after the vaccination, the fish were challenged intraperitoneally with either 1 x 10(2) or 1 x 10(3) tcid(50) of virus. fish that received the vhsg dna vaccine were highly protected against virus infection over a ...200616185791
two different types of hepcidins from the japanese flounder paralichthys olivaceus.the cysteine-rich peptide hepcidin is known to be an antimicrobial peptide and iron transport regulator that has been found in both fish and mammals. recently, we found two different types (designated hep-jf1 and hep-jf2) of hepcidin cdna in the japanese flounder, paralichthys olivaceus, by expressed sequence tag analysis. the identity of amino acid sequences between hep-jf1 and hep-jf2 was 51%. the hep-jf1 and hep-jf2 genes both consist of three exons and two introns, and both exist as single c ...200516218956
acute toxicity profile of cadmium revealed by proteomics in brain tissue of paralichthys olivaceus: potential role of transferrin in cadmium toxicity.an analytical approach using two-dimensional polyacrylamide gel electrophoresis (2d-page) separated proteins from the brain tissue of the fish paralichthys olivaceus. approximately 600 protein spots were detected from the brain sample when applying 600 microg protein to a 2d-page gel in the ph range 3.5-10.0. compared to a control sample, significant changes of 24 protein spots were observed in the fish tissue exposed to acute toxicity of seawater cadmium (scat) at 10 ppm for 24h. among these sp ...200616647146
purification and characterization of tributyltin-binding protein type 2 from plasma of japanese flounder, paralichthys olivaceus.we used gel filtration chromatography, anion-exchange chromatography and polyacrylamide gel electrophoresis to purify tributyltin-binding protein type 2 (tbt-bp 2) from plasma of japanese flounder (paralichthys olivaceus) injected intraperitoneally with tbt (5.0 mg/kg body weight). sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the molecular mass of tbt-bp 2 was approximately 48 kda, and isoelectric focusing-polyacrylamide gel electrophoresis indicated that the isoelect ...200717545251
differentiation of lymphocystis disease virus genotype by multiplex pcr.lymphocystis disease virus (lcdv) is the causative agent of lymphocystis disease. the viruses have been divided into three genotypes (genotype i for lcdv-1, ii for japanese flounder isolates, and iii for rockfish isolates) on the basis of major capsid protein (mcp) gene sequences. in this study, we developed a multiplex pcr primer set in order to distinguish these genotypes. we also analyzed the mcp gene of a new lcdv isolate from the sea bass (sb98yosu). comparison of sequence identities betwee ...200616728963
ultrastructural features of herpesvirus-infected cells in epidermal lesions in larvae of the japanese flounder paralichthys olivaceus.an electron microscopic study (em) was performed on herpesvirus-infected cells in epidermal lesions of larvae of the japanese flounder paralichthys olivaceus to document the ultrastructure of the lesions. epithelial cells were the primary cell type infected with the herpesvirus, and showed nuclei containing nucleocapsids, vesicles containing enveloped virions in the cytoplasm, and the release of enveloped virions through the cell membrane. herpesvirus-infected cells were highly vacuolated and ne ...200516231642
development and characterization of monoclonal antibody to the lymphocystis disease virus of japanese flounder paralichthys olivaceus isolated from china.lymphocystis disease virus (lcdv) can infect, both naturally and experimentally, about 100 different teleost fish species. in this study, lcdv was purified using differential and gradient centrifugation from skin tumours of japanese flounder paralichthys olivaceus. a panel of five monoclonal antibodies (mabs) against lcdv were produced by immunization of balb/c mice with purified virus preparations. analysed by the indirect enzyme-linked immunosorbent assay (elisa), indirect immunofluorescence a ...200616730805
the granulocyte colony-stimulating factors (csf3s) of fish and chicken.granulocyte colony-stimulating factor (csf3) is a glycoprotein cytokine, which influences the hematopoiesis of the phagocytic neutrophils and its precursors and was used extensively in cancer therapy and for the treatment of neutropenia in mammals. however, csf3 is yet to be identified in nonmammalian species mainly because of its rapid mutation. here, we report the first csf3 genes from three teleost fishes: japanese flounder (paralichthys olivaceus), fugu (takifugu rubripes), and green-spotted ...200616738939
transgenic zebrafish expressing chicken lysozyme show resistance against bacterial diseases.we established a transgenic zebrafish strain expressing chicken lysozyme gene under the control of the japanese flounder keratin gene promoter, and investigated its resistance to a pathogenic bacterial infection. to generate the lysozyme transgenic construct, japanese flounder keratin promoter was linked to both the hen egg white (hew) lyoszyme gene and green fluorescence protein (gfp) gene used as a selection marker for the transgenic strains, in a recombinant plasmid. the recombinant plasmid w ...200616779653
molecular identification, polymorphism, and expression analysis of major histocompatibility complex class iia and b genes of turbot (scophthalmus maximus).major histocompatibility complex (mhc) class ii has a central role in the adaptive immune system by presenting foreign peptides to the t-cell receptor. the full lengths of mhc class ii a and b cdna were cloned from turbot by homology cloning and rapid amplification of cdna ends polymerase chain reaction (race pcr), and genomic organization, molecular polymorphism, and expression of turbot class iib gene were examined to study the function of class iib gene in fish. the deduced amino acid sequenc ...200616832747
a genetic linkage map of atlantic halibut (hippoglossus hippoglossus l.).a genetic linkage map has been constructed for atlantic halibut on the basis of 258 microsatellites and 346 aflps. twenty-four linkage groups were identified, consistent with the 24 chromosomes seen in chromosome spreads. the total map distance is 1562.2 cm in the female and 1459.6 cm in the male with an average resolution of 4.3 and 3.5 cm, respectively. using diploid gynogens, we estimated centromere locations in 19 of 24 linkage groups. overall recombination in the female was approximately tw ...200717720928
cloning and expression of a novel serine protease from japanese flounder, paralichthys olivaceus.two different cdna clones of japanese flounder (types i-1 and i-2) with lengths of 1096 and 1572bp, respectively, were found to encode the same serine protease consisting of 244 identical amino acid residues with three putative n-glycosylation sites, an 18-amino acid signal peptide and a 2-amino acid activation peptide. the amino acid sequence of the japanese flounder serine protease shares 39-44% identity to known hematopoietic serine proteases. genomic analysis showed that two different clones ...200717084451
structural characterisation and expression analysis of toll-like receptor 2 gene from catfish.toll-like receptors (tlrs) are important components of innate immunity. they were found to recognise specific structures on pathogens termed pathogen-associated molecular patterns (pamps) and utilise conserved signaling pathways to activate pro-inflammatory cytokines and type-1 interferons. in spite of much understanding gained from the mammalian systems, many fish tlrs are unknown. recent studies in japanese flounder as well as in zebrafish suggested that the ligand binding and activation of in ...200717116407
molecular characterization of goose-type lysozyme homologue of large yellow croaker and its involvement in immune response induced by trivalent bacterial vaccine as an acute-phase protein.lysozyme acts as an innate immunity molecule against the invasion of bacterial pathogens. here, the cdna of a goose-type lysozyme (g-lysozyme) was cloned from large yellow croaker (pseudosciana crocea) by expressed sequence tags (est) and race-pcr techniques. the full-length cdna of large yellow croaker g-lysozyme (lycgl) is 716 nucleotides (nt) encoding a protein of 193 amino acids (aa), with a theoretical molecular weight of 21.3kda. the deduced lycgl possessed the typical structural features ...200717850883
molecular cloning and characterization of toll-like receptor 9 in japanese flounder, paralichthys olivaceus.toll-like receptor (tlr) 9 cdna and gene were cloned from japanese flounder, paralichthys olivaceus. the japanese flounder tlr9 cdna encodes 1065 amino acids. the leucine-rich domain (lrd) and the toll/interleukin-1 receptor (tir) domain found in other vertebrate tlr9s were conserved in japanese flounder tlr9. the gene is composed of three exons and two introns. the japanese flounder tumor necrosis factor (tnf) gene promoter was activated in japanese flounder tlr9-transformed hirame natural embr ...200717118454
cloning of atlantic halibut growth hormone receptor genes and quantitative gene expression during metamorphosis.to gain insight into the possible regulatory role of the growth hormone (gh)-insulin-like growth factor i (igf-i) system in flatfish metamorphosis, body ghr gene expression as well as igf-i protein content was quantified in larval atlantic halibut throughout metamorphosis (developmental stages 5-10). the cdna of the full-length gh receptor (hhghr) was cloned from adult liver and characterized. the hhghr shows common features of a ghr, including a (y/f)gefs motif in the extracellular domain, a si ...200717141236
anti-müllerian hormone (amh/amh) in the european sea bass: its gene structure, regulatory elements, and the expression of alternatively-spliced isoforms.in mammals, a multitude of studies have shown that anti-müllerian hormone (amh/amh), apart from inducing müllerian duct regression during male sexual differentiation, exerts inhibitory effects on male and female gonadal steroidogenesis and differentiation. however, in lower vertebrates like teleost fish, the function of amh/amh has been far less explored. as a first step to unravel its potential role in reproduction in teleost fish, we isolated and characterised the amh gene in the european sea ...200717157448
non-specific adsorption of fish immunoglobulin m (igm) to blocking reagents on elisa plate wells.enzyme-linked immunosorbent assay (elisa) is a popular technique for quantifiable detection of specific antibodies in warm-blooded animals, but it has not been accepted for detection of fish antibodies because of its low reproducibility, which is due in part to high background optical density (od) measurements. in the present study, we report that the high background of a fish antibody-detection elisa resulted from non-specific adsorption of fish immunoglobulin m (igm) to blocking reagents on th ...200718159673
molecular cloning, characterization, expression and functional analysis of japanese flounder paralichthys olivaceus fas ligand.we isolated and sequenced fas ligand cdna and its gene from japanese flounder (jf), paralichthys olivaceus. the jf-fas ligand cdna consisted of 1016 bp and encoded 230 amino acid residues. the identities of the deduced amino acid sequence of the jf-fas ligand to human fas ligand, tumor necrosis factor-alpha and lymphotoxin-alpha were 26.1%, 24.5% and 23.0%, respectively. a proline-rich domain (prd) that is important for localization of the protein was found in the n-terminal region, and two cyst ...200717197025
generation and characterization of monoclonal antibodies against the flounder paralichthys olivaceus rhabdovirus.two mabs (3c7 and 3c9) against flounder paralichthys olivaceus rhabdovirus (porv) were generated with hybridoma cell fusion technology and characterized by an indirect enzyme-linked immunosorbent assay, isotype test, western blot and immunodot analysis and immunofluorescence assay. isotyping tests demonstrated that both of the two mabs belonged to igm subclass. western blot analysis showed the mabs reacted with 42, 30, and 22 kda viral proteins, which were localized within the cytoplasm of porv- ...200818191468
a chlorophyll c2 analogue from the marine brown alga eisenia bicyclis inactivates the infectious hematopoietic necrosis virus, a fish rhabdovirus.we screened in vitro antiviral activity against a salmonid pathogenic virus, infectious hematopoietic necrosis virus (ihnv), from the extracts of a total of 342 species of marine algae collected from the japanese coastline. the anti-ihnv activity was found primarily in meoh extracts, and the extract from one marine brown alga in particular, eisenia bicyclis, showed high anti-ihnv activity. the anti-ihnv compound was isolated and purified as mc15 from the e. bicyclis extract, and the chemical str ...200717277903
dynamic expression pattern of the growth hormone receptor during early development of the chilean flounder.the entire cdna sequence of the growth hormone receptor (ghr) of the chilean flounder (paralichthys. adspersus) was cloned by rt-pcr and rna ligase rapid amplification of 5' and 3'ends. the deduced amino acid sequence contains 641 residues and codes for the ghr1 form. the receptor includes all the structural domains and motifs responsible for its interaction with the growth hormone and growth signal transduction. sequence comparison revealed 95 and 88% identity with other flat fish such as the j ...200818313961
profiles of alpha-melanocyte-stimulating hormone in the japanese flounder as revealed by a newly developed time-resolved fluoroimmunoassay and immunohistochemistry.profiles of alpha-melanocyte-stimulating hormone (alpha-msh) in the japanese flounder were examined by a newly developed time-resolved fluoroimmunoassay (tr-fia) and immunohistochemistry. a tr-fia for alpha-msh was newly developed, and its levels in the pituitary gland and plasma of japanese flounder reared in a white or black tank for 5 months were compared. a competitive assay using two antibodies was performed among secondary antibodies in the solid phase, alpha-msh antibodies, samples, and e ...200717286977
miamiensis avidus (ciliophora: scuticociliatida) causes systemic infection of olive flounder paralichthys olivaceus and is a senior synonym of philasterides dicentrarchi.the scuticociliate miamiensis avidus was isolated from olive flounder paralichthys olivaceus showing typical symptoms of ulceration and hemorrhages in skeletal muscle and fins. in an infection experiment, olive flounder (mean length: 14.9 cm; mean weight: 26.8 g) were immersion challenged with 2.0 x 10(3), 2.0 x 10(4) and 2.0 x 10(5) ciliates ml-1 of the cloned ys1 strain of m. avidus. cumulative mortalities were 85% in the 2.0 x 10(3) cells ml-1 treatment group and 100% in the other 2 infection ...200717330742
gene expression of leucocytes in vaccinated japanese flounder (paralichthys olivaceus) during the course of experimental infection with edwardsiella tarda.in this paper, we focused on the detection of differentially expressed genes in peripheral blood leucocytes (pbl) during the course of edwardsiella tarda infection in vaccinated and non-vaccinated japanese flounder (paralichthys olivaceus). cdna microarray analysis was performed to compare the gene expression patterns of the pbl between the vaccinated and non-vaccinated fish in response to e. tarda inoculation. fish were vaccinated twice, at a two-week interval and experimentally challenged with ...200717049272
cloning and expression analysis of myogenin from flounder (paralichthys olivaceus) and promoter analysis of muscle-specific expression.myogenin is a bhlh transcription factor of the myod family. it plays a crucial role in myoblast differentiation and maturation. we report here the isolation of flounder myogenin gene and the characterization of its expression patterns. sequence analysis indicated that flounder myogenin shared a similar structure and the conserved bhlh domain with other vertebrate myogenin genes. flounder myogenin gene contains 3 exons and 2 introns. sequence alignment and phylogenetic showed that flounder myogen ...200717336560
involvement of host recognition by oncomiracidia and post-larval survivability in the host specificity of heterobothrium okamotoi (monogenea: diclidophoridae).heterobothrium okamotoi, a monogenean gill parasite, shows high host specificity for tiger puffer, takifugu rubripes. in the present study, in vivo and in vitro experimental infections were conducted using various fish species, including t. rubripes, to understand the mechanisms of specificity. in in vivo experiments, t. rubripes, grass puffer, takifugu niphobles, olive flounder, paralichthys olivaceus, and red sea bream, pagrus major, were exposed to oncomiracidia of h. okamotoi labelled with a ...200717055510
identification and purification of a novel adhesion-associated protein in a new strain of lactobacillus, l15, from flounder (paralichthys olivaceus).adhesion is a crucial and prerequisite step for lactobacillus colonization in the digestive tract which subsequently confers its probiotic effects on the host. the aim of this report was to identify and purify a novel adhesion-associated protein which mediates the adherence of a new strain of lactobacillus, l15, to intestinal mucus from flounder. it was shown that surface proteins were involved in the adhesion of l15 to flounder mucus. the adhesion efficiencies of this strain were significantly ...200717350186
three different hepcidins from tilapia, oreochromis mossambicus: analysis of their expressions and biological functions.hepcidins are antimicrobial peptides that play important roles in resisting pathogenic infection. through hybridization of a phage library, the cdna sequences of three hepcidin-like antimicrobial peptides (named th1-5, th2-2, and th2-3) in tilapia, oreochromis mossambicus, were determined. the complete hepcidin cdna sequences of th1-5, th2-2, and th2-3 were respectively composed of 478, 533, and 583 bases, and contained a translated region of 88, 86, and 91 amino acids. an evolutionary assay of ...200717067680
difference in japanese flounder, paralichthys olivaceus gene expression profile following hirame rhabdovirus (hirrv) g and n protein dna vaccination.the glycoprotein (g protein) gene, but not other genes, of fish rhabdoviruses, when used as a dna vaccine was previously shown to be highly effective in inducing a protective immune response. in this study we used a dna microarray to examine differential gene expression in japanese flounder (paralichthys olivaceus) in response to a dna vaccine made from the genes of hirame rhabdovirus (rhabdovirus olivaceus) (hirrv) g protein (phrv-g) and nucleocapsid (n) protein (phrv-n). a high level of protec ...200717449275
teleostean il11b exhibits complementing function to il11a and expansive involvement in antibacterial and antiviral responses.interleukin 11 is a class-1 helical cytokine, having the four-helix bundle structure, possessing pleiotropic characteristics involved in physiological processes including blood production, bone formation and placentation. the interleukin 11 paralogues (il11a and il11b) have been identified in fish with only il11a from carp and trout have been characterized and analyzed for its expression thus far. here, we cloned and studied the structure and expression of il11b in japanese flounder (paralichthy ...200818538848
cloning and characterization of the ikappabalpha gene from japanese flounder, paralichthys olivaceus.we identified and characterized the japanese flounder (paralichthys olivaceus) inhibitor kappa b alpha (jfikba) cdna. the jfikba cdna contains an open reading frame of 960bp encoding 320 amino acid residues. jfikba contains 6 ankyrin repeats in the central coding region. expression studies by rt-pcr showed constitutive expression of the jfikba gene in several japanese flounder tissues (brain, muscle, gill, heart, kidney, liver, spleen and intestine). moreover, expression of jfikba mrna was induc ...200717478100
whitespotted puffer arothron hispidus, a new host for lymphocystis in qingdao aquarium of china.in april 2004 white nodular lesions were found on the fins of whitespotted puffer arothron hispidus (linnaeus). diagnostic studies were carried out to confirm the disease using light and electron microscopy, histochemical methods and pcr. the results revealed that the nodules were composed of giant cells up to 400 microm in diameter. these cells were surrounded by a periodic acid-schiff (pas)-positive hyaline capsule containing dot-shaped, feulgen-positive inclusion bodies in the cytoplasm and a ...200717523540
microarray analyses of gene expression in japanese flounder paralichthys olivaceus leucocytes during monogenean parasite neoheterobothrium hirame infection.in this study, the gene expression patterns of peripheral blood leucocytes (pbl) from japanese flounder paralichthys olivaceus were analyzed during the course of monogenean parasite neoheterobothrium hirame infection in order to select candidates for molecular biomarkers of infection. cdna microarray analysis was performed to compare the gene expression patterns of pbl between infected and non-infected fishes. among the 797 genes analyzed, 45 genes (5.6%) changed their expression levels. these g ...200717523546
identification of a novel c-type lectin gene in japanese flounder, paralichthys olivaceus.a novel c-type lectin of japanese flounder (jfctl) was identified among japanese flounder liver est. of 2083 of japanese flounder liver ests in the genbank database, 48 were identical to jfctl, suggesting that jfctl is abundantly produced in liver. jfctl cdna contains an open reading frame of 642 bp encoding 214 amino acids. a c-type lectin domain was conserved in the deduced amino acid sequence. the protein showed homology to transmembrane-type c-type lectin receptors rather than the soluble c- ...200717596963
response of neobenedenia girellae (monogenea) oncomiracidia to brightness and black-and-white contrast.neobenedenia girellae, a capsalid monogenean, is a significant pathogen due to both its ability to cause high mortality in fishes and its low host specificity. established control methods have both advantages and disadvantages. biological control measures with no unfavourable effects on the environment should be incorporated into the control strategy. the response of n. girellae oncomiracidia to brightness and black-and-white contrast was investigated to search for an alternative approach of dis ...200717599782
molecular cloning, promoter analysis and induced expression of the complement component c9 gene in the grass carp ctenopharyngodon idella.complement-mediated killing of pathogens through lytic pathway is an important effector mechanism of innate immune response. c9 is the ninth member of complement components, creating the membrane attack complex (mac). in the present study, a putative cdna sequence encoding the 650 amino acids of c9 and its genomic organization were identified in grass carp ctenopharyngodon idella. the deduced amino acid sequence of grass carp c9 (gcc9) showed 48% and 38.5% identity to japanese flounder and human ...200717604124
glyceraldehyde-3-phosphate dehydrogenase of edwardsiella tarda has protective antigenicity against vibrio anguillarum in japanese flounder.edwardsiella tarda glyceraldehyde-3-phosphate dehydrogenase (gapdh) may be an effective vaccine candidate against infection by e. tarda in japanese flounder paralichthys olivaceus. the gapdh of e. tarda is highly homologous to that of vibrio cholerae (91%), and therefore e. tarda gapdh may have protective antigenicity against vibrio species. in this study, we immunized japanese flounder with gapdh of e. tarda and infected the fish with v anguillarum. the result showed that gapdh prepared from e. ...200717629116
cloning and expression of partial japanese flounder (paralichthys olivaceus) igd.the cdna sequence of the japanese flounder (paralychthys olivaceus) igd has been previously reported (genbank accession no. ab052658) and this was followed by the detection of igd mrna expression in some flounder organ tissues. however, it has not been determined whether the flounder igd gene is virtually expressed into igd protein. to characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to igd ...200717669260
production, characterisation and applicability of monoclonal antibodies to immunoglobulin of japanese flounder (paralichthys olivaceus).immunoglobulin (ig) of japanese flounder (paralichthys olivaceus) was purified by a combination of salting-out and deae sepharose column chromatography. the purified immunoglobulin had an apparent molecular weight of 74 kda (heavy chain) and 24 kda (light chain) in sds-page. eighteen hybridomas secreting monoclonal antibodies (mabs) against japanese flounder ig were obtained by immunisation of balb/c mice with purified ig preparations, which were selected on the basis of the double indirect enzy ...200717719797
molecular cloning, expression, and functional analysis of caspase-10 from japanese flounder paralichthys olivaceus.we isolated and sequenced caspase-10 cdna and gene from japanese flounder, paralichthys olivaceus. the japanese flounder (jf)-caspase-10 cdna consisted of 2282 bp and encoded 495 amino acid residues. the characteristic death effector domains (deds) of caspases were observed in jf-caspase-10 as well as the three aspartic acid residues (d-186, -382 and -392), which are potential cleavage sites for the large and small subunit structures. the amino acid residue (his-325) and pentapeptide (qacqg), wh ...200717768069
antibiotic susceptibility and resistance of streptococcus iniae and streptococcus parauberis isolated from olive flounder (paralichthys olivaceus).the rates of antibiotic susceptibility and resistance were investigated in streptococcus iniae and streptococcus parauberis isolates obtained from diseased olive flounders (paralichthys olivaceus) collected from fish farms in jeju island, korea. isolates of s. iniae (n=65) were susceptible to cefotaxime, erythromycin, ofloxacin, penicillin, tetracycline and vancomycin, as demonstrated by the minimum inhibitory concentration (mic) test. isolates of s. parauberis (n=86) were highly resistant to er ...200919019569
characterization of thyroid hormones and thyroid hormone receptors during the early development of pacific bluefin tuna (thunnus orientalis).we studied the profiles of 3,5,3'-l-triiodothyronine (t3), thyroxine (t4), and thyroid hormone receptors (trs) in pacific bluefin tuna (thunnus orientalis) during embryonic and post-embryonic development. both t3 and t4 were detected in embryos just before hatching, and it was found that the levels of both were increased in postflexion fish. the thyroid follicles were increased in both size and number in postflexion fish compared with preflexion fish. a trbeta cdna clone was generated by race. t ...200817988667
growth differentiation factor 15, a novel acute phase response gene in japanese flounder, paralichthys olivaceus.the acute phase response, an important aspect of innate immunity, leads to the production of acute phase proteins (apps) in the liver which would consequently help restore homeostasis to the body. here, we identified a novel cytokine, growth differentiation factor 15 (gdf15) from japanese flounder. three out of the 384 est sequences derived from liver of japanese flounder treated with formalin-killed edwardsiella tarda showed significant homology with gdf of various species. after obtaining the ...200919059342
identification and characterization of the prepro-vasoactive intestinal peptide gene from the teleost paralichthys olivaceus.the gene encoding the prepro form of a vasoactive intestinal peptide (vip) was cloned, and its structural organization and expression profiles were determined in the teleost paralichthys olivaceus. the prepro-vip encodes both a vip and a peptide-histidine-isoleucine (phi). the vip gene comprises six exons with two distinct peptides encoded on separate exons, i.e., phi on exon iii and vip on exon iv. several elements involved in cytokine-mediated activation are highly conserved in an 806-bp segme ...200919084281
can the surface immobilization antigens of philasterides dicentrarchi (ciliophora: scuticociliatida) be used as target antigens to develop vaccines in cultured fish?in order to test whether immobilization antigens (i-antigens) of philasterides dicentrarchi could be suitable antigenic targets against scuticociliatosis, polyclonal olive flounder (paralichthys olivaceus) sera were raised against p. dicentrarchi by immunization with lysates of ciliates grown using chinook salmon epithelial (chse) cells, and the ability of the immune sera to kill the ciliates via classical complement pathway was analyzed in relation to agglutination activity. the immune sera sho ...200818023208
flatfish vitellogenin detection using optical waveguide lightmode spectroscopy-based immunosensor.a sensitive optical waveguide lightmode spectroscopy-based immunosensor was developed to detect vitellogenin in seawater flatfish (paralichthys olivaceus). for this purpose, anion-exchange column chromatography with de-52 resin was used to purify flatfish vitellogenin from flatfish serum containing vitellogenin that had been induced using an intraperitoneal 17beta-estradiol injection. the anti-flatfish vitellogenin antibody used as the biological component of the above immunosensor was prepared ...200718062221
in vitro and in vivo acute toxicity of fenpyroximate to flounder paralichthys olivaceus and its gill cell line fg.fenpyroximate, an acaricide, is widely used in the prevention of acarids (mites) in plants. in this study, the in vitro and in vivo acute toxicity of fenpyroximate was examined using the marine flounder paralichthys olivaceus and its gill cell line (fg). the 48h-ic(50) (95% confidence limits) values of fenpyroximate in the fg cells were 890 (790-990)nm, 950 (881-1019)nm and 1250 (1159-1341)nm, and 96h-ic(50) (95% confidence limits) were 480 (388-572)nm, 490 (454-526)nm and 510 (469-551)nm, for m ...200919185358
comparison of the responses of peritoneal macrophages from japanese flounder (paralichthys olivaceus) against high virulent and low virulent strains of edwardsiella tarda.in vivo infection studies in japanese flounder (paralichthys olivaceus) demonstrated that the number of viable cells of the virulent strain (nuf251) of edwardsiella tarda increased gradually in kidney and hepato-pancreas after intraperitoneal injection, but the low virulent strain (nuf194) did not. to gain insight into the virulence factors of e. tarda, in vitro responses of japanese flounder (p. olivaceus) peritoneal macrophages to these strains were compared in terms of phagocytosis, bacterici ...200818178102
[differential proteins revealed with proteomics in the brain tissue of paralichthys olivaceus under the stress of methyl parathion].the proteins from the brain tissue of the fish paralichtys olivaceus (po) were separated by the optimized two-dimensional polyacrylamide gel electrophoresis (2d-page). approximately 650 protein spots in each gel were detected from the brain sample when applying 200 mg protein to a 2d-page gel in the ph range of 3.0 - 10.0. the differential proteins from the po brain were separated and identified by proteomic technologies and they were further selected as the potential protein markers to monitor ...200819253540
evaluation of non-specific immune components from the skin mucus of olive flounder (paralichthys olivaceus).the innate immune system, particularly the external body surface, plays a frontier role in protecting fish under intensive aquaculture and at prolonged low temperatures from relevant infections due to inadequate adaptive immune responses. in the present study we aimed to understand the mucosal immunity of an economically important mariculture fish, olive flounder (paralichthys olivaceus) by evaluating the immune components from its skin mucus. the activities of lysozyme (233.33+/-171.82 units mg ...200818276162
formation and oral administration of alginate microspheres loaded with pdna coding for lymphocystis disease virus (lcdv) to japanese flounder.oral delivery of plasmid dna (pdna) is a desirable approach for fish immunization in intensive culture. however, its effectiveness is limited because of possible degradation of pdna in the fish's digestive system. in this report, alginate microspheres loaded with pdna coding for fish lymphocystis disease virus (lcdv) and green fluorescent protein were prepared with a modified oil containing water (w/o) emulsification method. yield, loading percent and encapsulation efficiency of alginate microsp ...200818328731
biological characterisation of a recombinant atlantic salmon type i interferon synthesized in escherichia coli.type i (alpha/beta) interferons (ifns) are a family of cytokines that stimulate the expression of numerous proteins that mediate an antiviral state in uninfected cells. two atlantic salmon (salmo salar) ifn-alpha (sasaifn-alpha1 & 2) genes have previously been cloned and both were found to contain a putative n-linked glycosylation site. recombinant sasaifn-alpha1 (rsasaifn-alpha1) produced in eukaryotic systems has repeatedly been shown to confer antiviral properties. however, different ifn-alph ...200818329900
cloning and characterization of phospholipase d from olive flounder (paralichthys olivaceus).the phospholipase d1 (pld1) cdna, designated popld, encoding a predicted protein of 1053 amino acids in olive flounder (paralichthys olivaceus) has been cloned. the deduced amino acid sequence shares high identity with that of pld1s and pld2 in human, rat and mouse. the phylogenic analysis and sequence comparison of popld with other pld isozymes were found to be closely related to the pld1 isozyme in primary structure. the tissue expression analysis of popld showed that the mrna of popld was pre ...200818329902
development and characterization of monoclonal antibodies to spring viraemia of carp virus.five monoclonal antibodies (mabs) against spring viraemia of carp (svcv0504, isolated from common carp in china) were produced from mice immunized with purified virus preparations. the virion of svcv contains five structural proteins, representing the nucleoprotein (n), phosphoprotein (p), matrix protein (m), glycoprotein (g) and rna-dependent rna polymerase (l). western blotting analysis revealed that three mabs (1h5, 1e10, and 1h7) recognized specifically to a single protein of 47kda (n), the ...200818378003
the formulation and immunisation of oral poly(dl-lactide-co-glycolide) microcapsules containing a plasmid vaccine against lymphocystis disease virus in japanese flounder (paralichthys olivaceus).nucleic acid-based immunotherapy is a new treatment option for fish immunisation in intensive culture. however, dna-based vaccines would be hydrolyzed or denaturized because of the existence of nucleases and severe gastrointestinal conditions. poly(dl-lactide-co-glycolide) (plga) microcapsules, loaded with plasmid dna (pdna) against lymphocystis disease virus (lcdv), were prepared by modified water in oil in water (w/o/w) double emulsion method in our laboratory. encapsulation efficiency, loadin ...200818442796
structural characterization of tn916-like element in streptococcus parauberis serotype ii strains isolated from diseased japanese flounder.to screen for the existence and determine the structure of tn916-like element in streptococcus parauberis serotype ii strains isolated from cultured japanese flounder in western japan.200919344360
genetic and phenotypic comparison of nocardia seriolae isolated from fish in japan.the phenotypic and genetic characterizations of 58 isolates of the fish pathogen nocardia seriolae, from amberjack, seriolae dumerili, yellowtail, seriola quinqueradiata, japanese flounder, paralichthys olivaceus, and chub mackerel, scomber japonicus, in japan from 1970-2005, were examined to investigate the epidemiological relationship between isolates. the phenotypic and genetic characterizations were determined by alpha-glucosidase activity and biased sinusoidal field gel electrophoresis (bsf ...200818482385
protection of japanese flounder (paralichthys olivaceus) against vibrio anguillarum with a dna vaccine containing the mutated zinc-metalloprotease gene.vibrio anguillarum is one of the causative agents of vibriosis, a systemic disease of fish characterized by acute hemorrhagic septicemia. the extracellular zinc metalloprotease (empa) is a putative virulence factor involved in pathogenicity of v. anguillarum. here we described the results of immunization against v. anguillarum with the plasmid expressing the mutated empa (m-empa7), which had no protelytic activity or cytotoxicity. in vitro protein expression of m-empa7 gene was determined by flu ...200919356619
genotoxic evaluation of the gh transgenic synechocystis using mice and turbot (scophthalmus maximus l.).in the present paper, the genotoxicity of transgenic synechocystis sp. pcc 6803 containing flounder (paralichthys olivaceus) growth hormone gene has been assessed using micronucleus tests in mice and fish. no micronucleus inductions in bone marrow erythrocyte of mice were observed at three doses of transgenic synechocystis (2.0, 5.0, and 10.0g/kg). the frequencies of micronucleus and other nuclear abnormalities in turbot (scophthalmus maximus l.) fed with 0.2%, 0.5% and 1.0% transgenic synechocy ...200818515177
effect of thyroid hormones on the stage-specific pigmentation of the japanese flounder paralichthys olivaceus.three experiments were carried out to clarify the effect of thyroid hormones on the pigmentation of larval japanese flounder paralichthys olivaceus. the first two experiments were conducted to investigate the critical concentration of thyroxine (t4) which causes abnormal pigmentation, and the third was to determine the sensitive stage of larval development at which albinism is induced by exogenous t4. the larvae treated with over 10 nm t4 showed a significant increase of albinism in experiments ...200018522465
altered behaviour and reduced survival of juvenile olive flounder, paralichthys olivaceus, infected by an invasive monogenean, neoheterobothrium hirame.neoheterobothriumhirame is a blood feeding monogenean of olive flounder paralichthys olivaceus. the parasite was first reported in the mid-1990s from the sea of japan and became epidemic within cultured and wild flounder populations after several years. infected fish often suffer from severe anaemia and thus the parasite is thought to have played an important role in the recent depletion of flounder populations in some areas of japan. however, the causal mechanism underlying the parasite epidemi ...200818561933
severe scuticociliate (philasterides dicentrarchi) infection in a population of sea dragons (phycodurus eques and phyllopteryx taeniolatus).scuticociliatosis is a disease of fish induced by ciliated parasites of the genus scuticociliatida. it has been described in sea horses (hippocampus sp.), flounders (paralichthys olivaceus), and turbots (scophthalmus maximus). here we present a case study of a population of sea dragons chronically infected with scuticociliates identified as philasterides dicentrarchi by histopathology and pcr. beginning in 2004, over a period of 19 months, 10 sea dragons (phycodurus eques and phyllopteryx taenio ...200818587104
expression, purification and structural characterization of recombinant hepcidin, an antimicrobial peptide identified in japanese flounder, paralichthys olivaceus.the cysteine-rich peptide hepcidin is an antimicrobial peptide and iron transport regulator that has been found in vertebrates including birds, fish and mammals. to elucidate the structure and biological function of fish hepcidin, which is difficult to produce synthetically, we have cloned several plasmid constructs encoding hepcidin from japanese flounder, paralichthys olivaceus, and tested expression of recombinant peptides, each with an n-terminal hexahistidine (6xhis) tag, in inclusion bodie ...200818595734
mhc polymorphism and disease resistance to vibrio anguillarum in 12 selective japanese flounder (paralichthys olivaceus) families.genetic variation in the major histocompatibility complex (mhc) class iib was tested in japanese flounder (paralichthys olivaceus) for survival after challenge with bacterial infection. the material consisted of 6000 japanese flounder from 60 families challenged with vibrio anguillarum, which causes significantly different mortality in flounder families. five individuals from each of six high-resistance (hr) and six low-resistance (lr) families were screened for their mhc class iib genotypes usi ...200818603444
distribution of marine birnavirus in cultured olive flounder paralichthys olivaceus in korea.surveys of marine birnavirus (mabv) were undertaken in cultured olive flounder paralichthys olivaceus from the south and west coastal areas and jeju in korea during the period january 1999 to april 2007. mabv was detected in all seasons from the fry, juveniles and adult fish from the areas examined. evident cytopathic effects of the virus including rounding and cell lysis were observed in chinook salmon embryo (chse-214) and rainbow trout gonad (rtg-2) cells, but not in fathead minnow (fhm) and ...200818604495
molecular cloning, tissue distribution and enzymatic characterization of cathepsin x from olive flounder (paralichthys olivaceus).in this study, we have cloned a cdna encoding for cathepsin x (poctx) from the olive flounder, paralichthys olivaceus. the presence of an hip motif, which is conserved in the unique cathepsin x family, poctx, clearly shows its relation to the cathepsin x group, apart from the cathepsin l or b subfamily. the results of rt-pcr and real-time pcr analyses revealed ubiquitous poctx expression in normal and lps-stimulated tissues. the cdna encoding for the proenzyme of poctx (propoctx) was expressed i ...200818674630
edwardsiella tarda t6ss component evpp is regulated by esrb and iron, and plays essential roles in the invasion of fish.edwardsiella tarda is a gram-negative pathogen for hemorrhagic septicemia in a broad range of hosts. the type vi secretion system (t6ss) has recently been dissected in e. tarda to secrete evpc, evpi and a novel effector protein evpp. in this study, sequencing and genetic alignments showed that evpp genes from different e. tarda isolates were highly similar and an evpp homolog was also found in aeromonas hydrophila 0865 isolated from a diseased eel, suggesting the possible lateral gene transfer o ...200919563898
duplication of phospholipase c-delta gene family in fish genomes.fishes possess more genes than other vertebrates, possibly because of a genome duplication event during the evolution of the teleost (ray-finned) fish lineage. to further explore this idea, we cloned five genes encoding phosphoinositide-specific phospholipase c-delta (plc-delta), designated respectively poplc-deltas, from olive flounder (paralichthys olivaceus), and we performed phylogenetic analysis and sequence comparison to compare our putative gene products (poplc-deltas) with the sequences ...200818722520
base changes in the flic gene of edwardsiella tarda: possible effects on flagellation and motility.edwardsiella tarda is a broad host-range pathogen infecting both animals and humans. e. tarda isolates from red sea bream pagrus major are non-motile, whereas isolates from japanese eel anguilla japonica and japanese flounder paralichthys olivaceus are motile with peritrichous flagella. we compared the flic gene coding for flagellin (flic) in motile and non-motile e. tarda strains isolated from diseased fish. twenty-two amino acid residues differed in the predicted flic amino acid sequences betw ...200717760384
identification of novel genes in japanese flounder (paralichthys olivaceus) head kidney up-regulated after vaccination with streptococcus iniae formalin-killed cells.streptococcosis caused by streptococcus iniae, a gram-positive bacterium, is one of the diseases often found in the culture of japanese flounder (paralichthys olivaceus). inactivated bacterial-whole cells of s. iniae have been used as a prophylactic treatment to prevent the disease. to understand the molecular mechanisms involved in activation of the fish immune system upon formalin-killed cell (fkc) treatment, we have investigated the gene transcription profile of japanese flounder head kidney ...200919038556
cdna cloning of japanese flounder stanniocalcin 2 and its mrna expression in a variety of tissues.stanniocalcin 1 (stc1) is a glycoprotein hormone important in the maintenance of calcium and phosphate homeostasis in both fishes and mammals. although two related stc genes, stc1 and stc2, were found to be expressed in multiple tissues as paracrine regulators in mammals, spatial expression pattern of stc2 mrna has not been elucidated in fishes in contrast to that of clearly described stc1. in the present study, we have cloned and characterized a full-length stc2 cdna from japanese flounder (par ...200919095074
modulation of the early immune response against viruses by a teleostean interferon regulatory factor-1 (irf-1).we investigated the role of a teleostean interferon regulatory factor-1 (irf-1) in the regulation of the fish immune system using japanese flounder, paralichthys olivaceus, as a model. fish were intramuscularly vaccinated with a recombinant plasmid expressing the japanese flounder irf-1 (jf irf-1) under the control of the cytomegalovirus immediate/early enhancer (cmv) promoter and were sampled at different days post-immunization. peripheral blood leukocytes (pbls) obtained from the jf irf-1-vacc ...200919101646
identification of vibrio anguillarum outer membrane vesicles related to immunostimulation in the japanese flounder, paralichthys olivaceus.we identified outer membrane vesicle (omv) production in vibrio anguillarum o1, a major fish pathogen that causes vibriosis, and characterized the omvs. they were produced during normal growth, and were appeared as spherical vesicle fractions. the protein profile of the omvs was similar to that of the outer membrane proteins, and the 38-kda major protein band of omv was identified as ompu. the omvs had enzyme activity of metalloprotease, hemolysin, and phospholipase, and stimulated the productio ...200919202277
phenotypic characteristics of streptococcus iniae and streptococcus parauberis isolated from olive flounder (paralichthys olivaceus).the etiological agents of streptococcosis were isolated from diseased olive flounder collected on the jeju island of korea. a total of 151 bacterial isolates were collected between 2003 and 2006. the isolates were examined using various phenotypic and proteomic analyses, including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page), immunoblotting, and glycoprotein assays. in addition, isolates were grown on blood agar to assess hemolytic activity, and biochemical assays were pe ...200919278524
identification and immunoprotective analysis of an in vivo-induced edwardsiella tarda antigen.edwardsiella tarda is a severe aquaculture pathogen that can infect many important fish species cultured worldwide. the aim of this study was to evaluate the vaccine potential of an e. tarda antigen, eta21, which was identified from a pathogenic e. tarda strain via the method of in vivo-induced antigen technology (iviat). eta21 is 510-amino acid in length and shares approximately 58% sequence identity with a putative peptidase of several bacterial species. eta21 was subcloned into escherichia co ...200919706328
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