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comparative membrane microviscosity of fish and mammalian rhabdoviruses studied by fluorescence depolarization.the microviscosity of the hydrophobic region of the membrane of infectious hematopoietic necrosis virus was determined using fluorescence depolarization analysis of the probe 1,6-diphenyl-1,3,5-hexatriene and was found to be much lower at 37 c than that of another rhabdovirus, vesicular stomatitis virus. however, the microviscosity of this fish virus at 18 c, the temperature at which it was grown, corresponded to the microviscosity of vesicular stomatitis virus at 37 c. data obtained with the fi ...1976181597
pathophysiology of infectious hematopoietic necrosis virus disease in rainbow trout: hematological and blood chemical changes in moribund fish.infectious hematopoietic necrosis (ihn) is a rhabdoviral disease of rainbow trout (salmo gairdneri). trout were injected with ihnv, and various hematological and biochemical measurements of clinically ill fish were compared to uninfected controls. infected fish had reduced corpuscular counts, hemoglobin, and packed cell volume, but normal mean corpuscular volume, mean corpuscular hemoglobulin, and mean corpuscular hemoglobin concentration. the percentage of immature erythrocytes was increased, b ...1975234912
partial purification and characterization of rtg-2 fish cell interferon.interferon produced by rainbow trout gonadal cells (rtg-2) was partially purified. the physical, chemical, and biological properties of this in vitro produced fish cell interferon were studied. purification was achieved by ultracentrifugation, molecular sieve gel chromatography, ion exchange chromatography, and polyacrylamide gel electrophoresis. the isoelectric point of rtg-2 interferon, as determined by cm-sephadex (c-50) chromatography, was 7.1. filtration through sephadex g-150 showed that r ...1975235493
fish rhabdoviruses: comparative study of protein structure.proteins from four fish rhabdoviruses have been studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the viruses were: trout viral hemorrhagic septicemia (vhs), infectious hematopoietic necrosis virus (ihn), spring viremia virus of carp (svc), and the pike fry rhabdovirus (pfr). for the two salmonid viruses (vhs-ihn), gel electrophoresis indicated the proteins, with molecular weights estimated to be 190,000, 80,000, 38,000, 25,000, and 19,000, respectively. the electrophoretic p ...19751171263
detection and transmission of infectious hematopoietic necrosis virus in rainbow trout.detection and transmission of infectious hematopoietic necrosis virus in rainbow trout (salmo gairdneri) was studied at a commercial trout hatchery. transmission of virus was demonstrated via water, feed and contaminated eggs. if eggs from carrier females were incubated several weeks in virus-free water, the resulting fry did not become infected. however, if fry subsequently became infected they were lifetime carriers. infectious virus was readily detectable in most tissues of moribund fish; in ...19751195486
neutralizing-enhancing monoclonal antibody recognizes the denatured glycoprotein of viral haemorrhagic septicaemia virus.two monoclonal antibodies (mabs), 1h10 and 1f10, have been selected against the denatured glycoprotein of the viral haemorrhagic septicaemia virus (vhsv) of salmonids by immunoblotting. three reference vhsv serotypes (f1, f2, 23.75) and five vhsv isolates from either different host species (trout, salmon, barbel) or geographical locations in spain reacted with both mabs by elisa. in vitro neutralization of all vhsv serotypes but not infectious haematopoietic necrosis virus (ihnv) was obtained on ...19921280943
staphylococcal coagglutination, a rapid method of identifying infectious hematopoietic necrosis virus.a staphylococcal coagglutination test was developed for the rapid detection of infectious hematopoietic necrosis virus (ihnv) in cell cultures and infected fish. the test could be completed in 15 min but required a minimum ihnv titer of 10(6) pfu/ml to obtain a positive reaction. all ihnv isolates, representing the five electropherotypes taken from a wide variety of species and different geographic ranges, caused coagglutination of staphylococcus aureus cells sensitized with rabbit polyclonal se ...19921539993
sensitive method for the detection of antibodies to infectious hematopoietic necrosis virus. 19921606268
nucleocapsid gene sequence of a north american isolate of viral haemorrhagic septicaemia virus, a fish rhabdovirus.viral haemorrhagic septicaemia is the most important viral disease of trout in europe. the causative agent, viral haemorrhagic septicaemia virus (vhsv), a member of the lyssavirus genus of the rhabdoviridae family, was formerly believed to be confined to portions of the european continent; however in 1988, vhsv was isolated from adult chinook (oncorhynchus tshawytscha) and coho (o. kisutch) salmon returning to two hatcheries in the northwestern part of the state of washington, u.s.a. initial fea ...19921634868
epitope mapping and characterization of the infectious hematopoietic necrosis virus glycoprotein, using fusion proteins synthesized in escherichia coli.a characterization of the antigenic determinants (epitopes) of the glycoprotein (g) of infectious hematopoietic necrosis virus was made by expressing different regions of the g gene in escherichia coli. a cdna copy of the g gene was divided into four fragments by taqi digestion, and the fragments were subcloned into path vectors, placing the expression of each g gene fragment under control of the trpe promoter. the resulting plasmids, pxl2, pxl3, and pxl7, encoded trpe-g fusion proteins subseque ...19911704930
the free nucleocapsids of the viral haemorrhagic septicaemia virus contain two antigenically related nucleoproteins.a protein of 34 kda (nx) was induced in vitro by the infection of fish cell cultures with the rhabdovirus agent of viral haemorrhagic septicaemia (vhs) of the trout. this protein only appeared as a major component in concentrated or intracellular labeled vhs virus but not in purified vhs or in the related infectious haematopoietic necrosis virus. that nx protein is antigenically related to the nucleoprotein of purified virus was shown by its reaction with four anti-nucleoprotein monoclonal antib ...19911877884
bacterially expressed nucleoprotein of infectious hematopoietic necrosis virus augments protective immunity induced by the glycoprotein vaccine in fish.the ribonucleoprotein gene of infectious hematopoietic necrosis virus (ihnv) has been expressed in escherichia coli as a trpe fusion protein. this viral protein does not induce protective immunity to lethal ihnv infection in fish, and virus-neutralizing antibodies do not react with this viral protein. however, when it was administered with a bacterial lysate containing a region of the ihnv glycoprotein, there was enhanced resistance in immunized fish to lethal virus infection.19912072459
fish cell lines: establishment and characterization of three new cell lines from grass carp (ctenopharyngodon idella).three new cell lines were established from tissues of the grass carp, ctenopharyngodon idella. derived from the fin, snout, and swim bladder of two apparently healthy diploid fry, these cell lines have been designated gcf, gcs-2, and gcsb, respectively. the cells grew at temperatures between 24 degrees and 36 degrees c with optimal growth at 32 degrees c and have been subcultured more than 50 times since their initiation in august 1986. two of the lines remained diploid or pseudodiploid after 38 ...19902108122
inhibitory effect of halocyamine, an antimicrobial substance from ascidian hemocytes, on the growth of fish viruses and marine bacteria.halocyamine a, an antimicrobial substance isolated from hemocytes of the solitary ascidian halocynthia roretzi, inhibited in vitro the growth of fish rna viruses (infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus). pretreatment of rna virus with halocyamine a reduced the infectivity of the virus toward host cells. the growth of marine bacteria, achromobacter aquamarinus and pseudomonas perfectomarinus, was also inhibited by halocyamine a but that of alteromonas put ...19902121517
cloning and sequencing the messenger rna of the n gene of viral haemorrhagic septicaemia virus.the mrna transcribed from the n gene of viral haemorrhagic septicaemia virus (vhsv) of salmonids has been cloned in escherichia coli and expressed. fusion proteins were recognized by monoclonal antibody directed against the n protein from the viral particle. a 1212 bp long open reading frame (orf) coding for 404 amino acids with a calculated mr of 44590 was deduced from the nucleotide sequence. the orf was preceded by a 93 bp segment including in position 42 the aacac pentanucleotide which is pr ...19902202782
antigen-capture elisa for viral haemorrhagic septicaemia virus serotype i.an antigen-capture elisa for viral haemorrhagic septicaemia virus serotype i (vhsv i) was developed. the assay employs two monoclonal antibodies (mab) directed against distinct epitopes of the viral envelope glycoprotein (gp). the antigen bound by the capture mab (a17) was detected by addition of a second mab (l7) conjugated to horseradish peroxidase, followed by addition of the enzyme substrate. the technique is highly sensitive, enabling detection of the virus at a protein concentration as low ...19902266147
expression of the glycoprotein gene from a fish rhabdovirus by using baculovirus vectors.a cdna fragment containing the gene encoding the glycoprotein of infectious hematopoietic necrosis virus was inserted into autographa californica baculovirus vectors under the control of the polyhedrin promoter. a 66-kilodalton protein, identical in size to the glycosylated glycoprotein of infectious hematopoietic necrosis virus, was expressed at high levels in spodoptera frugiperda cells infected with the recombinant viruses. the expressed protein reacted with antiserum to the glycoprotein on w ...19902403442
antiviral activities and the mechanism of 9-beta-d-ribofuranosyl-6-alkylthiopurines on several rna viruses from animals.a series of 9-beta-d-ribofuranosyl-6-alkylthiopurines (6-alkyl ti) were found to inhibit in vitro replication of infectious hematopoietic necrosis virus (ihnv), human influenza virus (ifv) and respiratory syncytial virus (rsv) with ic50 values of about 0.06 microgram/ml, 0.7-1.5 micrograms/ml and 1-3 micrograms/ml, respectively. viral rna synthesis in infected cells in the presence of actinomycin d was inhibited by treatment with the compounds dose-dependently. it was also found that the decreas ...19892608474
neutralizing antibodies for infectious hematopoietic necrosis virus in eggs of steelhead trout (salmo gairdneri).neutralizing antibodies specific for infectious hematopoietic necrosis virus (ihnv) were isolated from eggs of spawning steelhead trout (salmo gairdneri), using sodium sulfate precipitation. the isolated material was used in place of the primary antibody (rabbit anti-ihnv) in a protein immunoblotting assay to detect ihnv proteins specifically. the egg component that bound specifically to ihnv proteins was determined to be trout antibody by using antiserum toward trout immune globulins as the sec ...19892719385
the glycoprotein of infectious hematopoietic necrosis virus elicits neutralizing antibody and protective responses. 19892773597
detection of infectious hematopoietic necrosis virus in an invertebrate (callibaetis sp).infectious hematopoietic necrosis virus (ihnv) was detected in the common mayfly (callibaetis sp) by western immunoblot assay and was propagated in fish cells (chse-214) in culture. when propagated in cell culture, cytopathic effect characteristic of ihnv infection was observed. antibody specific for ihnv was used to detect all of the major proteins of ihnv in the western immunoblot assay. when the crude lysate was subjected to electron microscopy, bullet-shaped particles (84 nm x 194 nm) charac ...19892782712
nucleotide sequence of a cdna clone carrying the glycoprotein gene of infectious hematopoietic necrosis virus, a fish rhabdovirus.the nucleotide sequence of the mrna encoding the glycoprotein of infectious hematopoietic necrosis virus was determined from a cdna clone containing the entire coding region. the g-protein cdna is 1,609 nucleotides long (excluding the polyadenylic acid) and encodes a protein of 508 amino acids. the predicted amino acid sequence was compared with that of the glycoprotein of the indiana and new jersey serotypes of vesicular stomatitis virus and with the glycoprotein of rabies virus, using a comput ...19873033264
molecular filtration for recovery of waterborne viruses of fish.the effectiveness of tangential flow filtration (tff) for the recovery of infectious hematopoietic necrosis virus (ihnv) and infectious pancreatic necrosis virus (ipnv) from large volumes of water was evaluated. in laboratory studies, virus recovery from ihnv-seeded water following concentration by tff was approximately 13%. however, the addition of 0.1 and 1% fetal bovine serum to deionized water stabilized the virus, increasing virus recoveries to 95%. the addition of 0.03 and 0.3% beef extrac ...19883137867
isolation and identification of trout viruses in south africa.a total of 3,257 samples of diseased rainbow trout were examined for the presence of viruses from january 1983 to december 1987. a virus closely related to the vr 299 serotype of infectious pancreatic necrosis virus was isolated from 13 cases. an additional 7,228 viscera samples from asymptomatic fish were collected during the same period and a similar virus was isolated from 2 sites. during the same period 2,892 ovarian fluid samples were collected and a similar virus was isolated from 1 site. ...19883143095
the nucleocapsid gene of infectious hematopoietic necrosis virus, a fish rhabdovirus.the complete nucleotide sequence of the infectious hematopoietic necrosis virus (ihnv) nucleocapsid gene has been determined using cdna clones of genomic and messenger rnas. genomic clones were generated by using random dna oligomers to prime cdna synthesis and were mapped to their respective locations on the genome by the use of cdna probes derived from viral mrnas. interesting features of the ihnv nucleocapsid gene sequence elucidated by the sequencing of these clones include short homologies ...19883201758
the efficacy of amantadine and other antiviral compounds against two salmonid viruses in vitro.various compounds, with known clinical efficacy against human viruses, were evaluated for their ability to inhibit the growth of infectious hematopoietic necrosis virus (ihnv, a rhabdovirus), and infectious pancreatic necrosis virus (ipnv, a birnavirus), in rainbow trout cell cultures. amantadine inhibited the plaque forming ability of ihnv, at concentrations which did not affect cell growth or morphology, although it was not active against ipnv. metisazone and bis-benzimidazole were also effect ...19883228282
screening of bacteria with antiviral activity from fresh water salmonid hatcheries.bacteria isolated from two salmonid hatcheries were screened for antiviral activity against infectious hematopoietic necrosis virus (ihnv) to ascertain the presence of bacteria with anti-ihnv activity in the aquatic environment. out of 710 bacterial isolates from the water and sediment samples, 190 strains showed anti-ihnv activities of more than 50% plaque reduction. these antiviral activities were detected predominantly in pseudomonas, aeromonas/vibrio, and coryneforms. in one hatchery, the ba ...19883374405
transcription in vitro of infectious haematopoietic necrosis virus, a fish rhabdovirus.transcription in vitro by the rna polymerase of infectious haematopoietic necrosis virus (ihnv), a salmonid rhabdovirus, was investigated using different reaction conditions to maximize rna synthesis. the use of hepes buffer rather than tris buffer, and the addition of s-adenosyl-l-methionine to the reactions resulted in a sixfold increase in rna synthetic activity to 6400 pmol ump incorporated/mg viral protein/hour. the rna transcripts produced in this system contained polyadenylated species wh ...19873585284
the synergism of nucleoside antibiotics combined with guanine 7-n-oxide against a rhabdovirus, infectious hematopoietic necrosis virus (ihnv).guanine 7-n-oxide was shown to have synergistic activity in combination with neplanocin a against a rhabdovirus, infectious hematopoietic necrosis virus (ihnv), as reported previously. we examined further the antiviral activity of guanine 7-n-oxide in combination with other nucleoside antibiotics against ihnv. synergism was seen between guanine 7-n-oxide and d-eritadenine or cordycepin. it is considered that compounds inhibiting rna methylation show synergism with guanine 7-n-oxide.19863781928
occurrence of different types of infectious hematopoietic necrosis virus in fish.the virion protein patterns of 71 isolates of infectious hematopoietic necrosis virus (ihnv) from the pacific northwest were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [35s]-methionine-labeled virus. this analysis led to the classification of these virus isolates into four or more types. type 1 virus was characterized by a nucleocapsid protein with an approximate molecular weight of 40,500. type 2 and type 3 viruses have nucleocapsid proteins with molecular weights ...19863789723
molecular cloning of the six mrna species of infectious hematopoietic necrosis virus, a fish rhabdovirus, and gene order determination by r-loop mapping.plasmids carrying cdna sequences to the mrna species of infectious hematopoietic necrosis virus were constructed and cloned into escherichia coli. characterization of 21 cloned plasmids by hybridization to mrna blots identified sets of plasmids with homology to each of the six viral mrna species. r-loop mapping with these cdna plasmids determined that the gene order on the infectious hematopoietic necrosis virus genome is (3')n-m1-m2-g-nv-l(5').19853838192
antiviral activity and its mechanism of guanine 7-n-oxide on dna and rna viruses derived from salmonid.guanine 7-n-oxide produced by streptomyces sp. was found to inhibit in vitro the replication of herpes virus (oncorhynchus masou virus, omv), rhabdo virus (infectious hematopoietic necrosis virus, ihnv) and a bi-segmented double-strand virus (infectious pancreatic necrosis virus, ipnv) derived from salmonids with ic50 values of about 10 micrograms/ml, 20 micrograms/ml and 32 micrograms/ml, respectively. the agent was not toxic for the host cells (chinook salmon embryo, chse-214) at the ic50 conc ...19853841124
characterization of infectious hematopoietic necrosis virus mrna species reveals a nonvirion rhabdovirus protein.the genome rna and six mrna species of infectious hematopoietic necrosis virus were analyzed by denaturing gel electrophoresis. the following molecular weights were determined: genome rna, 3.7 x 10(6); mrna 1, 2.26 x 10(6); mrna 2, 5.63 x 10(5); mrna 3, 4.84 x 10(5); mrna 4 (containing two different mrna species), 3.00 x 10(5); and mrna 5, 1.95 x 10(5). densitometer analyses of gels were used to calculate the molar ratios of the intracellular mrna species: mrna 1, 0.02; mrna 2, 0.49; mrna 3, 1.0 ...19854038520
fish rhabdovirus replication in non-piscine cell culture: new system for the study of rhabdovirus-cell interaction in which the virus and cell have different temperature optima.the replication of three rhabdoviruses associated with diseases of fish has been demonstrated in cells of continuously cultivated non-piscine cell lines. spring viremia of carp (svc) virus and the salmonid fish viruses, egtved and infectious hematopoietic necrosis virus, all replicated in mammalian wi-38 (human diploid cell strain) and bhk/21 cells and in cells of one or more reptilian cell lines at the temperatures commonly used to propagate these viruses in fish cells. the infections were cyto ...19744210338
fish viruses: buffers and methods for plaquing eight agents under normal atmosphere.a universal procedure was sought for plaque assay of eight fish viruses (bluegill myxovirus, channel catfish virus, eel virus, egtved virus, infectious hematopoietic necrosis virus, infectious pancreatic necrosis virus, lymphocystis virus, and the agent of spring viremia of carp (rhabdovirus carpio), in dish cultures of various fish cells. eagle minimal essential medium with sodium bicarbonate-co(2) buffer (earle's salt solution) was compared with minimal essential medium buffered principally wi ...19734349252
further characterization of infectious hematopoietic necrosis virus of salmonid fish (oregon strain). 19744365605
autointerference in infectious hematopoietic necrosis virus of salmonid fish. 19744856418
in vitro and in vivo detection of infectious pancreatic necrosis virus in fish by enzyme-linked immunosorbent assay.a double antibody enzyme-linked immunosorbent assay (elisa) was used to detect infectious pancreatic necrosis virus (ipnv). the elisa detected vr299 strain of ipnv at a dose of 10 to 20 ng of purified ipnv protein or 10(4) tcid50 in tissue culture fluid. specificity of elisa was demonstrated by an elisa inhibition test. the elisa did not detect infectious hematopoietic necrosis virus. normal cell culture fluid and virus-non-inoculated rainbow trout (salmo gairdneri richardson) homogenate did not ...19846093652
comparison of in vitro growth characteristics of ten isolates of infectious haematopoietic necrosis virus.ten isolates of infectious haematopoietic necrosis from salmonid fishes of different locations on the west coast of north america from california to alaska were compared by plaque size, single-step growth curves at 15 and 18 degrees c, rate of appearance of cytopathic effects in cell cultures, and growth over a range of temperatures. all isolates were distinguishable on the basis of each growth characteristic examined. the co isolate from the sacramento river drainage of california was the most ...19846512504
appearance and quantification of infectious hematopoietic necrosis virus in female sockeye salmon (oncorhynchus nerka) during their spawning migration.the incidence and amount of infectious hematopoietic necrosis (ihn) virus was determined in 10 organs and body fluids from each of 100 female sockeye salmon (oncorhynchus nerka) before, during, and after their spawning migration into freshwater. virus was found in high concentrations only in fish sampled during and after spawning. infection rates increased from nil to 100 percent within 2 weeks. in spawning fish, incidences of ihn virus were high in all organs and fluids except brain and serum, ...19846539105
adsorption to fish sperm of vertically transmitted fish viruses.more than 99 percent of a vertically transmitted fish rhabdovirus, infectious hematopoietic necrosis virus, was removed from suspension in less than 1 minute by adsorption to the surface membrane of sperm from two genera of salmonid fishes. the vertically transmitted, infectious pancreatic necrosis virus adsorbed to a lesser degree, but no adsorption occurred with a second fish rhabdovirus that is not vertically transmitted. such adsorption may be involved in vertical transmission of these virus ...19846740314
plaquing procedure for infectious hematopoietic necrosis virus.a single overlay plaque assay was designed and evaluated for infectious hematopoietic necrosis virus. epithelioma papillosum carpio cells were grown in normal atmosphere with tris(hydroxymethyl)aminomethane- or hepes (n-2-hydroxyethylpiperazine-n'-2-ethanesulfonic acid)-buffered media. plaques were larger and formed more quickly on 1- to 3-day-old cell monolayers than on older monolayers. cell culture medium with a 10% addition of fetal calf serum (mem 10) or without serum (mem 0) were the most ...19806769393
thermal inactivation of infectious hematopoietic necrosis and infectious pancreatic necrosis viruses.a plaque assay was used to follow the inactivation kinetics of infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus in cell culture media at various temperatures. inactivation of infectious hematopoietic necrosis virus in a visceral organ slurry was compared with that in culture media.19817235707
recombinant infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus glycoprotein epitopes expressed in aeromonas salmonicida induce protective immunity in rainbow trout (oncorhynchus mykiss).fragments of the glycoprotein genes of viral hemorrhagic septicemia virus (vhsv) and infectious hematopoietic necrosis virus (ihnv) were cloned into a bacterial broad-host-range expression vector under the control of the plac promoter. western blot (immunoblot) analysis with monoclonal antibodies specific to the glycoproteins demonstrated the inducible expression of the fusion proteins in escherichia coli. aeromonas salmonicida is the causative agent of furunculosis in salmonid fish. it was conf ...19957486994
neutralization-resistant variants of infectious hematopoietic necrosis virus have altered virulence and tissue tropism.infectious hematopoietic necrosis virus (ihnv) is a rhabdovirus that causes an acute disease in salmon and trout. in this study, a correlation between changes in tissue tropism and specific changes in the virus genome appeared to be made by examining four ihnv neutralization-resistant variants (rb-1, rb-2, rb-3, and rb-4) that had been selected with the glycoprotein (g)-specific monoclonal antibody rb/b5. these variants were compared with the parental strain (rb-76) for their virulence and patho ...19947525991
distant strains of the fish rhabdovirus vhsv maintain a sixth functional cistron which codes for a nonstructural protein of unknown function.we used direct rna sequencing to determine the genomic organization of the region downstream from the g gene of viral hemorrhagic septicemia virus (vhsv), a fish rhabdovirus. this region contains a gene coding for a protein, identified as nonvirion protein (nv), and the gene coding for the rna polymerase (l). thus, vhsv genome organization was confirmed to be 3'-n-p-m-g-nv-l-5'. in both a virulent european (07-71) and an avirulent north american (makah) strain, the nv gene is transcribed into a ...19957571446
complete genomic sequence of the fish rhabdovirus infectious haematopoietic necrosis virus.the complete nucleotide sequence of the genome of the fish rhabdovirus infectious haematopoietic necrosis virus (ihnv) has been determined after cdna cloning of the viral genomic rna. sequence analysis showed the presence of six open reading frames encoding the nucleoprotein n, the matrix proteins m1 and m2, the glycoprotein g, a so-called non-structural protein nv, and the rna polymerase l. the genome organization is 3'n-m1-m2-g-nv-l 5'. the extreme 5' and 3' ends of the genome were sequenced a ...19957595355
the genetic diversity and epizootiology of infectious hematopoietic necrosis virus.infectious hematopoietic necrosis virus (ihnv) is a rhabdovirus which causes a serious disease in salmonid fish. the t1 ribonuclease fingerprinting method was used to compare the rna genomes of 26 isolates of ihnv recovered from sockeye salmon (oncorhynchus nerka), chinook salmon (o. tshawytscha), and steelhead trout (o. mykiss) throughout the enzootic portion of western north america. most of the isolates analyzed in this study were from a single year (1987) to limit time of isolation as a sour ...19957762287
detection of truncated virus particles in a persistent rna virus infection in vivo.infectious hematopoietic necrosis virus (ihnv) is a rhabdovirus which causes devastating epizootics of trout and salmon fry in hatcheries around the world. in laboratory and field studies, epizootic survivors are negative for infectious virus by plaque assay at about 50 days postexposure. survivors are considered virus free with no sequelae and, thus, are subsequently released into the wild. when adults return to spawn, infectious virus can again be isolated. two hypotheses have been proposed to ...19957884861
sequences of the n and m genes of the sigma virus of drosophila and evolutionary comparison.the genome of the sigma rhabdovirus of drosophila melanogaster consists of six genes in the order 3' n-2-3-4-g-l 5'. the nucleotide sequences of the n and of the fourth genes were determined from cdna clones. each gene contained a single long open reading frame encoding polypeptides with predicted mw of 50 and 25 kda, respectively. evidence that these genes encode the nucleocapsid n and the matrix m proteins were obtained using antibodies raised against recombinant proteins derived from the clon ...19948128620
molecular cloning and expression in escherichia coli of the glycoprotein gene of vhs virus, and immunization of rainbow trout with the recombinant protein.the gene encoding the envelope glycoprotein of a recent danish isolate of a salmonid rhabdovirus, viral haemorrhagic septicaemia virus (vhsv) has been cloned and sequenced at the cdna level. when compared with the deduced sequence of a french isolate of vhsv, it was noted that there were 13 amino acid substitutions in the danish virus. amino acid homologies with the glycoprotein of a north american salmonid rhabdovirus (infectious haematopoietic necrosis virus) indicate a high degree of structur ...19938468553
characterization of a rainbow trout mx gene.a full-length cdna clone of a rainbow trout (oncorhynchus mykiss) mx gene was obtained using race (rapid amplification of cdna ends) polymerase chain reaction (pcr) amplification of rna extracted from poly (i).(c)-induced rainbow trout gonad cells (rtg-2). mx was previously identified in rainbow trout by staeheli et al. by hybridization with a partial perch genomic mx probe to induced rainbow trout mrna. the 2.5 kb rainbow trout cdna clone contains an open reading frame of 1863 nt (nucleotides) ...19958528941
synthetic nucleosides and nucleotides. xxxv. synthesis and biological evaluations of 5-fluoropyrimidine nucleosides and nucleotides of 3-deoxy-beta-d-ribofuranose and related compounds.1-o-acetyl-2,5-di-o-p-chlorobenzoyl-3-deoxy-d-ribofuranose (1), derived from the antibiotic cordycepin was coupled with trimethylsilylated derivatives (2a-c) of n4-propionylcytosine, n4-p-toluoyl-5-fluorocytosine and 5-fluorouracil in the presence of trimethylsilyl trifluoromethanesulfonate (tms-triflate) to give fully acylated nucleosides (3a-b and 3d, respectively). selective removal of the n4-propionyl group of 3a by treatment with hydrazine hydrate gave 2',5'-di-o-p-chlorobenzoyl-3'-deoxycyt ...19958575037
differential diagnosis of fish pathogenic rhabdoviruses by reverse transcriptase-dependent polymerase chain reaction.reverse transcriptase-dependent polymerase chain reaction (rt-pcr) was applied to the detection and differentiation of viral hemorrhagic septicemia virus (vhsv) and infectious hematopoietic necrosis virus (ihnv) using primer pairs designed for the amplification of glycoprotein g-specific gene fragments of the two viruses. the products of 443 bp (vhs) and 548 bp (ihn), respectively, were amplified from the total rna extracts of rtg-2 cells infected with a total of 9 different strains of either vh ...19958576301
molecular epizootiology and evolution of the glycoprotein and non-virion protein genes of infectious hematopoietic necrosis virus, a fish rhabdovirus.infectious hematopoietic necrosis virus (ihnv) causes a highly lethal, economically important disease of salmon and trout. the virus is enzootic throughout western north america, and has been spread to asia and europe. the nucleotide sequences of the glycoprotein (g) and non-virion (nv) genes of 12 diverse ihnv isolates were determined in order to examine the molecular epizootiology of ihn, the primary structure and conservation of nv, and the evolution of the virus. the g and nv genes and their ...19958578856
the complete genome structure and phylogenetic relationship of infectious hematopoietic necrosis virus.infectious hematopoietic necrosis virus (ihnv), a member of the family rhabdoviridae, causes a severe disease with high mortality in salmonid fish. the nucleotide sequence (11,131 bases) of the entire genome was determined for the pathogenic wrac strain of ihnv from southern idaho. this allowed detailed analysis of all 6 genes, the deduced amino acid sequences of their encoded proteins, and important control motifs including leader, trailer and gene junction regions. sequence analysis revealed t ...19958578857
molecular biology of fish viruses: a review.the goal of this review is to present some of the recent molecular aspects in the fish virus studies. although more than 50 different fish virus have been isolated and tissue-culture adapted, very few of them have been molecularly cloned and sequenced. five virus families have been mostly studied: birnaviridae, the prototype being the infectious pancreatic necrosis virus (ipnv), and the channel catfish virus (ccv) belonging to the herpesviridae family. in the iridoviridae family, the fish lympho ...19958581005
immune response to synthetic peptides representing antigenic sites on the glycoprotein of infectious hematopoietic necrosis virus.monoclonal antibodies against infectious hematopoietic necrosis virus have been used to react with recombinant expression products in immunoblots and to select neutralization-resistant mutants for sequence analysis. these strategies identified neutralizing and non-neutralizing antigenic sites on the viral glycoprotein. synthetic peptides based upon the amino acid sequences of these antigenic sites were synthesized and were injected together with an adjuvant into rainbow trout. the constructs gen ...19958581009
comparison of the polymerases (l genes) of spring viremia of carp virus and infectious hematopoietic necrosis virus.we have determined the partial nucleotide sequences of the polymerase genes of the fish rhabdoviruses, spring viremia of carp virus (svcv) and infectious hematopoietic necrosis virus (ihnv). at this point we have deduced the amino acid sequences and analysed the first 1,400 amino acids comprising two thirds of the polymerase genes of svcv and ihnv. we have compared sequence similarities of svcv and ihnv polymerases with other rhabdovirus and paramyxovirus polymerases. the svcv polymerase showed ...19958581012
nucleotide sequence of the 2 matrix protein genes (m1 and m2) of hirame rhabdovirus (hrv), a fish rhabdovirus.we have cloned and sequenced the m1 and m2 protein genes of hirame rhabdovirus (hrv). the m1 protein gene was 684 nucleotides long, encoding 227 amino acids. sequence comparisons between the m1 protein genes of hrv and infectious hematopoietic necrosis virus (ihnv) or viral hemorrhagic septicemia virus (vhsv) revealed similarities of 65.1 and 40.8% at the nucleotide level and 81.1 and 44.8% at the amino acid level. the m2 protein gene was 582 nucleotides long, encoding 193 amino acids. sequence ...19958581014
characterization of ihnv isolates associated with neurotropism.rainbow trout with a 'neurotropic' form of infectious hematopoietic necrosis (ihn) exhibited behavioral abnormalities and had high virus concentrations localized in brain tissue. fish with 'hematopoietic' ihn exhibited typical disease signs with the highest concentrations of virus detected in kidney-spleen tissue. infectious hematopoietic necrosis virus (ihnv) isolates obtained from brain or kidney-spleen tissue were tested for serological relatedness and virulence and pathogenicity differences ...19958581018
aquareovirus interference mediated resistance to infectious hematopoietic necrosis virus.this study investigated the stimulation of specific host defenses to ihnv that are created by prior exposure to an avirulent reovirus (chum salmon reovirus; csv). preexposure of rainbow trout to csv for 1 h resulted in a relative percent survival that ranged from 68 to 100% when fish were challenged with ihnv over an 8 week period. the levels of serum neutralizing antibodies detected 28 d after the ihnv challenge were significantly lower among fish receiving prior exposures to csv than among non ...19958581022
the nv genes of fish rhabdoviruses: development of rnase protection assays for rapid assessment of genetic variation.the genomic rna of the fish rhabdovirus infectious hematopoietic necrosis virus (ihnv) contains a small nv gene which is not present in any other rhabdoviruses characterized to date. we have constructed a plasmid which carries a full length cdna copy of the ihnv nv gene between 2 rna polymerase promoters such that plus-sense and minus-sense rna transcripts of the ihnv nv gene can be synthesized. these were used to develop an rnase protection assay which was capable of detecting as little as 1-5 ...19958581026
serum neutralization test for epidemiological studies of salmonid rhabdoviroses in france.serological examination is not yet accepted as being a suitable diagnostic method for fish that are asymptomatic virus carriers. nevertheless, encouraging preliminary results using an end-point serum neutralization test (snt) in several french trout farm populations have demonstrated an excellent correlation between the snt and the previously established virus histories of the tested populations. following the isolation of infectious haematopoietic necrosis virus (ihnv) in france, serological sc ...19958581031
genetic immunization of rainbow trout (oncorhynchus mykiss) against infectious hematopoietic necrosis virus.plasmid vectors encoding the infectious hematopoietic necrosis virus (ihnv) nucleoprotein or glycoprotein gene under the control of a cytomegalovirus promoter were used to immunize rainbow trout (oncorhynchus mykiss) against ihnv. the plasmid dna was injected into the skeletal muscle of rainbow trout fry, and immunization was determined by the detection of virus-neutralizing and enzyme-linked immunosorbent assay antibody activity, and by protection against live virus challenge. fish injected wit ...19968680524
identification of the non-virion (nv) protein of fish rhabdoviruses viral haemorrhagic septicaemia virus and infectious haematopoietic necrosis virus.sequence analysis of a 795 nucleotide region of the fish rhabdovirus viral haemorrhagic septicaemia virus (vhsv) genome revealed one complete and one partial orf of 369 and 153 nucleotides, respectively. the latter orf probably encodes the amino-terminal part of the l (polymerase) protein. the former orf potentially encodes a 122 amino acid protein. the location of this orf as well as the size and deduced structure of the translation product indicate that it represents a homologue of the non-vir ...19968683214
monoclonal antibodies raised against infectious haematopoietic necrosis virus (ihnv) g protein and a cellular 90 kda protein neutralize ihnv infection in vitro.immune sera were obtained from four rainbow trout that had survived natural infection by infectious haematopoietic necrosis virus (ihnv), and five monoclonal antibodies (mabs) were prepared against a korean isolate of ihnv, ihnv-prt. these immune sera and mabs were characterized in terms of ihnv-neutralizing properties and reactivity in western blots with the viral proteins of ihnv-prt. all five mabs and four immune sera neutralized ihnv-prt to various extents. antibodies in these immune sera re ...19968760419
the structural proteins of infectious pancreatic virus are not glycosylated.the major capsid protein, vp2, of infectious pancreatic necrosis virus, a nonenveloped icosahedral virus, contains six n-glycosylation consensus sequences (asn-x-[thr/ser]). since vp2 contains the major virus-neutralizing epitopes, the possible role for glycosylation in capsid formation and antigenicity was examined. the carbohydrate content of the virion proteins was determined by chemical detection, pulse-chase experiments,[3h]mannose labeling, and alteration of protein migration on sodium dod ...19968794376
the glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: analysis of relationships with other rhabdoviruses.the nucleotide sequences of the glycoprotein genes and all of the internal gene junctions of the fish pathogenic rhabdoviruses spring viremia of carp virus (svcv) and hirame rhabdovirus (hirrv) have been determined from cdna clones generated from viral genomic rna. the svcv glycoprotein gene sequence is 1588 nucleotides (nt) long and encodes a 509 amino acid (aa) protein. the hirrv glycoprotein gene sequence comprises 1612 nt, coding for a 508 aa protein. in sequence comparisons of 15 rhabdoviru ...19968806175
mother to fry, successful transfer of immunity against infectious haematopoietic necrosis virus infection in rainbow trout.we have tested whether immunity can be transferred from a mother fish to its fry. rainbow trout mother fish were inoculated against infectious haematopoietic necrosis virus (ihnv) by intraperitoneal injection of a fragment of the ihnv glycoprotein spanning amino acids 31 to 310. this protein fragment was obtained by isolating the specific cdna from japanese ihnv strain hv7601 and expressing it in escherichia coli. fry from immunized and control fish were exposed to ihnv at various intervals afte ...19968887476
mapping the neutralizing epitopes on the glycoprotein of infectious haematopoietic necrosis virus, a fish rhabdovirus.twelve neutralizing monoclonal antibodies (mabs) against the fish rhabdovirus, infectious haematopoietic necrosis virus (ihnv), were used to select 20 mab escape mutants. the nucleotide sequence of the entire glycoprotein (g) gene was determined for six mutants representing differing cross-neutralization patterns and each had a single nucleotide change leading to a single amino acid substitution within one of three regions of the protein. these data were used to design nested pcr primers to ampl ...19969000094
linker mutagenesis of the caulobacter crescentus s-layer protein: toward a definition of an n-terminal anchoring region and a c-terminal secretion signal and the potential for heterologous protein secretion.linker insertion mutagenesis was used to modify the paracrystalline surface layer (s-layer) protein (rsaa) of the gram-negative bacterium caulobacter crescentus. eleven unique bamhi linker insertions in the cloned rsaa gene were identified; at the protein level, these linker insertions introduced 4 to 6 amino acids at positions ranging from the extreme n terminus to the extreme c terminus of the 1,026-amino-acid rsaa protein. all linker-peptide insertions in the rsaa n terminus caused the secret ...19979006010
distribution and variation of nv genes in fish rhabdoviruses.the fish rhabdovirus infectious haematopoietic necrosis virus (ihnv) contains a non-virion (nv) gene between the glycoprotein (g) and polymerase (l) genes on its rna genome. the present study investigated three other fish rhabdovirus genomes and found that the nv gene of hirame rhabdovirus is closely related to the nv of ihnv, whereas the viral haemorrhagic septicemia nv gene showed evidence of significant divergence. most importantly, spring viraemia of carp virus, the only vesiculovirus-like f ...19979010293
a novel 90-kda stress protein induced in fish cells by fish rhabdovirus infection.a 90 kda cellular protein in a fish cell, chse-214, showed increased expression by the infection of infectious hematopoietic necrosis virus (ihnv), heat shock, 2-mercaptoethanol, copper sulfate, and cadmium sulfate, and was detected in various kinds of cells such as human, rat, and mouse cells. the molecular mass of the 90 kda protein was different from those of the hsp90 and grp94. in addition, all the anti-stress protein mabs did not react with the 90 kda protein. finally, the subcellular dist ...19979144531
detection of the infectious hematopoietic necrosis virus directly from infected fish tissues by dot blot hybridization with a non-radioactive probe.a method was developed for the rapid diagnosis of the infectious hematopoietic necrosis virus (ihnv) based on dot blot hybridization with a non-radioactive probe. when the assay was developed through a color reaction both biotin- and alkaline phosphatase-labeled probes were highly specific for ihnv and the sensitivity reached to 20 pg of viral rna. when the alkaline phosphatase-labeled probe was developed by a chemiluminiscent reaction, the sensitivity showed a five-fold increase to 4 pg of vira ...19979186951
cloning of the rainbow trout (oncorhynchus mykiss) mx2 and mx3 cdnas and characterization of trout mx protein expression in salmon cells.two rainbow trout (oncorhynchus mykiss) mx cdnas were cloned by using race (rapid amplification of cdna ends) pcr and were designated rbtmx2 and rbtmx3. the deduced rbtmx2 and rbtmx3 proteins were 636 and 623 amino acids in length with molecular masses of 72 and 70.8 kda, respectively. these proteins, along with the previously described rbtmx1 protein (g. d. trobridge and j. a. leong, j. interferon cytokine res. 15:691-702, 1995), have between 88.7 and 96.6% identity at the amino acid level. all ...19979188599
immunization with viral antigens: infectious haematopoietic necrosis.infectious haematopoietic necrosis (ihn) is one of the most important viral diseases of salmonids, especially among juvenile fish where losses can be high. for over 20 years, researchers have tested a variety of preparations for control of ihn. early vaccines consisted of killed virus and were effective when delivered by injection, but too costly to be practical on a large scale. attenuated vaccines were developed by serial passage in cell culture and by monoclonal antibody selection. these offe ...19979270850
fish vaccine antigens produced or delivered by recombinant dna technologies.current efforts to develop vaccines, particularly for aquacultured species, have turned largely to biotechnology because it provides the means to inexpensively produce sufficient quantities of the immunoprotective antigen. these efforts have resulted in several prototype vaccines for fish and the publication of a large number of articles on the subject. however, there are only a few recombinant dna-based vaccines for aquaculture in the licensing pipeline. continued funding of research on recombi ...19979270855
sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral haemorrhagic septicaemia virus, a fish rhabdovirus.to evaluate the genetic diversity of viral haemorrhagic septicaemia virus (vhsv), the sequence of the glycoprotein genes (g) of 11 north american and european isolates were determined. comparison with the g protein of representative members of the family rhabdoviridae suggested that vhsv was a different virus species from infectious haemorrhagic necrosis virus (ihnv) and hirame rhabdovirus (hirrv). at a higher taxonomic level, vhsv, ihnv and hirrv formed a group which was genetically closest to ...19979367370
rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells.a reverse transcriptase-polymerase chain reaction (rt-pcr) assay was developed and applied to the detection and differentiation of viral haemorrhagic septicaemia virus (vhsv) and infectious hematopoietic necrosis virus (ihnv) in organ samples and cultured cells, regardless of the serotype. this method was developed by selecting primer sets corresponding to highly conserved regions of the glycoprotein g-gene sequences of the 2 viruses. the very fast rna extraction, reverse transcription and pcr p ...19989789975
combined dna immunization with the glycoprotein gene of viral hemorrhagic septicemia virus and infectious hematopoietic necrosis virus induces double-specific protective immunity and nonspecific response in rainbow trout.glycoprotein (g) of viral hemorrhagic septicemia virus (vhsv) and infectious hematopoietic necrosis virus (ihnv) contains several neutralizing epitopes. however, recombinant g protein never matches intact viral particles for immunogenicity. dna immunization offers the possibility to deliver the antigen through the cellular machinery, thus mimicking natural infection. we constructed pcdna gvhs and pcdna gihn plasmids with the g gene of vhsv and ihnv under the control of the cmv promoter, and we t ...19989791021
location of neutralizing epitopes on the g protein of bovine ephemeral fever rhabdovirus.the surface glycoprotein g is the major neutralizing and protective antigen of bovine ephemeral fever rhabdovirus (befv). twelve neutralizing mabs against befv strain bb7721 were used to select 33 neutralization escape mutants. the mutants had been classified previously into three major antigenic sites (g1-g3) based on their cross-neutralization patterns. the nucleotide sequence of the entire extracellular domain of the g protein gene was determined for all mutants. each contained a single nucle ...19989820132
truncated particles produced in fish surviving infectious hematopoietic necrosis virus infection: mediators of persistence?infectious hematopoietic necrosis virus (ihnv) is a rhabdovirus that produces an acute, lethal infection in rainbow trout (oncorhynchus mykiss). fish that survive infection cease to produce detectable infectious virus at approximately 46 days after infection, yet there is evidence that survivor fish continue to harbor virus particles (b. s. drolet, p. p. chiou, j. heidel, and j. c. leong, j. virol. 69:2140-2147, 1995). in an effort to determine the biological function of these particles, the kid ...19999847400
vibrio spp., the dominant flora in shrimp hatchery against some fish pathogenic viruses.two strains of vibrio spp., nica 1030 and nica 1031, which are the dominant flora in shrimp hatcheries, were studied for antiviral activity by exposing them to the infectious hematopoietic necrosis virus (ihnv) and oncorhynchus masou virus (omv) before being inoculated to the chse-214 cells. antiviral activities were exhibited as plaque reduction percentages of 98 and 92% for the nica 1030 and of 99 and 62% for the nica 1031, respectively.19989852624
[detection of viral hemorrhagic septicemia virus (vhsv) in rainbow trout (oncorhynchus mykiss) by reverse transcription followed by polymerase chain reaction. diagnostic validation].viral haemorrhagic septicaemia virus (vhsv) is a fish pathogen that attacks rainbow trout (oncorhynchus mykiss) farming. the only diagnostic method recognised by the european community for vhs is based on the detection of the viral agent in cell culture followed by an immunological identification of the pathogen. reverse transcription followed by a double amplification of the polymerase chain reaction (rt-pcr) for the gene encoding the viral glycoprotein g is proposed here as an alternative meth ...199910081112
virus susceptibility of the fish cell line saf-1 derived from gilt-head seabream.the recently reported saf-1 cell line from fins of gilt-head seabream was evaluated for susceptibility to lymphocystis disease virus (ldv) and to several salmonid fish viruses, such as infectious haematopoietic necrosis virus (ihnv), viral haemorrhagic septicemia virus (vhsv) and several strains of infectious pancreatic necrosis virus (ipnv). ldv, vhsv and ihnv replicated well in the cultured fin cells as demonstrated by cell lysis and increases in viral titer. the potential use of this cell lin ...199910092979
temperature dependent characteristics of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells.a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein) was produced in insect cells using a baculovirus vector (autographa californica nuclear polyhedrosis virus). characteristics of this protein were evaluated in relation to native viral g protein. a full-length (1.6 kb) cdna copy of the glycoprotein gene of ihnv was inserted into the baculovirus vector under control of the polyhedrin promoter. high levels of g protein (approximately 0.5 microgram/1 x 10(5) cells) ...199910349547
immunogenicity of a recombinant infectious hematopoietic necrosis virus glycoprotein produced in insect cells.a recombinant infectious hematopoietic necrosis virus (ihnv) glycoprotein (g protein), produced in spodoptera frugiperda (sf9) cells following infection with a baculovirus vector containing the full-length (1.6 kb) glycoprotein gene, provided very limited protection in rainbow trout oncorhynchus mykiss challenged with ihnv. fish were injected intraperitoneally (i.p.) with sf9 cells grown at 20 degrees c (recglow) or 27 degrees c (recghigh) expressing the glycoprotein gene. various antigen (ag) p ...199910349553
viral coinfection in salmonids: infectious pancreatic necrosis virus interferes with infectious hematopoietic necrosis virus.coinfection of farm-reared salmonids involving two viruses has been described, but there is no report on the interactions between viruses. here we examine whether infectious pancreatic necrosis virus (ipnv) strain sp interferes with the growth of infectious hematopoietic necrosis virus (ihnv) strain s46, a coinfected isolate from rainbow trout. when bf-2 cell culture was inoculated with s46 the infective titer of the ihnv fraction decreased by 3 log10 units compared to the growth curve of ihnv i ...199910365159
establishment and characterization of 13 cell lines from a green turtle (chelonia mydas) with fibropapillomas.thirteen cell lines were established and characterized from brain, kidney, lung, spleen, heart, liver, gall bladder, urinary bladder, pancreas, testis, skin, and periorbital and tumor tissues of an immature male green turtle (chelonia mydas) with fibropapillomas. cell lines were optimally maintained at 30 degrees c in rpmi 1640 medium supplemented with 10% fetal bovine serum. propagation of the turtle cell lines was serum dependent, and plating efficiencies ranged from 13 to 37%. the cell lines, ...199910462202
nested pcr improves detection of infectious hematopoietic necrosis virus in cells coinfected with infectious pancreatic necrosis virus.a nested assay using the reverse transcription-polymerase chain reaction (rt-pcr) was developed for the detection of infectious hematopoietic necrosis virus (ihnv) in cell cultures coinfected with infectious pancreatic necrosis virus (ipnv). two pairs of primers were designed: one for the amplification of glycoprotein g-specific gene rna from ihnv (or 1512 bp fragment), and the other for the amplification of an inner 753 bp fragment using the cdna from the g gene as substrate. direct rt-pcr was ...199910488755
inter-laboratory comparison of cell lines for susceptibility to three viruses: vhsv, ihnv and ipnv.eleven european national reference laboratories participated in an inter-laboratory comparison of the susceptibility of 5 selected cell lines to 3 fish pathogenic viruses. the test included viral hemorrhagic septicaemia virus (vhsv); infectious hematopoietic necrosis virus (ihnv) and infectious pancreatic necrosis virus (ipnv), and the cell lines derived from bluegill fry (bf-2), chinook salmon embryo (chse-214), epithelioma papulosum cyprini (epc), fathead minnow (fhm) and rainbow trout gonad ( ...199910494498
complete genomic sequence of viral hemorrhagic septicemia virus, a fish rhabdovirus.the complete nucleotide sequence of the fish rhabdovirus viral hemorrhagic septicemia virus (vhsv) has been determined. the genome comprises 11158 bases and contains six long open reading frames encoding the nucleoprotein n, phosphoprotein p, matrix protein m, glycoprotein g, nonstructural viral protein nv, and polymerase l. genes are arranged in the order 3'-n-p-m-g-nv-l-5'. the exact 3' and 5' ends were determined after rna-oligonucleotide ligation or race. they show inverse complementarity as ...199910499451
molecular characterization of the glycoproteins from two warm water rhabdoviruses: snakehead rhabdovirus (shrv) and rhabdovirus of penaeid shrimp (rps)/spring viremia of carp virus (svcv).we have determined the complete coding sequences for the glycoprotein (g) genes from two rhabdoviruses that infect warm water aquatic animals, the snakehead rhabdovirus (shrv) and rhabdovirus of penaeid shrimp (rps). surprisingly, the g nucleotide sequence from rps, a virus which has been isolated from diseased shrimp in hawaii on numerous occasions, was over 99% identical to the g nucleotide sequence from spring viremia of carp virus (svcv), a fish virus from europe and asia. this is the first ...199910518707
multiplex reverse transcriptase pcr assay for simultaneous detection of three fish viruses.a multiplex reverse transcriptase (rt)-pcr assay was developed for the simultaneous detection of three different fish viruses: infectious pancreatic necrosis virus (ipnv), infectious hematopoietic necrosis virus (ihnv), and viral hemorrhagic septicemia virus (vhsv). the sensitivity levels of the multiplex rt-pcr assay were 100, 1, and 32 50% tissue culture infective doses/ml for ipnv, ihnv, and vhsv, respectively.199910565946
naked dna vaccination of atlantic salmon salmo salar against ihnv.a naked plasmid dna encoding the glycoprotein (pcmv4-g) of a 1976 isolate of infectious hematopoietic necrosis virus (ihnv) obtained from steelhead oncorhynchus mykiss was used to vaccinate atlantic salmon salmo salar against ihnv. eight weeks post-vaccination the fish were challenged with a strain of ihnv originally isolated from farmed atlantic salmon undergoing an epizootic. fish injected with the glycoprotein-encoding plasmid were significantly (p < 0.05) protected against ihnv by both immer ...199910686669
mx mrna expression and rflp analysis of rainbow trout oncorhynchus mykiss genetic crosses selected for susceptibility or resistance to ihnv.three interferon-inducible mx genes have been identified in rainbow trout oncorhynchus mykiss and their roles in virus resistance have yet to be determined. in mice, expression of the mx1 protein is associated with resistance to influenza virus. we report a study to determine whether there was a correlation between the expression of mx in rainbow trout and resistance to a fish rhabdovirus, infectious hematopoietic necrosis virus (ihnv). a comparison of mx mrna expression was made between differe ...200010785857
nanogram quantities of a dna vaccine protect rainbow trout fry against heterologous strains of infectious hematopoietic necrosis virus.the efficacy of a dna vaccine containing the glycoprotein gene of infectious hematopoietic necrosis virus (ihnv), a rhabdovirus affecting trout and salmon, was investigated. the minimal dose of vaccine required, the protection against heterologous strains, and the titers of neutralizing antibodies produced were used to evaluate the potential of the vaccine as a control pharmaceutical. results indicated that a single dose of as little as 1-10 ng of vaccine protected rainbow trout fry against wate ...200010812224
genetic diversity and epidemiology of infectious hematopoietic necrosis virus in alaska.forty-two infectious hematopoietic necrosis virus (ihnv) isolates from alaska were analyzed using the ribonuclease protection assay (rpa) and nucleotide sequencing. rpa analyses, utilizing 4 probes, n5, n3 (n gene), gf (g gene), and nv (nv gene), determined that the haplotypes of all 3 genes demonstrated a consistent spatial pattern. virus isolates belonging to the most common haplotype groups were distributed throughout alaska, whereas isolates in small haplotype groups were obtained from only ...200010843554
comparison of infectious haematopoietic necrosis virus (ihnv) isolation on monolayers and in suspended cells.a cell culture virus isolation procedure for infectious haematopoietic necrosis virus (ihnv) in the epithelioma papulosum cyprini cell line (epc) is described. ovarian fluid samples were collected from fish and tested for ihnv at 9 farms. the samples were inoculated in parallel on 24 h old epc cell monolayers and in freshly trypsinized cells. the titre of the initial virus isolation and of first passages were compared using the 2 methods for each sample. titres were consistently higher in suspen ...200010843561
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