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complete genome analysis of the mandarin fish infectious spleen and kidney necrosis iridovirus.the nucleotide sequence of the infectious spleen and kidney necrosis virus (isknv) genome was determined and found to comprise 111,362 bp with a g+c content of 54.78%. it contained 124 potential open reading frames (orfs) with coding capacities ranging from 40 to 1208 amino acids. the analysis of the amino acid sequences deduced from the individual orfs revealed that 35 of the 124 potential gene products of isknv show significant homology to functionally characterized proteins of other species. ...200111878882
sequence analysis of the complete genome of an iridovirus isolated from the tiger frog.we have isolated a tiger frog virus (tfv) from diseased tiger frogs, rana tigrina rugulosa. the genome was a linear double-stranded dna of 105,057 basepairs in length with a base composition of 55.01% g+c. about 105 open reading frames were identified with coding capacities for polypeptides ranging from 40 to 1294 amino acids. computer-assisted analyses of the deduced amino acid sequences revealed that 39 of 105 putative gene products showed significant homology to functionally characterized pro ...200211878922
cladistic analysis of iridoviruses based on protein and dna sequences.cladograms of iridoviruses were inferred from bootstrap analysis of molecular data sets comprising all published protein and dna sequences of the major capsid protein, atpase and dna polymerase genes of members of the iridoviridae family iridovirus. all data sets yielded cladograms supporting the separation of the iridovirus, ranavirus and lymphocystivirus genera, and the cladogram based on data derived from major capsid proteins further divided both the iridovirus and ranavirus genera into two ...200314579177
genomic sequence of a ranavirus (family iridoviridae) associated with salamander mortalities in north america.disease is among the suspected causes of amphibian population declines, and an iridovirus and a chytrid fungus are the primary pathogens associated with amphibian mortalities. ambystoma tigrinum virus (atv) and a closely related strain, regina ranavirus (rrv), are implicated in salamander die-offs in arizona and canada, respectively. we report the complete sequence of the atv genome and partial sequence of the rrv genome. sequence analysis of the atv/rrv genomes showed marked similarity to other ...200314599794
complete genomic dna sequence of rock bream iridovirus.iridovirus is a causative agent of epizootics among cultured rock bream (oplegnathus fasciatus) in korea. here, we report the complete genomic sequence of rock bream iridovirus (rbiv). the genome of rbiv was 112080 bp long and contained at least 118 putative open reading frames (orfs), and its genome organization was similar to that of infectious spleen and kidney necrosis virus (isknv). of the rbiv's 118 orfs, 85 orfs showed 60-99% amino acid identity to those of isknv. phylogenetic analysis of ...200415246274
complete genome sequence analysis of an iridovirus isolated from the orange-spotted grouper, epinephelus coioides.orange-spotted grouper iridovirus (osgiv) was the causative agent of serious systemic diseases with high mortality in the cultured orange-spotted grouper, epinephelus coioides. here we report the complete genome sequence of osgiv. the osgiv genome consists of 112,636 bp with a g+c content of 54%. 121 putative open reading frames (orf) were identified with coding capacities for polypeptides varying from 40 to 1168 amino acids. the majority of osgiv shared homologies to other iridovirus genes. phy ...200515964605
phylogenetic analysis of the major capsid protein gene of iridovirus isolates from cultured flounders paralichthys olivaceus in korea.in 2003, 13 isolates of iridovirus were obtained from cultured flounders paralichthys olivaceus during epizootics in korea. the full open reading frames (orfs) encoding the major capsid protein (mcp) (1362 bp) from the 13 flounder iridoviruses (flivs) were sequenced and the deduced amino acid sequences were phylogenetically analyzed. phylogenetic analysis of the mcp revealed that all 13 flivs were the same species as rock bream iridovirus (rbiv), red sea bream iridovirus (rsiv), and infectious s ...200515997817
the molecular epidemiology of iridovirus in murray cod (maccullochella peelii peelii) and dwarf gourami (colisa lalia) from distant biogeographical regions suggests a link between trade in ornamental fish and emerging iridoviral diseases.iridoviruses have emerged over 20 years to cause epizootics in finfish and amphibians in many countries. they may have originated in tropical asia and spread through trade in farmed food fish or ornamental fish, but this has been difficult to prove. consequently, mcp, atpase and other viral genes were sequenced from archival formalin-fixed, paraffin-embedded tissues from farmed murray cod (maccullochella peelii peelii) that died during an epizootic in 2003 and from diseased gouramis that had bee ...200616697343
ring finger proteins of infectious spleen and kidney necrosis virus (isknv) function as ubiquitin ligase enzymes.infectious spleen and kidney necrosis virus (isknv) is the etiological agent that causes a pandemic and severe disease in fish characteristic of enlarged and damaged spleen and kidney. to identify viral proteins involved in infection and pathogenesis, we characterized five open reading frames (orfs) of the isknv genome, orf12, orf65, orf66, orf99 and orf111, which encode ring finger proteins (rfps). we assessed the ubiquitin ligase (e3) activity of these recombinant rfps fused to maltose-binding ...200717049660
molecular epidemiology and phylogenetic analysis of a marine fish infectious spleen and kidney necrosis virus-like (isknv-like) virus.infectious spleen and kidney necrosis virus-like (isknv-like) virus causes a serious systemic disease with high morbidity and mortality of freshwater and marine fishes. based on the isknv putative major capsid protein (mcp), the vascular endothelial growth factor (vegf), the mrna capping enzyme (capping), and the tumor necrosis factor receptor-associated protein (tnfr) genes, primers were designed and used in pcr to determine the host range of isknv-like viruses. from the sampling of >1600 marin ...200717131065
differential gene expression profile in spleen of mandarin fish siniperca chuatsi infected with isknv, derived from suppression subtractive hybridization.to study the interaction between an invading virus and its host, we investigated differential gene expression in mandarin fish siniperca chuatsi experimentally infected with infectious spleen and kidney necrosis virus (isknv). subtractive cdna libraries were constructed by suppression subtractive hybridization (ssh) from spleens of mock- and isknv-infected fish. both forward- and reverse-subtracted libraries were generated. in the forward library, genes of the ubiquitin-proteasome proteolytic pa ...200617260830
tumor necrosis factor-alpha gene from mandarin fish, siniperca chuatsi: molecular cloning, cytotoxicity analysis and expression profile.in this report, we cloned a cdna and the gene of mandarin fish (siniperca chuatsi) tumor necrosis factor-alpha (tnf-alpha). the length of tnf-alpha cdna was 1461 bp, contained an open reading frame (orf) of 762 bp, which encoded 253 amino acids. the positions of cysteine residues, transmembrane sequence, and protease cleavage site were similar with other reported fish tnf-alpha and mammalian tnf-alpha. mandarin fish tnf-alpha gene has a length of 1940 bp, and consists of four exons and three int ...200717482265
characterization of a membrane protein (vp001l) from infectious spleen and kidney necrosis virus (isknv).infectious spleen and kidney necrosis virus (isknv) is the type species of megalocytivirus, iridoviridae. a novel membrane protein corresponding to the first open reading frame (orf001l) of isknv genome was identified. this 378-residue protein, termed the vp001l protein, has a high content of hydrophobic sequences and contains 10-11 putative transmembrane domains, indicating it may be a membrane protein. the vp001l mrna start site was extended 433 bp upstream of the start codon and the temporal ...200818046636
infectious spleen and kidney necrosis virus orf48r functions as a new viral vascular endothelial growth factor.infectious spleen and kidney necrosis virus (isknv) causes a pandemic and serious disease in fish. infection by isknv causes epidermal lesions, in which petechial hemorrhages and abdominal edema are prominent features. isknv orf48r contains a domain similar to that of the platelet-derived growth factor and vascular endothelial growth factor (vegf) families of proteins. isknv orf48r showed higher similarity to the vegfs encoded by megalocytivirus and parapoxvirus than to those encoded in fish and ...200818305039
outbreaks and risks of infectious spleen and kidney necrosis virus disease in freshwater ornamental fishes.we examined the distribution of iridoviruses in 10 freshwater ornamental fish species hatched in korea and imported from other asian countries using both 1-step and 2-step polymerase chain reation (pcr). none of the 10 fish species analyzed were free of iridovirus as shown by 2-step pcr positive results, and 3 species yielded 1-step pcr positive results with associated mortality. cloned pcr amplicons of the adenosine triphosphatase (atpase) and major capsid protein (mcp) genes in genomic dna of ...200818380219
a zebrafish (danio rerio) model of infectious spleen and kidney necrosis virus (isknv) infection.zebrafish is a model animal for studies of genetics, development, toxicology, oncology, and immunology. in this study, infectious spleen and kidney necrosis virus (isknv) was used to establish an infection in zebrafish, and the experimental conditions were established and characterized. mortality of adult zebrafish infected with isknv by intraperitoneal (i.p.) injection exceeded 60%. isknv can be passed stably in zebrafish for over ten passages. the ailing zebrafish displayed petechial hemorrhag ...200818423508
development of a mandarin fish siniperca chuatsi fry cell line suitable for the study of infectious spleen and kidney necrosis virus (isknv).infectious spleen and kidney necrosis virus (isknv) is a typical species of the genus megalocytivirus in the family iridoviridae. however, until recently, no suitable piscine cell line is stably susceptible to isknv. here, a continuous cell culture derived from the mandarin fish fry (mff-1) was developed and has been subcultured over 60 passages during a period of 18 months. mff-1 consists predominantly of epithelial-like cells and grows well in dmem supplemented with 10% fetal bovine serum. mff ...200818485510
molecular cloning of two c1q-like cdnas in mandarin fish siniperca chuatsi.c1q, a subunit of the c1 complex, plays a key role in the recognition of immune complexes to initiate the classical complement pathway. in this study, we reported two c1q-like cdnas from mandarin fish (siniperca chuatsi), mc1q-like-1 (mc1ql1) and mc1q-like-2 (mc1ql2). the full-length cdna of mc1ql1was 990bp, containing a 71bp 5'-untranslated region (utr), an open reading frame (orf) of 723bp, and a 196bp long 3'-utr. mc1ql2 cdna was 1193bp, containing a 100bp 5'-utr, followed by an orf of 756bp ...200818571244
development of an rt-pcr detection method for mud crab reovirus.mud crab reovirus (mcrv) causes high mortality in the cultivated mud crab. to control better an outbreak of this virus, a rapid, specific and sensitive detection method based on rt-pcr was developed. the mcrv detection method was designed based on the one-step and two-step rt-pcr which resulted in the amplification of predicted products of 433 and 304 bp. the method is specific as no cross-reaction was observed between grass carp hemorrhage virus (gchv), white spot syndrome virus (wssv), tiger f ...200818572256
cloning, characterization and expression analysis of a cxcr1-like gene from mandarin fish siniperca chuatsi.in this study we cloned and characterized a cxcr1-like gene (mfcxcr1) from mandarin fish (siniperca chuatsi). the full-length cdna of mfcxcr1 is 2,173 bp and contains a 1,056 bp open reading frame (orf) that encodes a protein of 351 amino acids. the 5' and 3' untranslated regions (utr) are 57 and 1,080 bp in length, respectively. the coding region of the mfcxcr1 gene consists of a single exon with a 734 bp intron that is two nucleotides upstream of the atg start codon in the 5' utr. the mfcxcr1 ...200919093223
cloning of irak1 and its upregulation in symptomatic mandarin fish infected with isknv.interleukin-1 receptor activated kinases (iraks) play crucial roles in the toll-like receptor (tlr) mediated signal transduction pathways that control host innate immune responses. here we report the cloning of an irak1 cdna (named scirak1) from the mandarin fish. the predicted scirak1 peptide contains a death domain and a serine/threonine-specific kinase domain. quantitative rt-pcr showed that scirak1 mrna was primarily expressed in blood cells and posterior kidney. seven days following infecti ...200919336221
the alpha inhibitor of nf-kappab (ikappabalpha) from the mandarin fish binds with p65 nf-kappab.the nf-kappab/ikappabalpha pathway plays an important role in the regulation of immune and inflammatory responses. ikappabalpha is an inhibitory molecule that sequesters transcription activator nf-kappab dimer in the cytoplasm of unstimulated cells. here, we isolated the full-length cdnas of the mandarin fish (siniperca chuatsi) alpha inhibitor of nf-kappab (scikappabalpha) and p65 nf-kappab (scp65). multiple sequence alignments showed that the amino acid sequences of both scikappabalpha and scp ...200919340946
pcr amplification and sequence analysis of the major capsid protein gene of megalocytiviruses isolated in taiwan.viruses belonging to the genus megalocytivirus in the family iridoviridae are one of the major agents causing mass mortalities in marine and freshwater fish in asian countries. outbreaks of iridovirus disease have been reported among various fish species in taiwan. however, the genotypes of these iridoviruses have not yet been determined. in this study, seven megalocytivirus isolates from four fish species: king grouper, epinephelus lanceolatus (bloch), barramundi perch, lates calcarifer (bloch) ...200919460084
detection of red-spotted grouper nervous necrosis virus by loop-mediated isothermal amplification.red-spotted grouper nervous necrosis virus (rgnnv) causes high mortality in marine fish larvae cultured in china. to control better an outbreak of this virus, a rapid, specific and sensitive detection method based on loop-mediated isothermal amplification (lamp) was developed. a set of four primers, two outer and two inner, was designed from rgnnv genome rna. the lamp reaction mix was optimized. the method was specific as no cross-reaction was observed between white spot syndrome virus, koi herp ...201019755132
development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus.a loop-mediated isothermal amplification (lamp) assay was developed for the detection of lymphocystis disease virus (lcdv). a set of five specific primers, two inner and two outer primers and a loop primer, were designed on the basis of the major capsid protein gene of lcdv. the reaction time and temperatures were optimized for 60 min at 63 degrees c, respectively. lamp amplification products were detected by a ladder-like appearance on agarose gel electrophoresis or a naked-eye inspection of a ...201019891986
a new marine megalocytivirus from spotted knifejaw, oplegnathus punctatus, and its pathogenicity to freshwater mandarinfish, siniperca chuatsi.megalocytivirus is a newly defined piscine iridovirus and has been shown to be an important causative agent of viral diseases in fish. here, a new megalocytivirus strain, designated skiv-zj07, was isolated from spotted knifejaw (oplegnathus punctatus) using a mandarinfish fry cell line (mff-1). phylogenetic analysis of the major capsid protein and atpase genes showed that skiv-zj07 was most similar to the orange-spotted grouper iridovirus (osgiv) from china and a u1 strain red sea bream iridovir ...201019895861
rapid and sensitive detection of infectious spleen and kidney necrosis virus by loop-mediated isothermal amplification combined with a lateral flow dipstick.loop-mediated isothermal amplification (lamp) allows rapid amplification of nucleic acids under isothermal conditions. in this report, a 20-min lamp amplification of the dpol gene of infectious spleen and kidney necrosis virus (isknv) using a biotin-labeled primer was combined with lateral flow dipstick (lfd) chromatography for rapid and simple visual detection of isknv-specific amplicons. the lfd process involves a 5-min specific hybridization with an fitc-labeled dna probe to confirm the prese ...201020107846
administration of recombinant ifn1 protects zebrafish (danio rerio) from isknv infection.in the present study, we used the infectious spleen and kidney necrosis virus (isknv) and zebrafish model system to investigate the inhibitory effect of recombinant zebrafish interferon 1 (zfrifn1) on acute viral infection and the impact of time of zfrifn1 administration on its protective efficacy. in vivo experiments showed that administration of recombinant zfrinf1 up-regulated expression of several ifn-stimulated genes within 24 h of injection, and expression levels of these genes dropped to ...201020438850
identification of the vp92r gene from infectious spleen and kidney necrosis virus.megalocytiviruses, which belong to the family iridoviridae, are among the most harmful viruses to cultured fishes. infectious spleen and kidney necrosis virus (isknv) is the type species of the megalocytivirus genus. the genome of isknv was sequenced and annotated by he et al. (virology 291:126-139, 2001) in 2001 and re-annotated by eaton et al. (virol. j. 4:11, 2007) in 2007. the orf092r of isknv was identified in the annotations of he et al. but was not reported by eaton et al. in this study, ...201020577793
complete genome sequence of a megalocytivirus (family iridoviridae) associated with turbot mortality in china.turbot reddish body iridovirus (trbiv) causes serious systemic diseases with high mortality in the cultured turbot, scophthalmus maximus. we here sequenced and analyzed the complete genome of trbiv, which was identified in shandong province, china.201020630106
characteristics of the interferon regulatory factor pairs zfirf5/7 and their stimulation expression by isknv infection in zebrafish (danio rerio).the interferon regulatory factor (irf) family plays critical roles in a host's virus infection responses. in this study, two irf family members, zfirf5 and zfirf7, are identified in zebrafish. the zfirf5 protein encodes 297 amino acids without the carboxyl irf3 domain. we suggest that zfirf5 is a new kind of splicing variant, following the nine other kinds of irf5 splicing variants found in mammals. the zfirf7 protein is identified as a member of the irf7 family, compared to the human irf7 prote ...201020637800
vp23r of infectious spleen and kidney necrosis virus mediates formation of virus-mock basement membrane to provide attaching sites for lymphatic endothelial cells.putative open reading frames (orfs) encoding laminin-like proteins are found in all members of the genus megalocytivirus, family iridoviridae. this is the first study that identified the vp23r protein encoded by orf23r of the infectious spleen and kidney necrosis virus (isknv), a member of these genes of megalocytiviruses. the vp23r mrna covering the isknv genomic coordinates 19547 to 22273 was transcribed ahead of the major capsid protein. immunofluorescence analysis demonstrated that vp23r was ...201020810728
detection of megalocytivirus from imported tropical ornamental fish, paradise fish macropodus opercularis.megalocytivirus was detected from paradise fish macropodus opercularis imported from indonesia. four of 11 fish (36%) in 2006 and 40 of 117 fish (34%) in 2008 were found to be pcr-positive for megalocytivirus. phylogenetic analysis based on partial major capsid protein (mcp) gene nucleotide sequences revealed that the sequences detected in paradise fish were classified as genotype ii, which includes freshwater fish isolates from southeast asian countries, closely related to infectious spleen and ...201020815332
proteomic analysis of zebrafish (danio rerio) infected with infectious spleen and kidney necrosis virus.iridovirus infections remain a severe problem in aquaculture industries worldwide. infectious spleen and kidney necrosis virus (isknv), the type species of the genus megalocytovirus in the family iridoviridae, has caused significant economic losses among freshwater fish in different asian countries. to investigate the molecular mechanism of iridoviral pathogenesis, we analyzed the differential proteome from the spleen of isknv-infected zebrafish through two-dimensional gel electrophoresis (2-de) ...201021075138
genotype and host range analysis of infectious spleen and kidney necrosis virus (isknv).infectious spleen and kidney necrosis virus (isknv) is the causative agent of a disease causing high mortality in mandarin fish, siniperca chuatsi. in this study, complete major capsid protein (mcp) genes of nine isknv isolates were sequenced and compared with other known megalocytiviruses to evaluate genetic variation and host range of the viruses. comparison of nucleotide sequences of mcp gene revealed 92.6-100% identity among nine isknv isolates. a phylogenetic tree revealed that 33 megalocyt ...201021107672
global landscape of structural proteins of infectious spleen and kidney necrosis virus.infectious spleen and kidney necrosis virus (isknv), the type species of the genus megalocytivirus in the family iridoviridae, causes severe damage to mandarin fish cultures in china. little is known about the proteins of isknv virions. in this study, a total of 38 isknv virion-associated proteins were identified by four different workflows with systematic and comprehensive proteomic approaches. among the 38 identified proteins, 21 proteins were identified by the gel-based workflows (one-dimensi ...201121209107
involvement of caveolin-1 in the jak-stat signaling pathway and infectious spleen and kidney necrosis virus infection in mandarin fish (siniperca chuatsi).caveolae, the major source of caveolin-1 protein, are specialized invaginated microdomains of the plasma membrane that act as organizing centers for signaling molecules in the immune system. in the present study, we report the cloning and characterization of caveolin-1 (mcav-1) from mandarin fish (siniperca chuatsi) and study on the roles of mcav-1 in the fish jak-stat signaling pathway and in virus infection. the cdna sequence of mcav-1 was 707bp in size, encoding a protein of 181 amino acids, ...201121296425
cloning, characterization, and biological function analysis of the sidt2 gene from siniperca chuatsi.the systemic rna interference defective protein (sid)-1 plays an important role in dsrna uptake in cells. we identified the scsidt2 gene from the mandarin fish (siniperca chuatsi), which is the first-studied sid-1 homolog in fish. scsidt2 mrna is 3380 bp long and contains a 2568-bp open reading frame that encodes an 855-amino-acid protein with an n-terminal signal peptide and ten putative transmembrane domains. tissue distribution profile in healthy fish and expression profiles of scsidt2 in inf ...201121334374
the viral ankyrin repeat protein (orf124l) from infectious spleen and kidney necrosis virus attenuates nf-{kappa}b activation and interacts with ikk{beta}the ankyrin (ank) repeat is one of the most common protein-protein interaction motifs predominantly found in eukaryotes and bacteria, but the functions of ank repeat are rarely researched in animal viruses with the exception of poxviruses. infectious spleen and kidney necrosis virus (isknv) is a typical species of the genus megalocytivirus in the family iridoviridae and a causative agent of epizootics in fish. the genome of isknv contains four putative viral ankyrin (vank) repeat proteins, and t ...201121471317
genetic analysis of fish iridoviruses isolated in taiwan during 2001-2009.to investigate the genetic relationships between field strains of iridoviruses gathered from various fish species in taiwan, viruses that were collected from 2001 to 2009 were analyzed. open reading frames encoding the viral major capsid protein (mcp) and adenosine triphosphatase (atpase) were sequenced for phylogenetic analysis. our results indicated that iridoviruses from taiwan aquaculture fishes could be classified into two groups: prior to 2005, the viruses were closely related to members o ...201121603939
Antigenic identification of virion structural proteins from infectious spleen and kidney necrosis virus.Infectious spleen and kidney necrosis virus (ISKNV), belonging to the genus Megalocytivirus in the family Iridoviridae, is one of the major agents causing mortality and economic losses to the freshwater fish culture industry in Asian countries. Currently, little information regarding the antigenic properties of Megalocytivirus (especially ISKNV) is available. Our previous study using four different workflows with systematic and comprehensive proteomic approaches led to the identification of 38 I ...201121888976
infectious spleen and kidney necrosis virus (a fish iridovirus) enters mandarin fish fry cells via caveolae-dependent endocytosis.infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus from the family iridoviridae. megalocytiviruses have been implicated in more than 50 fish species infections and currently threaten the aquaculture industry, causing great economic losses in china, japan, and southeast asia. however, the cellular entry mechanisms of megalocytiviruses remain largely uncharacterized. in this study, the main internalization mechanism of isknv is investigated in mand ...201122171272
a novel viral socs from infectious spleen and kidney necrosis virus: interacts with jak1 and inhibits ifn-α induced stat1/3 activation.interferon (ifn)-induced janus kinase (jak)/signal transducer and activator of transcription (stat) pathway is important in controlling immune responses and is negatively response-regulated by the suppressor of cytokine signaling (socs) proteins. however, several viruses have developed various strategies to inhibit this pathway to circumvent the anti-viral immunity of the host. the infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus in the family ...201222844427
display of isknv orf086 protein on the surface of aeromonas hydrophila and its immunogenicity in chinese perch (siniperca chuatsi).co-infection with infectious spleen and kidney necrosis virus (isknv) and aeromonas hydrophila is becoming ever more widespread in chinese perch (siniperca chuatsi) aquaculture industry, so that it's necessary to develop the combined vaccine against isknv and a. hydrophila disease. the surface display of heterologous on bacteria using anchoring motifs from outer membranes proteins has already been explored as an effective delivery system of viral antigens. in present study, the isknv orf086 gene ...201627436517
susceptibility of a number of australian freshwater fishes to dwarf gourami iridovirus (infectious spleen and kidney necrosis virus).megalocytiviruses cause high mortality diseases that have seriously impacted aquaculture, with the most frequent outbreaks occurring in east and south-east asia. the international trade of juvenile fish for food and ornamental aquaculture has aided the spread of these viruses, which have spread to europe and australia and other regions. australian freshwater fishes were examined for susceptibility to infection with the exotic megalocytivirus, dwarf gourami iridovirus (dgiv), which belongs to a g ...201727334576
detection of dwarf gourami iridovirus (infectious spleen and kidney necrosis virus) in populations of ornamental fish prior to and after importation into australia, with the first evidence of infection in domestically farmed platy (xiphophorus maculatus).the movement of ornamental fish through international trade is a major factor for the transboundary spread of pathogens. in australia, ornamental fish which may carry dwarf gourami iridovirus (dgiv), a strain of infectious spleen and kidney necrosis virus (isknv), have been identified as a biosecurity risk despite relatively stringent import quarantine measures being applied. in order to gain knowledge of the potential for dgiv to enter australia, imported ornamental fish were sampled prior to e ...201526452601
megalocytiviruses.the genus megalocytivirus, represented by red sea bream iridovirus (rsiv), the first identified and one of the best characterized megalocytiviruses, infectious spleen and kidney necrosis virus (isknv), the type species of the genus, and numerous other isolates, is the newest genus within the family iridoviridae. viruses within this genus are causative agents of severe disease accompanied by high mortality in multiple species of marine and freshwater fish. to date outbreaks of megalocytivirus-ind ...201222590684
early protein orf086 is an effective vaccine candidate for infectious spleen and kidney necrosis virus in mandarin fish siniperca chuatsi.infectious spleen and kidney necrosis virus (isknv) has caused significant loss in the mandarin fish (siniperca chuatsi) aquaculture industry. vaccination is an important measure to prevent fatal isknv infection. in this study, the orf086 gene encoding an early protein helicase of isknv was cloned into the prokaryotic pet32a (+) and eukaryotic pcdna3.1 (+) expression vectors and designated as pet086 and pcdna086, respectively. a recombinant 36 kda protein was detected in escherichia coli bl21 (h ...201526099219
protein encoded by orf093 is an effective vaccine candidate for infectious spleen and kidney necrosis virus in chinese perch siniperca chuatsi.infectious spleen and kidney necrosis virus (isknv) is the causative agent of a disease causing high mortality and economic losses in chinese perch, siniperca chuatsi in china. little information about the antigenicity of isknv proteins is available. in this study the orf093 gene of isknv was cloned into the prokaryotic expression vector pet32a(+) and eukaryotic expression vector pcdna3.1(+), and designated as pet-093 and pcdna-093, respectively. a recombinant 51-kda protein was detected in esch ...201525462463
application and development of a taqman real-time pcr for detecting infectious spleen and kidney necrosis virus in siniperca chuatsi.infectious spleen and kidney necrosis virus (isknv) is one of the major epidemiological agents that had caused great economic loss in chinese perch (siniperca chuatsi). in this study, a specific taqman real-time pcr was developed using a pair of primers and a taqman probe specific to the orf007 gene of isknv to rapidly detect isknv copies in chinese perch samples. this assay was optimized to produce linearity from 8.75 × 10(8) to 8.75 × 10(1) copies in standard curve with an efficiency of 98% an ...201728323153
tc38, a teleost tfpi-2 peptide that kills bacteria via penetration of the cell membrane and interaction with nucleic acids.tissue factor pathway inhibitor 2 (tfpi-2) is an analog of tfpi-1 and a potent endogenous inhibitor of tissue factor (tf)-mediated blood coagulation. recent reports have proven that the c-terminal of tfpi-2 peptides in humans and several other vertebrates possesses antibacterial activity against gram-positive and gram-negative bacteria. in our previous study, we reported that the tfpi-2 peptide, tc38 in tongue sole (cynoglossus semilaevis) was active against micrococcus luteus. in this study, we ...201728263880
detection of infectious spleen and kidney necrosis virus (isknv) and turbot reddish body iridovirus (trbiv) from archival ornamental fish samples.although infections caused by megalocytiviruses have been reported from a wide range of finfish species for several decades, molecular characterisation of the viruses involved has been undertaken only on more recent cases. sequence analysis of the major capsid protein and adenosine triphosphatase genes is reported here from formalin-fixed, paraffin-embedded material from 2 archival ornamental fish cases from 1986 and 1988 in conjunction with data for a range of genes from fresh frozen tissues fr ...201628000602
glutamine and glutaminolysis are required for efficient replication of infectious spleen and kidney necrosis virus in chinese perch brain cells.viruses rely on host cellular metabolism for energy and macromolecule synthesis during their replication. infectious spleen and kidney necrosis virus (isknv) causes significant economic losses in the chinese perch (siniperca chuatsi) industry worldwide. however, little is known about the relationship between isknv replication and cellular metabolism. using transcriptomic analysis, we observed that glutamine metabolism in chinese perch brain (cpb) cells is altered during isknv infection. moreover ...201727911855
megalocytivirus infection in cultured nile tilapia oreochromis niloticus.megalocytiviruses, such as infectious spleen and kidney necrosis virus (isknv), induce lethal systemic diseases in both ornamental and food fish species. in this study, we investigated an epizootic affecting nile tilapia oreochromis niloticus cultured in the us midwest. diseased fish displayed lethargy, gill pallor, and distension of the coelomic cavity due to ascites. histopathological examination revealed a severe systemic abundance of intravascular megalocytes that were especially prominent i ...201627225209
first outbreak of an infection with infectious spleen and kidney necrosis virus (isknv) in ornamental fish in germany.in 2014, infectious spleen and kidney necrosis virus (isknv), a member of the genus megalocytivirus, was detected for the first time in ornamental fish in germany. since 2013, angelfish pterophyllum spp. originating from colombia have experienced significant epizootics in a number of german retailers' facilities. the diseased fish showed symptoms such as increased ventilation, swollen gills, and ulcerations of the skin. in 2014, diseased angelfish p. altum and platys xiphophorus maculatus mainta ...201627225207
detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue-loop-mediated isothermal amplification assay.infectious spleen and kidney necrosis virus (isknv) has recently been recognized as a causative agent of serious systemic disease in tilapia. our objective was to establish a new colorimetric loop-mediated isothermal amplification (lamp) assay with pre-addition of hydroxynapthol blue (blue-lamp) to investigate isknv transmission in tilapia.201627138064
a microrna from infectious spleen and kidney necrosis virus modulates expression of the virus-mock basement membrane component vp08r.infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus, family iridoviridae. infection of isknv is characterized by a unique pathological phenomenon in that the infected cells are attached by lymphatic endothelial cells (lecs). isknv mediates the formation of a virus-mock basement membrane (vmbm) structure on the surface of infected cells to provide attaching sites for lecs. the viral protein vp08r is an important component of vmbm. in this study, a ...201626896933
differential autophagic effects triggered by five different vertebrate iridoviruses in a common, highly permissive mandarinfish fry (mff-1) cell model.autophagy of five vertebrate iridoviruses, including one megalocytivirus (infectious spleen and kidney necrosis virus, isknv) and four ranaviruses (chinese giant salamander iridovirus, cgsiv; tiger frog virus, tfv; grouper iridovirus, giv; and largemouth bass virus, lmbv) were investigated in a common, highly permissive mandarinfish fry (mff-1) cell model. the results showed marked autophagosome formation in giv- and lmbv-infected cells but not in isknv-, cgsiv- and tfv-infected mff-1 cells. str ...201626748344
molecular confirmation of infectious spleen and kidney necrosis virus (isknv) in farmed and imported ornamental fish in australia.viruses of the genus megalocytivirus have not been detected in wild populations of fish in australia but circulate in imported ornamental fish. in 2012, detection of a megalocytivirus in healthy platys xiphophorus maculatus was reported from a farm in australia during surveillance testing as part of a research project undertaken at the university of sydney. confirmatory testing of the original samples at the aahl fish diseases laboratory verified the presence of an infectious spleen and kidney n ...201526480913
transcriptomic analysis of mandarin fish brain cells infected with infectious spleen and kidney necrosis virus with an emphasis on retinoic acid-inducible gene 1-like receptors and apoptosis pathways.infectious spleen and kidney necrosis virus (isknv) has caused significant economic losses in the cultured mandarin fish (siniperca chuatsi) industry. the molecular mechanisms that underlie the pathogenesis of the viral infection remain poorly understood. in this study, deep rna sequencing technique was used to analyze the transcriptomic profiles of mandarin fish brain cells (cpb) at progressive time points after isknv infection. a total of 96,206,040 clean data from 98,235,240 sequence reads we ...201525982401
molecular cloning and expression analysis of an apoptosis-associated gene daxx from zebrafish, danio rerio.the death domain-associated protein daxx exerts many functions including the induction and inhibition of apoptosis, regulation of chromatin remodeling and gene transcription. in this report, we have cloned and characterized a daxx ortholog from the zebrafish, danio rerio. the bioinformatics analysis results indicated that the open reading frame (orf) of zebrafish daxx is 2,151bp long and encodes a putative protein of 716 amino acids containing daxx domain. though quantitative pcr analyses, daxx ...201525862973
a novel fish cell line derived from the brain of chinese perch siniperca chuatsi: development and characterization.in this study, a continuous cell line (named as cpb) was established from siniperca chuatsi brain and has been subcultured >140 times. cpb cell line predominantly consisted of fibroblast-like cells that could grow better in leibovitz's l-15 supplemented with 10% foetal bovine serum at 28° c. polymerase chain reaction amplification of 18s recombinant (r)rna confirmed the origin of this cell line from s. chuatsi. the cpb cell line was cryopreserved at different passage levels and revived successfu ...201525376532
interaction of infectious spleen and kidney necrosis virus orf119l with pinch leads to dominant-negative inhibition of integrin-linked kinase and cardiovascular defects in zebrafish.infectious spleen and kidney necrosis virus (isknv) is the type species of the megalocytivirus genus, iridoviridae family, causing a severe systemic disease with high mortality in mandarin fish (siniperca chuatsi) in china and southeast asia. at present, the pathogenesis of isknv infection is still not fully understood. based on a genome-wide bioinformatics analysis of isknv-encoded proteins, we found that isknv open reading frame 119l (orf119l) is predicted to encode a three-ankyrin-repeat (3an ...201525355883
protective immunity against infectious spleen and kidney necrosis virus induced by immunization with dna plasmid containing mcp gene in chinese perch siniperca chuatsi.infectious spleen and kidney necrosis virus (isknv) is the causative agent of a disease leading to high mortality and economic losses in chinese perch, siniperca chuatsi. there is an urgent need to develop an effective vaccine against this fatal disease. in this study, the mcp gene encoding the major capsid protein, the predominant structural component of the iridovirus particles, was cloned into a eukaryotic expression vector pcdna3.1+, and the recombinant plasmid, designated as pcmcp, was cons ...201425038286
orf005l from infectious spleen and kidney necrosis virus is located in the inner mitochondrial membrane and induces apoptosis.infectious spleen and kidney necrosis virus (isknv) belongs to the genus megalocytivirus in the family iridoviridae. this virus is the etiological agent of a serious and pandemic disease in fish. cells infected with isknv undergo apoptosis. in this study, the isknv orf005l gene was characterized and functionally investigated. bioinformatics analysis revealed that the orf005l protein contains a region similar to the catalytic domain of ctd-like phosphatases. real-time quantitative-pcr results sho ...201424862228
vp08r from infectious spleen and kidney necrosis virus is a novel component of the virus-mock basement membrane.infectious spleen and kidney necrosis virus (isknv), the type species of the genus megalocytivirus, family iridoviridae, brings great harm to fish farming. in infected tissues, isknv infection is characterized by a unique phenomenon, in that the infected cells are attached by lymphatic endothelial cells (lecs), which are speculated to wall off the infected cells from host immune attack. a viral membrane protein, vp23r, binds and recruits the host nidogen-1 protein to construct a basement membran ...201424599992
disease resistance and immune-relevant gene expression in golden mandarin fish, siniperca scherzeri steindachner, infected with infectious spleen and kidney necrosis virus-like agent.infectious spleen and kidney necrosis virus (isknv), family iridoviridae, genus megalocytivirus, may cause high mortality rates such as those seen in mandarin fish, siniperca chuatsi. isknv has attracted much attention due to the possible environmental threat and economic losses it poses on both cultured and wild populations. we have investigated the pathogenicity of isknv-like agent megalocytivirus, isolated from infected pearl gourami, in golden mandarin fish, siniperca scherzeri - a member of ...201424111797
field trial tests of fkc vaccines against rsiv genotype megalocytivirus in cage-cultured mandarin fish (siniperca chuatsi) in an inland reservoir.megalocytiviruses are one of the most important causative agents in finfish industry in china, japan and south east asia. the viruses are mainly composed of isknv, rsiv and trbiv genotypes. among them, isknv genotype isolate is the most important causative agent in mandarin fish industry in south china. since its first occurrence in mid-1990s in china, no effective drug has been developed to prevent and control this virus until our recent work. in this study, unusual rsiv genotype megalocytiviru ...201324035751
detection and molecular characterization of infectious spleen and kidney necrosis virus from major ornamental fish breeding states in peninsular malaysia.'gold standard' oie reference pcr assay was utilized to detect the presence of infectious spleen and kidney necrosis virus (isknv) in freshwater ornamental fish from malaysia. from total of 210 ornamental fish samples representing 14 species, isknv was detected in 36 samples representing 5 fish species. all positive cases did not show any clinical signs of isknv. three restriction enzymes analyses showed that the fish were infected by identical strains of the same virus species within megalocyti ...201423952914
the first case of infectious spleen and kidney necrosis virus (isknv) infection in aquarium-maintained mandarin fish, siniperca chuatsi (basilewsky), in japan. 201423763499
a truncated danio rerio pkz isoform functionally interacts with eif2α and inhibits protein synthesis.a protein kinase containing z-dna binding domains (pkz), which resembles protein kinase r (pkr) in domain organization, was recently discovered to be a member of the eif2α kinase family in fish. pkr has roles in antiviral immunity through inhibiting protein synthesis and activating nf-κb; therefore, it is thought that pkz may have a similar role in fish antiviral immunity. in the present study, the roles of two danio rerio pkz isoforms (drpkz-a and drpkz-b) in eif2α phosphorylation and protein s ...201323742890
the potential role of microfilaments in host cells for infection with infectious spleen and kidney necrosis virus infection.infectious spleen and kidney necrosis virus (isknv) belongs to the genus megalocytivirus from the family iridoviridae. megalocytivirus causes severe economic losses to tropical freshwater and marine culture industry in asian countries and is devastating to the mandarin fish farm industry in china particularly.201323497248
mandarin fish caveolin 1 interaction with major capsid protein of infectious spleen and kidney necrosis virus and its role in early stages of infection.infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus from the family iridoviridae. isknv is one of the major agents that cause mortality and economic losses to the freshwater fish culture industry in asian countries, particularly for mandarin fish (siniperca chuatsi). in the present study, we report that the interaction of mandarin fish caveolin 1 (mcav-1) with the isknv major capsid protein (mcp) was detected by using a virus overlay assay and con ...201323283951
efficacy of a formalin-killed cell vaccine against infectious spleen and kidney necrosis virus (isknv) and immunoproteomic analysis of its major immunogenic proteins.infectious spleen and kidney necrosis virus (isknv), the type species of genus megalocytivirus in the family iridoviridae, is the most important etiological agents in mandarin fish industry in china. since its first occurrence in china in the early 1990s, there is no effective method to prevent and control this virus. here we report the successful development of a formalin-killed cell-cultured (fkc) vaccine again isknv. immuno-protection experiments showed that greater than 90% of fish immunized ...201323177910
immunological evaluation of vibrio alginolyticus, vibrio harveyi, vibrio vulnificus and infectious spleen and kidney necrosis virus (isknv) combined-vaccine efficacy in epinephelus coioides.combined vaccines are immunological products intended for immunization against multifactorial infectious diseases caused by different types or variants of pathogens. in this study, the effectiveness of vibrio alginolyticus (va), vibrio harveyi (vh), vibrio vulnificus (vv) and infectious spleen and kidney necrosis virus (isknv), an iridovirus, combined-vaccine (vibrio and isknv combined vaccines, vicv), va+vh+vv inactive vaccine (viv) and isknv whole cell inactive vaccine (iwciv) in epinephelus c ...201223010220
protective immunity against iridovirus disease in mandarin fish, induced by recombinant major capsid protein of infectious spleen and kidney necrosis virus.infectious spleen and kidney necrosis virus (isknv) is the causative agent of a disease causing high mortality and economic losses in mandarin fish, siniperca chuatsi in china. but little information about vaccine development against isknv disease is available. in this study the gene encoding the major capsid protein (mcp), which is predominant structural component of the iridovirus particles, was cloned into a temperature induction prokaryotic expression vector pbv220 and a recombinant protein ...201222971336
the viral traf protein (orf111l) from infectious spleen and kidney necrosis virus interacts with tradd and induces caspase 8-mediated apoptosis.infectious spleen and kidney necrosis virus (isknv) is the type species of the megalocytivirus genus of the iridoviridae family. it causes a serious and potentially pandemic disease in wild and cultured fishes. isknv infection induces evident apoptosis in mandarin fish (siniperca chuatsi) and zebrafish (danio renio). however, the mechanism is still unknown. after a genome-wide bioinformatics analysis of isknv-encoded proteins, the isknv open reading frame 111l (orf111l) shows a high similarity t ...201222615868
high reduced/oxidized glutathione ratio in infectious spleen and kidney necrosis virus-infected cells contributes to degradation of vp08r multimers.infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus, family iridoviridae. the isknv-infected cells in fish tissues are attached by lymphatic endothelial cells (lecs), which is a unique pathological phenomenon of isknv infection. the viral proteins vp23r and vp08r and the host protein nidogen-1 constitute the virus-mock basement membrane (vmbm) on the membrane of infected cells to provide attaching sites for lecs. vp08r can form cross-linked multim ...201728757023
infectious spleen and kidney necrosis disease (isknd) outbreaks in farmed barramundi (lates calcarifer) in vietnam.emergence of a disease with clinical signs resembling megalocytivirus infection seriously affected large-scale barramundi farms in vietnam in 2012-2014 with estimated losses reaching $435,810 per year. an oil-based, inactivated vaccine against red sea bream iridovirus (rsiv) was applied in one farm for disease prevention without analysis of the causative agent, and the farmer reported inadequate protection. here we describe histological and molecular analysis of the diseased fish. pcr targeting ...201728663128
a loop-mediated isothermal amplification method for the detection of members of the genus ranavirus.a loop-mediated isothermal amplification (lamp) method was developed for detection of members of the genus ranavirus. the optimum reaction mixture contained 2.5 μl of each inner primer, rv-fip (20 pmol/μl) and rv-bip (20 pmol/μl), 0.5 μl of each outer primer, rv-f3 (10 pmol/μl) and rv-b3 (10 pmol/μl), 1.25 μl of each loop primer, rv-lf (20 pmol/μl) and rv-lb (20 pmol/μl), 3.5 μl dntp mix (10 mm each), 8 μl mgso4 (25 mm), 1 μl of bst dna polymerase (8 u/ml, large fragment; new england biolabs inc ...201323665768
vp15r from infectious spleen and kidney necrosis virus is a non-muscle myosin-ii-binding protein.vp15r protein, encoded by the 15th open reading frame of infectious spleen and kidney necrosis virus (isknv), was identified. vp15r is a 263-residue protein that is first transcribed within 12 h post-infection. the vp15r mrna is transcribed beginning at isknv genomic coordinate 12111, extending 167 bp upstream of the initiation codon. no signal peptides, transmembrane fragments, or nuclear localization signal sequences were predicted in the vp15r sequence. the 102-202 sequence of vp15r is homolo ...201020882305
tetraodon nigroviridis as a nonlethal model of infectious spleen and kidney necrosis virus (isknv) infection.infectious spleen and kidney necrosis virus (isknv) is the type species of the genus megalocytivirus, family iridoviridae. we have previously established a high mortality isknv infection model of zebrafish (danio rerio). in this study, a nonlethal tetraodon nigroviridis model of isknv infection was established. isknv infection did not cause lethal disease in tetraodon but could infect almost all the organs of this species. electron microscopy showed isknv particles were present in infected tissu ...201020674952
development of cross-priming amplification coupled with vertical flow visualization for rapid detection of infectious spleen and kidney necrosis virus (isknv) in mandarin fish, siniperca chuatsi.infectious spleen and kidney necrosis virus (isknv) has been recognized as the causative agent of the most serious disease in cultured mandarin fish, siniperca chuatsi, in china. disease outbreaks have resulted in substantial losses to the aquaculture industry. currently, reliable laboratory detection and identification methods are available for this virus. however, rapid detection methods applicable for on-site diagnosis of this infectious agent are unavailable. to address this need, a nearly i ...201729288074
complete genome sequence and phylogenetic analysis of megalocytivirus rsiv-ku: a natural recombination infectious spleen and kidney necrosis virus.megalocytiviruses are classified into three genotypes, infectious spleen and kidney necrosis virus (isknv), red seabream virus (rsiv), and turbo reddish body iridovirus (trbiv), based on the major capsid protein and atpase genes. however, only a few complete genome sequences have been obtained. this paper reports the complete genome sequence and phylogenetic analysis of an rsiv-ku strain megalocytivirus. the genome sequence comprises 111,154 bp, has 132 putative open reading frames, and is homol ...201729282546
to17: a teleost antimicrobial peptide that induces degradation of bacterial nucleic acids and inhibits bacterial infection in red drum, sciaenops ocellatus.tissue factor pathway inhibitor (tfpi)-1 is well known for its role as an inhibitor of blood coagulation. several studies have demonstrated that the c-terminal peptides of tfpi-1 are active against a broad spectrum of microorganisms. in a previous study, we found that to17 (with 17 amino acids), a tfpi-1 c-terminal peptide from red drum (sciaenops ocellatus), was active against edwardsiella tarda. in the present study, we investigated further the antimicrobial spectrum, action mode, as well as t ...201829183811
composition and transcription of all interferon regulatory factors (irfs), irf1‒11 in a perciform fish, the mandarin fish, siniperca chuatsi.interferon regulatory factors (irfs) are a family of mediators in various biological processes including immune modulation of interferon (ifn) and proinflammatory cytokine expression. however, the data on the complete composition of irfs is rather limited in teleost fish. in the present study, all irf members, i.e. irf1‒11 with two irf4, irf4a and irf4b have been characterised in an aquaculture species of fish, the mandarin fish, siniperca chuatsi, in addition to the previous report of irf1, irf ...201829180032
co-infections of infectious spleen and kidney necrosis virus and siniperca chuatsi rhabdovirus in chinese perch (siniperca chuatsi).in spite of the quite common co-infections of viruses in the cultured fish, most of the previous studies have just simply focused on the infection of a single pathogen. in this report, we observed that about 13% of cultured chinese perch have been co-infected by infectious spleen and kidney necrosis virus (isknv) and siniperca chuatsi rhabdovirus (scrv). furthermore, chinese perch could co-infected by isknv and scrv by intraperitoneally injection with the two viruses. interestingly, we revealed ...201728890148
the core ubiquitin system of mandarin fish, siniperca chuatsi, can be utilized by infectious spleen and kidney necrosis virus.the process of ubiquitination regulates various cellular processes. the ubiquitin-proteasome system (ups) in fish, which is important for the generation of innate and adaptive immune responses to pathogens, is the target of aquatic viruses to achieve immune evasion. we cloned and characterized three genes, namely, a ubiquitin-activating enzyme (sce1), a ubiquitin-conjugating enzyme (sce2), and a hect-type ubiquitin ligase (sce3) of mandarin fish siniperca chuatsi. the genes were expressed in all ...201728889013
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