ribonucletic acid polymerase activity in purified infectious pancreatic necrosis virus of trout. 1975164186
the structure of infectious pancreatic necrosis virus rna.the rna from infectious pancreatic necrosis virus has been purified and had a sedimentation velocity of 14s on sucrose gradients, a buoyant density of 1-60 g/ml in cs2so4 and pyrimidine to purine ratios near unity. the rna had the appearance of a linear double stranded molecule with an average length of 0-92 mum and a standard deviation of 0-07 mum when observed under the electron microscope using the kleinschmidt protein film technique. this would correspond to a mol. wt. of 2-4 +/- 0-2 x 10(6) ...1977557073
isolation and characterization of infectious pancreatic necrosis virus from pike (esox lucius).a nonenveloped icosahedral virus measuring 60 (+/-5)nm in diameter has been isolated from pike fry (esox lucius). the isolate is stable to heat, lipid solvent and acid. iododeoxyuridine has no effect on its replication. serologically, the virus related to the major european strains of infectious pancreatic necrosis virus (ipn) sp and ab. the isolate is neutralized by normal trout serum and is not pathogenic for trout fry or for young pike. a line of chse-214 cells persistently infected with the ...1978697575
[infectious pancreatic necrosis virus: acquired susceptibility to trout serum neutralizing factor after repeated passage in cell culture].it has been shown previously that in normal sera from ipn free rainbow trout a "6 s molecule" neutralized sp type ipn viruses routinely used in european fish diseases laboratories. these viruses were unable to reproduce the disease under experimental conditions. after a survey of trout farms, a sp type pathogenic virus was isolated (isolate 31/75). after one passage on rtg2 cells this virus was fed to 7 weeks old fry and led to a 70% mortality. in contrast, a routinely passed and cloned virus (2 ...1975815032
relationship of a virus from tellina tenuis to infectious pancreatic necrosis virus.the physicochemical and serological properties of a virus isolated from the bivalve mollusc, tellina tenuis, have been examined. the virus has a diam. of 59 nm, sediments at 430s in sucrose gradients and bands at a density of i-32 g/ml in cscl. the virus contains rna with a mol. wt. about 2-8 x 10(6) as extimated by polyacrylamide gel electrophoresis but in sucrose gradients the rna sediments at 14s. the virus rna is resistant to ribonuclease under conditions in which ribosomal rna and the singl ...1977886305
[initial isolation and identification of the infectious pancreatic necrosis virus in salmonidae in the federal republic of germany]. 19751091261
infectious pancreatic necrosis virus internalization and endocytic organelles in chse-214 cells.the early events that take place during the internalization of infectious pancreatic necrosis virus (ipnv) into chinook salmon embryo cells (chse-214) were analyzed ultrastructurally. endocytic tracers were employed in order to characterize the organization of endocytic organelles in chse-214 cells, as well its relation to the ipnv penetration. results demonstrate that ipnv appear internalized within vesicular compartments which are located peripherally in chse-214 cells. despite the high rate o ...19921358463
evidence of genomic variations between infectious pancreatic necrosis virus strains determined by restriction fragment profiles.infectious pancreatic necrosis virus (ipnv) is the aetiological agent of an important disease in hatchery-reared salmonid fish in north america, europe and japan. it belongs to the family birnaviridae and shows a high degree of antigenic heterogeneity. however, genomic variations between the 10 identified serotypes have not yet been studied. in order to correlate genomic heterogeneity with the different serotypes, oligonucleotides were synthesized according to the published sequence of the jaspe ...19921359001
inoculation of infectious pancreatic necrosis virus serotype sp did not cause pancreas disease in atlantic salmon (salmo salar l.).atlantic salmon were selected from a fish farm with no previous record of pancreas disease (pd) or infectious pancreatic necrosis virus (ipnv) infection. groups of fish were inoculated with either ipnv (strain sp) from cell culture, organ material from fish with pd or control material as phosphate-buffered saline (pbs). virological, histological and immunohistochemical examinations were carried out throughout the experiment. none of the fish died or showed clinical symptoms of pd. histological e ...19911667971
rapid serotyping of infectious pancreatic necrosis virus by one-step enzyme-linked immunosorbent assay using monoclonal antibodies.a one-step elisa has been developed for detection and serotyping of infectious pancreatic necrosis virus (ipnv) in infected cell cultures using monoclonal antibodies (mab) raised against strains representing the sp, ab and vr 299 serotypes of ipnv. this assay uses a serotype-specific mab as capture and a mab directed against a common epitope as detector antibody. avidin-peroxidase was employed for amplification. the assay was performed in 90 min by simultaneous incubation of the samples, the bio ...19911707894
immunohistochemical identification of infectious pancreatic necrosis virus in paraffin-embedded tissues of atlantic salmon (salmo salar).infectious pancreatic necrosis virus serotype sp was identified by immunohistochemistry in formaldehyde-fixed and paraffin-embedded tissue of atlantic salmon (salmo salar). the immunoreaction was present in degenerating and necrotic cells in exocrine pancreatic cells. cross reactions were observed with rabbit antisera against serotypes sp, ab, and vr-299 in neutralization tests and western blotting. immunohistochemically, only sp antiserum produced positive immunostaining to sp antigens, whereas ...19901965633
characterization of the vpg-dsrna linkage of infectious pancreatic necrosis the use of strong denaturing agents, a genome-linked protein (vpg)-rna complex was purified from infectious pancreatic necrosis virus. ribonuclease treatment of 125i-labelled vpg-rna released a 90k polypeptide identical to the minor structural polypeptide vp1 (the putative rna polymerase), as determined by peptide mapping. the polypeptide is linked to the rna by a serine-5' gmp phosphodiester bond. the results identify birnaviruses as the only dsrna viruses with a vpg, the size of which is th ...19911919532
sequence analysis of infectious pancreatic necrosis virus genome segment b and its encoded vp1 protein: a putative rna-dependent rna polymerase lacking the gly-asp-asp motif.the genome segment b sequence of infectious pancreatic necrosis virus was determined for both the jasper and sp serotypes. the sequences are 2784 and 2630 bp long, respectively, and contain a single large open reading frame encoding the vp1 protein, the putative rna-dependent rna polymerase (rdrp) of ipnv. the proteins exhibit an 88% homology with each other, but only 41% with infectious bursal disease virus (ibdv) vp1, another member of the birnaviridae. despite the low overall homology between ...19911901682
detection of infectious pancreatic necrosis virus (ipnv) rna by hybridization with an oligonucleotide dna probe.marine farming of atlantic salmon (salmo salar) is growing rapidly in norway, and fish diseases have now become one of the biggest obstacles for this new industry. infectious pancreatic necrosis virus (ipnv) is commonly found in fish from norwegian sea farms. diagnosis of ipnv infection is, at present, mainly based on virus isolation in cell cultures and identification by neutralization tests (nt). a dna-rna hybridization assay was developed using a 24 base dna oligonucleotide probe. this is hom ...19902169675
fish viral infections in northwest of spain.during a three years survey, a total of 149 samples from 20 farms of rainbow trout, salmon and turbot were examined for the presence of virus with the purpose to study the viral infections affecting cultured fish and their incidence in the fishfarms of northwestern spain. infectious pancreatic necrosis virus (ipnv) was the only viral agent isolated from salmonid fish. fry and fingerlings of trout showed the highest infection rate (24%). this virus was not detected in broodstock or embryonated eg ...19902169768
method for the concentration of infectious pancreatic necrosis virus from hatchery water.a method was developed for concentrating infectious pancreatic necrosis virus from hatchery water using positively charged 1-mds filters. the method consists of passing large volumes (ca. 1001) of hatchery water through 1-mds microporous filter followed by the elution of the adsorbed virus using a high ph buffer. the virus adsorbed efficiently to 1-mds filters when the ph of the water was 5.5 and was eluted optimally with 3% beef extract solution (ph 10). this procedure permitted the processing ...19911864906
genome cloning and analysis of the large rna segment (segment a) of a naturally avirulent serotype 2 infectious bursal disease virus.the genome of infectious bursal disease virus (ibdv) of serotype 2 (strain oh) has been cloned, and 3171 nucleotides of genome segment a cdna sequence have been determined for the first time. sequence homology of oh-ibdv with the most distant serotype 1 ibdv at the nucleotide level is 83.1%, and the amino acid sequence homology of the polyprotein is 89.6%. alignment of the polyprotein amino acid sequences showed the hypervariable region in vp2 to be 151-152 amino acid residues long in ibdv. a se ...19911651602
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
antigenic and genomic differences of two jasper strains of infectious pancreatic necrosis virus.strains of infectious pancreatic necrosis virus (ipnv-jasper) obtained from two different laboratories were compared serologically with polyclonal and monoclonal antibodies. nucleotide sequence and restriction endonuclease patterns of 359-bp fragment of genome segment a cdna were also compared. substantial differences were found in both analyses that will support the fact that the two jasper strains are not identical.19921308002
studies on infectious pancreatic necrosis virus interactions with rtg-2 and fhm cells: selection of a variant virus-type in fhm cells.evidence is presented that adaptation of ipn virus (strain vr 299) to fhm cells entails the selection of a variant virus type that differs significantly from the parental, and most representative, rtg-2 virus type in being able to adsorb efficiently to, and form plaques in, fhm cells. the plaque titre of fhm-non-adapted virus stocks (rtg-2 viruses) was reduced by at least 99-99% in fhm cells, while fhm-adapted virus stocks (fhm viruses) produced plaques at equally high titres in both rtg-2 and f ...19751171932
size and structure of the genome of infectious pancreatic necrosis virus.the genome of infectious pancreatic necrosis virus consists of two segments of dsrna, in equimolar amounts, with molecular weights of 2.5 x 10(6) and 2.3 x 10(6) daltons, as determined by polyacrylamide gel electrophoresis and autoradiography. the viral rna was resistant to ribonuclease, and in sucrose gradient it co-sedimented at 14s with rnase resistant rna from virus infected cells. upon denaturation in 98% formamide, the viral genome sedi-mented at 24s in formamide sucrose gradient and becam ...1976987579
comparison of rnas and polypeptides of infectious pancreatic necrosis virus isolates from eel and rainbow trout.thirteen isolates of infectious pancreatic necrosis virus (ipnv) from eel and rainbow trout in taiwan were compared with the selected serotypes ab, sp and vr-299 by page analysis of their rna genomes and early polypeptides. all the ipnv isolates from eels (1984 to 1986) from lu kang and ping tung, and from rainbow trout eggs and fry from dan sway (1985 to 1986) were most closely related to the ab serotype. however four rainbow trout isolates from dan sway (1984) had a similar rna pattern to that ...19892769237
quantitative analysis of defective interfering particles in infectious pancreatic necrosis virus preparations.infectious pancreatic necrosis virus exhibited an interference phenomenon that resulted in the survival of the infected cell with one hit kinetics. the responsible factor was found to co-purify with standard virus through a purification regime that employed two cscl gradients and a sucrose gradient. this result suggested that a defective interfering (di) viral particle was involved. it was possible to estimate the number of di particles by a statistical method using the poisson distribution that ...1978566090
effect of exogenous corticosteroids on circulating virus and neutralizing antibodies in striped bass (morone saxatilis) infected with infectious pancreatic necrosis virus.corticosteroids have been reported to induce immunosuppression in fish exposed to many types of bacterial antigens. we document a similar phenomenon in fish exposed to infectious pancreatic necrosis virus (ipnv). fingerling striped bass that were injected with the steroid triamcinolone acetonide (100 mg/kg body weight) 24 hours before receiving intraperitoneal inoculation with ipnv became viremic 3 days post inoculation (dpi) and virus was still detected in the buffy coat cells 14 dpi. in contra ...19863020770
effect of temperature on survival and growth of infectious pancreatic necrosis virus.infectious pancreatic necrosis virus was stable for 10 days at 4 c in stream and well water, after which the virus had a half-life of 7.5 days. at 15 c, the virus was stable for 5 days, and then had a half-life between 5 and 6 days. viral antigen in infected cells developed much more slowly at 4 c than at 20 c. infected cells released infectious viral particles at temperatures as low as 4 c. nutrition had a greater effect on the production of infectious virus at 4 c than at 20 c.1975806530
peptide map comparison of infectious pancreatic necrosis virus-specific investigation of virus-specific protein synthesis in infectious pancreatic necrosis virus (ipnv)-infected rainbow trout gonad cells was undertaken to find a relationship between the coding capacity of the virus genome (two segments of double-stranded rna of 2.5 x 10(6) and 2.3 x 10(6) molecular weight) and the sizes and relative amounts of virus-specific proteins. using polyacrylamide slabgel electrophoresis and autoradiography, eight distinct virus-specific polypeptides were detected in infe ...1977563479
[isolation and identification of the infectious pancreatic necrosis virus in trout].the first isolation and identification of the infectious pancreatic necrosis virus in trout in czechoslovakia is described. the rtg-2, pg and chse cell lines were used for isolation and the identification was made by the methods of electron microscopy, cross virus-neutralizing test, and immunofluorescence. as demonstrated, all the isolates in the czechoslovak territory are serologically identical and are closely related (or perhaps congruent) with the sp reference strain which has the highest fr ...19863083571
oligonucleotide fingerprints of the rnas from infectious pancreatic necrosis virus. 1977562144
health status of salmonids in river systems in natal. ii. isolation and identification of viruses.a total of 678 fish from 26 different sites along 8 river systems was examined for the presence of trout viruses. no isolations of any viruses were made from any fish collected from these systems in natal during this study. no antibodies against any of the trout viruses were detected in the serum collected from these fish. it would appear from this study that the river systems in natal are free of any of the known viruses of trout. infectious pancreatic necrosis virus was, however, isolated from ...19911881657
characterization of a new serotype of infectious pancreatic necrosis virus isolated from atlantic salmon.virus particles isolated from hatchery reared fish with infectious pancreatic necrosis (ipn) were neutralized by homologous immune sera but not by immune sera raised against ipn virus serotype 1, 2, and 3. this virus isolate, called the n1 strain, was detected in one year old atlantic salmon (salmo salar) during an outbreak with histopathological lesions of ipn and slightly increased mortality. the polypeptide pattern of n1 virus differed markedly from that of the three classical ipn virus serot ...19883214272
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
a physical map of the viral genome for infectious pancreatic necrosis virus sp: analysis of cell-free translation products derived from viral cdna clones.the two segments of double-stranded rna from infectious pancreatic necrosis virus sp were cloned into the plasmid vector puc8. two sets of overlapping clones were identified by restriction enzyme and southern blot analyses. each of these sets was shown by northern blot analysis to be exclusively related to either segment a or b of the genomic rna. the entire lengths of the cloned segments were estimated to be 2.9 and 2.6 kilobases, respectively. sequences from the two segments of viral cdna were ...19863023652
infectious pancreatic necrosis virus persistently infects chinook salmon embryo cells independent of interferon. 1979442542
rapid identification of infectious pancreatic necrosis virus in infected cell cultures by immunoperoxidase and indirect immunoperoxidase (ip) techniques were evaluated for their potential in identifying infectious pancreatic necrosis (ipn) virus. both techniques were shown to offer a relatively simple, rapid and efficient means for the specific identification of ipn virus in infected cells. the direct ip method resulted in less nonspecific staining; however, the indirect method was clearly specific and utilized commercially available reagents.1978368352
biophysical studies of infectious pancreatic necrosis virus.the molecular weight of infectious pancreatic necrosis virus (ipnv) has been determined by analytical ultracentrifugation and dynamic light scattering. the sedimentation coefficient of the virus was found to be 435s. the average value for molecular weight is (55 +/- 7) x 106. the virus genome consists of two segments of double-stranded rna (molecular weights, 2.5 x 106 and 2.3 x 106), which represents 8.7% of the virion mass. the capsid protein moiety of ipnv consists of four species of polypept ...1977558343
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
monoclonal antibodies to the sp strain of infectious pancreatic necrosis virus.twelve hybrids secreting antibody to the sp serotype of infectious pancreatic necrosis virus (ipnv) were isolated from the fusion of murine myeloma cells and spleen cells from mice immunized with pelleted virus. all of the monoclonal antibodies possessed the kappa (k) light chain isotype. nine contained the mu (m), two had the gamma 2a (g2a), and one had the gamma 1 (g1) heavy chain isotype. using an enzyme-linked immunosorbent assay (elisa), 10 antibodies were found to be broadly reactive again ...19863765359
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
biochemical studies with infectious bursal disease virus: comparison of some of its properties with infectious pancreatic necrosis virus.infectious bursal disease (ibd) virus was purified from the bursae of infected chickens. two morphologically indistinguishable populations of virus particles were separated in sucrose gradients and possessed sedimentation coefficients of 295s and 460s. both populations contained rna and had identical polypeptide compositions. ibd virus banded at a density of 1.31 g/ml in cscl and at 1.24 g/ml in sodium potassium tartrate. ibd virus contained two rna segments with mol. wts. of 2.4x10(6) and 2.2x1 ...1979228637
the persistence of infectious pancreatic necrosis virus in atlantic salmon.atlantic salmon leucocytes from infectious pancreatic necrosis virus (ipnv) carriers showed a suppressed response to phytohemagglutinin stimulation compared with uninfected controls. a significant degree of inhibition of dna synthesis was observed using 3h-thymidine incorporation. ipnv was isolated from 41% of the stimulated leucocyte cultures supernatants, while only 6% of the unstimulated cultures were found to be positive.19863765357
biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded rna genomes.infectious pancreatic necrosis virus of fish, infectious bursal disease virus of chickens, tellina virus and oyster virus of bivalve molluscs, and drosophila x virus of drosophila melanogaster are naked icosahedral viruses with an electron microscopic diameter of 58 to 60 nm. the genome of each of these viruses consists of two segments of double-stranded rna (molecular weight range between 2.6 x 10(6) and 2.2 x 10(6), and the virion, capsid proteins fall into three size class categories (large, ...1979228080
inheritance and longevity of infectious pancreatic necrosis virus in the zebra fish, brachydanio rerio (hamilton-buchanan).zebra fish (brachydanio rerio) were injected with infectious pancreatic necrosis virus (ipnv) and then spawned to determine whether the virus was passed on to the eggs, and if it was, how long it remained in the free-swimming f1. the mating variations included parents receiving one or two injections of virus, and within these categories, matings in which both parents were treated or only one parent was treated. the results showed that transmission of ipnv to the egg did occur, and that this tran ...1977196549
[experimental data on the remanence of the virus causing the infectious pancreatic necrosis in salmonidae in water environment (author's transl)].to study the virus persistence in water environment, a purified suspension of the infectious pancreatic necrosis virus was submitted to different conditions of conservation: temperature of 4 degrees c and 1.4 degrees c in a mineral water of mean mineralization, temperature of 4 degrees c in a river water with a higher minerlization, filtered or not, with or without mud and vegetation. the cyto-infectious power of the virus subsists at least during 300 days at + 4 degrees c and 60 days at + 14 de ...1976185935
experimental induction of the carrier state in yearling brook trout: a model challenge protocol for ipnv immunization.brook trout fry (salvelinus fontinalis) were not protected from infectious pancreatic necrosis virus (ipnv) challenge by immersion vaccination with inactivated, purified virus at concentrations of 10(7) to 10(9) pfu/ml. mortalities in vaccinated groups were higher than for the unvaccinated control group and appeared to be dose-dependent. a challenge protocol for adult brook trout was developed for future vaccine trials. a single intraperitoneal injection of virulent, purified virus was sufficien ...19863765358
virus-specific protein synthesis in cells infected by infectious pancreatic necrosis virus.a study of virus-specific protein synthesis in infectious pancreatic necrosis virus-infected rtg-2 cells was undertaken to find a relationship between the coding capacity of virus genome (two segments of double-stranded rna of 2.5 x10(6) and 2.3 x 10(6) molecular weight) and the sizes and relative amounts of polypeptides in the virion and in infected cells. the time course of virus-specific protein synthesis was followed by pulse labeling infected uv-irradiated cells with [35s]methionine and ana ...1977556780
intracellular replication of infectious pancreatic necrosis virus. 19724113706
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
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
cell-free translational analysis of the processing of infectious pancreatic necrosis virus polyprotein.a cdna clone of the large genomic segment of infectious pancreatic necrosis virus (ipnv) was inserted into transcription vectors and used for production of rna transcripts of various lengths. these rna transcripts were used to prime the synthesis of virus-specific polypeptides in a rabbit reticulocyte translation system. full-length transcripts resulted in the synthesis of processed viral proteins pvp2, ns, and vp3. transcripts which were deleted in the vp2 coding region did not affect the proce ...19902145691
electron microscopical and biochemical characterization of infectious pancreatic necrosis electron microscopical and biochemical examination of the properties of infectious pancreatic necrosis virus (ipn) and of its ribonucleic acid (rna) was made. the buoyant density of ipn in cscl was found to be 1.33 g/cm(3). electron microscopical examination of the banded virus revealed structures similar in size (74 nm) and shape to reoviruses but lacking a characteristic inner capsid structure. polyacrylamide gel electrophoretic analysis of ipn-rna revealed a single non-segmented component ...19724117351
physico-chemical and morphological features of infectious pancreatic necrosis virus. 19734128635
infectious pancreatic necrosis virus--temperature and age factors in mortality. 19714361130
[infectious pancreatic necrosis virus: a study of the influence of temperature and host-cell on virus replication and virus-induced interferon synthesis (author's transl)]. 19744478747
isolation of infectious pancreatic necrosis virus from white suckers (catastomus commersoni). 19724553697
further analysis on the structural proteins of infectious pancreatic necrosis virus.the structural proteins of infectious pancreatic necrosis virus (ipnv) have been analyzed. two-dimensional gel electrophoresis showed that ipnv proteins are slightly acidic with apparent pis ranging from 5.8 to 6.6. to identify the ipnv surface-located proteins, purified virus was labelled either with fluorescein isothiocyanate (fitc) or with na 125i. after analysis by sds-page, only the major viral protein, vp2, was labelled by either procedure. the accessibility of vp2 to these reagents sugges ...19902107647
nucleotide sequence analysis of genome segment a of infectious bursal disease virus.the nucleotide sequence of genome segment a cdna of the stc strain of infectious bursal disease virus (ibdv) was determined and compared with sequences of the homologous genome segment of the 002-73 strain of ibdv and the jasper strain of infectious pancreatic necrosis virus (ipnv). the stc-ibdv genome segment a was determined to be 3262 base pairs (bp), which is close to the estimated total length of 3300 bp for genome segment a in ibdv, although there is no proof that it is the real length of ...19902155995
morphology of infectious pancreatic necrosis virus. 19654957052
multiplication of infectious pancreatic necrosis virus. 19654957053
salmonid viruses: infectious pancreatic necrosis virus. morphology, pathology and serology of first european isolations. 19715005507
infectious pancreatic necrosis virus-induced pancreatic lesions in swiss albino mice: electron microscopy. 19725039522
effect of actinomycin d on the multiplication of the infectious pancreatic necrosis virus of trout. 19715134315
infectious pancreatic necrosis virus--temperature and age factors in mortality. 19715165970
neutralization tests of infectious pancreatic necrosis virus with polyvalent antiserum. 19734701178
infectious pancreatic necrosis virus: lyophilization and subsequent stability in storage at 4 c.infectious pancreatic necrosis virus (atcc strain vr # 299) was lyophilized with different additives and stored at 4 c. ampoules were assayed at 2 days, at 1, 3, and 6 months, and at 1, 3, and 4 years. processing losses were least and subsequent maintenance of infectivity was best in skim milk, lactalbumin hydrolysate, and lactose. quantitative data are given in tabular form.19695819010
expression in escherichia coli of the large genomic segment of infectious pancreatic necrosis virus.a complete cdna clone of the larger a segment of the genome of infectious pancreatic necrosis virus (ipnv) was expressed in escherichia coli in an effort to develop a vaccine for ipnv in fish. when the cdna insert was positioned in the correct orientation to the puc19 lacz promoter, the viral proteins vp2, ns, and vp3 were synthesized and processed as observed in infected cells. when the insert was placed in the opposite orientation, vp3 and a 38-kda virus-specific polypeptide were also synthesi ...19902219718
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
identification of a double-stranded rna virus by using polymerase chain reaction and magnetic separation of the synthesized dna segments.a double-nested polymerase chain reaction assay (pcr), followed by magnetic separation of the pcr-synthesized dna segments, was developed to detect a double-stranded rna virus, infectious pancreatic necrosis virus from salmonid fish. viral rna was extracted from cell cultures and used for cdna synthesis. the cdna produced was used as a template in a double pcr. the sensitivity of this double pcr was approximately 0.8 pg of template double-stranded rna. the dna segment produced from the first pcr ...19902229352
antigen-capture elisa for viral haemorrhagic septicaemia virus serotype 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
sequence of the large double-stranded rna segment of the n1 strain of infectious pancreatic necrosis virus: a comparison with other birnaviridae.the cdna sequence of the large dsrna segment (segment a) of the n1 strain of infectious pancreatic necrosis virus (ipnv) has been determined. the nucleotide and deduced amino acid sequences were compared to the sequences of segment a of the jasper strain of ipnv and to the sequences of segments a and b (5' and 3' flanking regions) of the 002-73 strain of infectious bursal disease virus (ibdv). the comparison demonstrated that the precursor protein of the major structural polypeptide, pvp2, is hi ...19902307963
[occurrence of infectious pancreatic necrosis virus (ipn) in different fish species]. 19806248021
monoclonal antibodies to infectious pancreatic necrosis virus: analysis of viral epitopes and comparison of different isolates.a panel of five monoclonal antibodies (mabs) produced against the west buxton isolate of infectious pancreatic necrosis virus was used to investigate viral antigens and to compare different aquatic birnavirus isolates antigenically. reciprocal blocking elisa and neutralization assays indicated that these mabs identified four, and possibly five, structurally and/or functionally different epitopes on the virion. western immunoblot analysis demonstrated that one epitope was present on vp2, the larg ...19862428927
rapid detection of infectious pancreatic necrosis virus (ipnv) by the enzyme-linked immunosorbent assay (elisa).the enzyme-linked immunosorbent assay (elisa) was used to demonstrate the presence of infectious pancreatic necrosis virus (ipnv) antigen in cell cultures and in fish. virus antigen could be detected in infected cell cultures before visible cytopathic effect (c.p.e.) was evident and cell cultures showing complete viral c.p.e. produced intense colour reactions. virus antigen was detected in infected fry during and immediately after an epizootic, but although ipnv-carrier fish could be detected by ...19836300287
the molecular biology of infectious pancreatic necrosis virus: a review. 19836303539
infectious pancreatic necrosis virus: isolation from asymptomatic wild arctic char (salvelinus alpinus l.). 19846530721
expression in escherichia coli of the major outer capsid protein of infectious pancreatic necrosis virus.the outer capsid polypeptide, vp2, represents the major neutralizing antigen of infectious pancreatic necrosis virus (ipnv). a 926-bp viral cdna, encoding an n-terminal truncated vp2, was cloned into the pwr590 expression plasmid family resulting in a c-terminal extension of a truncated escherichia coli beta-galactosidase (beta gal) under the control of the lac promoter. when cells transformed by in-phase hybrid plasmids were induced by isopropylthiogalactoside, high levels of the 100-kda beta g ...19892507397
natural anti-tnp antibodies from rainbow trout interfere with viral infection in vitro.normal and viral-infected rainbow trout (rt) were tested for serum antibody activity against self and nonself antigens. particularly high titres of anti-trinitrophenyl (tnp) antibodies were noted, as in other fish species. to analyse this, the anti-tnp antibodies were isolated by affinity chromatography and their capacity to interfere with viral infection in vitro was studied. we selected rt fibroblasts as target cells, and two common pathogenic viruses in trout, a rhabdovirus, viral haemorrhagi ...19892595079
enzyme-linked immunosorbent assay for identification of infectious pancreatic necrosis virus.the sensitivity, reproducibility, and specificity of an enzyme-linked immunosorbent assay for the identification of infectious pancreatic necrosis virus was determined. it was shown that 10(5.5) tissue culture infectious doses could be assayed by this method. the assay provided significant advantages in terms of time and ease of performance over the routinely employed serological tests. when an antiserum against a single isolate was used, it was possible to identify nine of ten north american is ...19826752187
a host range restriction in infectious pancreatic necrosis virus maps to the large rna segment and involves virus attachment to the cell surface. 19826758322
mapping of the large rna genome segment of infectious pancreatic necrosis virus by hybrid arrested translation.segment a, the larger dsrna segment of ipnv which encodes three of the four virus-coded polypeptides (prevp2, vp3, and ns) was cloned and physically mapped. the plus and minus rna strands of the virus genome were separated and the a+ and b+ rna strands were identified. a nested set of cdna subclones, coterminal with the 5' end of a+ rna, were used in hybrid arrested translation experiments. hybrid arrest conditions which blocked the 5' two-thirds of a+ rna allowed the in vitro synthesis of only ...19872953102
isolation of a bisegmented double-stranded rna virus from thirlmere reservoir.a novel bisegmented double-stranded rna virus has been isolated from water processed from thirlmere reservoir. the virus is icosahedral, 58 nm in diam., has a buoyant density of 1.32 g/ml in cscl, has an s value of 400 and a rna/protein ratio of 0.087. the two linear segments of rna have approx. mol. wt. of 2.26 x 10(6) and 2.09 x 10(6). the virus contains six polypeptides. the virus was isolated in drosophila melanogaster cells and fails to replicate in other insect, amphibian, avian, piscine, ...19826815302
three new strains of infectious pancreatic necrosis virus isolated in canada.three new samples of infectious pancreatic necrosis virus (ipnv) isolated in canada were compared with known serotypes by phenotypic properties, immunofluorescence, and protein and rna molecular weights. the three new isolates were each found to be unique by these parameters. the most convincing physical difference used to distinguish the new isolates from known serotypes was their rna molecular weights. thus, the new cb, nasrc, and char isolates of ipnv represent three new strains of the virus.19836831312
biochemical characterization of eel virus european.the polypeptide and rna compositions of purified virions of eel virus european were compared to those of three strains (vr-299, sp and ab) of infectious pancreatic necrosis virus (ipnv) isolated from trout. all three ipnv strains could be distinguished by the relative mobilities of either the virion polypeptides or the double-stranded rna genome segments. the eel virus had a similar polypeptide profile to strain ab ipnv but differences between the two viruses in the migration of genome segments ...19836854276
immunosuppressive effect of infectious pancreatic necrosis virus on rainbow trout (oncorhynchus mykiss).the infectivity of infectious pancreatic necrosis virus (ipnv) for rainbow trout (oncorhynchus mykiss) mononuclear leukocyte subpopulations was investigated to determine the mechanisms of immunosuppression caused by the virus. ipnv was recovered from nylon wool-adherent, surface immunoglobulin (ig)-positive leukocytes of head kidney, spleen and peripheral blood collected from virus-inoculated fish with higher titers than non-adherent, ig-negative cells. non-adherent cell population showed mitoge ...19902177807
messenger rna of infectious pancreatic necrosis virus is polycistronic. 19827078640
immunofluorescent cell assay of infectious pancreatic necrosis immunofluorescent cell (ifc) assay technique was developed for the quantification of infectious pancreatic necrosis (ipn) virus of salmonid fishes. cover slip cultures of rainbow trout gonad (rtg-2) cells were infected with diluted virus preparations. after incubation to permit antigen development, the cells were stained with antiviral fluorescent antibody, and the number of fluorescing (infected) cells was counted. optimal conditions for the ifc assay procedure are: (i) the use of rtg-2 cell ...19744596390
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
detection of infectious pancreatic necrosis virus using an inhibition enzyme-linked immunosorbent inhibition enzyme-linked immunosorbent assay was used to detect infectious pancreatic necrosis (ipn) virus. in this assay the presence of virus was determined by measuring the decrease in titer of a known antiserum after incubation with a sample suspected to contain virus. the titer of the antiserum was measured with an indirect enzyme-linked assay. compared to the double antibody sandwich method this assay required fewer reagents (only one anti-ipn serum was required). this assay was also se ...19863027134
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
effect of ammonium chloride on the multiplication of infectious pancreatic necrosis virus.the multiplication of ipnv in chse cells is inhibited by ammonium chloride. this inhibition is complete if nh4cl is added early in the infective cycle. immunofluorescence analysis shows that nh4cl reduces the number of virus producing cells and thus suggests it may act by inhibiting virus internalization. in addition, nh4cl can produce partial inhibition of ipnv multiplication if added later in the replicative cycle. analysis of 3h-uridine incorporation into viral rna shows that this later effec ...19883348747
synthesis of the infectious pancreatic necrosis virus polyprotein, detection of a virus-encoded protease, and fine structure mapping of genome segment a coding regions.full-length and truncated genome segment a-specific infectious pancreatic necrosis virus cdna was subcloned into plasmid transcription vectors, and runoff transcripts were produced in vitro. these transcripts were translated in cell-free rabbit reticulocyte lysates and the translation products were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. virus-specific polypeptides were gel purified and mapped by partial proteolysis with n-chlorosuccinimide and sodium dodecyl sulfa ...19873316706
the polypeptides of infectious pancreatic necrosis virus. 19744818278
[a study of the cause of massive mortality among marine-cultured rainbow trout (oncorrhynchus mykiss, walbaum) in a fish farm in the southwestern netherlands].the present study investigated the mass mortality of marine cultured young rainbow trout (oncorhynchus mykiss, walbaum) in the netherlands. the course of the disease, the clinical symptoms, and bacteriological and virological investigations lead to the diagnosis: 'primary infectious pancreatic necrosis virus (ipnv) infection followed by secondary vibriosis'. treatment with flumequine seemed to be effective. the trout were possible infected with ipnv at the trout hatchery and nursery from which t ...19957482471
attachment and entry of infectious pancreatic necrosis virus (ipnv) into chse-214 cells.infectious pancreatic necrosis virus (ipnv) attaches to chse-214 cells through two types of cell components: specific and non-specific ones. competition experiments with inactivated ipnv showed that ipnv requires specific components to productively infect cells. just a low amount of adsorbed ipnv enters the cell. after 20 minutes, part of the adsorbed ipnv was internalized into acid compartments. also, the viruses adsorbed on the cell surface require similar periods of time to escape from the ne ...19957503682
the nucleotide sequence of infectious pancreatic necrosis virus (ipnv) dsrna segment a reveals one large orf encoding a precursor polyprotein. 19863737418
infectious pancreatic necrosis: detection of virus and antibodies in rainbow trout ipnv-carrier (salmo gairdneri). 19863811691
nucleotide sequence analysis of the serotype-specific epitope of infectious pancreatic necrosis virus.the cdna nucleotide sequences of the infectious pancreatic necrosis virus (ipnv) serotype-specific epitope of the major structural protein vp 2 were determined for a scottish strain (sp serotype), and an ipnv field isolate from shetland. comparison of the sequence data indicated that the field isolate was of the sp serotype. the scottish sp strain and the field isolate showed less than 3% difference in nucleotide sequence over this region compared to the norwegian n 1 strain, and all three europ ...19937682403
polyacrylamide gel electrophoresis of viral genomic rna as a diagnostic method for infectious pancreatic necrosis virus detection.a rapid and simple technique for the diagnosis of ipnv from cell culture and infected fish tissues has been developed. it is based on the observation of ipnv bisegmented double-stranded rna genome in silver stained polyacrylamide gels after electrophoresis. the method is highly specific and can detect as little as 1 ng of viral rna, which corresponds to 1 x 10(5) pfu, making it possible to visualize the viral genome as soon as the initial cythopatic effect appears. furthermore, the rna viral gen ...19947714046
neutralizing activity against infectious pancreatic necrosis virus in striped bass, morone saxatilis, from the chesapeake bay. 19873820417
capsid symmetry of viruses of the proposed birnavirus group.the capsid architecture of three members of the tentatively formed group of bisegmented ds rna containing animal viruses (birnavirus group) was studied in comparison with the fine structure of reovirus type 1 using high resolution negative staining and shadowing techniques followed by rotational enhancement analysis. ipnv of the trout, ibdv of the chicken, and eel virus (berlin) are single-shelled particles with a capsid diameter of 61-65 nm and thus clearly distinguishable from the double-shell ...19853994515
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