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new insights into the evolution of ifns: zebrafish group ii ifns induce a rapid and transient expression of ifn-dependent genes and display powerful antiviral activities.the ifns and their receptors have existed in early chordates for approximately 500 million years and represent the early elements in innate and adaptive immunity. both types i and ii ifns have been discovered in fish, and type i has recently been classified into two groups based on their primary protein sequences. however, the biological activities of fish ifns and their roles in infection are largely unknown. using the zebrafish and manageable bacterial (streptococcus iniae) and viral (spring v ...200919265122
an immune responsive complement factor d/adipsin and kallikrein-like serine protease (podak) from the olive flounder paralichthys olivaceus.the cdna encoding of a complement factor d/adipsin and kallikrein-like serine protease, designated podak, was isolated from the olive flounder paralichthys olivaceus. podak cdna encodes a polypeptide with 277 amino acids containing conserved catalytic triad residues of serine proteases. the amino acid sequence of podak showed high similarity to the kallikrein-like protein of medaka, mammalian adipsin/complement factor d and tissue kallikrein homolog, kt-14 of trout, complement factor d of zebraf ...200919591942
host-directed evolution of a novel lactate oxidase in streptococcus iniae isolates from barramundi (lates calcarifer).in streptococcus iniae, lactate metabolism is dependent upon two proteins, lactate permease that mediates uptake and lactate oxidase, a flavin mononucleotide-dependent enzyme that catalyzes oxidation of alpha-hydroxyacids. a novel variant of the lactate oxidase gene, lcto, in australian isolates of s. iniae from diseased barramundi was found during a diagnostic screen using lox-1 and lox-2 primers, yielding amplicons of 920 bp instead of the expected 869 bp. sequencing of the novel gene variant ...200919270123
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
streptococcal cellulitis following preparation of fresh raw seafood.we describe three zoonotic streptococcal soft tissue infections resulting from fresh seafood contact. one was a localized thumb infection with streptococcus iniae in an immunocompetent healthy young male resulting from a puncture wound from a crab pincer. the other two were cases of ascending upper limb cellulitis associated with bacteraemia in mastectomy patients. one of these infections was caused by s. iniae while the other was caused by streptococcus dysgalactiae subsp. dysgalactiae, a speci ...200919309483
adult zebrafish model of streptococcal infection.streptococcal pathogens cause a wide array of clinical syndromes in humans, including invasive systemic infections resulting in high mortality rates. many of these pathogens are human specific, and therefore difficult to analyze in vivo using typical animal models, as these models rarely replicate what is observed in human infections. this unit describes the use of the zebrafish (danio rerio) as an animal model for streptococcal infection to analyze multiple disease states. this model closely mi ...200919412913
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
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
enhanced mortality in nile tilapia oreochromis niloticus following coinfections with ichthyophthiriasis and streptococcosis.ichthyophthirius multifiliis fouquet (ich) and streptococcus iniae are 2 major pathogens of cultured nile tilapia oreochromis niloticus (l). currently there is no information available for the effect of coinfection by ich and s. iniae on fish. the objective of this study was to determine the effects of parasite load and ich development size on fish mortality following s. iniae infection. low mortality (< or =20%) was observed in tilapia exposed to ich or s. iniae alone. mortalities increased fro ...200919750806
towards control of streptococcus iniae.streptococcus iniae is an emerging zoonotic pathogen; such infections generally occur through injuries associated with preparing whole fresh fish for cooking. those infected to date have been of asian descent, are usually elderly (average age 68 years), and have had >/=1 underlying conditions that may predispose them to infection. studies of the foundations of growth characteristics of s. iniae and its interactions with piscine host cells have recently been complemented by molecular studies. adv ...200919961667
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
evaluation of optimal culture conditions for recombinant ghost bacteria vaccine production with the antigen of streptococcus iniae gapdh.for the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in escherichia coli dh5alpha. ghost bacteria vaccine production from escherichia coli dh5alpha/phce-inan-gapdh-ghost 37 sdm (sig) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. the maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase (od600=2.0) with ...200919809256
determination of florfenicol dose rate in feed for control of mortality in nile tilapia infected with streptococcus iniae.a dose titration study was conducted to determine the dosage of florfenicol (ffc) in feed to control streptococcus iniae-associated mortality in nile tilapia oreochromis niloticus. six tanks were assigned to each of five treatments: (1) not challenged with s. iniae and fed unmedicated feed; (2) challenged with s. iniae by injection and fed unmedicated feed; (3) challenged with s. iniae and given ffc at 5 mg/kg of body weight (bw) in medicated feed; (4) challenged with s. iniae and given 10 mg ff ...201021192545
the novel polysaccharide deacetylase homologue pdi contributes to virulence of the aquatic pathogen streptococcus iniae.the aquatic zoonotic pathogen streptococcus iniae represents a threat to the worldwide aquaculture industry and poses a risk to humans who handle raw fish. because little is known about the mechanisms of s. iniae pathogenesis or virulence factors, we established a high-throughput system combining whole-genome pyrosequencing and transposon mutagenesis that allowed us to identify virulence proteins, including pdi, the polysaccharide deacetylase of s. iniae, that we describe here. using bioinformat ...201019762441
identification and immunoprotective analysis of a streptococcus iniae subunit vaccine candidate.streptococcus iniae is a gram-positive bacterium and a severe aquaculture pathogen that can infect a wide range of farmed fish species. in the summer of 2006, an epidemic broke out in a fish farm in north china, and examination of moribund fish (japanese flounder) identified the possible etiological agent of the outbreak as a strain named sf1, which exhibited apparent virulence in a japanese flounder infection model and conforms to the description of s. iniae by 16s rrna sequence analysis and ap ...201020096393
dried leaves of rosmarinus officinalis as a treatment for streptococcosis in tilapia.dietary application of dried rosmarinus officinalis (rosemary) leaves as a treatment for streptococcal infection was studied in tilapia, oreochromis sp. feeding with dried rosemary leaves significantly reduced mortality following infection with streptococcus iniae: 44% mortality in the group fed 8% rosemary, similar to oxytetracycline treatment (43% mortality), and significantly lower than the control (65%). dietary administration of 16% rosemary significantly reduced mortality because of strept ...201020158579
identification and molecular analysis of a stress-inducible hsp70 from sciaenops ocellatus.hsp70 proteins are a family of molecular chaperones that are involved in many aspects of protein homeostasis. in this study, an hsp70 homologue (sohsp70) was identified from red drum sciaenops ocellatus and analyzed at molecular level. the open reading frame of sohsp70 is 1920 bp and intronless, with a 5'-untranslated region (utr) of 399 bp and a 3'-utr of 241 bp. the deduced amino acid sequence of sohsp70 shares 84-92% overall identities with the hsp70s of a number of fish species. in silico an ...201020541016
lactobacillus sakei bk19 enriched diet enhances the immunity status and disease resistance to streptococcosis infection in kelp grouper, epinephelus bruneus.the effect of lactobacillus sakei bk19 (10(8) cells g(-1)) supplemented diet fed to kelp grouper, epinephelus bruneus against streptococcosis caused by streptococcus iniae and streptococcus parauberis with reference to the innate immune response and disease resistance was evaluated at 1, 2, and 4 weeks. maximum reduction in mortalities was observed in kelper feeding the probiotic diet for two weeks after challenged with the pathogens when compared to the infected group fed with basal diet; simil ...201020816809
differential regulation of sciaenops ocellatus viperin expression by intracellular and extracellular bacterial pathogens.viperin is an antiviral protein that has been found to exist in diverse vertebrate organisms and is involved in innate immunity against the infection of a wide range of viruses. however, it is largely unclear as to the potential role played by viperin in bacterial infection. in this study, we identified the red drum sciaenops ocellatus viperin gene (sovip) and analyzed its expression in relation to bacterial challenge. the complete gene of sovip is 2570 bp in length and contains six exons and fi ...201020420911
molecular cloning and expression analysis of interferon regulatory factor 10 (irf10) in japanese flounder, paralichthys olivaceus.interferons (ifns) are important for the defense against intracellular pathogens. interferon regulatory factor 10 (irf10) is a transcript factor involved in regulating ifn signaling induced by virus infections in birds, but little is know of its role in the immune response in non-avian vertebrates. here, we identified and characterized the irf10 gene and examined its expression pattern in a teleost fish, japanese flounder, paralichthys olivaceus. japanese flounder irf10 (poirf10) gene is a singl ...201020883793
characterization and gene expression of transcription factors, pu.1 and c/ebpα driving transcription from the tumor necrosis factor α promoter in japanese flounder, paralichthys olivaceus.both pu.1 and c/ebpα transcription factors play important roles in myeloid development and inflammatory response. these transcripts were cloned from the japanese flounder (paralichthys olivaceus) and were highly conserved with those of other vertebrates. pu.1 mrna was mainly expressed in lymphoid tissues while c/ebpα mrna was widely expressed in all tissues examined. higher levels of pu.1 mrna were expressed in the igm(+) cells of both pbl and kl, while c/ebpα expression was higher only in the i ...201020951726
a commercial rapid optical immunoassay detects streptococcus agalactiae from aquatic cultures and clinical specimens.the biostar strep b optical immunoassay (strep b oia) (biostar oia strep b assay kit; biostar incorporation, louisville, co, usa), commonly used for diagnosis of human maternal group b streptococcus (gbs) colonization, was evaluated for its diagnostic and analytical sensitivity and specificity to aquatic animal gbs isolates, cross-reactivity, and diagnosis and recovery of gbs directly from clinically- infected fish swabs. strep b oia identified 25 known fish and dolphin gbs isolates. thirteen no ...201020430538
dietary administration of nucleotides to enhance growth, humoral immune responses, and disease resistance of the rainbow trout (oncorhynchus mykiss) fingerlings.a feeding study was conducted to determine the effect of dietary nucleotides (nt) on growth performance, immune responses and resistance of rainbow trout fingerlings to streptococcus iniae. a basal diet supplemented with 0 (control), 0.5, 1, 1.5 and 2 g nt kg(-1) to formulate five experimental diets. each diet was randomly allocated to triplicate groups of fish with initial average weight of approximately 23 g. after 8 weeks of feeding trial, fish fed diets with 1.5-2 g nt kg(-1) had the highest ...201020955799
mtsb, a hydrophobic membrane protein of streptococcus iniae, is an effective subunit vaccine candidate.streptococcus iniae is a major bacterium that causes invasive disease in cultured fish worldwide. the protection relies mainly on anti-microbial compounds and vaccines, and there is much interest in developing s. iniae vaccine based on conserved protein immunogens. subcellular localization of protein has important influence on its immunogenicity. the surface and extracellular proteins of pathogenic bacteria can be easily recognized by the infected host compare to intracellular proteins, which ar ...201021111784
immunomodulatory effect of sodium alginate enriched diet in kelp grouper epinephelus brneus against streptococcus iniae.the effect of diets containing sodium alginate at 0, 0.5, 1.0, and 2.0 g kg⁻¹ following challenge with streptococcus iniae in kelp grouper epinephelus bruneus were assessed with reference to survival rate and innate immune parameters such as alternative complement, lysozyme, natural haemagglutination, respiratory burst, superoxide dismutase, and phagocytic activities on week 1, 2, and 4. fish fed with sodium alginate containing diet at 1.0 and 2.0 g kg⁻¹ after being challenged with s. iniae had ...201021130885
characterization and expression analysis of a goose-type lysozyme from the rock bream oplegnathus fasciatus, and antimicrobial activity of its recombinant protein.lysozyme (muramidase) represents an important defense molecule of the fish innate immune system. known for its bactericidal properties, lysozyme catalyzes the hydrolysis of β-(1,4)-glycosidic bonds between the n-acetyl glucosamine and n-acetyl muramic acid in the peptidoglycan layer of bacterial cell walls. in this study, the complete coding sequence of a g-type lysozyme (rbglyz) was identified in the oplegnathus fasciatus rock bream fish genome by means of multi-tissue normalized cdna pyroseque ...201021167286
molecular cloning and characterization of lps-binding protein/bactericidal permeability-increasing protein (lbp/bpi) from olive flounder, paralichthys olivaceus.a lipopolysaccharide (lps)-binding protein/bactericidal permeability-increasing protein (lbp/bpi) homolog was isolated from peripheral blood leukocytes cdna library of olive flounder paralichthys olivaceus. the isolated lbp/bpi cdna is 2806bp in length with a 1419bp open reading frame (orf) that encodes a protein of 472 amino acid residues. the lps-binding domain is well conserved in the n-terminal barrel, showing high sequence identities with other teleost lbp/bpi as well as those of mammals. r ...201019698997
dietary administration of zooshikella sp. enhance the innate immune response and disease resistance of paralichthys olivaceus against sreptococcus iniae.we report the growth, innate immune response, and disease resistance in olive flounder (paralichthys olivaceus) challenged with streptococcus iniae after feeding with diet enriched with zooshikella sp. strain je-34 three different concentration i.e. low (3.4 x 10(4), n = 50), medium (3.5 x 10(6), n = 50), and high (3.4 x 10(8), n = 50) cfu ml(-1) supplemented diets, the changes were monitored on weeks 1, 2, 4, 8, 12, and 16. with all diets the innate immune parameters, such as superoxide anion p ...201020206273
protection against heterologous streptococcus iniae isolates using a modified bacterin vaccine in nile tilapia, oreochromis niloticus (l.).streptococcus iniae is a significant pathogen impacting aquaculture production worldwide. the objectives of this study were to determine whether a developed modified s. iniae (ars-98-60) bacterin vaccine is efficacious in nile tilapia, oreochromis niloticus (l.), against challenge with heterologous isolates from diverse geographical locations and to evaluate protein and antigenic variability among the isolates tested. two groups of tilapia (approximately 5 g) were intraperitoneally (ip) vaccinat ...201020298447
construction and analysis of an experimental streptococcus iniae dna vaccine.streptococcus iniae is a severe aquaculture pathogen that can also infect humans and animal. a putative secretory antigen, sia10, was identified from a pathogenic s. iniae strain by in vivo-induced antigen technology. using turbot as an animal model, the immunoprotective effect of sia10 was examined as a dna vaccine in the form of plasmid psia10, which expresses sia10 under the cytomegalovirus immediate-early promoter. in fish vaccinated with psia10, transcription of sia10 was detected in muscle ...201020394724
evaluation of streptococcus iniae killed bacterin and live attenuated vaccines in hybrid striped bass through injection and bath immersion.streptococcus iniae poses a serious threat to finfish aquaculture operations worldwide. stringent regulatory standards limit the use of antibiotics to treat s. iniae infections; improved vaccination strategies are thus of great interest. we investigated the potential for efficient, non-injectable batch vaccination via the use of live attenuated vaccines. three attenuated s. iniae strains with genetic mutations eliminating the production of virulence factors--capsular polysaccharide (delta cpsd), ...201020402229
a pivotal role for the streptococcus iniae extracellular polysaccharide in triggering proinflammatory cytokines transcription and inducing death in rainbow trout.streptococcus iniae is a major pathogen of fish, causing considerable economic losses in israel, the united states and the far east. containment of mortalities through vaccination was recently compromised due to the emergence of novel vaccine-escape strains that are distinguished from previous strains by their ability to produce large amounts of extracellular polysaccharide (eps) that is released to the medium. in vitro and in vivo data now indicate that the eps is a major virulence factor, capa ...201020199577
effectiveness of aquaflor (50% florfenicol) to control mortality associated with streptococcus iniae in freshwater-reared subadult sunshine bass.we conducted a field trial to evaluate the effectiveness of aquaflor (50% florfenicol) for controlling mortality associated with streptococcus iniae in freshwater-reared subadult sunshine bass (female white bass morone chrysops x male striped bass m. saxatilis). bacterial samples collected from moribund fish representing a reference population were presumptively identified microbiologically and were later confirmed to be s. iniae by biochemical characterization and polymerase chain reaction. the ...201021413510
first report of streptococcus iniae in red porgy (pagrus pagrus, l.).this work describes the first isolation of streptococcus iniae in red porgy, pagrus pagrus (l.), and the first european isolation of this pathogen in gilthead seabream, sparus aurata (l.). in both farmed fish species, infection resulted in lethargy, anorexia, abnormal swimming, exophthalmia and sudden death, with mortality rates of over 25% in red porgy and 10% in gilthead seabream. beta-haemolytic gram-positive cocci, catalase negative and oxidase negative, were isolated in pure culture from in ...201021504081
identification and molecular analysis of a novel c-type lectin from scophthalmus maximus.c-type lectins are calcium-dependent carbohydrate-binding proteins that play important roles in innate immunity. in this study, a c-type lectin homologue (smlec1) was identified from turbot (scophthalmus maximus) and analyzed at expression and functional levels. the open reading frame of smlec1 is 504 bp, with a 5'-untranslated region (utr) of 101 bp and a 3'-utr of 164 bp. the deduced amino acid sequence of smlec1 shares 34%-38% overall identities with the c-type lectins of several fish species ...201020226863
a solute-binding protein for iron transport in streptococcus iniae.streptococcus iniae (s. iniae) is a major pathogen that causes considerable morbidity and mortality in cultured fish worldwide. the pathogen's ability to adapt to the host affects the extent of infection, hence understanding the mechanisms by which s. iniae overcomes physiological stresses during infection will help to identify potential virulence determinants of streptococcal infection. grow s. iniae under iron-restricted conditions is one approach for identifying host-specific protein expressi ...201021122131
in vitro antibacterial activities of ethanol extract of iranian propolis (eeip) against fish pathogenic bacteria (aeromonas hydrophila, yersinia ruckeri & streptococcus iniae).the "in vitro" antibacterial activity of ethanol extract of propolis (eeip) from urmia, iran was investigated against three prevalent species of fish bacterial pathogens including: aeromonas hydrophila lmg 3770, yersinia ruckeri lmg 3279 and streptococcus iniae lmg 14520. in this study two standard susceptibility testing techniques (micro-broth dilution method and agar-well diffusion method) were used to evaluation of the antibacterial activity of eeip against the mentioned micro-organisms. also ...201024031591
molecular epidemiology of zoonotic streptococcosis/lactococcosis in rainbow trout (oncorhynchus mykiss) aquaculture in iran.streptococcosis/lactococcosis is a hyperacute systemic disease that can occur in marine and fresh waters of many species of fish. the aim of this work was to study the disease outbreak in the major rainbow trout (oncorhynchus mykiss) production of iran.201022347573
molecular identification and expression analysis of a natural killer cell enhancing factor (nkef) from rock bream oplegnathus fasciatus and the biological activity of its recombinant protein.natural killer cell enhancing factor (nkef) belongs to the defined peroxiredoxin (prx) family. rock bream nkef cdna was identified by expressed sequence tag (est) analysis of rock bream liver that was stimulated with the lps. the full-length rbnkef cdna (1062 bp) contained an open reading frame (orf) of 594 bp encoding 198 amino acids. rbnkef was significantly expressed in the gill, liver, and intestine. mrna expression of nkef in the head kidney was examined under viral and bacterial challenge ...201124371552
molecular characterisation and expression analysis of a fish-egg lectin in rock bream, and its response to bacterial or viral infection.a fish-egg lectin, rbfel, was identified from rock bream (oplegnathus fasciatus) and its expression analysed. in both vertebrates and invertebrates, carbohydrate-binding proteins (lectins) play an important role in innate immunity against microbial invasion. here, we report the cloning of a fish-egg lectin from rock bream using a combination of expression sequence tag (est) analyses. the full-length cdna of rbfel is composed of 1512-bp with a 780-bp orf that encodes 259 amino acid residues. the ...201122056500
salt effect of nisin z isolated from a marine fish on the growth inhibition of streptococcus iniae, a pathogen of streptococcosis.a bacteriocin-producing lactococcus lactis subsp. lactis was isolated from the intestine of olive flounder. the bacteriocin was identified as nisin z. it was active against gram-positive bacteria. nisin z at 3,200 arbitrary units (au) was more effective in seawater than in pbs; growth of streptococcus iniae was completely inhibited within 3 h. nisin z preparations with 3.5% (w/v) nacl was the most effective against s. iniae being similar to nisin z in seawater. nisin z is thus a good alternative ...201122042302
Efficacy of QCDCR formulated CpG ODN 2007 in Nile tilapia against Streptococcus iniae and identification of upregulated genes.The potential of using a QCDCR (quilA:cholesterol:dimethyl dioctadecyl ammonium bromide:carbopol:R1005 glycolipid) formulated CpG oligodeoxynucleotide (ODN), ODN 2007, to confer protection in Nile tilapia against Streptococcus iniae infection was evaluated in this study. At two days post treatment, QCDCR formulated ODN 2007 elicited significant (P<0.05) protection to Nile tilapia, with relative percent survival of 63% compared to fish treated by QCDCR alone. To understand the molecular mechanism ...201122129787
Molecular cloning and functional analysis of nucleotide-binding oligomerization domain 1 (NOD1) in olive flounder, Paralichthys olivaceus.The gene encoding nucleotide-binding oligomerization domain 1 (NOD1) was cloned from olive flounder (Paralichthys olivaceus) and the role played by NOD1 during Edwardsiella tarda infection was evaluated. The complete open reading frame of NOD1 was 2820bp in length, encoding a 939-amino acid polypeptide. The NOD1 protein contains three conserved domain structures including C-terminal LRRs, a central NACHT motif, and an N-terminal CARD domain, which show similarities of 49-74% to those of other ve ...201122142819
Distinct signaling mechanisms mediate neutrophil attraction to bacterial infection and tissue injury.The signals that guide neutrophils to sites of tissue injury or infection remain elusive. H(2) O(2) has been implicated in neutrophil sensing of tissue injury and transformed cell, however its role in neutrophil recruitment to infection has not been explored. Here, using a pharmacological inhibitor of NADPH oxidases, diphenyleneiodonium (DPI), and genetic depletion of an epithelial specific NADPH oxidase, we show that H(2) O(2) is not required for neutrophil detection of localized infection wi ...201122188170
The Streptococcus iniae transcriptional regulator CpsY is required for protection from neutrophil-mediated killing and proper growth in vitro.The ability of a pathogen to metabolically adapt to the local environment for optimal expression of virulence determinants is a continued area of research. Orthologs of the Streptococcus iniae LysR family regulator CpsY have been shown to regulate methionine biosynthesis and uptake pathways but appear to influence expression of several virulence genes as well. An S. iniae mutant with an in-frame deletion of cpsY (?cpsY mutant) is highly attenuated in a zebrafish infection model. The ?cpsY mutant ...201121911465
immunoproteomic analysis of the antibody response obtained in nile tilapia following vaccination with a streptococcus iniae vaccine.streptococcus iniae is one of the most economically important gram-positive pathogens in cultured fish species worldwide. the usda-ars aquatic animal health research unit developed a modified (contains concentrated culture supernatant) s. iniae bacterin that has been demonstrated to be efficacious, and protection is mediated by specific anti-s. iniae antibodies. although effective, the specific vaccine components important for efficacy are not known. in the present study, an immunoproteomic appr ...201121601381
a holistic view of the dynamisms of teleost igm: a case study of streptococcus iniae vaccinated rainbow trout (oncorhynchus mykiss).to date, little is known about how trout igm, the primary antibody of fish, varies in titer, specificity, disulfide cross-linking, and affinity following immunization with a pathogen. work using defined antigens has demonstrated that the disulfide cross-linking structure of igm becomes increasingly more polymerized during an immune response, coinciding with an increase in affinity, but it is unknown if this has relevance to aquatic pathogens. understanding how igm varies following vaccination wi ...201121641928
molecular cloning and characterization of toll-like receptor 14 in japanese flounder, paralichthys olivaceus.toll-like receptors (tlrs) are essential for activation of the innate immune system in response to invading pathogens. tlr14, which is unique to fish, has been identified in several fish species, but its function is unclear. in this study, japanese flounder (paralichthys olivaceus) tlr14 gene (jftlr14) was cloned and its expression profiles were analyzed after infection with viral hemorrhagic septicemia virus, gram-positive streptococcus iniae and gram-negative edwardsiella tarda. the coding reg ...201120728538
identification and expression profile of multiple genes in nile tilapia in response to formalin killed streptococcus iniae vaccination.twenty-eight expressed sequence tags (ests) were isolated from a nile tilapia (oreochromis niloticus) vaccinated vs non-vaccinated subtractive library at 12-h post injection of a formalin killed streptococcus iniae ars-98-60 vaccine. the 28 ests were classified in terms of their putative functions. half of the ests identified were unknown proteins. of the remaining half ests, 17% have putative functions in protein biosynthesis and 11% have putative functions in immunity, energy production, and s ...201121640389
rapid and sensitive detection of streptococcus iniae by loop-mediated isothermal amplification (lamp). 201121401641
molecular characterization, expression and functional analysis of a nuclear oligomerization domain proteins subfamily c (nlrc) in japanese flounder (paralichthys olivaceus).pattern recognition receptors (prrs) are involved in the effective innate defense against several microbes. here, we identified a nucleotide-oligomerization domain (nod)-like receptor subfamily c (nlrc) from japanese flounder (paralichthys olivaceus). full-length transcript of jfnlrc is composed of 3976bp encoding a protein of 1175 deduced amino acid residues. the presence of a signature nucleotide-binding domain (nacht) and leucine-rich repeated domain (lrr) suggested that the protein is a memb ...201121642003
rapid identification of streptococcus iniae by specific pcr assay utilizing genetic markers in its rdna.the 16s-23s intergenic spacers (its) of ribosomal dna from ten independent isolates of streptococcus iniae and one reference strain atcc29178 were sequenced, aligned and used to design a polymerase chain reaction (pcr) primer set for rapid and specific detection and identification of s. iniae. this primer set amplified a 377-bp dna fragment specifically from s. iniae, but not from other common bacterial pathogens of fish or from non-fish pathogens. the pcr conditions were optimized to allow dete ...201121294750
soft tissue infections caused by marine bacterial pathogens: epidemiology, diagnosis, and management.skin and soft tissue infections (sstis) are one of the most common infection syndromes and may be caused by a large number of microorganisms. some principles of aquatic injuries are different than those of land-based trauma. wounds sustained in marine environment are exposed to a milieu of bacteria rarely encountered in different settings. these include vibrio spp., aeromonas spp., shewanella spp., erysipelothrix rhusiopathiae, mycobacterium marinum, streptococcus iniae, and other microbes. fail ...201121785929
development of loop-mediated isothermal amplification method for rapid detection of streptococcus iniae, the causative agent of streptococcicosis in fish.streptococcus iniae is a major pathogen that causes sever economic losses in tilapia aquaculture. a set of four specific primers was designed by targeting lcto gene. with bst dna polymerase, the target dna can be clearly amplified for 60 min at 64 °c in a simple water bath. the sensitivity of the lamp assay for the detection of s. iniae is about 12.4 cells per reaction in both of pure cultures and added fish tissues cultures. lamp products could be judged with agar gel or naked eye after additio ...201121780152
molecular identification and expression analysis of cathepsins o and s from rock bream, oplegnathus fasciatus.cathepsins are lysosomal cysteine proteases belonging to the papain family, members of which play important roles in normal metabolism for maintenance of cellular homeostasis. rock bream (oplegnathus fasciatus) cathepsin o and s (rbctso and rbctss, respectively) cdnas were identified by expressed sequence tag (est) analysis of a lipopolysaccharide (lps)-stimulated rock bream liver cdna library. the full-length rbctso cdna (1698 bp) contained an open reading frame (orf) of 1017 bp encoding 338 am ...201121767649
a multivalent killed whole-cell vaccine induces effective protection against edwardsiella tarda and vibrio anguillarum.edwardsiella tarda, vibrio anguillarum, streptococcus iniae, and vibrio harveyi are important aquaculture pathogens that affect a wide range of fish species. in this study, we examined in a japanese founder (paralichthys olivaceus) model the possibility of developing simple, killed whole-cell vaccines with protective effect against two or more of the above pathogens. for this purpose, we utilized four pathogenic fish isolates: e. tarda tx1, v. anguillarum c312, s. iniae sf1, and v. harveyi t4d. ...201121763772
differential expression and intrachromosomal evolution of the sghc1q genes in zebrafish (danio rerio).the secreted globular head c1q (sghc1q) genes can be characterized as a family of genetic loci encoding signal peptides followed by single complement component 1q globular (gc1q) motifs. members of this family have been referred to as precerebellin-like (cblnl), c1q-like or ovary specific c1q-like factors, and are transcribed in response to infection and/or during early development. this study was primarily undertaken to identify the zebrafish sghc1q (or drsghc1q) genes that increase their trans ...201121683091
identification and expression profiles of multiple genes in nile tilapia in response to bacterial infections.to understand the molecular mechanisms involved in response of nile tilapia (oreochromis niloticus) to bacterial infection, suppression subtractive cdna hybridization technique was used to identify upregulated genes in the posterior kidney of nile tilapia at 6h post infection with aeromonas hydrophila. a total of 31 unique expressed sequence tags (ests) were identified from 192 clones of the subtractive cdna library. quantitative pcr revealed that nine of the 31 ests were significantly (p<0.05) ...201121840065
development and efficacy of a novobiocin-resistant streptococcus iniae as a novel vaccine in nile tilapia (oreochromis niloticus).a novel attenuated streptococcus iniae vaccine was developed from a virulent strain of streptococcus iniae (iset0901) through selection for novobiocin resistance (named isno). the safety of isno was then evaluated in nile tilapia (oreochromis niloticus) through intraperitoneal (ip) injection. when male tilapia (average weight 10g) were ip injected with 2+ù10(7) colony-forming units (cfu) of the attenuated s. iniae vaccine strain, no fish died. however, when the same age and size matched tilapia ...201121708205
molecular cloning and expression analysis of two distinct f-type lectins from the rock bream, oplegnathus fasciatus.several lectin families characterized by distinct signature sequence motifs and structural folds, such as c-type, peptidoglycan recognition protein, ficolin, pentraxins, and most recently galectins, have been implicated in immune surveillance. in this study, two distinct f-type lectins rbftl-1 and rbftl-2, from the rock bream (oplegnathus fasciatus), were identified and their expression was analyzed. the full-length cdna of rbftl-1 was composed of 1204bp with a 945-bp open reading frame (orf) th ...201121683734
pathological changes in cultured channel catfish ictalurus punctatus spontaneously infected with streptococcus iniae.the pathological changes present in channel catfish ictalurus punctatus spontaneously infected by streptococcus iniae are described. the most consistent gross findings were marked petechial hemorrhages of the skin and congestion of internal organs, particularly the liver, spleen and kidneys. other features included color fading at the edge of fin rays, enteritis and ascites. histological examination showed oedema, degeneration and necrotic changes in many organs. further, hepatitis, splenitis, i ...201121932531
intraspecific and interspecific interactions among proteins regulating exopolysaccharide synthesis in streptococcus thermophilus, streptococcus iniae, and lactococcus lactis subsp. cremoris and the assessment of potential lateral gene transfer.using the yeast two-hybrid system, intraspecific protein interactions were detected in streptococcus iniae and lactococcus lactis subsp. cremoris between the transmembrane activation protein (cpsc and epsa, respectively) and the protein tyrosine kinase (cpsd and epsb, respectively), between two protein tyrosine kinases, and between the protein tyrosine kinase and the phosphotyrosine phosphatase (cpsb and epsc, respectively). for each of these intraspecific interactions, interspecific interac ...201122107596
molecular cloning and characterisation of the rock bream, oplegnathus fasciatus, fas (cd95/apo-1), and its expression analysis in response to bacterial or viral infection.fas belongs to the tumour necrosis factor (tnf) receptor superfamily and can transmit a death signal leading to apoptosis. in the present study, we isolated the full-length cdna for rock bream (oplegnathus fasciatus) fas (rbfas). the full-length rbfas cdna was 1770 bp long and contained an open reading frame of 957 bp that encoded 319 amino acid residues with a predicted molecular mass of 35.1 kda. the 319 amino-acid predicted rbfas sequence is homologous to other fas sequences, contains three c ...201124371547
construction and comparative study of monovalent and multivalent dna vaccines against streptococcus iniae.streptococcus iniae is an important fish pathogen with a broad host range that includes both marine and freshwater fish species. with an aim to develop effective vaccines against s. iniae, we in this study constructed three monovalent dna vaccines, i.e., psagf, psagg, and psagi, based on sagf, g, and i, which are components of the streptolysin s cluster. the immunoprotective potentials of these vaccines were examined in a model of japanese flounder (paralichthys olivaceus). the results showed th ...201223063784
effects of dietary supplementation of inosine monophosphate on growth performance, innate immunity and disease resistance of olive flounder (paralichthys olivaceus).this study was investigated to examine the effects of dietary inosine monophosphate (imp) supplementation on growth performance, feed utilization, innate immunity, hematological parameters and disease resistance of juvenile olive flounder. five experimental diets were formulated to contain imp at levels of 0, 0.1, 0.2, 0.4 and 1.0%. all diets were maintained isonitrogenous (48% crude protein) and isocaloric (20.7 mj/kg diet). triplicate groups of olive flounder (initial body weight, 7.5 ± 0.02 g ...201222986588
a divalent dna vaccine based on sia10 and ompu induces cross protection against streptococcus iniae and vibrio anguillarum in japanese flounder.streptococcosis and vibriosis caused by streptococcus iniae and vibrio anguillarum respectively have affected fish culture industries around the world. previous studies have indicated that the s. iniae antigen sia10 and the v. anguillarum outer membrane protein ompu, when used as dna vaccines, induce protection in turbot (scophthalmus maximus) and asian seabass (lates calcarifer) respectively. in this study, with an effort to develop effective vaccines against s. iniae and v. anguillarum, we con ...201222480661
streptococcus iniae outbreaks in brazilian nile tilapia (oreochromis niloticus l:) farms.this is the first report of outbreaks of streptococcus iniae in nile tilapia farms in south america. seven isolates were identified by biochemical, serological and molecular tests. their 16s rrna gene sequences showed 100% similarity with s. iniae atcc 29178 and two distinct pfge patterns were observed for brazilian isolates.201224031866
cpsy influences streptococcus iniae cell wall adaptations important for neutrophil intracellular survival.the ability of a pathogen to evade neutrophil phagocytic killing mechanisms is critically important for dissemination and establishment of a systemic infection. understanding how pathogens overcome these innate defenses is essential for the development of optimal therapeutic strategies for invasive infections. cpsy is a conserved transcriptional regulator previously identified as an important virulence determinant for systemic infection of streptococcus iniae. while orthologs of cpsy have been a ...201222354020
evolution of the capsular operon of streptococcus iniae in response to vaccination.streptococcus iniae causes severe septicemia and meningitis in farmed fish and is also occasionally zoonotic. vaccination against s. iniae is problematic, with frequent breakdown of protection in vaccinated fish. the major protective antigens in s. iniae are the polysaccharides of the capsule, which are essential for virulence. capsular biosynthesis is driven and regulated by a 21-kb operon comprising up to 20 genes. in a long-term study, we have sequenced the capsular operon of strains that hav ...201223001668
in vitro study of adherence, invasion, and persistence of streptococcus iniae in fibroblastic-like fish cell line saf-1.streptococcus iniae is a major fish pathogen producing invasive infections that result in economic losses in aquaculture. gentamicin protection assays were used to investigate the ability of different s. iniae strains to invade and adhere to fibroblastic-like fish cell line saf-1. all strains tested were detected intracellularly using both techniques, with variable internalization degrees between strains. the experiments carried out at 4°c demonstrated that active cell metabolism is necessary fo ...201222897319
channel catfish (ictalurus punctatus) protein disulphide isomerase, pdia6: molecular characterization and expression regulated by bacteria and virus inoculation.protein disulfide isomerases (pdis) are thought to aid protein folding and assembly by catalyzing formation and shuffling of cysteine disulfide bonds in the endoplasmic reticulum (er). currently, increasing evidence suggests pdis play an important role in host cell invasion and they are relevant targets for the host immune response. however the roles of specific pdis in teleosts are little known. here, we characterized the protein disulfide isomerase family a, member 6 (pdia6) from channel catfi ...201222561356
genetic characterization of streptococcus iniae in diseased farmed rainbow trout (onchorhynchus mykiss) in iran.genetic characterization of strains of streptococcus iniae recovered from morbidity and mortality of farmed rainbow trout in different provinces of iran were studied. the gram-positive cocci isolates were obtained from the kidney tissues of diseased rainbow trout on blood agar at 25°c for 72 h. the grown bacteria were then characterized using biochemical and molecular works. the identified 26 isolates of s. iniae producing a 513 bp in pcr procedure were then compared using random amplified polym ...201222550467
Phenoloxidase in the scallop Chlamys farreri: Purification and antibacterial activity of its reaction products generated in vitro.Phenoloxidase (PO) was purified from hemocytes of the scallop Chlamys farreri using native-PAGE and gel permeation column chromatography, and then substrate specificity and antibacterial activity generated from reaction products of purified PO were analyzed. The results showed purified PO had a molecular mass of 576 kDa in native-PAGE and 53 kDa in denatured PAGE, and could catalyze the substrates L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, catechol and hydroquinone suggesting it is a type ...201222119576
simultaneous detection of multiple fish pathogens using a naked-eye readable dna microarray.we coupled 16s rdna pcr and dna hybridization technology to construct a microarray for simultaneous detection and discrimination of eight fish pathogens (aeromonas hydrophila, edwardsiella tarda, flavobacterium columnare, lactococcus garvieae, photobacterium damselae, pseudomonas anguilliseptica, streptococcus iniae and vibrio anguillarum) commonly encountered in aquaculture. the array comprised short oligonucleotide probes (30 mer) complementary to the polymorphic regions of 16s rrna genes for ...201222736973
cloning and localization of mcdef, a defensin from manila clams (ruditapes philippinarum).a defensin-like peptide was previously detected in hemocytes of manila clams (ruditapes philippinarum). in the current study, we cloned and characterized this defensin, designated mcdef. cloning produced a full-length gene sequence of 201 bp predicted to encode a 66-amino-acid precursor protein maturing to a 44-amino-acid residue. amino acid sequence analysis showed that mcdef is similar to defensins from marine mollusks and ticks. phylogenetic analysis suggested that mcdef is closely related to ...201221945146
identification of bacillus strains for biological control of catfish pathogens.bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against edwardsiella ictaluri and aeromonas hydrophila, the causative agents of enteric septicemia of catfish (esc) and motile aeromonad septicaemia (mas), respectively. twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including ed ...201223029244
purification and characterization of yfgap, a gapdh-related novel antimicrobial peptide, from the skin of yellowfin tuna, thunnus albacares.a 3.4 kda of antimicrobial peptide was purified from an acidified skin extract of the yellowfin tuna, thunnus albacares, by preparative acid-urea-polyacrylamide gel electrophoresis and c(18) reversed-phase hplc. a comparison of the n-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the n-terminus of glyceraldehyde-3-phosphate dehydrogenase (gapdh); thus, this peptide was designated as the yellowfin tuna gapdh-related antimicro ...201222771964
purification and characterization of an antimicrobial histone h1-like protein and its gene from the testes of olive flounder, paralichthys olivaceus.an approximately 21 kda antimicrobial protein was purified from an acidified testis extract of olive flounder, paralichthys olivaceus, by ion-exchange and c(18) reversed-phase hplc. a comparison of the n-terminal amino acid sequence with those of other known antimicrobial polypeptides revealed high homology between this antimicrobial protein and other histone h1 molecules; thus, it was designated flounder histone h1-like protein (fh1lp). fh1lp showed potent antimicrobial activity against gram-po ...201222538352
Expressed sequence tags (ESTs) based identification of genes and expression analysis of leukocyte cell-derived chemotaxin-2 (LECT2) from Epinephelus bruneus.The kelp grouper, Epinephelus bruneus, is an economically important intensively cultured species in Southeast Asia. Despite the insatiable demand its large-scale production has been hindered by problems associated with water quality, nutrition, and diseases especially due to increased rearing density. It is generally accepted that in fish both innate and adaptive immune system provide protection from diseases. In the present study a cDNA library of Streptococcus iniae-challenged kelp grouper was ...201221983421
streptococcus hongkongensis sp. nov., isolated from a patient with an infected puncture wound and from a marine flatfish.a bacterium, hku30(t), was isolated from the infected tissue of a patient with wound infection after puncture by a fish fin. cells are facultative anaerobic, non-spore-forming, non-motile, gram-positive cocci arranged in chains. colonies were non-haemolytic. the strain was catalase, oxidase, urease and voges-proskauer test negative. it reacted with lancefield's group g antisera and was resistant to optochin. it grew on bile aesculin agar and in 5 % nacl. it was unidentified by three commercial i ...201223264498
expression profiles of nods in channel catfish (ictalurus punctatus) after infection with edwardsiella tarda, aeromonas hydrophila, streptococcus iniae and channel catfish hemorrhage reovirus.nlrs are a large family of intracellular pathogen recognition receptors (prrs) that recognize pathogen associated molecular patterns (pamps). in the previous study, the identification of nlrs subfamily a (nods) and gene expression was carried out in channel catfish (ictalurus punctatus). however, the gene expression profiles of channel catfish nods (nod1, nod2, nlrc3, nlrc5 and nlrx1) after infection with various bacteria and virus are still unclear. in this study, expression of five nods genes ...201222796486
isolation and identification of flavobacterium columnare and streptococcus iniae antibacterial compounds from the terrestrial plant atraphaxis laetevirens.columnaris disease, enteric septicemia of catfish, and streptococcosis are common bacterial diseases of certain freshwater fish and are caused by flavobacterium columnare , edwardsiella ictaluri , and streptococcus iniae , respectively. during the process of evaluating several species of plants to isolate and identify compounds with toxicity against these bacteria, a promising extract from the aerial parts of the terrestrial plant atraphaxis laetevirens (ledeb.) jaub. et spach (polygonaceae juss ...201223030835
interferon regulatory factors 4 and 8 in rock bream, oplegnathus fasciatus: structural and expressional evidence for their antimicrobial role in teleosts.the interferon regulatory factor (irf) members irf4 and irf8 contribute to b-lymphocyte development and can act as regulators of immunoglobulin (ig) light chain gene transcription. these two irfs are closely interrelated and are expressed at high levels in the lymphoid and myeloid cells of the immune system. in this study, the complete cdna and genomic sequences of rock bream irf4 (rbirf4) and irf8 (rbirf8) were identified by homology screening of a multi-tissue normalized cdna library and a bac ...201222885028
a streptococcus iniae dna vaccine delivered by a live attenuated edwardsiella tarda via natural infection induces cross-genus protection.edwardsiella tarda and streptococcus iniae are important fish pathogens. we have reported previously a live e. tarda vaccine based on the attenuated strain tx5rm and a s. iniae dna vaccine based on the antigen sia10. in this study, we examined the possibility of constructing a cross-genus vaccine by taking advantage of the residual infectivity of tx5rm and using it as a carrier host for the natural delivery of a s. iniae dna vaccine. for this purpose, the recombinant tx5rm, tx5rms10, was created ...201222967257
a teleostean angiotensinogen from oplegnathus fasciatus responses to immune and injury challenges.angiotensinogen (agt) is the precursor of the renin-angiotensin system and contributes to osmoregulation, acute-phase and immune responses. a full-length cdna of the agt (2004 bp with a 1389 bp coding region) was isolated from rock bream (rb), oplegnathus fasciatus. the encoded polypeptide of 463 amino acids had a predicted molecular mass of 51.6 kda. rbagt possessed a deduced signal peptide of 22 residues upstream of a putative angiotensin i sequence ((23)nrvyvhpfhl(32)). rbagt possessed a spec ...201222300787
class ii, major histocompatibility complex, transactivator (ciita) in channel catfish: identification and expression patterns responding to different pathogens.nlrs are a large family belonging to pattern recognition receptors which could recognize pathogen associated molecular patterns. class ii, major histocompatibility complex, transactivator (ciita) is a member of nlr family. it is a critical transcription factor which could regulate the expression of mhc class ii. in this study, a full-length cdna of ciita was cloned from channel catfish according to ten sequenced ests. this cdna contains a 5'-utr of 71 bp, a 3'-utr of 238 bp and an orf of 3,210 b ...201223073771
edwardsiella tarda dnak: expression, activity, and the basis for the construction of a bivalent live vaccine against e. tarda and streptococcus iniae.edwardsiella tarda and streptococcus iniae are important aquaculture pathogens that affect many species of farmed fish. in this study, we analyzed the expression, activity, and immunoprotective potential of e. tarda heat shock protein dnak. we found that dnak expression was upregulated under conditions of heat shock, oxidative stress, and infection of host cells. recombinant dnak (rdnak) purified from escherichia coli exhibited atpase activity and induced protection in japanese flounder (paralic ...201222281608
genomic characterization and expression analysis of complement component 9 in rock bream (oplegnathus fasciatus).the complement component 9 (c9) is a single-chain glycoprotein that mediates formation of the membrane attack complex (mac) on the surface of target cells. full-length c9 sequence was identified from a cdna library of rock bream (oplegnathus fasciatus), and its genomic sequence was obtained by screening and sequencing of a bacterial artificial chromosome (bac) genomic dna library of rock bream. the rock bream complement component 9 (rb-c9) gene contains 11 exons and 10 introns and is composed of ...201222796422
molecular identification and expression analysis of two distinct bpi/lbps (bactericidal permeability-increasing protein/lps-binding protein) from rock bream, oplegnathus fasciatus.we identified two cdnas designated as rbbpi/lbp-1 and rbbpi/lbp2, respectively, which were identified by expressed sequence tag (est) analysis of a lipopolysaccharide (lps)-stimulated rock bream liver cdna library. the two cdna displayed 36.9% identity at the translated amino acid level. despite the low level of identity between the two genes, high conservation was seen in the bpi/lbp/cetp n-terminal, lps-binding, the proline-rich central and the bpi/lbp/cetp c-terminal domains. the full-length ...201222521422
birc7 gene in channel catfish (ictalurus punctatus): identification and expression analysis in response to edwardsiella tarda, streptococcus iniae and channel catfish hemorrhage reovirus.a family member of inhibitor of apoptosis protein (iap) termed baculoviral iap repeat-containing 7 (birc7) from channel catfish (ictalurus punctatus) was identified, the full length cdna sequence of channel catfish birc7 (ccbirc7) was 1686 bp, containing a 5'utr of 93 bp, a 3'utr of 399 bp with a poly (a) tail and an orf of 1194 bp encoding a putative protein of 398 amino acids. the putative ccbirc7 protein contains two bir super-family conservative domains and a c-terminal ring finger motif. ph ...201222510211
molecular cloning and functional characterization of two duplicated two-cysteine containing type i interferon genes in rock bream oplegnathus fasciatus.two type i interferon (ifn) genes, designated as rbifn1 and rbifn2, have been cloned and characterized in rock bream. they are both comprised of 5 exons and 4 introns, and are closely linked on the rock bream chromosome in a unique head-to-head configuration. both genes encode 183 amino acid (aa) precursor with a putative 17 aa signal peptide in the n-terminal. only one amino acid divergence is present between two ifns. compared with the type i ifns in higher vertebrates, two rock bream ifns pos ...201222889848
a novel acute phase reactant, serum amyloid a-like 1, from oplegnathus fasciatus: genomic and molecular characterization and transcriptional expression analysis.acute phase response is a significant component of innate immunity, playing a vital role in the signaling processes and elimination of invading pathogens. acute phase proteins are synthesized in liver and secreted into the blood for transportation to an infection site, where the defense function is exerted. serum amyloid a (saa) and c-reactive proteins are the major positive acute phase proteins. in this study, we have identified and characterized a novel saa related gene from rock bream (oplegn ...201222504166
japanese flounder (paralichthys olivaceus) hsp70: adjuvant effect and its dependence on the intrinsic atpase activity.heat shock protein (hsp) 70 is a molecular chaperone that plays an important role in protein folding and transport. in addition, hsp70 is also involved in regulation of innate and adaptive immune response. in this study, we examined the biological activity and the immunomodulatory property of an hsp70 homologue, pohsp70, from japanese flounder (paralichthys olivaceus). recombinant pohsp70 purified from escherichia coli exhibits apparent atpase activity; however, a mutant pohsp70, pohsp70m, that ...201222898664
molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, epinephelus coioides.lysozymes are key proteins of the host innate immune system against pathogen infection. in this study, a c-type lysozyme gene (ec-lysc) was cloned and characterized from orange-spotted grouper, epinephelus coioides. the full-length ec-lysc cdna is composed of 533 bp and encodes a polypeptide of 144-residue protein with 94% identity to lysc of kelp grouper, epinephelus bruneus. the genomic dna of ec-lysc consists of 4 exons and 3 introns, with a total length of 1897 bp. amino acid sequence alignm ...201222579550
pcr detection and pfge genotype analyses of streptococcal clinical isolates from tilapia in china.large-scale streptococcal outbreaks occurred continuously in tilapia farms of china from 2009 to 2011. the objective of this study was to characterize the prevalent strains of tilapia streptococci from the main cultured areas of china through species specific pcr and pulse field gel electrophoresis (pfge). a total of 105 prevalent strains were isolated from guangdong, guangxi, hainan and fujian provinces between 2006 and 2011, 85 of which were identified as streptococcus agalactiae while the res ...201222677479
evaluation of colorimetric loop-mediated isothermal amplification assay for visual detection of streptococcus agalactiae and streptococcus iniae in tilapia.streptococcus agalactiae and strep. iniae are bacterial pathogens that cause streptococcosis in many fish species. an accelerated colorimetric loop-mediated isothermal amplification (lamp) assay with pre-addition of calcein was established, and the transmission and detection of strep. agalactiae and strep. iniae in tilapia under natural aquatic environment were investigated. a positive reaction was observed by a colour change from orange to green through the naked eyes after completion at 63°c f ...201323746201
evaluation of reference genes for quantitative real-time rt-pcr analysis of gene expression in nile tilapia (oreochromis niloticus).quantitative real-time reverse-transcriptase polymerase chain reaction (rt-qpcr) has been used frequently to study gene expression related to fish immunology. in such studies, a stable reference gene should be selected to correct the expression of the target gene. in this study, seven candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase (gadph), ubiquitin-conjugating enzyme (ubce), 18s ribosomal rna (18s rrna), beta-2-microglobulin (b2m), elongation factor 1 alpha (ef1a), tubulin ...201323792389
lactococcus lactis bfe920 activates the innate immune system of olive flounder (paralichthys olivaceus), resulting in protection against streptococcus iniae infection and enhancing feed efficiency and weight gain in large-scale field studies.the protective effect of a food-grade lactic acid bacterium lactococcus lactis bfe920 against disease of olive flounder (paralichthys olivaceus) cultivated on a large scale was studied. initially, antimicrobial activity of l. lactis against several fish pathogens was evaluated in vitro; the probiotic showed strong antibacterial activity against streptococcus iniae, streptococcus parauberis and enterococcus viikkiensis, and moderate activity against lactococcus garviae. when olive flounders were ...201324041843
molecular characterisation and biological activity of a novel cxc chemokine gene in rock bream (oplegnathus fasciatus).chemokines are chemoattractant cytokines defined by the presence of four conserved cysteine residues. in mammals, these cytokines can be divided into four subfamilies depending on the arrangement of the first two conserved cysteines in the sequence, and include the cxc(α), cc(β), c(γ), and cx3c(δ) classes. we identified cxc chemokine cdna, designated rbcxc, isolated using expressed sequence tag analysis of a lipopolysaccharide (lps)-stimulated rock bream liver cdna library. the full-length rbcxc ...201323376472
molecular identification and expression analysis of the cc chemokine gene in rock bream (oplegnathus fasciatus) and the biological activity of the recombinant protein.we identified the cc chemokine cdna designated as rbcc1 (cc chemokine 1 in rock bream, oplegnathus fasciatus), which was isolated using expressed sequence tag (est) analysis of a lipopolysaccharide (lps)-stimulated rock bream liver cdna library. the full-length rbcc1 cdna (850 bp) contained an open reading frame (orf) of 366 bp encoding 122 amino acids. results from our phylogenetic analysis demonstrated that the rbcc1 was closest relationship to the orange-spotted grouper and mi-iyu croaker cc ...201323357024
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