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[comparison of characteristics of svcv strains isolated in china and in europe].in the nationwide epidemiological investigation, svcv-741 was for the first time isolated in beijing region, china in 2003, and designated as svcv asian strain. in this paper, we compared svcv-741 (asian strains isolated in china) with svcv-10/3 (europe reference strain) on their physico-chemical, biological and morphological characteristics. the results indicated that there were no distinct differences between two svcv strains on phycico-chemical and morphological characteristics. the main exis ...200919437886
cost analysis of multiplex pcr testing for diagnosing respiratory virus infections.we performed a cost analysis study using decision tree modeling to determine whether the use of multiplex pcr testing for respiratory viruses (xtag rvp test) is a more or less costly strategy than the status quo testing methods used for the diagnosis of respiratory virus infections in pediatric patients. the decision tree model was constructed by using four testing strategies for respiratory virus detection, viz., direct fluorescent-antibody staining (dfa) alone, dfa plus shell vial culture (svc ...200919571025
phylogenetic analysis of spring viraemia of carp virus isolates from austria indicates the existence of at least two subgroups within genogroup id.genetic relationships between 22 spring viraemia of carp virus (svcv) isolates from austria collected between 1994 and 2007 were determined based on the partial nucleotide sequence of the glycoprotein gene (g gene). phylogenetic analyses located all austrian isolates except one in genogroup id. one isolate collected in 2007 was placed within the svcv ia genogroup. more importantly, the study also revealed 3 distinct clusters within genogroup id, designated id1, id2 and id3. existence of subgroup ...200919593931
characterization of the complete genome sequence of pike fry rhabdovirus.the complete genome sequence of pike fry rhabdovirus (pfrv), consisting of 11,097 nucleotides, was determined. the genome contains five genes, encoding the nucleoprotein (n), phosphoprotein (p), matrix protein (m), glycoprotein (g), and rna-dependent rna polymerase (l) protein in the order 3'-n-p-m-g-l-5'. 3' leader- and 5' trailer-sequences in the pfrv genome show inverse complementarity. the pfrv proteins share the highest homology to the proteins of spring viremia of carp virus (svcv), rangin ...200919603256
molecular analysis of spring viraemia of carp virus in china: a fatal aquatic viral disease that might spread in east asian.spring viraemia of carp (svc) is a fatal viral disease for cyprinid fish, which is caused by spring viraemia of carp virus (svcv). to date, no svc outbreak has been reported in china. between 1998 and 2002, outbreaks of svc were reported in ornamental and wild fish in europe and america, imported from multiple sources including china. based on phylogenetic analysis, the viral strain isolated from america was shown to be originated from asia. these outbreaks not only resulted in huge economic los ...200919623265
cloning of common carp socs-3 gene and its expression during embryogenesis, gh-transgene and viral infection.as a member of a newly discovered protein family, the suppressor of cytokine signalling 3 (socs-3) has been shown to regulate the responses of many immune cytokines in a negative auto-regulatory manner. the full-length cdna of common carp socs-3 was 1603 bp and contained a 630 bp open reading frame (orf) coding for a protein of 209 amino acids. carp socs-3 molecule was well conserved especially in the src homology 2 (sh2) and the socs box. the kinase inhibitory region (kir) and ess domains, upst ...201020025976
zebrafish larvae are unable to mount a protective antiviral response against waterborne infection by spring viremia of carp virus.interferons (ifns) and their receptors exist in all classes of vertebrates, where they represent early elements in innate and adaptive immunity. both types i and ii ifns have been discovered in fish and type i ifn has recently been classified into two groups based on their primary protein sequences and biological activities. thus, although groups i and ii zebrafish ifn show powerful antiviral activities, only group i (ifnphi1) is able to protect the fish against bacterial infection. in addition, ...201020045026
prospective follow-up of patients with acute hepatitis c virus infection in brazil.the natural outcome of infection with hepatitis c virus (hcv) varies substantially among individuals. however, little is known about host and viral factors associated with a self-limiting or chronic evolution of hcv infection.201020235831
assessing the impact of climate change on disease emergence in freshwater fish in the united kingdom.a risk framework has been developed to examine the influence of climate change on disease emergence in the united kingdom. the fish immune response and the replication of pathogens are often correlated with water temperature, which manifest as temperature ranges for infection and clinical diseases. these data are reviewed for the major endemic and exotic disease threats to freshwater fish. increasing water temperatures will shift the balance in favour of either the host or pathogen, changing the ...201020561287
development and evaluation of a sensitive and quantitative assay for hirame rhabdovirus based on quantitative rt-pcr.the aim of the present work was to develop a quantitative reverse-transcription polymerase chain reaction (qrt-pcr) assay using a taqman probe to detect and quantify hirame rhabdovirus (hrv). the results demonstrated that the assay had a detection limit of 100 copies of rna per reaction and a log-linear range up to 10(8) copies of hrv rna. regression analysis demonstrated a significant correlation with an r(2) value of 0.9963 and a slope of -3.18 between the mean c(t) values and hrv crna. this a ...201020723563
a rational design for hepatitis b virus x protein refolding and bioprocess development guided by second virial coefficient studies.the hepatitis b virus x (hbx) protein is well known for its role in hepatitis b virus infection that often leads to hepatocellular carcinoma. despite the clinical importance of hbx, there is little progress in anti-hbx drug development strategies due to shortage of hbx from native sources. consistent expression of hbx as insoluble inclusion bodies within various expression systems has largely hindered hbx manufacturing via economical biosynthesis routes. confronted by this roadblock, this study ...201121212947
identification and functional characterization of dicer2 and five single vwc domain proteins of litopenaeus vannamei.dicer (dcr) is the key protein of the rna interference (rnai) pathway. to investigate the role of the rnai pathway in shrimp anti-viral immunity, litopenaeus vannamei dcr2 (designated as lvdcr2) was identified and characterized. the full-length cdna of lvdcr2 was 5513bp long, with an open reading frame encoding a putative protein of 1502 amino acids. in addition, five proteins homologous to the single von willebrand factor type c (vwc) domain protein (svc) were also identified in l. vannamei and ...201121256150
a comparative review on european-farmed finfish rna viruses and their vaccines.the diseases causing the highest ecological and socio-economical impacts in european farmed finfish are produced by rna viruses. salmon, trout, sea bream, sea bass, carp and turbot, suffer viral nervous necrosis produced by betanodaviruses (vnnv), infectious pancreatic necrosis produced by aquabirnaviruses (ipnv), viral haemorrhagic septicemia (vhsv) and infectious haematopoietic necrosis (ihnv) produced by novirhabdoviruses, spring viremia of carp produced by vesicular-like rhabdoviruses (svcv) ...201121320546
genes and hepatitis c: susceptibility, fibrosis progression and response to treatment.hepatitis c virus contact and infection show three different phenotypes: spontaneous viral clearance (svc), chronic hepatitis c (chc) and sustained virological response (svr) following antiviral treatment. many factors, including genetics, influence the evolution of these three phenotypes. we performed a literature search (pubmed) up to 31 january 2010 without language restriction to identify relevant studies on genes and hepatitis c. additional studies were sought by reviewing the reference lis ...201121382156
establishment and characterization of a new cell line derived from kidney of grouper, epinephelus akaara (temminck & schlegel), susceptible to singapore grouper iridovirus (sgiv).a marine fish cell line derived from the kidney of red-spotted grouper, epinephelus akaara, designated as eagk was established and characterized. the eagk cells multiplied well in leibovitz's l-15 medium containing 10% foetal bovine serum at 25ôçâ-¦c and have been subcultured for more than 90 passages. karyotyping, chromosomal typing and ribosomal rna (rrna) genotyping analysis revealed that eagk had a modal diploid chromosome number of 82 and was a fibroblast cell line originated from grouper. ...201121838711
antibody recognition of the glycoprotein g of viral haemorrhagic septicemia virus (vhsv) purified in large amounts from insect larvae.abstract:201121693048
Down-regulation of heme oxygenase-1 by SVCV infection.Heme oxygenase (HO-1) is a cytoprotective enzyme that plays a critical role in defending the body against oxidant-induced injury during inflammatory processes. In mammalian systems, viral infection or antigen expression can down-regulate the expression of HO-1. In turn, the induction of HO-1 or overexpression of HO-1 results in potent and direct antiviral activity that targets the replication of several mammalian viruses. In this study, the HO-1 gene of Cyprinus carpio was cloned, and the expres ...201222146702
first detection of pike fry-like rhabdovirus in barbel and spring viraemia of carp virus in sturgeon and pike in aquaculture in the czech republic.rapid antigen detection enzyme-linked immunosorbent assay (elisa) testing of cell cultures with organ homogenate from fish, collected from farms with a predominance of common carp or in natural aquaculture in the czech republic between 1995 and 2008, identified piscine vesiculovirus in 27 of 178 samples. using reverse transcription semi-nested pcr, targeting a 550 nucleotide region of the glycoprotein (g) gene, piscine vesiculovirus was confirmed in 23 of the 27 organ samples diagnosed by elisa ...201121848117
genetic diversity of perch rhabdoviruses isolates based on the nucleoprotein and glycoprotein genes.despite the increasing impact of rhabdoviruses in european percid farming, the diversity of the viral populations is still poorly investigated. to address this issue, we sequenced the partial nucleoprotein (n) and complete glycoprotein (g) genes of nine rhabdoviruses isolated from perch (perca fluviatilis) between 1999 and 2010, mostly from france, and analyzed six of them by immunofluorescence antibody test (ifat). using two rabbit antisera raised against either the reference perch rhabdovirus ...201121927897
development and validation of a reverse transcription quantitative pcr for universal detection of viral hemorrhagic septicemia virus.viral hemorrhagic septicemia virus (vhsv) infects over 70 fish species inhabiting marine, brackish or freshwater environments throughout the northern hemisphere. over its geographic range, 4 vhsv genotypes and multiple subtypes exist. here, we describe the development and validation of a rapid, sensitive and specific real-time reverse transcription quantitative pcr assay (rt-qpcr) that amplifies sequence from representative isolates of all vhsv genotypes (i, ii, iii and iv). the pan-specific vhs ...201121848118
molecular evolutionary and epidemiological dynamics of a highly pathogenic fish rhabdovirus, the spring viremia of carp virus (svcv).spring viremia of carp virus (svcv), which causes an acute hemorrhagic and highly contagious disease in cyprinids, was first described in europe and subsequently reported in parts of asia and north america. svcv can be classified into four genogroups: ia, ib, ic, and id. while ia and id have wide circulation and are reported to cause outbreaks in north america and europe, respectively, ib and ic were last reported in the 1980s. we used a bayesian framework to determine the nucleotide substitutio ...201122033044
Molecular diagnosis of respiratory virus infections.The appearance of eight new respiratory viruses, including the SARS coronavirus in 2003 and swine-origin influenza A/H1N1 in 2009, in the human population in the past nine years has tested the ability of virology laboratories to develop diagnostic tests to identify these viruses. Nucleic acid based amplification tests (NATs) for respiratory viruses were first introduced two decades ago and today are utilized for the detection of both conventional and emerging viruses. These tests are more sensit ...201122185616
a higher correlation of hcv core antigen with cd4+ t cell counts compared with hcv rna in hcv/hiv-1 coinfected patients.development of hcv infection is typically followed by chronic hepatitis c (chc) in most patients, while spontaneous hcv viral clearance (svc) occurs in only a minority of subjects. compared with the widespread application of hcv rna testing by quantitative rt-pcr technique, hcv core antigen detection may be an alternative indicator in the diagnosis of hepatitis c virus infections and in monitoring the status of infectious individuals. however, the correlation and differences between these two in ...201121858166
long-term outcome of hepatitis c virus infections acquired after pediatric liver transplantation.the outcomes and characterization of hepatitis c virus (hcv) infections after pediatric liver transplantation (lt) have rarely been reported. we describe our experience with hcv infections after pediatric lt. ten of 207 children (4.8%) who underwent lt at our institution (1985-2010) developed previously undiagnosed hcv disease. eight received a liver graft before blood product and donor screening for hcv became available. the mean age at transplantation was 8.9 ± 4.3 years, and the median time f ...201121932378
establishment, characterization and viral susceptibility of a new cell line derived from goldfish, carassius auratus (l.), tail fin.a continuous cell line [goldfish tail fin (gftf)] derived from a goldfish tail fin, carassius auratus, was established and characterized. gftf cells predominantly consist of fibroblast-like cells that were maintained and subcultured more than 50 times over a period of 15 months. cells grew at temperatures between 15 and 37°c, with an optimum temperature of 25°c. the growth rate of gftf cells increased proportionally with the foetal bovine serum (fbs) concentration (5-20%), with optimum growth at ...201121916901
characterization and virus susceptibility of a skin cell line from red-spotted grouper (epinephelus akaara).a red-spotted grouper epinephelus akaara skin (rgs) cell line was established and characterized. rgs cells had a normal diploid chromosome number of 2n = 48, the morphology of which was fibroblastic-like in 3 days and epithelial-like over 5 after 16 passages. the cells multiplied well in dulbecco's modified eagle's medium supplemented with 10% of fetal bovine serum at 25°c. susceptibilities of rgs and grass carp ovary (gco) cells to two viruses were tested, and the results showed that the titer ...201222252337
concentration of infectious aquatic rhabdoviruses from freshwater and seawater using ultrafiltration.infectious hematopoietic necrosis virus (ihnv), viral hemorrhagic septicemia virus, and spring viremia of carp virus were concentrated and detected from freshwater and seawater samples by using hollow-fiber ultrafiltration. within 60 min, virus in a 50-l freshwater or saltwater sample was concentrated more than 70-fold, and virus retention efficiencies were consistently greater than 88%. retention efficiency was highly dependent upon concentrations of column blocking and sample stabilization sol ...201122372250
enhanced arbovirus surveillance with deep sequencing: identification of novel rhabdoviruses and bunyaviruses in australian mosquitoes.viral metagenomics characterizes known and identifies unknown viruses based on sequence similarities to any previously sequenced viral genomes. a metagenomics approach was used to identify virus sequences in australian mosquitoes causing cytopathic effects in inoculated mammalian cell cultures. sequence comparisons revealed strains of liao ning virus (reovirus, seadornavirus), previously detected only in china, livestock-infecting stretch lagoon virus (reovirus, orbivirus), two novel dimarhabdov ...201324314645
melanoma differentiation-associated gene 5 in zebrafish provoking higher interferon-promoter activity through signalling enhancing of its shorter splicing variant.melanoma differentiation-associated gene 5 (mda5) is one of the three members in the retinoic acid-inducible gene i-like receptor (rlr) family, which are cytoplasmic pathogen recognition receptors recognizing intracellular viruses. in the present study, mda5 and its spliced shorter forms, named as mda5a and mda5b, were identified in zebrafish. mda5a and mda5b can be up-regulated in cell lines following the infection of a negative ssrna virus, the spring viraemia of carp virus (svcv), and an intr ...024116956
establishment of caudal fin cell lines from tropical ornamental fishes puntius fasciatus and pristolepis fasciata endemic to the western ghats of india.two new cell lines, pff and cff were established from the caudal fin of the puntius fasciatus, and pristolepis fasciata respectively. since their initiation, these cell lines (pff and cff) have been subcultured in l-15 medium with 10% fetal bovine serum for more than 35 passages at 28°c and both the cell lines were characterized. karyotyping analysis of pff and cff cells at 25th passage indicated that the modal chromosome number was 2n=50 and 2n=48 respectively. the cell line was cryopreserved i ...201323933168
establishment and characterization of a heart-derived cell line from goldfish (carassius auratus).the goldfish carassius auratus, a freshwater fish in the family cyprinidae, was one of the earliest fish to be domesticated for ornamental purposes. a cell line was established from goldfish heart (gh) tissue to create a biological monitoring tool for viral diseases. the gh cell line was optimally maintained at 25 °c in m199 medium supplemented with 10-20% fetal bovine serum. a chromosomal analysis indicated that the cell line remained diploid, with a mean chromosomal count of 100. in viral inoc ...201728097494
evaluation of serial urine viral cultures for the diagnosis of cytomegalovirus infection in neonates and infants.cytomegalovirus (cmv) is the most common cause of congenital infection worldwide. urine viral culture is the standard for cmv diagnosis in neonates and infants. the objectives of this study were to compare the performance of serial paired rapid shell vial cultures (svc) and routine viral cultures (rvc), and to determine the optimal number of cultures needed to detect positive cases. from 2001 to 2011, all paired cmv svc and rvc performed on neonates and infants less than 100 days of age were rec ...201424617645
emergence of carp edema virus in cultured ornamental koi carp, cyprinus carpio koi, in india.a disease outbreak was reported in adult koi, cyprinus carpio koi, from a fish farm in kerala, india, during june 2015. the clinical signs were observed only in recently introduced adult koi, and an existing population of fish did not show any clinical signs or mortality. microscopic examination of wet mounts from the gills of affected koi revealed minor infestation of dactylogyrus sp. in a few koi. in bacteriological studies, only opportunistic bacteria were isolated from the gills of affected ...201627902384
proteomic analysis of cytoskeleton proteins in fish.in this chapter, we describe laboratory protocols for rearing fish and a simple and efficient method of extracting and identifying pathogen and host proteins that may be involved in entry and replication of commercially important fish viruses. we have used the common carp (cyprinus carpio l.) and goldfish (cyprinus auratus) as a model system for studies of proteins involved in viral entry and replication. the chapter describes detailed protocols for maintenance of carp, cell culture, antibody pu ...201626498797
recombinant lactobacillus expressing g protein of spring viremia of carp virus (svcv) combined with orf81 protein of koi herpesvirus (khv): a promising way to induce protective immunity against svcv and khv infection in cyprinid fish via oral vaccination.spring viremia of carp virus (svcv) and koi herpesvirus (khv) are highly contagious and pathogenic to cyprinid fish, causing enormous economic losses in aquaculture. although dna vaccines reported in recent years could induce protective immune responses in carps against these viruses via injection, there are a number of consequences and uncertainties related to dna vaccination. therefore, more effective and practical method to induce protective immunity such as oral administration would be highl ...201525981489
spring viraemia of carp (svc) in the uk: the road to freedom.spring viraemia of carp (svc) is a disease of international importance that predominantly affects cyprinid fish and can cause significant mortality. in the united kingdom (uk), svc was first detected in 1977 with further cases occurring in fisheries, farms, wholesale and retail establishments throughout england and wales (but not scotland, where few cyprinid populations exist, nor northern ireland where svc has never been detected) over the subsequent 30 years. following a control and eradicatio ...201323597621
a model to approximate lake temperature from gridded daily air temperature records and its application in risk assessment for the establishment of fish diseases in the uk.ambient water temperature is a key factor controlling the distribution and impact of disease in fish populations, and optimum temperature ranges have been characterised for the establishment of a number important aquatic diseases exotic to the uk. this study presents a simple regression method to approximate daily average surface water temperature in lakes of 0.5-15 ha in size across the uk using 5 km(2) gridded daily average air temperatures provided by the uk meteorological office. a geographi ...201322862828
interferon type i responses to virus infections in carp cells: in vitro studies on cyprinid herpesvirus 3 and rhabdovirus carpio infections.interferons (ifns) are secreted mediators that play a fundamental role in the innate immune response against viruses among all vertebrate classes. common carp is a host for two highly contagious viruses: spring viraemia of carp virus (rhabdovirus carpio, svcv) and the cyprinid herpesvirus 3 (cyhv-3), which belong to rhabdoviridae and alloherpesviridae families, respectively. both viruses are responsible for significant losses in carp aquaculture. in this paper we studied the mrna expression prof ...201222683518
comparison of routes for achieving parenteral access with a focus on the management of patients with ebola virus disease.dehydration is an important cause of death in patients with ebola virus disease (evd). parenteral fluids are often required in patients with fluid requirements in excess of their oral intake. the peripheral intravenous route is the most commonly used method of parenteral access, but inserting and maintaining an intravenous line can be challenging in the context of evd. therefore it is important to consider the advantages and disadvantages of different routes for achieving parenteral access (e.g. ...201525914907
higher antiviral response of rig-i through enhancing rig-i/mavs-mediated signaling by its long insertion variant in zebrafish.as an intracellular pattern recognition receptor (prr), the retinoic acid-inducible gene-i (rig-i) is responsible for the recognition of cytosolic viral nucleic acids and the production of type i interferons (ifns). in the present study, an insertion variant of rig-i with 38 amino acids inserted in the n-terminal card2 domain, as well as the typical type, named as rig-ia and rig-ib respectively were identified in zebrafish. rig-ia and rig-ib were all up-regulated following the infection of a neg ...201525524497
monoclonal antibody against m protein of spring viremia of carp virus.m protein of spring viremia of carp virus (svcv) was expressed in escherichia coli and then used to immunize balb/c mice. one monoclonal antibody (5a1) against m protein was generated by fusion of mouse myeloma cell line sp2/0 and spleen lymphocytes from immunized mice. the characterizations of this mab were confirmed by elisa (enzyme linked immunosorbent assay), ifa (immunofluorescent assay), and western blot analysis. all results indicate that mab 5a1 was specific to svcv m protein.201525897611
monoclonal antibodies against g protein of spring viremia of carp virus.part of the g protein (3094-4170 bp) of spring viremia of carp virus (svcv) was expressed in escherichia coli and purified by dialysis in our study. two clones of monoclonal antibodies (mabs 1h11 and 4b8) against g protein were generated by fusion of mouse myeloma cell line sp2/0 and spleen lymphocytes from part of g protein (3094-4170 bp) immunized mice. the results of elisa (enzyme-linked immunosorbent assay), ifa (indirect immunofluorescent assay), and western blot assay further demonstrated ...201425358003
card and tm of mavs of black carp play the key role in its self-association and antiviral ability.mitochondrial antiviral signaling protein (mavs) is an adaptor protein of the innate immune system of higher vertebrate. in this paper, the transcription profile of black carp mavs (bcmavs) in host cells in response to spring viremia of carp virus (svcv) and grass carp reovirus (gcrv) infection was identified. epc cells expressing bcmavs possessed obviously enhanced antiviral activity against both svcv and gcrv. immunofluorescence (if) staining data demonstrated that bcmavs molecules were redist ...201728232279
in vitro antiviral efficacy of moroxydine hydrochloride and ribavirin against grass carp reovirus and giant salamander iridovirus.moroxydine hydrochloride (mor) and ribavirin (rib) have been reported to exhibit multi-antiviral activities against dna and rna viruses, but their antiviral activities and pharmacologies have seldom been studied in aquaculture. this paper has selected 3 aquatic viruses including a double-stranded rna virus (grass carp reovirus, gcrv), a single-stranded rna virus (spring viraemia of carp virus, svcv) and a dna virus (giant salamander iridovirus, gsiv) for antiviral testing. the results showed tha ...201627786157
development of an ussuri catfish pseudobagrus ussuriensis skin cell line displaying differential cytopathic effects to three aquatic animal viruses.an ussuri catfish pseudobagrus ussuriensis skin (ucs) cell line was developed and subcultured for more than 60 passages. ucs cells consisted of mostly epithelial-like cells and multiplied well in tc199 medium supplemented with 10% fetal bovine serum at 25°c. chromosome analysis revealed that most ucs cells had a normal diploid karyotype with 2n=52. ucs cells showed differential cytopathic effects (cpes) after inoculation of spring viremia of carp virus (svcv, a negative-strand rna virus), grass ...201424794839
identification of drei as an antiviral factor regulated by rlr signaling pathway.retinoic acid-inducible gene i (rig-i)-like receptors (rlrs) had been demonstrated to prime interferon (ifn) response against viral infection via the conserved rlr signaling in fish, and a novel fish-specific gene, the grass carp reovirus (gcrv)-induced gene 2 (gig2), had been suggested to play important role in host antiviral response.201222412872
hepatitis c virus genotype analyses in chronic hepatitis c patients and individuals with spontaneous virus clearance using a newly developed serotyping assay.we developed a novel hcv serotyping assay and detected the genotypes in chronic hepatitis c (chc) patients and individuals with spontaneous viral clearance (svc).201727292225
vertically acquired hepatitis c virus infection: correlates of transmission and disease progression.the worldwide prevalence of hepatitis c virus (hcv) infection in children is 0.05%-0.4% in developed countries and 2%-5% in resource-limited settings, where inadequately tested blood products or un-sterile medical injections still remain important routes of infection. after the screening of blood donors, mother-to-child transmission (mtct) of hcv has become the leading cause of pediatric infection, at a rate of 5%. maternal hiv co-infection is a significant risk factor for mtct and anti-hiv ther ...201626819507
predictors of spontaneous viral clearance and outcomes of acute hepatitis c infection.this study evaluated the predictors of spontaneous viral clearance (svc), as defined by two consecutive undetectable hepatitis c virus (hcv) rna tests performed ≥ 12 weeks apart, and the outcomes of acute hepatitis c (ahc) demonstrating svc or treatment-induced viral clearance.201425548743
detection of infectious pancreatic necrosis virus from the leeches hemiclepsis marginata and hirudo medicinalis.leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (ihnv), and viral hemorrhagic septicemia virus (vhsv), and also may contain blood protozoa. in the present study, leeches were collected from water bodies located in kurdistan province, iran. the specimens were tested for ihnv, vhsv, and infectious pancreatic necrosis virus (ipnv) using the pcr method. the results showed that two different species ...201627687675
targeting and stimulation of the zebrafish (danio rerio) innate immune system with lps/dsrna-loaded nanoliposomes.herein we report the use of immunostimulant-loaded nanoliposomes (called nlcliposomes) as a strategy to protect fish against bacterial and/or viral infections. this work entailed developing a method for in vivo tracking of the liposomes administered to adult zebrafish that enables evaluation of their in vivo dynamics and characterisation of their tissue distribution. the nlc liposomes, which co-encapsulate poly(i:c) and lps, accumulate in immune tissues and in immunologically relevant cells such ...201424837767
inhibition of spring viraemia of carp virus replication in an epithelioma papulosum cyprini cell line by rnai.spring viraemia of carp virus (svcv) is an aetiological agent of a serious disease affecting carp farms in europe and is a member of the rhabdoviridae family of viruses. the genome of svcv codes for five proteins: nucleoprotein (n), phosphoprotein (p), matrix protein (m), glycoprotein (g) and rna-dependent rna polymerase (l). rna-mediated interference (rnai) by small interfering rnas (sirnas) is a powerful tool to inhibit gene transcription and is used to study genes important for viral replicat ...201524460815
establishment of a new cell line from the heart of giant grouper, epinephelus lanceolatus (bloch), and its application in toxicology and virus susceptibility.a new marine fish cell line, derived from the heart of giant grouper, epinephelus lanceolatus (bloch), was established and characterized. the cell line was designated as elgh and subcultured with more than 60 passages. the elgh cells were mainly composed of fibroblast-like cells and multiplied well in leibovitz's l-15 medium supplemented with 10% foetal bovine serum (fbs) at 28 °c. chromosome analysis indicated that the modal chromosome number was 48. the fluorescent signals were detected in elg ...201524372271
medaka haploid embryonic stem cells are susceptible to singapore grouper iridovirus as well as to other viruses of aquaculture fish species.viral infection is a challenge in high-density aquaculture, as it leads to various diseases and causes massive or even complete loss. the identification and disruption of host factors that viruses utilize for infection offer a novel approach to generate viral-resistant seed stocks for cost-efficient and sustainable aquaculture. genetic screening in haploid cell cultures represents an ideal tool for host factor identification. we have recently generated haploid embryonic stem (es) cells in the la ...201323828270
zebrafish foxo3b negatively regulates antiviral response through suppressing the transactivity of irf3 and irf7.forkhead box o (foxo)3, a member of the foxo family of transcription factors, plays key roles in various cellular processes, including development, longevity, reproduction, and metabolism. recently, foxo3 has also been shown to be involved in modulating the immune response. however, how foxo3 regulates immunity and the underlying mechanisms are still largely unknown. in this study, we show that zebrafish (danio rerio) foxo3b, an ortholog of mammalian foxo3, is induced by polyinosinic-polycytidyl ...201627815423
zebrafish stat6 negatively regulates ifnφ1 production by attenuating the kinase activity of tank-binding kinase 1.the aquatic spring viremia of carp virus (svcv) causes significant mortality in common carp (cyprinus carpio), and tbk1 plays a crucial role in the retinoic acid-inducible gene i (rig-i)-like receptor (rlr) system by phosphorylating its substrates to induce type i interferons (ifns) and cellular antiviral responses. in this study, we report that zebrafish stat6 is induced during svcv infection and reduces ifnφ1 expression by suppressing tbk1 phosphorylation. a typical ifn stimulatory response el ...201727743998
molecular characterization, tissue distribution and expression, and potential antiviral effects of trim32 in the common carp (cyprinus carpio).tripartite motif-containing protein 32 (trim32) belongs to the tripartite motif (trim) family, which consists of a large number of proteins containing a ring (really interesting new gene) domain, one or two b-box domains, and coiled coil motif followed by different c-terminal domains. the trim family is known to be implicated in multiple cellular functions, including antiviral activity. however, it is presently unknown whether trim32 of common carp (cyprinus carpio) has the antiviral effect. in ...201627735853
the p protein of spring viremia of carp virus negatively regulates the fish interferon response by inhibiting the kinase activity of tank-binding kinase 1.spring viremia of carp virus (svcv) is an efficient pathogen causing high mortality in the common carp. fish interferon (ifn) is a powerful cytokine enabling host cells to establish an antiviral response; therefore, the strategies that svcv uses to avoid the cellular ifn response were investigated. here, we report that the svcv p protein is phosphorylated by cellular tank-binding kinase 1 (tbk1), which decreases ifn regulatory factor 3 (irf3) phosphorylation and suppresses ifn production. first, ...201627654289
detection of spring viraemia of carp virus in imported amphibians reveals an unanticipated foreign animal disease threat.global translocation of plants and animals is a well-recognized mechanism for introduction of pathogens into new regions. to mitigate this risk, various tools such as preshipment health certificates, quarantines, screening for specific disease agents and outright bans have been implemented. however, such measures only target known infectious agents and their hosts and may fail to prevent translocation of even well-recognized pathogens if they are carried by novel host species. in a recent exampl ...201627599472
spring viraemia of carp virus enters grass carp ovary cells via clathrin-mediated endocytosis and macropinocytosis.spring viraemia of carp virus (svcv) is the causative pathogen of the outbreaks of an acute haemorrhagic and contagious viraemia responsible for the significant mortality in several cyprinid species. however, the endocytic pathway(s) and their regulatory molecules have not been characterized for svcv. here, using a combination of specific pharmacological inhibitors, transmission electron microscopy, immunofluorescence microscopy and real-time quantitative pcr, we found that svcv entered grass ca ...201627590028
zebrafish irf1, irf3, and irf7 differentially regulate ifnφ1 and ifnφ3 expression through assembly of homo- or heteroprotein complexes.in mammals, ifn regulatory factor (irf)1, irf3, and irf7 are three critical transcription factors that are pivotal for cooperative regulation of the type i ifn response. in this study, we explored the relative contribution of zebrafish (danio rerio) irf1 (drirf1), irf3 (drirf3), and irf7 (drirf7) (drirf1/3/7) to zebrafish ifnφ1 (drifnφ1) and ifnφ3 (drifnφ3) (drifnφ1/3) activation. following spring viremia of carp virus infection, drifnφ1/3 and drirf1/3/7 transcripts are significantly induced in ...201627496972
emergence of a new rhabdovirus associated with mass mortalities in eelpout (zoarces viviparous) in the baltic sea.we report the first description of a new rhabdoviridae tentatively named eelpout rhabdovirus (eprv genus perhabdovirus). this virus was associated with mass mortalities in eelpout (zoarces viviparous, linnaeus) along the swedish baltic sea coast line in 2014. diseased fish showed signs of central nervous system infection, and brain lesions were confirmed by histology. a cytopathogenic effect was observed in cell culture, but elisas for the epizootic piscine viral haemorrhagic septicaemia virus ( ...201727416895
toll-like receptors and interferon associated immune factors responses to spring viraemia of carp virus infection in common carp (cyprinus carpio).pattern recognition receptor (prr) toll-like receptors (tlrs), antiviral agent interferon (ifn) and the effector ifn stimulated genes (isgs) play a fundamental role in the innate immune response against viruses among all vertebrate classes. common carp is a host for spring viraemia of carp virus (rhabdovirus carpio, svcv), which belong to rhabdoviridae family. the present in-vivo experiment was conducted to investigate the expression of these innate immune factors in early phase as well as durin ...201627263115
restrictions on the importation of zebrafish into canada associated with spring viremia of carp virus.the zebrafish model system is helping researchers improve the health and welfare of people and animals and has become indispensable for advancing biomedical research. as genetic engineering is both resource intensive and time-consuming, sharing successfully developed genetically modified zebrafish lines throughout the international community is critical to research efficiency and to maximizing the millions of dollars in research funding. new restrictions on importation of zebrafish into canada b ...201627248438
spring viremia of carp virus n protein suppresses fish ifnφ1 production by targeting the mitochondrial antiviral signaling protein.for a virus to replicate efficiently, it must try and inhibit host ifn expression because ifn is an important host defense at early stages after viral infection. for aquatic viruses, the mechanisms used to escape the hosts ifn system are still unclear. in this study, we show that the n protein of spring viremia of carp virus (svcv) inhibits zebrafish ifnφ1 production by degrading the mitochondrial antiviral signaling protein (mavs). first, the upregulation of ifnφ1 promoter activity stimulated b ...201626994222
spring viraemia of carp virus: recent advances.spring viraemia of carp is an environmentally and economically important disease affecting cyprinids, primarily common carp (cyprinus carpio). the causative agent of this disease is spring viraemia of carp virus (svcv) - a member of the genus vesiculovirus of the family rhabdoviridae. the disease is presently endemic in europe, america and several asian countries, where it causes significant morbidity and mortality in affected fish. svcv infection is generally associated with exophthalmia; abdom ...201626905065
genetic diversity of spring viraemia of carp virus isolates based on the glycoprotein gene. 201626791607
an n-targeting real-time pcr strategy for the accurate detection of spring viremia of carp virus.spring viremia of carp virus (svcv) is a highly pathogenic agent of several economically important cyprinidae fish species. currently, there are no effective vaccines or drugs for this virus, and prevention of the disease mostly relies on prompt diagnosis. previously, nested rt-pcr and rt-qpcr detection methods based on the glycoprotein gene g have been developed. however, the high genetic diversity of the g gene seriously limits the reliability of those methods. compared with the g gene, phylog ...201626717888
pathogenesis of spring viremia of carp virus in emerald shiner notropis atherinoides rafinesque, fathead minnow pimephales promelas rafinesque and white sucker catostomus commersonii (lacepede).spring viremia of carp (svc) is a reportable disease to the world organization of animal health (oie) as it is known to cause significant international economic impact. in canada, the first and only isolation of svc virus (svcv) was in 2006, from common carp cyprinus carpio l., at hamilton harbour, lake ontario. the susceptibility of fathead minnow pimephales promelas rafinesque, emerald shiner notropis atherinoides rafinesque and white sucker catostomus commersonii (lacepede) to intraperitoneal ...201626411333
ifna of black carp is an antiviral cytokine modified with n-linked glycosylation.type i interferons (ifns) play an important role in the antiviral immune response in teleost fish. in this study, one type i interferon (bcifna) of black carp (mylopharyngodon piceus) has been cloned and characterized. the full-length cdna of bcifna gene consists of 783 nucleotides and the predicted bcifna protein contains 185 amino acids. semi-quantitative rt-pcr analysis demonstrated that bcifna mrna transcription level in all the selected tissues of black carp was greatly increased at 33 h po ...201526208754
development of an improved rt-pcr for specific detection of spring viraemia of carp virus.spring viraemia of carp (svc) is a rhabdovirus infection, which has a significant economic impact in pond cultures of carp in europe and western independent states of the former soviet union. the causative agent of svc, spring viraemia of carp virus (svcv), has been divided into four subgroups, ia, ib, ic and id, on the basis of glycoprotein (g) protein gene sequences. in this study, a new primer set was designed from a g gene sequence of svcv to identify the four subtypes of svcv by reverse tra ...201625832786
establishment and characterization of fin cell lines from diploid, triploid, and tetraploid oriental weatherfish (misgurnus anguillicaudatus).continuous fin cell lines from diploid, triploid, and tetraploid oriental weatherfish, misgurnus anguillicaudatus, were established and characterized. the cell lines, designated dimf, trmf, and temf, respectively, were subcultured more than 80 times since initiation in october 2012 and were preserved at the china center for type culture collection as sample numbers c2013109, c2013110, c2013111, respectively. the cell lines consist predominantly of fibroblast-like cells. at the 50th passage, the ...201525721100
characterization of two thymosins as immune-related genes in common carp (cyprinus carpio l.).prothymosin alpha (protα) and thymosin beta (tβ) belong to thymosin family, which consists of a series of highly conserved peptides involved in stimulating immune responses. protα b and tβ are still poorly studied in teleost. here, the full-length cdnas of protα b and tβ-like (tβ-l) were cloned and identified in common carp (cyprinus carpio l.). the expressions of carp protα b and tβ-l exhibited rise-fall pattern and then trended to be stable during early development. after spring viraemia of ca ...201525596145
zebrafish irf1 regulates ifn antiviral response through binding to ifnϕ1 and ifnϕ3 promoters downstream of myd88 signaling.in mammals, type i ifns (mainly ifn-α/β) are primarily regulated by transcription factors of the ifn regulatory factor (irf) family. fish ifns do not show a one-to-one orthologous relationship with mammalian type i ifn homologues. using a bacterial one-hybrid reporter screening system and an overexpression approach to explore the molecular mechanism underlying fish ifn induction, we identified zebrafish danio rerio irf (drirf)1 as a positive regulator of the fish ifn antiviral response. among 12 ...201525535281
spring viraemia of carp virus induces autophagy for necessary viral replication.outbreaks of spring viraemia of carp virus (svcv) in several carp species and other cultivated fish can cause significant mortality and jeopardize the billion-dollar worldwide fish industry. spring viraemia of carp virus, also known as rhabdovirus carpio, is a bullet-shaped rna virus that enters and amplifies in gill epithelium and later spreads to internal organs. young fish under stressed conditions (spring cold water, etc.) are more vulnerable to svcv-induced lethality because of their lack o ...201525376386
transcriptome analysis of epithelioma papulosum cyprini cells after svcv infection.spring viraemia of carp virus (svcv) has been identified as the causative agent of spring viraemia of carp (svc) and it has caused significant losses in the cultured common carp (cyprinus carpio) industry. the molecular mechanisms that underlie the pathogenesis of the disease remain poorly understood. in this study, deep rna sequencing was used to analyse the transcriptome and gene expression profile of epc cells at progressive times after svcv infection. this study addressed the complexity of v ...201425344771
microrna profile analysis of epithelioma papulosum cyprini cell line before and after svcv infection.micrornas (mirnas) play significant roles in regulating almost all of the biological processes in eukaryotes. an accumulating body of evidence shows that mirnas are associated with cellular changes following viral infection. spring viremia of carp virus (svcv) is the pathogen of spring viremia of carp (svc), which results in heavy losses in the cultured common carp (cyprinus carpio) industry in many countries. to study the involvement of mirnas during svcv infection, we adopted the solexa sequen ...201525291211
vhsv g glycoprotein major determinants implicated in triggering the host type i ifn antiviral response as dna vaccine molecular adjuvants.we have recently identified the two major determinants of the glycoprotein g of the viral hemorrhagic septicaemia rhabdovirus (gpgvhsv), peptides p31 and p33 implicated in triggering the host type i ifn antiviral response associated to these rhabdoviral antigens. with the aim to investigate the properties of these viral glycoprotein regions as dna molecular adjuvants, their corresponding cdna sequences were cloned into a plasmid (pmcv1.4) flanked by the signal peptide and transmembrane sequences ...201425203447
up-regulation of nuclear factor e2-related factor 2 upon svcv infection.nuclear factor e2 - related factor 2 (nrf2) is a crucial transcription factor that regulates the basal and inducible expression of many antioxidant response element (are)-dependent genes, including heme oxygenase-1 (ho-1) and superoxide dismutase 1 (sod1). the nrf2/are pathway has been regarded as a critical switch in the initiation of cellular defence systems for surviving oxidative insults and viral infection. in this study, the nrf2 gene of epc cells, which is originally derived from pimephal ...201425038284
ifn regulatory factor 10 is a negative regulator of the ifn responses in fish.ifn regulatory factor (irf) 10 belongs to the irf family and exists exclusively in birds and fish. most irfs have been identified as critical regulators in the ifn responses in both fish and mammals; however, the role of irf10 is unclear. in this study, we identified irf10 in zebrafish (danio rerio) and found that it serves as a negative regulator to balance the innate antiviral immune responses. zebrafish irf10 (drirf10) was induced by intracellular polyinosinic:polycytidylic acid in zf4 (zebra ...201424958903
genomic evidence of homologous recombination in spring viremia of carp virus: a negatively single stranded rna virus.a new strain of spring viraemia of carp virus, denominated svcv-265, was isolated from an ornamental common carp (cyprinus carpio) in shanghai, china, 2013. the isolate could produce obvious cytopathic effects on epc cells, while was shown to be of low virulence for juvenile koi. complete genome sequencing revealed the genome of the svcv-265 strain is 11,029 nucleotides in length and phylogenetic analysis showed the isolate was clustered within asia clade but was divergent from chinese a1, a2 an ...201424954789
selection and characterization of single-chain recombinant antibodies against infectious haematopoietic necrosis virus from mouse phage display library.six single-chain fragment variable (scfv) antibodies against infectious haematopoietic necrosis virus (ihnv) were selected from an antibody phage display library by phage display technology. the soluble scfv antibodies showed a molecular weight 32kda by western blot. dot blot analysis revealed that the six scfv antibodies could recognize ihnv. for enzyme linked immunosorbent assay (elisa), four scfv antibodies (p1a4, p1a12, p1d5 and p3e2) showed cross-reactivity with spring viraemia of carp viru ...201424747010
caspian white fish (rutilus frisii kutum) as a host for spring viraemia of carp virus.rutilus frisii kutum is a fish of the cyprinidae family which is native in caspian sea and commercially cultured in iran. this study was conducted to investigate susceptibility of caspian white fish to spring viraemia of carp virus (svcv) infection and to evaluate influence of different challenge routes on virulence of the virus. fingerlings were infected by immersion, intra-peritoneal (i.p.) injection, cohabitation and orally. dead and surviving fish were collected for histological examination ...201424685241
codon usage bias of the phosphoprotein gene of spring viraemia of carp virus and high codon adaptation to the host.in this study, we calculated the relative synonymous codon usage (rscu) value and the effective number of codons (enc) value to carry out principal component analysis (pca) and correlation analysis of the codon usage pattern of the phosphoprotein gene (p gene) of spring viraemia of carp virus (svcv). the synonymous codon usage pattern in p genes is geography-specific, based on pca analysis. the high correlation between (g + c)1,2 % and (g + c)3 % suggests that mutational pressure rather than nat ...201424519460
experimental susceptibility of caspian white fish, rutilus frisii kutum to spring viraemia of carp virus.caspian white fish (rutilus frisii kutum) is a fish of the family cyprinidae, which is commercially harvested from the caspian sea. experimental infection with spring viraemia of carp virus (svcv) was conducted in order to examine susceptibility of caspian white fish and clinical impacts of infection. fingerling fish were injected intra-peritoneally or waterborne-exposed with svcv and were monitored daily for 7 weeks. dead fish and those survived at the end of experimental period were collected ...201424426310
selection and characterization of single-chain recombinant antibodies against spring viraemia of carp virus from mouse phage display library.antibody-displaying phage library was selected after three rounds of panning against spring viraemia of carp virus (svcv) by phage display technology. eight positive clones which could produce soluble single-chain fragment variable (scfv) antibody induced by isopropyl-beta-d-thiogalactopyranoside (iptg) were obtained. dot blot results showed that the eight scfv antibodies could recognize svcv. the soluble scfv antibodies showed a molecular weight 29 kd by western blot. all scfv antibodies could ...201323994147
proteomic analysis of epithelioma papulosum cyprini cells infected with spring viremia of carp virus.spring viremia of carp (svc), caused by spring viremia of carp virus (svcv) is an important disease due to its drastic effects on carp fisheries in many countries. to better understand molecular responses to svcv infection, two dimensional electrophoresis (2-de) and maldi-tof/tof were performed to investigate altered proteins in epithelioma papulosum cyprini cells (epcs). differentially expressed proteins in mock-infected epcs and svcv-infected epcs were compared. a total of 54 differentially ex ...201323583725
spring viremia of carp virus in minnehaha creek, minnesota.spring viremia of carp virus (svcv) causes a highly contagious and serious disease of freshwater cyprinid fishes, generating significant economic and ecological impacts throughout the world. the svcv is therefore listed as a notifiable pathogen by the international organization for animal health. in june 2011, a significant mortality event of wild common carp cyprinus carpio occurred in minnehaha creek near its confluence with mississippi river pool 2 in minneapolis, minnesota. clinical signs of ...201223072656
direct detection of unamplified spring viraemia of carp virus rna using unmodified gold nanoparticles.spring viraemia of carp (svc) is a viral disease that mainly affects carp cyprinus carpio and other cyprinid fish, causing severe economic losses. rapid detection and identification of spring viraemia of carp virus (svcv) is crucial for effective disease management. recent advances in nanoscience are having a significant impact on many scientific fields, especially biodiagnostics, where a number of nanoparticle-based assays have been introduced for biomolecular detection. single- and double-stra ...201222885508
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