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Sample records for septicaemia virus vhsv

  1. Development of a monoclonal antibody against viral haemorrhagic septicaemia virus (VHSV) genotype IVa.

    PubMed

    Ito, T; Olesen, N J; Skall, H F; Sano, M; Kurita, J; Nakajima, K; Iida, T

    2010-02-24

    The viral haemorrhagic septicaemia virus (VHSV) comprises 4 major genotypes and a number of subtypes with, in most cases, distinct geographical distribution. A quick and simple detection method that can discriminate the different genotypes is desirable for a quick and more efficient prevention of the spread of genotypes to new geographical areas. A monoclonal antibody (MAb) against VHSV genotype IVa was produced, with the aim of providing a simple method of discriminating this genotype from the other VHSV genotypes (I, II, III and IVb). Balb/c mice were injected with purified VHSV-JF00Ehil (genotype IVa) from diseased farmed Japanese flounder. Ten hybridoma clones secreting monoclonal antibodies (MAbs) against VHSV were established. One of these, MAb VHS-10, reacted only with genotype IVa in indirect fluorescent antibody technique (IFAT) and ELISA. Using cell cultures that were transfected with each of the viral protein genes, it was shown that the MAb VHS-10 recognizes a nonlinear genotype IVa-specific epitope on the VHSV N-protein.

  2. Virulence of viral haemorrhagic septicaemia virus (VHSV) genotype III in rainbow trout.

    PubMed

    Ito, Takafumi; Kurita, Jun; Mori, Koh-ichiro; Olesen, Niels J

    2016-01-08

    In general, viral haemorrhagic septicaemia virus (VHSV) isolates from marine fish species in European waters (genotypes GIb, GII and GIII) are non- to low virulent in rainbow trout. However, a VHSV isolation was made in 2007 from a disease outbreak in sea farmed rainbow trout in Norway. The isolate, named NO-2007-50-385, was demonstrated to belong to GIII. This isolate has attracted attention to assess which of the viral genome/proteins might be associated with the virulence in rainbow trout. In this study, we describe the difference of virulence in rainbow trout between the NO-2007-50-385 and 4p168 isolates as representatives of virulent and non-virulent GIII isolates, respectively. Rainbow trout were bath challenged with VHSV NO-2007-50-385 for 1 and 6 h, resulting in cumulative mortalities of 5 and 35%, respectively. No mortality was observed in the rainbow trout groups immersed with the genotype III VHSV isolate 4p168 for 1 and 6 h. The viral titre in organs from fish challenged with NO-2007-50-385 for 6 h increased more rapidly than those exposed for 1 h. By in vitro studies it was demonstrated that the final titres of VHSV DK-3592B (GI), NO-2007-50-385 and 4p168 inoculated on EPC cells were very similar, whereas when inoculated on the rainbow trout cell line RTG-2 the titre of the non-virulent 4p168 isolate was 3-4 logs below the two other VHSV isolates. Based on a comparative analysis of the entire genome of the genotype III isolates, we suggest that substitutions of amino acids in positions 118-123 of the nucleo-protein are candidates for being related to virulence of VHSV GIII in rainbow trout.

  3. Monthly occurrence and prevalence of viral haemorrhagic septicaemia virus (VHSV) in whiting Merlangius merlangus.

    PubMed

    Altuntaş, C; Ogut, H

    2010-01-25

    A field survey was carried out to determine the occurrence and spread of viral haemorrhagic septicaemia virus (VHSV) in whiting Merlangius merlangus (L.), a key prey species in the southeastern Black Sea region. In March, April, September and December 2007 and monthly in 2008, 70 to 105 M. merlangus captured by gill nets were examined in pools of 7 fish per pool (total number of fish = 1603). After determining the seasonal occurrence of VHSV in whiting, 43, 80 and 88 whiting were sampled individually in January, February and March 2009, respectively, to determine age and size of whiting commonly infected with VHSV. The samples were tested in duplicate using BF-2 and EPC cell lines for virus isolation. Positive cultures (those demonstrating cytopathic effect, [CPE]), were tested further with a commercial ELISA kit to confirm that the observed CPE was caused by VHSV. VHSV was detected in M. merlangus only between February and April following a drop in the surface water temperature to 9 degrees C. All infected fish from the sampled whiting were asymptomatic carriers with low levels of virus. The estimated prevalence from pooled samples ranged between 1.35% (2.5 to 97.5 percentiles; 0.04 to 7.32%) to 3.14% (0.36 to 10.95%), whereas prevalence estimated from individual sampling was 7, 5 and 1% in January, February and March, respectively. Individual sampling also revealed that prevalence was higher among young fish (1 to 1.5 yr old) and decreased with increasing age. This is the first report of VHSV infection in whiting in the Black Sea.

  4. Identifying potential virulence determinants in viral haemorrhagic septicaemia virus (VHSV) for rainbow trout.

    PubMed

    Campbell, S; Collet, B; Einer-Jensen, K; Secombes, C J; Snow, M

    2009-11-09

    We identified viral haemorrhagic septicaemia virus (VHSV) isolates classified within Genotype Ib which are genetically similar (>99.4% glycoprotein amino acid identity) yet, based on their isolation history, were suspected to differ in virulence in juvenile rainbow trout. The virulence of an isolate recovered in 2000 from a viral haemorrhagic septicaemia disease episode in a marine rainbow trout farm in Sweden (SE-SVA-1033) was evaluated in juvenile rainbow trout via intraperitoneal injection and immersion challenge alongside 3 isolates recovered from wild-caught marine fish (DK-4p37, DK-5e59 and UKMLA98/6HE1) suspected of being of low pathogenicity to trout. Mortality data revealed that isolate SE-SVA-1033 caused VHSV-specific mortality in both intraperitoneal and immersion challenges (75.0 and 15.4%, respectively). The remaining Genotype Ib isolates caused significantly lower mortalities using the same experimental infection routes (<35.0 and <2.0%, respectively). Having identified VHSV isolates with clear differences in their pathogenicity, coding and inter-genic non-coding regions of 2 isolates (SE-SVA-1033 and DK-4p37) were determined and compared in order to identify potential markers responsible for the observed differences in virulence. Only 4 predicted amino acid substitutions were identified across the genome sequenced; these occurred in the N (R46G), G (S113G), NV (L12F) and L (S56A) proteins. These findings form the basis for further studies aimed at determining the biological significance of these mutations and suggest that small changes at the molecular level can cause significant changes in the virulence properties of VHSV isolates.

  5. Risks associated with commodity trade: transmission of viral haemorrhagic septicaemia virus (VHSV) to rainbow trout fry from VHSV-carrying tissue-homogenates.

    PubMed

    Oidtmann, B; Joiner, C; Reese, R A; Stone, D; Dodge, M; Dixon, P

    2011-06-01

    Movements of commodity fish present a potential risk of transferring pathogens. Within a study to estimate the risk from imported rainbow trout Oncorhynchus mykiss carcases, fry were exposed to tissue homogenates from market size rainbow trout infected experimentally with viral haemorrhagic septicaemia (VHS) by waterborne exposure to VHS virus (VHSV, isolate of genotype Ia). Tissues were collected from fish that showed clinical signs and from recent mortalities. Homogenates of (i) internal organs, (ii) brain/gills and (iii) muscle tissue were prepared and added to tanks holding the fry. Virus transmission occurred from all tissues tested, causing high mortality of the fry. The results underline the potential risk of introduction of VHSV through the trade of fish products.

  6. Investigation of wild caught whitefish, Coregonus lavaretus (L.), for infection with viral haemorrhagic septicaemia virus (VHSV) and experimental challenge of whitefish with VHSV.

    PubMed

    Skall, H F; Kjaer, T E; Olesen, N J

    2004-07-01

    Abstract One hundred and forty-eight wild whitefish, Coregonus lavaretus (L.), were caught by electrofishing and sampled for virological examination in December 1999 and 2000, during migration from the brackish water feeding grounds to the freshwater spawning grounds, where the whitefish may come into contact with farmed rainbow trout. All samples were examined on cell cultures. No viruses were isolated. Three viral haemorrhagic septicaemia virus (VHSV) isolates of different origin were tested in infection trials by immersion and intraperitoneal (IP) injection, using 1.5 g farmed whitefish: an isolate from wild caught marine fish, a farmed rainbow trout isolate with a suspected marine origin and a classical freshwater isolate. The isolates were highly pathogenic by IP injection where 99-100% of the whitefish died. Using an immersion challenge the rainbow trout isolates were moderately pathogenic with approximately 20% mortality, whereas the marine isolate was virtually non-pathogenic. At the end of the experiment it was possible to isolate VHSV from survivors infected with the marine and suspected marine isolates. Because of the low infection rate in wild whitefish in Denmark, the role of whitefish in the spread of VHSV in Denmark is probably not significant. The experimental studies, however, showed that whitefish are potential carriers of VHSV as they suffer only low mortality after infection but continue to carry virus. Copyright 2004 Blackwell Publishing Ltd

  7. Transcriptome analysis of rainbow trout in response to non-virion (NV) protein of viral haemorrhagic septicaemia virus (VHSV).

    PubMed

    Chinchilla, Blanca; Encinas, Paloma; Estepa, Amparo; Coll, Julio M; Gomez-Casado, Eduardo

    2015-02-01

    The non-virion (NV) protein of viral haemorrhagic septicaemia virus (VHSV), an economically important fish novirhabdovirus, has been implicated in the interference of some host innate mechanisms (i.e. apoptosis) in vitro. This work aimed to characterise the immune-related transcriptome changes in rainbow trout induced by NV protein that have not yet been established in vivo. For that purpose, immune-targeted microarrays were used to analyse the transcriptomes from head kidney and spleen of rainbow trout (Oncorhynchus mykiss) after injection of recombinant NV (rNV). Results showed the extensive downregulation (and in some cases upregulation) of many innate and adaptive immune response genes not related previously to VHSV infection. The newly identified genes belonged to VHSV-induced genes (vigs), tumour necrosis factors, Toll-like receptors, antigen processing and presentation, immune co-stimulatory molecules, interleukins, macrophage chemotaxis, transcription factors, etc. Classification of differentially downregulated genes into rainbow trout immune pathways identified stat1 and jun/atf1 transcription factor genes as the most representative of the multipath gene targets of rNV. Altogether, these results contribute to define the role and effects of NV in trout by orchestrating an immunosuppression of the innate immune responses for favouring viral replication upon VHSV infection. Finally, these transcriptome results open up the possibility to find out new strategies against VHSV and better understand the interrelationships between some immune pathways in trout.

  8. Molecular epidemiology of viral haemorrhagic septicaemia virus (VHSV) in British Columbia, Canada, reveals transmission from wild to farmed fish.

    PubMed

    Garver, Kyle A; Traxler, Garth S; Hawley, Laura M; Richard, Jon; Ross, Jay P; Lovy, Jan

    2013-05-27

    Viral haemorrhagic septicaemia virus (VHSV) is a fish pathogen found throughout the Northern Hemisphere and is capable of infecting and causing mortality in numerous marine and freshwater hosts. In the coastal waters of British Columbia, Canada, the virus has been detected for 20 yr with many occurrences of mass mortalities among populations of Pacific herring Clupea pallasii (Valenciennes) and sardine Sardinops sagax as well as detections among cultured Atlantic Salmo salar and Chinook Oncorhynchus tshawytscha salmon. We compared nucleotide sequence of the full glycoprotein (G) gene coding region (1524 nt) of 63 VHSV isolates sampled during its recorded presence from 1993 to 2011 from 6 species and a total of 29 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVa within the major VHSV genetic group IV. Of the 63 virus isolates, there were 42 unique sequences, each of which was ephemeral, being repeatedly detected at most only 1 yr after its initial detection. Multiple sequence types were revealed during single viral outbreak events, and genetic heterogeneity was observed within isolates from individual fish. Moreover, phylogenetic analysis revealed a close genetic linkage between VHSV isolates obtained from pelagic finfish species and farmed salmonids, providing evidence for virus transmission from wild to farmed fish.

  9. Susceptibility of various Japanese freshwater fish species to an isolate of viral haemorrhagic septicaemia virus (VHSV) genotype IVb.

    PubMed

    Ito, Takafumi; Olesen, Niels Jørgen

    2013-11-25

    Genotype IVb of viral haemorrhagic septicaemia virus (VHSV) was isolated for the first time in the Great Lakes basin in 2003, where it spread and caused mass mortalities in several wild fish species throughout the basin. In order to prevent further spreading of the disease and to assess risks of new genotypes invading new watersheds, basic microbiological information such as pathogenicity studies are essential. In this study, experimental infections were conducted on 7 indigenous freshwater fish species from Japan by immersion with a VHSV genotype IVb isolate. In Expt 1, cumulative mortalities in bluegill Lepomis macrochirus used as positive controls, Japanese fluvial sculpin Cottus pollux, and iwana Salvelinus leucomaenis pluvius were 50, 80 and 0%, respectively. In Expt 2, cumulative mortalities of 100, 100 and 10% were observed in Japanese fluvial sculpin C. pollux, Japanese rice fish Oryzias latipes and yoshinobori Rhinogobius sp., respectively. No mortality was observed in honmoroko Gnathopogon caerulescens, akaza Liobagrus reini or Japanese striped loach Cobitis biwae. VHSV was detected by RT-PCR from samples of kidney, spleen, and brain from all dead fish, and virus re-isolation by cell culture was successful from all dead fish. We detected the virus in the brain from a few surviving bluegill 50 d post exposure by both cell culture and RT-PCR. These results revealed that VHSV IVb could become a serious threat to wild freshwater fish species in Japan, and that some surviving fish might become healthy carriers of the virus.

  10. Protection and antibody response induced by intramuscular DNA vaccine encoding for viral haemorrhagic septicaemia virus (VHSV) G glycoprotein in turbot (Scophthalmus maximus).

    PubMed

    Pereiro, P; Martinez-Lopez, A; Falco, A; Dios, S; Figueras, A; Coll, J M; Novoa, B; Estepa, A

    2012-06-01

    Turbot (Scophthalmus maximus) is a high-value farmed marine flatfish with growing demand and production levels in Europe susceptible to turbot-specific viral haemorrhagic septicaemia virus (VHSV) strains. To evaluate the possibility of controlling the outbreaks of this infectious disease by means of DNA vaccination, the gpG of a VHSV isolated from farmed turbot (VHSV(860)) was cloned into an expression plasmid containing the human cytomegalovirus (CMV) promoter (pMCV1.4-G(860)). In our experimental conditions, DNA immunised turbots were more than 85% protected against VHSV(860) lethal challenge and showed both VHSV-gpG specific and neutralizing antibodies. To our knowledge this is the first report showing the efficacy of turbot genetic immunisation against a VHSV. Work is in progress to determine the contribution of innate and adaptive immunity to the protective response elicited by the immunization.

  11. Susceptibility of goldsinny wrasse, Ctenolabrus rupestris L. (Labridae), to viral haemorrhagic septicaemia virus (VHSV) genotype III: Experimental challenge and pathology.

    PubMed

    Matejusova, I; Noguera, P A; Hall, M; McBeath, A J A; Urquhart, K; Simons, J; Fordyce, M J; Lester, K; Ho, Y-M; Murray, W; Bruno, D W

    2016-04-15

    Cleaner fish, such as wrasse, are being increasingly used to combat the sea lice infestation of Atlantic salmon (Salmo salar L.) in many European countries. To determine susceptibility of the goldsinny wrasse (Ctenolabrus rupestris L.) and pathogenesis of the viral haemorrhagic septicaemia virus (VHSV) genotype III isolate 12-654, previously associated with VHSV infection in the Shetland Islands in 2012, fish were experimentally challenged by intraperitoneal injection (IP), bath immersion and cohabitation routes. Cumulative proportion of moribund wrasse reached 17% following the virus immersion challenge while by the IP-route moribunds exceeded 50% within 14days post-challenge. Typical signs of VHS as reported in rainbow trout (Oncorhynchus mykiss), were not observed in moribund goldsinny wrasse. The most pronounced histopathological changes, consistent regardless of the route of infection, were observed within the heart and included atrium myofibril degeneration, focal infiltration and multifocal necrosis, with prominent swelling of the endocardium and occasional detachment. Pathological changes in the atrium were associated with presence of the viral antigen as confirmed by a positive immunohistochemical staining. Virus clearance and heart tissue recovery were noted although further experiments are required to confirm these observations. The results of a cohabitation experiment confirmed that goldsinny wrasse shed viable virus and therefore represent a risk of virus transmission to other VHSV susceptible species. Similarities between the pathology in goldsinny wrasse induced through the controlled experimental challenges and that of wrasse spp. from an infection occurrence in Shetland are discussed.

  12. Interferon response following infection with genetically similar isolates of viral haemorrhagic septicaemia virus (VHSV) exhibiting contrasting virulence in rainbow trout.

    PubMed

    Campbell, S; McBeath, A; Secombes, C; Snow, M; Collet, B

    2011-01-01

    Isolates of viral haemorrhagic septicaemia virus (VHSV) were identified which are genetically similar yet, based on their isolation history were considered likely to differ in virulence in juvenile rainbow trout. An experimental infection study was performed in order to verify this hypothesis and provide an experimental infectivity model with which to investigate the basis for susceptibility of rainbow trout to this commercially important virus. Significant differences in mortality were obtained following both intraperitoneal (IP) injection and immersion challenges with an early marine (DK-M.Rhabdo) and early rainbow trout VHSV isolate (DK-F1) respectively. Expression of Type I IFN, Mx1 (an IFN-inducible protein), and viral genes (encoding nucleo-, phospho-, matrix, glyco- and non-viron proteins) was studied in sequential tissue samples using real-time quantitative PCR (QPCR). Resulting data revealed a significant increase in IFN and Mx1 expression detected in fish challenged by IP injection with both isolates. Expression levels of these genes were directly related to the degree of viral replication as measured by the expression of VHSV RNAs. In immersion-challenged fish a significant increase in Mx1 was observed only when using the virulent isolate DK-F1; however no elevated host response was detectable in fish challenged with the marine isolate DK-M.Rhabdo. Quintessentially the inability to detect any virus in trout challenged with the marine isolate via immersion suggests the virus was incapable of establishing infection. The mechanisms for this appear to be more related to initial cellular entry and replication rather than due to the overcoming of initial infection via an elevated host innate immune response.

  13. Development of a sensitive and controlled real-time RT-PCR assay for viral haemorrhagic septicaemia virus (VHSV) in marine salmonid aquaculture.

    PubMed

    Matejusova, Iveta; McKay, Paul; McBeath, Alastair J A; Collet, Bertrand; Snow, Michael

    2008-07-07

    A survey was undertaken to determine the potential distribution of viral haemorrhagic septicaemia virus (VHSV) in marine cage-based salmonid farms in Scotland. A rapid, accurate and sensitive quantitative real-time RT-PCR (qRT-PCR) assay was developed, targeting a conserved region of the nucleoprotein (N) gene of the virus. The qRT-PCR assay was shown to be more sensitive than the conventional VHSV RT-PCR. A validation protocol included several different virus isolates as the target and confirmed that the assay could detect all European VHSV genotypes (I, II and III). Both endogenous and exogenous controls were designed to control for integrity of template and distinguish between true VHSV positives and contamination with the positive control material. In total, the universal European VHSV qRT-PCR assay with exogenous positive control was applied to screen 2040 individual Atlantic salmon Salmo salar and 150 individual rainbow trout Oncorhynchus mykiss. No evidence of the presence of VHSV in association with either salmonid species in Scottish marine farms was detected. However, both marine Atlantic salmon and rainbow trout are still considered possible carriers of VHSV, which remains a potential threat to freshwater farming. Therefore, a continued surveillance of these species in marine environment is recommended.

  14. Microarray analysis of gene expression in olive flounder liver infected with viral haemorrhagic septicaemia virus (VHSV).

    PubMed

    Cho, Hyun Kook; Kim, Julan; Moon, Ji Young; Nam, Bo-Hye; Kim, Young-Ok; Kim, Woo-Jin; Park, Jung Youn; An, Cheul Min; Cheong, Jaehun; Kong, Hee Jeong

    2016-02-01

    The most fatal viral pathogen in olive flounder Paralichthys olivaceus, is viral hemorrhagic septicemia virus, which afflicts over 48 species of freshwater and marine fish. Here, we performed gene expression profiling on transcripts isolated from VHSV-infected olive flounder livers using a 13 K cDNA microarray chip. A total of 1832 and 1647 genes were upregulated and down-regulated over two-fold, respectively, after infection. A variety of immune-related genes showing significant changes in gene expression were identified in upregulated genes through gene ontology annotation. These genes were grouped into categories such as antibacterial peptide, antigen-recognition and adhesion molecules, apoptosis, cytokine-related pathway, immune system, stress response, and transcription factor and regulatory factors. To verify the cDNA microarray data, we performed quantitative real-time PCR, and the results were similar to the microarray data. In conclusion, these results may be useful for the identification of specific genes or for the diagnosis of VHSV infection in flounder.

  15. Viral replication in excised fin tissues (VREFT) corresponds with prior exposure of Pacific herring, Clupea pallasii (Valenciennes), to viral haemorrhagic septicaemia virus (VHSV).

    PubMed

    Grady, C A; Gregg, J L; Wade, R M; Winton, J R; Hershberger, P K

    2011-01-01

    Procedures for a viral replication in excised fin tissue (VREFT) assay were adapted to Pacific herring, Clupea pallasii, and optimized both to reduce processing time and to provide the greatest resolution between naïve herring and those previously exposed to viral haemorrhagic septicaemia virus (VHSV), Genogroup IVa. The optimized procedures included removal of the left pectoral fin from a euthanized fish, inoculation of the fin with >10(5) plaque-forming units (PFU) mL(-1) VHSV for 1 h, rinsing the fin in fresh medium six times to remove unadsorbed virions, incubation of the fin in fresh medium for 4 days and enumeration of the viral titre in a sample of the incubation medium by plaque assay. The optimized VREFT assay was effective at identifying the prior exposure history of laboratory-reared Pacific herring to VHSV. The geometric mean VREFT value was significantly greater (P < 0.01) among naïve herring (1.2 × 10(3) PFU mL(-1) ) than among groups that survived exposure to VHSV (1.0-2.9 × 10(2) PFU mL(-1) ); additionally, the proportion of cultures with no detectable virus was significantly greater (P = 0.0002) among fish that survived exposure to VHSV (39-47%) than among naïve fish (3.3%). The optimized VREFT assay demonstrates promise for identifying VHSV exposure history and forecasting disease potential in populations of wild Pacific herring.

  16. Viral replication in excised fin tissues (VREFT) corresponds with prior exposure of Pacific herring, Clupea pallasii (Valenciennes), to viral haemorrhagic septicaemia virus (VHSV)

    USGS Publications Warehouse

    Grady, C.A.; Gregg, J.L.; Wade, R.M.; Winton, J.R.; Hershberger, P.K.

    2011-01-01

    Procedures for a viral replication in excised fin tissue (VREFT) assay were adapted to Pacific herring, Clupea pallasii, and optimized both to reduce processing time and to provide the greatest resolution between na??ve herring and those previously exposed to viral haemorrhagic septicaemia virus (VHSV), Genogroup IVa. The optimized procedures included removal of the left pectoral fin from a euthanized fish, inoculation of the fin with >105 plaque-forming units (PFU) mL-1 VHSV for 1 h, rinsing the fin in fresh medium six times to remove unadsorbed virions, incubation of the fin in fresh medium for 4 days and enumeration of the viral titre in a sample of the incubation medium by plaque assay. The optimized VREFT assay was effective at identifying the prior exposure history of laboratory-reared Pacific herring to VHSV. The geometric mean VREFT value was significantly greater (P < 0.01) among na??ve herring (1.2 ?? 103 PFU mL-1) than among groups that survived exposure to VHSV (1.0-2.9 ?? 102 PFU mL-1); additionally, the proportion of cultures with no detectable virus was significantly greater (P = 0.0002) among fish that survived exposure to VHSV (39-47%) than among na??ve fish (3.3%). The optimized VREFT assay demonstrates promise for identifying VHSV exposure history and forecasting disease potential in populations of wild Pacific herring. ?? 2010 Blackwell Publishing Ltd.

  17. Viral haemorrhagic septicaemia virus (VHSV) genotype II isolated from European river lamprey Lampetra fluviatilis in Finland during surveillance from 1999 to 2008.

    PubMed

    Gadd, Tuija; Jakava-Viljanen, Miia; Einer-Jensen, Katja; Ariel, Ellen; Koski, Perttu; Sihvonen, Liisa

    2010-02-17

    We examined the occurrence of viral haemorrhagic septicaemia virus (VHSV) in the main spawning stocks of wild European river lamprey Lampetra fluviatilis in the rivers of Finland from 1999 to 2008. Pooled samples of internal organs (kidney, liver and heart or brain) from 2621 lampreys were examined for the presence of VHSV by standard virological techniques. VHSV was isolated from 5 samples from the rivers Lestijoki and Kalajoki, which flow from Finland into the Bothnian Bay of the Baltic Sea. The presence of VHSV was confirmed by immunofluorescent antibody technique (IFAT), ELISA and RT-PCR. Phylogenetic analysis based on the full-length VHSV glycoprotein (G) gene sequence revealed that the isolates were most closely related to the VHSV strain isolated in 1996 from herring Clupea harengus and sprat Sprattus sprattus in the Eastern Gotland Basin of the Baltic Sea, and were therefore assigned to VHSV genotype II. The partial G gene sequences obtained (nt 1 to 672-1129) of all 5 lamprey VHSV isolates were identical, and so were the entire G genes (nt 1 to 1524) of 2 isolates sequenced. The virulence of one of the lamprey isolates was evaluated by an experimental infection trial in rainbow trout Oncorhynchus mykiss fry. No mortality was induced postinfection by waterborne and intraperitoneal challenge, respectively, while 2 genotype Id isolates originating from Finnish rainbow trout caused marked mortality under the same conditions. The infection in the European river lamprey is thought to be independent from the epidemic in farmed rainbow trout in Finnish brackish waters, because the isolates from rainbow trout were of a different genotype. This is the first report of VHSV found in the European river lamprey. The role of wild river lampreys in maintaining the infection in the marine environment remains unclear.

  18. Microarray-based identification of differentially expressed genes in families of turbot (Scophthalmus maximus) after infection with viral haemorrhagic septicaemia virus (VHSV).

    PubMed

    Díaz-Rosales, P; Romero, A; Balseiro, P; Dios, S; Novoa, B; Figueras, A

    2012-10-01

    Viral haemorrhagic septicaemia virus (VHSV) is one of the major threats to the development of the aquaculture industry worldwide. The present study was aimed to identify genes differentially expressed in several turbot (Scophthalmus maximus) families showing different mortality rates after VHSV. The expression analysis was conducted through genome-wide expression profiling with an oligo-microarray in the head kidney. A significant proportion of the variation in the gene expression profiles seemed to be explained by the genetic background, indicating that the mechanisms by which particular species and/or populations can resist a pathogen(s) are complex and multifactorial. Before the experimental infections, fish from resistant families (low mortality rates after VHSV infection) showed high expression of different antimicrobial peptides, suggesting that their pre-immune state may be stronger than fish of susceptible families (high mortality rates after VHSV infection). After infection, fish from both high- and low-mortality families showed an up-modulation of the interferon-induced Mx2 gene, the IL-8 gene and the VHSV-induced protein 5 gene compared with control groups. Low levels of several molecules secreted in the mucus were observed in high-mortality families, but different genes involved in viral entrance into target cells were down-regulated in low-mortality families. Moreover, these families also showed a strong down-modulation of marker genes related to VHSV target organs, including biochemical markers of renal dysfunction and myocardial injury. In general, the expression of different genes involved in the metabolism of sugars, lipids and proteins were decreased in both low- and high-mortality families after infection. The present study serves as an initial screen for genes of interest and provides an extensive overview of the genetic basis underlying the differences between families that are resistant or susceptible to VHSV infection.

  19. Viral hemorrhagic septicaemia virus (VHSV) up-regulates the cytotoxic activity and the perforin/granzyme pathway in the rainbow trout RTS11 cell line.

    PubMed

    Ordás, M C; Cuesta, A; Mercado, L; Bols, N C; Tafalla, C

    2011-08-01

    A survey of immune-relevant genes that might be up-regulated in response to viral hemorrhagic septicaemia virus (VHSV) in the rainbow trout monocyte-macrophage cell line, RTS11, unexpectedly revealed an increased expression of perforin (PRF) and granzyme (GRZ) genes, which represent components of the major cytotoxic pathway. The natural killer-enhancing factor (NKEF), also known to modulate cytotoxic activity, was up-regulated at the gene but strikingly down-regulated at protein level. The expression of these genes was not affected in head kidney leukocytes (HKLs) infected with VHSV, leading us to evaluate the potential cytotoxic activity of RTS11 and HKLs. For the first time, the cytotoxic activity of RTS11 against xenogeneic targets has been demonstrated, although this was modest relative to HKLs. Yet the activity in RTS11 was significantly increased by VHSV, as in HKLs. This cytotoxic activity elicited by viral infection appeared to require viral gene expression because inactivated VHSV failed to increase RTS11 cytotoxic activity. As for other immune functions, RTS11 cells provide a model for further studying cytotoxic activities of fish monocyte-macrophages.

  20. In vivo virulence of viral haemorrhagic septicaemia virus (VHSV) in rainbow trout Oncorhynchus mykiss correlates inversely with in vitro Mx gene expression.

    PubMed

    Cano, Irene; Collet, Bertrand; Pereira, Clarissa; Paley, Richard; Aerle, Ronny van; Stone, David; Taylor, Nick G H

    2016-05-01

    The in vitro replication of viral haemorrhagic septicaemia virus (VHSV) isolates from each VHSV genotype and the associated cellular host Mx gene expression were analysed. All the isolates were able to infect RTG-2 cells and induce increased Mx gene expression (generic assay detecting isoforms 1 and 3 [Mx1/3]). A trout pathogenic, genotype Ia isolate (J167), showing high replication in RTG-2 cells (by infective titre and N gene expression) induced lower Mx1/3 gene expression than observed in VHSV isolates known to be non-pathogenic to rainbow trout: 96-43/8, 96-43/10 (Ib); 1p49, 1p53 (II); and MI03 (IVb). Paired co-inoculation assays were analysed using equal number of plaque forming units per ml (PFU) of J167 (Ia genotype) with other less pathogenic VHSV genotypes. In these co-inoculations, the Mx1/3 gene expression was significantly lower than for the non-pathogenic isolate alone. Of the three rainbow trout Mx isoforms, J167 did not induce Mx1 up-regulation in RTG-2 or RTgill-W1 cells. Co-inoculating isolates resulted in greater inhibition of Mx in both rainbow trout cell lines studied. Up-regulation of sea bream Mx in SAF-1 cells induced by 96-43/8 was also lower in co-inoculation assays with J167. The RTG-P1 cell line, expressing luciferase under the control of the interferon-induced Mx rainbow trout gene promoter, showed low luciferase activity when inoculated with pathogenic strains: J167, DK-5131 (Ic), NO-A-163/68 (Id), TR-206239-1, TR-22207111 (Ie), 99-292 (IVa), and CA-NB00-01 (IVc). Co-inoculation assays showed a J167-dose dependent inhibition of the luciferase activity. The data suggest that virulent VHSV isolates may interfere in the interferon pathways, potentially determining higher pathogenicity. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  1. Application of a sensitive, specific and controlled real-time PCR assay to surveillance indicates a low prevalence of viral haemorrhagic septicaemia virus (VHSV) in wild herring, Clupea harengus L., in Scottish waters.

    PubMed

    Matejusova, I; McKay, P; Bland, F; Snow, M

    2010-10-01

    Surveillance data on the distribution of viral haemorrhagic septicaemia virus (VHSV) in the North Sea (UK), targeting Atlantic herring in areas with previous virus detection, were obtained from research cruises conducted during 2005. The sensitive molecular approach of real-time RT-PCR (qRT-PCR) was applied alongside a newly developed endogenous positive control assay specific for herring (elongation factor 1α) to ensure integrity of template. Three hundred and five pools from 1937 individual herring were tested, and no evidence of VHSV in association with wild Atlantic herring was detected. Samples were obtained from Scottish waters where marine aquaculture is conducted. The results confirm that previous tissue culture studies have most likely not significantly underestimated the prevalence of carrier herring in this area. The significance of migratory species such as herring as a reservoir species for VHSV, with the potential to translocate virus genotypes between geographical areas, is discussed.

  2. Temperature-dependent viral replication and antiviral apoptotic response in viral haemorrhagic septicaemia virus (VHSV)-infected olive flounder (Paralichthys olivaceus).

    PubMed

    Avunje, Satheesha; Kim, Wi-Sik; Oh, Myung-Joo; Choi, Ilsu; Jung, Sung-Ju

    2012-06-01

    The olive flounder (Paralichthys olivaceus) shows a high rate of mortality to viral haemorrhagic septicaemia virus (VHSV) in the winter and spring but has zero mortality over 20 °C. In this experiment, we studied the effect of rearing temperature on viral replication, viral transcription and antiviral apoptotic immune response in VHSV-infected olive flounder by real-time polymerase chain reaction. Olive flounder were given intra-peritoneal injections of VHSV (10(7.8) TCID(50)/ml) and were reared at 15 °C or 20 °C. Five fish were randomly sampled for head kidney at 3, 6 and 12 h post-infection (hpi) and 1, 2, 4 and 7 days post-infection (dpi). Total RNA extracted from the tissue was reverse transcribed and used as template for real-time PCR. In the 15 °C group, the number of viral gRNA copies peaked after 2 dpi and remained high through 7 dpi, while in the 20 °C group, the copy number was at the highest at 1 dpi but drastically declined at later stages. Viral mRNA levels in the 15 °C group gradually increased starting at 3 hpi to reach their maximum value at 12 hpi and remained high until 2 dpi, whereas the other group showed much lower copy numbers that were undetectably low at 4 and 7 dpi. Type II IFN expression increased as the viral copies increased and the 20 °C group showed quicker and stronger expression than the 15 °C group. The MHC class I and CD8 expression was high in both the groups at early stage of infection (3-6 hpi) but at later stages (2-7 dpi) in 15 °C group expression reduced below control levels, while they expressed higher to control in 20 °C group. The expression of granzyme in 15 °C fish showed a single peak at 2 dpi, but was consistently expressing in 20 °C fish. Individuals expressed very high levels of perforin expressed very high levels of caspase 3. In 15 °C fish, TNFα, FasL and p53 expressed significantly higher than 20 °C only at initial stages of infection (3-6 hpi). Caspase 3 expression found to be low in 15 °C fish

  3. Differential effects of viral hemorrhagic septicaemia virus (VHSV) genotypes IVa and IVb on gill epithelial and spleen macrophage cell lines from rainbow trout (Oncorhynchus mykiss).

    PubMed

    Pham, P H; Lumsden, J S; Tafalla, C; Dixon, B; Bols, N C

    2013-02-01

    The two most prominent genotypes of viral hemorrhagic septicemia virus (VHSV) are -I in the Northeastern Atlantic region and -IV in North America, but much more is known about the cellular pathogenesis of genotype -I than -IV. VHSV genotype -IV is divided into -IVa from the Northeast Pacific Ocean and -IVb from the Great Lakes and both of which are less virulent to rainbow trout than genotype -I. In this work, infections of VHSV-IVa and -IVb have been studied in two rainbow trout cell lines, RTgill-W1 from the gill epithelium, and RTS11 from spleen macrophages. RTgill-W1 produced infectious progeny of both VHSV-IVa and -IVb. However, VHSV-IVa was more infectious than -IVb toward RTgill-W1: -IVa caused cytopathic effect (CPE) at a lower viral titre, elicited CPE earlier, and yielded higher titres. By contrast, no CPE and no increase in viral titre were observed in RTS11 cultures infected with either genotype. Yet in RTS11 all six VHSV genes were expressed and antiviral genes, Mx2 and Mx3, were up regulated by VHSV-IVb and -IVa. However, replication appeared to terminate at the translational stage as viral N protein, presumably the most abundant of the VSHV proteins, was not detected in either infected RTS11 cultures. In RTgill-W1, Mx2 and Mx3 were up regulated to similar levels by both viral genotypes, while VHSV-IVa induced higher levels of IFN1, IFN2 and LGP2A than VHSV-IVb. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Identification of the functional regions of the viral haemorrhagic septicaemia virus (VHSV) NV protein: Variants that improve function.

    PubMed

    Chinchilla, Blanca; Gomez-Casado, Eduardo

    2017-09-05

    Non-virion (NV) protein is essential for an efficient replication increasing the pathogenicity of the Salmonid novirhabdovirus (formerly IHNV), Piscine novirhabdovirus (formerly VHSV), and Hirame novirhabdovirus (HIRV). The interferon system, apoptosis, and other immune-related genes are modulated by NV to finally induce a deficient antiviral state in the cell. However, little is known about the VHSV NV regions involved in function and location. Here, eight different NV 07.71 fragments and eleven NV 07.71 mutants derived from the region between the two first α-helices have been studied in order to establish the mx and il8 transcript levels in ZF4 cells and the subcellular location. As a result, we determined that the N-terminal part of NV preserves the same ability as the wild-type (wt) NV in mx/il8 modulation and it also shares the subcellular location. Among NV mutants, some induced mx upregulation (N34A, C35A, D38A, and S40A) but maintained the il8 levels stable when compared to wt-NV in ZF4. Four NV mutants (D28A, N31A, L33A, and F37A) were not affected by the mutation and showed mx and il8 transcript levels similar to wt-NV. Surprisingly, mutants D36A, R39A, and D41A induced a stronger downregulation of both mx and il8 transcript levels than wt-NV, suggesting that a more stable structure and an improved interaction with ligands could be achieved through these mutations. Amino acids at positions 36 and 39 are conserved among known VHSV NV proteins whereas at position 41 two different amino acids have been described. To date, no natural NV proteins with alanine at positions 36, 39, and 41 have been found. In addition, wt-NV, all NV mutants, and one N-terminal NV fragment were located at cytoplasm with a characteristic pattern, which might support that cytoplasm is the site for interaction with candidate ligands such as PPM1Bb. Taken together, the data presented in this work indicated that NV function relies on the first part of the molecule and is dependent on

  5. The immunogenicity of viral haemorragic septicaemia rhabdovirus (VHSV) DNA vaccines can depend on plasmid regulatory sequences.

    PubMed

    Chico, V; Ortega-Villaizan, M; Falco, A; Tafalla, C; Perez, L; Coll, J M; Estepa, A

    2009-03-18

    A plasmid DNA encoding the viral hemorrhagic septicaemia virus (VHSV)-G glycoprotein under the control of 5' sequences (enhancer/promoter sequence plus both non-coding 1st exon and 1st intron sequences) from carp beta-actin gene (pAE6-G(VHSV)) was compared to the vaccine plasmid usually described the gene expression is regulated by the human cytomegalovirus (CMV) immediate-early promoter (pMCV1.4-G(VHSV)). We observed that these two plasmids produced a markedly different profile in the level and time of expression of the encoded-antigen, and this may have a direct effect upon the intensity and suitability of the in vivo immune response. Thus, fish genetic immunisation assays were carried out to study the immune response of both plasmids. A significantly enhanced specific-antibody response against the viral glycoprotein was found in the fish immunised with pAE6-G(VHSV). However, the protective efficacy against VHSV challenge conferred by both plasmids was similar. Later analysis of the transcription profile of a set of representative immune-related genes in the DNA immunized fish suggested that depending on the plasmid-related regulatory sequences controlling its expression, the plasmid might activate distinct patterns of the immune system. All together, the results from this study mainly point out that the selection of a determinate encoded-antigen/vector combination for genetic immunisation is of extraordinary importance in designing optimised DNA vaccines that, when required for inducing protective immune response, could elicit responses biased to antigen-specific antibodies or cytotoxic T cells generation.

  6. Isolation of viral haemorrhagic septicaemia virus from mummichog, stickleback, striped bass and brown trout in eastern Canada.

    PubMed

    Gagné, N; Mackinnon, A-M; Boston, L; Souter, B; Cook-Versloot, M; Griffiths, S; Olivier, G

    2007-04-01

    Viral haemorrhagic septicaemia virus (VHSV) was isolated from mortalities occurring in populations of mummichog, Fundulus heteroclitus, stickleback, Gasterosteus aculeatus aculeatus, brown trout, Salmo trutta, and striped bass, Morone saxatilis, in New Brunswick and Nova Scotia, Canada. The isolated viral strains produced a cytopathic effect on the epithelioma papillosum cyprini cell line. Serum neutralization indicated the virus was VHSV and sequencing identified the rhabdovirus isolates as the North American strain of VHSV. Phylogenetic analysis indicated that the isolates are closely related and form a distinguishable subgroup of North American type VHSV. To our knowledge, this is the first report of VHSV in mummichog and striped bass.

  7. Potential distribution of the viral haemorrhagic septicaemia virus in the Great Lakes region.

    PubMed

    Escobar, L E; Kurath, G; Escobar-Dodero, J; Craft, M E; Phelps, N B D

    2017-01-01

    Viral haemorrhagic septicaemia virus (VHSV) genotype IVb has been responsible for large-scale fish mortality events in the Great Lakes of North America. Anticipating the areas of potential VHSV occurrence is key to designing epidemiological surveillance and disease prevention strategies in the Great Lakes basin. We explored the environmental features that could shape the distribution of VHSV, based on remote sensing and climate data via ecological niche modelling. Variables included temperature measured during the day and night, precipitation, vegetation, bathymetry, solar radiation and topographic wetness. VHSV occurrences were obtained from available reports of virus confirmation in laboratory facilities. We fit a Maxent model using VHSV-IVb reports and environmental variables under different parameterizations to identify the best model to determine potential VHSV occurrence based on environmental suitability. VHSV reports were generated from both passive and active surveillance. VHSV occurrences were most abundant near shore sites. We were, however, able to capture the environmental signature of VHSV based on the environmental variables employed in our model, allowing us to identify patterns of VHSV potential occurrence. Our findings suggest that VHSV is not at an ecological equilibrium and more areas could be affected, including areas not in close geographic proximity to past VHSV reports.

  8. Potential distribution of the viral haemorrhagic septicaemia virus in the Great Lakes region

    USGS Publications Warehouse

    Escobar, Luis E.; Kurath, Gael; Escobar-Dodero, Joaquim; Craft, Meggan E.; Phelps, Nicholas B.D.

    2017-01-01

    Viral haemorrhagic septicaemia virus (VHSV) genotype IVb has been responsible for large-scale fish mortality events in the Great Lakes of North America. Anticipating the areas of potential VHSV occurrence is key to designing epidemiological surveillance and disease prevention strategies in the Great Lakes basin. We explored the environmental features that could shape the distribution of VHSV, based on remote sensing and climate data via ecological niche modelling. Variables included temperature measured during the day and night, precipitation, vegetation, bathymetry, solar radiation and topographic wetness. VHSV occurrences were obtained from available reports of virus confirmation in laboratory facilities. We fit a Maxent model using VHSV-IVb reports and environmental variables under different parameterizations to identify the best model to determine potential VHSV occurrence based on environmental suitability. VHSV reports were generated from both passive and active surveillance. VHSV occurrences were most abundant near shore sites. We were, however, able to capture the environmental signature of VHSV based on the environmental variables employed in our model, allowing us to identify patterns of VHSV potential occurrence. Our findings suggest that VHSV is not at an ecological equilibrium and more areas could be affected, including areas not in close geographic proximity to past VHSV reports.

  9. Differentially expressed genes after viral haemorrhagic septicaemia virus infection in olive flounder (Paralichthys olivaceus).

    PubMed

    Hwang, Jee Youn; Kwon, Mun-Gyeong; Seo, Jung Soo; Do, Jung Wan; Park, Myoung-Ae; Jung, Sung-Hee; Ahn, Sang Jung

    2016-09-25

    A strain of viral haemorrhagic septicaemia virus (VHSV) was isolated from cultured olive flounder (Paralichthys olivaceus) during epizootics in South Korean. This strain showed high mortality to olive flounder in in vivo challenge experiment. The complete genomic RNA sequences were determined and phylogenetic analysis of the amino acid sequences of glycoprotein revealed that this isolate was grouped into genotype IVa of genus Novirhabdovirus. Expression profile of genes in olive flounder was analyzed at day 1 and day3 after infection with this VHSV isolate by using cDNA microarray containing olive flounder 13K cDNA clones. Microarray analysis revealed 785 up-regulated genes and 641 down-regulated genes by at least two-fold in virus-infected fish compared to healthy control groups. Among 785 up-regulated genes, we identified seven immune response-associated genes, including the interferon (IFN)-induced 56-kDa protein (IFI56), suppressor of cytokine signaling 1 (SOCS1), interleukin 8 (IL-8), cluster of differentiation 83 (CD83), α-globin (HBA), VHSV-induced protein-6 (VHSV6), and cluster of differentiation antigen 9 (CD9). Our results confirm previous reports that even virulent strain of VHSV induces expression of genes involved in protective immunity against VHSV. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Survey of viral haemorrhagic septicaemia virus in wild fishes in the southeastern Black Sea.

    PubMed

    Ogut, H; Altuntas, C

    2014-05-13

    Species diversity in the Black Sea ecosystem has been declining rapidly over the last 2 decades. To assess the occurrence and distribution of viral haemorrhagic septicaemia virus (VHSV) in various wild fish species, a wild marine fish survey was carried out in 2009, 2010, and 2011. The pooled or individual samples of kidney, liver, and spleen of 5025 specimens, belonging to 17 fish species, were examined virologically using cell culture. The cells showing cytopathic effects (CPE) were subjected to ELISA and multiplex reverse transcriptase polymerase chain reaction (RT-mPCR), for VHSV and infectious pancreatic necrosis virus (IPNV), after blind passaging to determine the virus species causing CPE. The virus species and possibility of co-infection with IPNV were verified by the RT-mPCR developed in this study. Twelve species of fish (pontic shad Alosa immaculata, red mullet Mullus barbatus, three-bearded rockling Gaidropsarus vulgaris, black scorpionfish Scorpaena porcus, Mediterranean horse mackerel Trachurus mediterraneus, whiting Merlangius merlangus euxinus, stargazer Uranoscopus scaber, pilchard Sardina pilchardus, garfish Belone belone, round goby Neogobius melanostomus, thornback ray Raja clavata, and anchovy Engraulis encrasicolus) tested positive for VHSV Genotype Ie (VHSV-Ie). Except whiting, pilchard, and round goby, the rest are new host records for VHSV. The extent and spread of VHSV-Ie was significantly higher among bottom fish than among pelagic fish. Sensitivity and specificity of the RT-mPCR developed was sufficiently high, suggesting that this assay may be used for both diagnostic and surveillance testing. According to the RT-mPCR results, IPNV was not present in wild fish. These results support the hypothesis that the VHSV-Ie genotype, highly prevalent among fish species in the Black Sea, may have a serious impact on the population dynamics of wild fish stocks.

  11. Analysis of the nucleoprotein gene identifies three distinct lineages of viral haemorrhagic septicemia virus (VHSV) within the European marine environment

    USGS Publications Warehouse

    Snow, M.; Cunningham, C.O.; Melvin, W.T.; Kurath, G.

    1999-01-01

    A ribonuclease (RNase) protection assay (RPA) has been used to detect nucleotide sequence variation within the nucleoprotein gene of 39 viral haemorrhagic septicaemia virus (VHSV) isolates of European marine origin. The classification of VHSV isolates based on RPA cleavage patterns permitted the identification of ten distinct groups of viruses based on differences at the molecular level. The nucleotide sequence of representatives of each of these groupings was determined and subjected to phylogenetic analysis. This revealed grouping of the European marine isolates of VHSV into three genotypes circulating within distinct geographic areas. A fourth genotype was identified comprising isolates originating from North America. Phylogenetic analyses indicated that VHSV isolates recovered from wild caught fish around the British Isles were genetically related to isolates responsible for losses in farmed turbot. Furthermore, a relationship between naturally occurring marine isolates and VHSV isolates causing mortality among rainbow trout in continental Europe was demonstrated. Analysis of the nucleoprotein gene identifies distinct lineages of viral haemorrhagic septicaemia virus within the European marine environment. Virus Res. 63, 35-44. Available from: 

  12. Inter-laboratory comparison of cell lines for susceptibility to three viruses: VHSV, IHNV and IPNV.

    PubMed

    Lorenzen, E; Carstensen, B; Olesen, N J

    1999-07-30

    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 (RTG-2). The results showed that for isolation of VHSV, BF-2 and RTG-2 cells performed equally well and had higher sensitivity compared to the other cell lines. For IHNV, EPC and FHM cells gave the best results, and for IPNV it was BF-2 and CHSE-214 cells. FHM cells showed the largest variability among laboratories, whereas EPC was the cell line showing the smallest variability.

  13. Epidemiological aspects of viral haemorrhagic septicaemia virus genotype II isolated from Baltic herring, Clupea harengus membras L.

    PubMed

    Gadd, T; Jakava-Viljanen, M; Tapiovaara, H; Koski, P; Sihvonen, L

    2011-07-01

    This study was carried out to clarify the role of wild fish, especially Baltic herring, Clupea harengus membras L., in the epidemiology of viral haemorrhagic septicaemia virus (VHSV) in brackish water in Finland. Baltic herring with no visible signs of disease were collected from the Archipelago Sea, the Gulf of Bothnia and the eastern Gulf of Finland. In total, 7580 herring were examined by virus isolation as 758 pooled samples and 3029 wild salmonid broodfish as pooled samples during 2004-2006. VHSV was isolated from 51 pooled herring samples in bluegill fibroblast-2 cells, but not in epithelioma papulosum cyprini cells. The majority of isolations were from the coastal archipelago and from fish caught during the spawning season. Based on glycoprotein (G) gene sequences, the virus was classified as a member of genotype II of VHSV. Pairwise comparisons of the G gene regions of herring isolates revealed that all the isolates were closely related, with 98.8-100% nucleotide homology. Phylogenetic analyses revealed that they were closely related to the strains isolated previously from herring and sprat, Sprattus sprattus (L.), in Gotland and to the VHSV isolates from European river lamprey, Lampetra fluviatilis (L.), in the rivers that flow into the Bothnian Bay. The infection in Baltic herring is likely to be independent of the VHSV Id epidemic in farmed rainbow trout, Oncorhynchus mykiss (Walbaum). © 2011 Blackwell Publishing Ltd.

  14. Do imports of rainbow trout carcasses risk introducing viral haemorrhagic septicaemia virus into England and Wales?

    PubMed

    Pearce, F M; Oidtmann, B C; Thrush, M A; Dixon, P F; Peeler, E J

    2014-06-01

    A qualitative import risk assessment was undertaken to assess the likelihood of introduction and establishment of viral haemorrhagic septicaemia virus (VHSV) genotype 1a in England and Wales (E&W), via the processing of imported rainbow trout (Oncorhynchus mykiss) carcasses from continental Europe. The likelihood was estimated for one import from an infected farm. Four main routes by which susceptible populations could be exposed to VHSV via processing waste were considered: (i) run-off from solid waste to watercourses, (ii) contamination of birds or rodents with VHSV by scavenging solid waste, (iii) discharge of liquid waste to mains drainage, and (iv) discharge of liquid waste directly to watercourses. Data on the biophysical characteristics of VHSV, its epidemiology, fish processing practices and waste management were collected. Likelihoods for each step of the four pathways were estimated. Pathway 4 (discharge of liquid waste to a watercourse) was judged as the most likely to result in infection of susceptible individuals. Levels of virus entering the aquatic environment via pathways 1-3 were judged to be many times lower than pathway 4 due mainly to the treatment of solid waste (pathways 1 and 2) and high levels of dilution (pathways 1, 2 and 3). Thirty-four trout farms process fish, of which seven have imported carcasses for processing. Compared with other processing facilities, on-farm processing results in a higher likelihood of VHSV exposure and establishment via all four pathways. Data availability was an issue; the analysis was particularly constrained by a lack of data on the prevalence of VHSV in Europe, volume of trade of carcasses into the UK and processing practices in E&W. It was concluded that the threat of VHSV introduction into E&W could be reduced by treatment of liquid effluent from processing plants and by sourcing carcasses for on-farm processing only from approved VHSV free areas. © 2012 Crown copyright. Transboundary and Emerging Diseases

  15. A mortality event in wrasse species (Labridae) associated with the presence of viral haemorrhagic septicaemia virus.

    PubMed

    Munro, E S; McIntosh, R E; Weir, S J; Noguera, P A; Sandilands, J M; Matejusova, I; Mayes, A S; Smith, R

    2015-04-01

    Viral haemorrhagic septicaemia (VHS) is an infectious disease of farmed and wild fish and has an extensive host range in both freshwater and marine environments. In December 2012, a wrasse population consisting of ballan, Labrus bergylta (Ascanius), corkwing, Symphodus melops (L.), cuckoo, Labrus mixtus L., goldsinny, Ctenolabrus rupestris (L.), and rock cook, Centrolabrus exoletus (L.), held at a marine hatchery in the Shetland Isles, Scotland, experienced a mortality event. Approximately 10 000 wrasse were being held at the facility on behalf of an Atlantic salmon, Salmo salar L., aquaculture company prior to being deployed for the biological control of parasites on marine pen Atlantic salmon, aquaculture sites. Fish Health Inspectors from Marine Scotland Science initiated a diagnostic investigation, and subsequent diagnostic testing confirmed the site to be VHSV positive by qRT-PCR and virus isolation followed by ELISA. A VHSV genotype-specific qRT-PCR assay revealed that the isolates belonged to genotype III, the European marine strain of the virus. The virus genotype was further confirmed by nucleic acid sequencing of the partial nucleoprotein (N) and glycoprotein (G) genes followed by BLAST nucleotide searches. This study reports for the first time the detection of VHSV within multiple wrasse species and highlights the need for a comprehensive risk-based approach to the use of wrasse and other finfish species as biological controls within the aquaculture industry. © 2014 Crown copyright.

  16. Genotyping of the fish rhabdovirus, viral haemorrhagic septicaemia virus, by restriction fragment length polymorphisms

    USGS Publications Warehouse

    Einer-Jensen, Katja; Winton, James R.; Lorenzen, Niels

    2005-01-01

    The aim of this study was to develop a standardized molecular assay that used limited resources and equipment for routine genotyping of isolates of the fish rhabdovirus, viral haemorrhagic septicaemia virus (VHSV). Computer generated restriction maps, based on 62 unique full-length (1524 nt) sequences of the VHSV glycoprotein (G) gene, were used to predict restriction fragment length polymorphism (RFLP) patterns that were subsequently grouped and compared with a phylogenetic analysis of the G-gene sequences of the same set of isolates. Digestion of PCR amplicons from the full-lengthG-gene by a set of three restriction enzymes was predicted to accurately enable the assignment of the VHSV isolates into the four major genotypes discovered to date. Further sub-typing of the isolates into the recently described sub-lineages of genotype I was possible by applying three additional enzymes. Experimental evaluation of the method consisted of three steps: (i) RT-PCR amplification of the G-gene of VHSV isolates using purified viral RNA as template, (ii) digestion of the PCR products with a panel of restriction endonucleases and (iii) interpretation of the resulting RFLP profiles. The RFLP analysis was shown to approximate the level of genetic discrimination obtained by other, more labour-intensive, molecular techniques such as the ribonuclease protection assay or sequence analysis. In addition, 37 previously uncharacterised isolates from diverse sources were assigned to specific genotypes. While the assay was able to distinguish between marine and continental isolates of VHSV, the differences did not correlate with the pathogenicity of the isolates.

  17. Absence of viral hemorrhagic septicemia virus (VHSV) and infectious hemorrhagic necrosis virus (INHV) in a Tunisian fish farm: a case study.

    PubMed

    Cherif, N; Hammami, S

    2012-01-01

    Beyond the obvious problems related to the sustainable management of wild stocks, current fish farming practices in the Mediterranean area entail important environmental risks and potential outbreaks of fish diseases linked to massive translocations across regional boundaries. Viral haemorrhagic septicaemia (VHS) and infectious hematopoietic necrosis (IHN) are well-known fish diseases caused by the VHSV and IHNV viruses, and positive cases are subject to obligatory reporting to the World Organisation for Animal Health (OIE). In August 2004, the OIE published the first record of a VHS outbreak in a sea bass (Dicentrarchus labrax) aquaculture facility on the coast of the Mediterranean Sea. However, D. labrax is not considered as a susceptible host species for viral hemorrhagic septicaemia according to OIE's International Aquatic Animal Health Code (2009) and VHSV was not previously reported in the Mediterranean. In this sense and given the high risk of disease translocation associated with farmed fish in marine aquaculture, the present study was aimed at investigating the presence of VHSV and IHNV in stocks of sea bass and sea bream (Sparus aurata) reared inside a Tunisian coastal fish farm. Cell culture, IFAT and RT-PCR were applied to screen for both VHSV and IHNV in 69 pooled samples of sea bass and 24 pooled samples of sea bream. All three techniques showed the absence of both viruses within fish at the selected site.

  18. Virus isolation vs RT-PCR: which method is more successful in detecting VHSV and IHNV in fish tissue sampled under field conditions?

    PubMed

    Knüsel, R; Bergmann, S M; Einer-Jensen, K; Casey, J; Segner, H; Wahli, T

    2007-09-01

    This study compared the results of reverse transcription-polymerase chain reaction (RT-PCR) and traditional virus isolation on cell culture in detection of viral haemorrhagic septicaemia virus (VHSV) and infectious haematopoietic necrosis virus (IHNV). RT-PCR was used for 172 tissue sample pools (total of 859 fish) originating from a field survey on the occurrence of VHSV and IHNV in farmed and wild salmonids in Switzerland. These samples represented all sites with fish that were either identified as virus-positive by means of virus isolation (three sites, four positive tissue sample pools) and/or demonstrated positive anti-VHSV-antibody titres (83 sites, 121 positive blood samples) in a serum plaque neutralization test (SPNT). The RT-PCR technique confirmed the four VHSV-positive tissue sample pools detected by virus isolation and additionally identified one VHSV-positive sample that showed positive anti-VHSV-AB titres, but was negative in virus isolation. With IHNV, RT-PCR detected two positive samples not identified by virus isolation while in these fish the SPNT result had been questionable. One of the IHNV-positive samples represents the first detection of IHNV-RNA in wild brown trout in Switzerland. Compared to SPNT, the RT-PCR method detected, as with virus isolation, a much lower number of positive cases; reasons for this discrepancy are discussed. Our results indicate that RT-PCR can not only be successfully applied in field surveys, but may also be slightly more sensitive than virus isolation. However, in a titration experiment under laboratory conditions, the sensitivity of RT-PCR was not significantly higher when compared with virus isolation.

  19. Antibody recognition of the glycoprotein g of viral haemorrhagic septicemia virus (VHSV) purified in large amounts from insect larvae

    PubMed Central

    2011-01-01

    Background There are currently no purification methods capable of producing the large amounts of fish rhabdoviral glycoprotein G (gpG) required for diagnosis and immunisation purposes or for studying structure and molecular mechanisms of action of this molecule (ie. pH-dependent membrane fusion). As a result of the unavailability of large amounts of the gpG from viral haemorrhagic septicaemia rhabdovirus (VHSV), one of the most dangerous viruses affecting cultured salmonid species, research interests in this field are severely hampered. Previous purification methods to obtain recombinant gpG from VHSV in E. coli, yeast and baculovirus grown in insect cells have not produced soluble conformations or acceptable yields. The development of large-scale purification methods for gpGs will also further research into other fish rhabdoviruses, such as infectious haematopoietic necrosis virus (IHNV), spring carp viremia virus (SVCV), hirame rhabdovirus (HIRRV) and snakehead rhabdovirus (SHRV). Findings Here we designed a method to produce milligram amounts of soluble VHSV gpG. Only the transmembrane and carboxy terminal-deleted (amino acid 21 to 465) gpG was efficiently expressed in insect larvae. Recognition of G21-465 by ß-mercaptoethanol-dependent neutralizing monoclonal antibodies (N-MAbs) and pH-dependent recognition by sera from VHSV-hyperimmunized or VHSV-infected rainbow trout (Oncorhynchus mykiss) was demonstrated. Conclusions Given that the purified G21-465 conserved some of its most important properties, this method might be suitable for the large-scale production of fish rhabdoviral gpGs for use in diagnosis, fusion and antigenicity studies. PMID:21693048

  20. Generation and characterization of NV gene-knockout recombinant viral hemorrhagic septicemia virus (VHSV) genotype IVa.

    PubMed

    Kim, Min Sun; Kim, Dong Soo; Kim, Ki Hong

    2011-11-03

    A recombinant viral hemorrhagic septicemia virus (rVHSV-deltaNV-EGFP) containing the enhanced green fluorescent protein (EGFP) gene instead of the NV gene was produced using the reverse-genetics method. For use as a positive control, another recombinant virus (rVHSV-wild) was also generated, which had an identical nucleotide sequence to the wild-type VHSV genome except for a few artificially replaced nucleotides. The rVHSVs were rescued using a system controlled by T7 RNA polymerase supplied by a retroviral vector. Generation of rVHSV-deltaNV-EGFP and rVHSV-wild was confirmed by sequencing of RT-PCR products, and rescue of infectious rVHSVs was confirmed by observation of plaque formation. Replication efficiency of rVHSV-wild was distinctly lower than that of wild-type VHSV, suggesting that the artificially replaced nucleotides, especially when immediately preceding the G or NV gene start codons, might affect the replication of the virus. Replication of rVHSV-deltaNV-EGFP was slightly lower than that of rVHSV-wild when epithelioma papulosum cyprini cells were infected with multiplicity of infection (MOI) 1.0, but much lower when cells were infected with MOI 0.00001. These results suggest that the NV gene plays an important role in VHSV replication through interactions with host-cell responses, and the lower replication ability of rVHSV-wild compared to wild-type VHSV might be caused by replaced nucleotides just before the NV gene open reading frame (ORF) rather than the G gene ORF. In olive flounder Paralichthys olivaceus, rVHSV-wild produced slower-progressing mortalities than wild-type VHSV, whereas rVHSV-deltaNV-EGFP pathogenesis was highly attenuated. These results suggest that the NV protein of VHSV may play an important role not only in viral replication but also in viral pathogenesis.

  1. Oral immunization of olive flounder (Paralichthys olivaceus) with recombinant live viral hemorrhagic septicemia virus (VHSV) induces protection against VHSV infection.

    PubMed

    Kim, Min Sun; Kim, Dong Soo; Kim, Ki Hong

    2011-08-01

    A recombinant viral hemorrhagic septicemia virus (rVHSV-ΔNV-EGFP) that has enhanced green fluorescent protein (EGFP) gene instead of NV gene was previously generated using reverse genetics technology. In this study, potential of the rVHSV-ΔNV-EGFP to be used as a live oral vaccine candidate was assessed. The presence of the recombinant virus in internal organs of orally administered olive flounder (Paralichthys olivaceus) was analyzed by semi-quantitative RT-PCR. Although the recombinant VHSV-specific band was detected only when the number of PCR cycle was increased to 35, the band was detected from internal organs, such as kidney, spleen, and liver of fish that were reared at either 15 °C or 20 °C till even 20 days, suggesting that a few orally administered rVHSV-ΔNV-EGFP might be transported to internal organs, and might keep weak replication ability in the organs. VHSV-neutralizing activity was induced by oral immunization of olive flounder with the NV gene knock-out recombinant VHSV not only in skin and intestinal mucus but also in serum, suggesting that mucosal and systemic adaptive immune responses were elicited by oral immunization. In challenge experiment, groups of fish immunized with 10⁴, 10⁵, and 2 × 10⁵ PFU of rVHSV-ΔNV-EGFP/fish showed 25%, 50%, and 70% of relative percent survival (RPS), respectively. The RPSs were elevated to 60%, 75%, and 90% by a boost immunization in fish boost immunized with 10⁴, 10⁵, and 2 × 10⁵ PFU of rVHSV-ΔNV-EGFP, respectively. The cumulative mortality of fish in the control groups was 100%. Conclusionly, the present results demonstrate that the NV gene knock-out recombinant VHSV administered orally to olive flounder can induce dose- and boosting-dependent VHSV-neutralizing antibody in mucus and serum, and can provide a high protection in olive flounder against a virulent VHSV challenge.

  2. The Role of Viral Hemorrhagic Septicemia Virus (VHSV) Matrix (M) Protein in Suppressing Host Transcription

    USDA-ARS?s Scientific Manuscript database

    Viral Hemorrhagic Septicemia virus (VHSV) is a pathogenic fish rhabdovirus found in discrete locales throughout the northern hemisphere. VHSV infection of fish cells leads to upregulation of the host's virus detection response, but the virus quickly suppresses interferon (IFN) production and antivir...

  3. Restricted replication of viral hemorrhagic septicemia virus (VHSV) in a birnavirus-carrier cell culture.

    PubMed

    Parreño, Ricardo; Almagro, Lucía; Belló-Pérez, Melissa; Medina-Gali, Regla M; Estepa, Amparo; Perez, Luis

    2017-04-01

    Viral hemorrhagic septicemia virus (VHSV) and infectious pancreatic necrosis virus (IPNV) are economically important pathogens of the salmonid aquaculture industry. In previous work we demonstrated that a cell line persistently infected with IPNV (EPC(IPNV)) exhibited antiviral activity against superinfection with the heterologous virus VHSV. This work extends our study by analyzing the replication of VHSV in the IPNV-persistently infected cells. At early and late stages of infection VHSV RNA synthesis, as well as VHSV-induced syncytia formation, were examined in EPC(IPNV) cultures. During the course of VHSV infection the accumulation of VHSV RNA is inhibited in EPC(IPNV) cells. Typical VHSV-induced membrane fusion at the late stages of infection is also absent in the IPNV carrier cultures. VHSV binding and fusion to EPC(IPNV) cells did not appear to be impaired, but a potent inhibitory effect on VHSV RNA synthesis is exerted at early times of infection in the IPNV carrier culture. In conclusion, the EPC(IPNV) cells are considered to be a useful system to study viral interference as well to analyze the mechanisms underlying the phenomenon of superinfection exclusion.

  4. Factors controlling the early stages of viral haemorrhagic septicaemia epizootics: Low exposure levels, virus amplification and fish-to-fish transmission

    USGS Publications Warehouse

    Hershberger, P.K.; Gregg, J.L.; Grady, C.A.; Hart, L.M.; Roon, S.R.; Winton, J.R.

    2011-01-01

    Viral haemorrhagic septicaemia virus, Genogroup IVa (VHSV), was highly infectious to Pacific herring, Clupea pallasii (Valenciennes), even at exposure doses occurring below the threshold of sensitivity for a standard viral plaque assay; however, further progression of the disease to a population-level epizootic required viral amplification and effective fish-to-fish transmission. Among groups of herring injected with VHSV, the prevalence of infection was dose-dependent, ranging from 100%, 75% and 38% after exposure to 19, 0.7 and 0.07 plaque-forming units (PFU)/fish, respectively. Among Pacific herring exposed to waterborne VHSV (140PFUmL-1), the prevalence of infection, geometric mean viral tissue titre and cumulative mortality were greater among cohabitated herring than among cohorts that were held in individual aquaria, where fish-to-fish transmission was prevented. Fish-to-fish transmission among cohabitated herring probably occurred via exposure to shed virus which peaked at 680PFUmL-1; shed virus was not detected in the tank water from any isolated individuals. The results provide insights into mechanisms that initiate epizootic cascades in populations of wild herring and have implications for the design of VHSV surveys in wild fish populations. ?? Published 2011. This article is a US Government work and is in the public domain in the USA.

  5. Sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral haemorrhagic septicaemia virus, a fish rhabdovirus.

    PubMed

    Benmansour, A; Basurco, B; Monnier, A F; Vende, P; Winton, J R; de Kinkelin, P

    1997-11-01

    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 the genus Lyssavirus. Compared with each other, the G genes of VHSV displayed a dissimilar overall genetic diversity which correlated with differences in geographical origin. The multiple sequence alignment of the complete G protein, showed that the divergent positions were not uniformly distributed along the sequence. A central region (amino acid position 245-300) accumulated substitutions and appeared to be highly variable. The genetic heterogeneity within a single isolate was high, with an apparent internal mutation frequency of 1.2 x 10(-3) per nucleotide site, attesting the quasispecies nature of the viral population. The phylogeny separated VHSV strains according to the major geographical area of isolation: genotype I for continental Europe, genotype II for the British Isles, and genotype III for North America. Isolates from continental Europe exhibited the highest genetic variability, with sub-groups correlated partially with the serological classification. Neither neutralizing polyclonal sera, nor monoclonal antibodies, were able to discriminate between the genotypes. The overall structure of the phylogenetic tree suggests that VHSV genetic diversity and evolution fit within the model of random change and positive selection operating on quasispecies.

  6. Development and validation of a novel Taqman-based real-time RT-PCR assay suitable for demonstrating freedom from viral haemorrhagic septicaemia virus.

    PubMed

    Jonstrup, S P; Kahns, S; Skall, H F; Boutrup, T S; Olesen, N J

    2013-01-01

    Viral haemorrhagic septicaemia (VHS) is a serious disease in several fish species. VHS is caused by the rhabdovirus viral haemorrhagic septicaemia virus (VHSV). To prevent spreading of the pathogen, it is important to use a fast, robust, sensitive and specific diagnostic tool to identify the infected fish. Traditional diagnosis based on isolation in cell culture followed by identification using, for example, ELISA is sensitive and specific but slow. By switching to RT-PCR for surveillance and diagnosis of VHS the time needed before a correct diagnosis can be given will be considerably shortened and the need for maintaining expensive cell culture facilities reduced. Here we present the validation, according to OIE guidelines, of a sensitive and specific Taqman-based real-time RT-PCR. The assay detects all isolates in a panel of 79 VHSV isolates covering all known genotypes and subtypes, with amplification efficiencies of approximately 100%. The analytical and diagnostic specificity of the real-time RT-PCR is close to 1, and the analytical and diagnostic sensitivity is comparable with traditional cell-based methods. In conclusion, the presented real-time RT-PCR assay has the necessary qualities to be used as a VHSV surveillance tool on par with cell culture assays. © 2012 Blackwell Publishing Ltd.

  7. RNA aptamers inhibit the growth of the fish pathogen viral hemorrhagic septicemia virus (VHSV).

    PubMed

    Punnarak, Porntep; Santos, Mudjekeewis D; Hwang, Seong Don; Kondo, Hidehiro; Hirono, Ikuo; Kikuchi, Yo; Aoki, Takashi

    2012-12-01

    Viral hemorrhagic septicemia virus (VHSV) is a serious disease impacting wild and cultured fish worldwide. Hence, an effective therapeutic method against VHSV infection needs to be developed. Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we aimed at selecting RNA aptamers that can specifically bind to and inhibit the growth of a strain of fish VHSV both in vitro and in vivo. Three VHSV-specific RNA aptamers (F1, F2, and C6) were selected from a pool of artificially and randomly produced oligonucleotides using systematic evolution of ligands by exponential enrichment. The three RNA aptamers showed obvious binding to VHSV in an electrophoretic mobility shift assay but not to other tested viruses. The RNA aptamers were tested for their ability to inhibit VHSV in vitro using hirame natural embryo (HINAE) cells. Cytopathic effect and plaque assays showed that all aptamers inhibited the growth of VHSV in HINAE cells. In vivo tests using RNA aptamers produced by Rhodovulum sulfidophilum showed that extracellular RNA aptamers inhibited VHSV infection in Japanese flounder. These results suggest that the RNA aptamers are a useful tool for protection against VHSV infection in Japanese flounder.

  8. Detection of Viral Hemorrhagic Septicemia virus (VHSV) from the leech Myzobdella lugubris Leidy, 1851.

    PubMed

    Faisal, Mohamed; Schulz, Carolyn A

    2009-09-28

    The leech Myzobdella lugubris is widespread in the Lake Erie Watershed, especially Lake St. Clair. However, its role in pathogen transmission is not fully understood. In this same watershed, several widespread fish mortalities associated with the Viral Hemorrhagic Septicemia virus (VHSV) were recorded. Viral Hemorrhagic Septicemia is an emerging disease in the Great Lakes Basin that is deadly to the fish population, yet little is known about its mode of transmission. To assess the potential role of M. lugubris in VHSV transmission, leeches were collected from Lake St. Clair and Lake Erie and pooled into samples of five. Cell culture and reverse transcriptase polymerase chain reaction (RT-PCR) were used to determine the presence of the virus and its identity. Results showed that 57 of the 91 pooled leech samples were positive by cell culture for VHSV and 66 of the 91 pooled leech samples were positive by RT-PCR for the VHSV. Two representative virus isolates were sequenced for further genetic confirmation and genotype classification. VHSV detected within M. lugubris was homologous to the Great Lakes strain of VHSV genotype IVb. This is the first record of the VHSV being detected from within a leech, specifically M. lugubris, and suggests the potential of M. lugubris being involved in VHSV transmission.

  9. Detection of Viral Hemorrhagic Septicemia virus (VHSV) from the leech Myzobdella lugubris Leidy, 1851

    PubMed Central

    Faisal, Mohamed; Schulz, Carolyn A

    2009-01-01

    The leech Myzobdella lugubris is widespread in the Lake Erie Watershed, especially Lake St. Clair. However, its role in pathogen transmission is not fully understood. In this same watershed, several widespread fish mortalities associated with the Viral Hemorrhagic Septicemia virus (VHSV) were recorded. Viral Hemorrhagic Septicemia is an emerging disease in the Great Lakes Basin that is deadly to the fish population, yet little is known about its mode of transmission. To assess the potential role of M. lugubris in VHSV transmission, leeches were collected from Lake St. Clair and Lake Erie and pooled into samples of five. Cell culture and reverse transcriptase polymerase chain reaction (RT-PCR) were used to determine the presence of the virus and its identity. Results showed that 57 of the 91 pooled leech samples were positive by cell culture for VHSV and 66 of the 91 pooled leech samples were positive by RT-PCR for the VHSV. Two representative virus isolates were sequenced for further genetic confirmation and genotype classification. VHSV detected within M. lugubris was homologous to the Great Lakes strain of VHSV genotype IVb. This is the first record of the VHSV being detected from within a leech, specifically M. lugubris, and suggests the potential of M. lugubris being involved in VHSV transmission. PMID:19785752

  10. Isolation of viral haemorrhagic septicaemia virus from muskellunge, Esox masquinongy (Mitchill), in Lake St Clair, Michigan, USA reveals a new sublineage of the North American genotype

    USGS Publications Warehouse

    Elsayed, E.; Faisal, M.; Thomas, M.; Whelan, G.; Batts, W.; Winton, J.

    2006-01-01

    Viral haemorrhagic septicaemia virus (VHSV) was isolated from muskellunge, Esox masquinongy (Mitchill), caught from the NW portion of Lake St Clair, Michigan, USA in 2003. Affected fish exhibited congestion of internal organs; the inner wall of the swim bladder was thickened and contained numerous budding, fluid-filled vesicles. A virus was isolated using fish cell lines inoculated with a homogenate of kidney and spleen tissues from affected fish. Focal areas of cell rounding and granulation appeared as early as 24 h post-inoculation and expanded rapidly to destroy the entire cell sheet by 96 h. Electron microscopy revealed virions that were 170-180 nm in length by 60-70 nm in width having a bullet-shaped morphology typical of rhabdoviruses. The virus was confirmed as VHSV by reverse transcriptase-polymerase chain reaction. Sequence analysis of the entire nucleoprotein and glycoprotein genes revealed the virus was a member of the North American genotype of VHSV; however, the isolate was sufficiently distinct to be considered a separate sublineage, suggesting its origin may have been from marine species inhabiting the eastern coastal areas of the USA or Canada.

  11. The role of viral hemorrhagic septicemia virus (VHSV) NV gene in TNF-α- and VHSV infection-mediated NF-κB activation.

    PubMed

    Kim, Min Sun; Kim, Ki Hong

    2013-05-01

    The role of viral hemorrhagic septicemia virus (VHSV) NV gene in nuclear factor-κB (NF-κB) activation was investigated. Epithelioma papulosum cyprini (EPC) cells pre-treated with tumor necrosis factor (TNF)-α showed a strong resistance against VHSV infection, but cells treated with TNF-α after VHSV infection showed no resistance, suggesting that immediate early TNF-α-mediated responses inhibit VHSV replication. Activation of NF-κB is a key step in TNF-α-mediated immunomodulatory pathways. In this study, activation of NF-κB by TNF-α exposure was inhibited in EPC cells harboring NV gene expressing vectors, indicating that the NV gene of VHSV can suppress TNF-α-mediated NF-κB activation. Furthermore, the NV gene knock-out recombinant VHSV (rVHSV-ΔNV-EGFP) induced significantly higher NF-κB activity in EPC cells than wild-type VHSV, suggesting that VHSV adopted a strategy to suppress early activation of NF-κB in host cells through and NV gene.

  12. Over-passage of epithelioma papulosum cyprini (EPC) cells increased viral hemorrhagic septicemia virus (VHSV) replication.

    PubMed

    Kim, Min Sun; Choi, Seung Hyuk; Kim, Ki Hong

    2016-11-01

    Vaccines based on inactivated or attenuated viruses can be a way to prevent viral hemorrhagic septicemia virus (VHSV) disease, and the efficiency of viral production is a critical factor that can determine the practical use of developed vaccines in aquaculture farms. To know the effects of epithelioma papulosum cyprini (EPC) cells over-subculture on VHSV replication, the VHSV titer produced from high-passage EPC cells (subcultured more than 200 times in our laboratory) was compared to the titer produced from low-passage EPC cells (subcultured 5-15 times). Furthermore, to know whether immune factors are involved in VHSV titers, differences not only in the expression of Mx1 and ISG15 genes but also in the apoptosis progression by VHSV infection between high- and low-passage EPC cells were analyzed. The VHSV titers from high-passage EPC cells were significantly higher than titers from low-passage EPC cells, suggesting that the changed properties of EPC cells by over-subculture were favorable for VHSV proliferation. The DNA laddering of high-passage EPC cells by VHSV infection took a longer time than that of low-passage EPC cells, suggesting that over-subculture might delay apoptosis in VHSV infected EPC cells, and the delay of apoptosis by over-subculture can be thought as one of the factors that increased VHSV titers in high-passage EPC cells. The increased folds of Mx1 and ISG15 genes in high-passage EPC cells were significantly lower than those in low-passage EPC cells when exposed to either poly (I:C) or VHSV. However, the expression levels of Mx1 and ISG15 genes of high-passage EPC cells that were not stimulated with poly I:C or VHSV were almost equal to or higher than the expression levels of low-passage EPC cells that were exposed to poly (I:C) or VHSV. This result suggests that high-passage EPC cells were already in an excited state in type I interferon responses without any stimulants. The full open reading frame (ORF) sequences of Mx1 gene between high- and

  13. Comparison of treatments to inactivate viral hemorrhagic septicemia virus (VHSV-IVb) in frozen baitfish.

    PubMed

    Phelps, Nicholas B D; Goodwin, Andrew E; Marecaux, Emily; Goyal, Sagar M

    2013-02-28

    Current US state and federal fish health regulations target the spread of viral hemorrhagic septicemia virus-IVb (VHSV-IVb) through movement restrictions of live fish; however, they largely ignore the potential for the virus to be spread through commercial distribution and use of frozen baitfish from VHSV-IVb-positive regions. Some state laws do require treatment of frozen baitfish to inactivate VHSV, and additional methods have been proposed, but few scientific studies have examined the efficacy of these treatments. In this study, bluegills Lepomis macrochirus were challenged with VHSV-IVb and frozen to represent standard industry methods, disinfected by various treatments, and tested for infectious VHSV-IVb using virus isolation. The virus was isolated from 70% of fish subjected to 3 freeze/thaw cycles. All other treatment methods were effective in inactivating the virus, including treatment with isopropyl alcohol, mineral oil, salt and borax, and dehydration. Dehydration followed by rehydration is rapid and effective, and therefore, seems to be the best option for inactivating VHSV-IVb present in frozen baitfish while maintaining their usefulness as bait.

  14. Molecular characterization of the Great Lakes viral hemorrhagic septicemia virus (VHSV) isolate from USA

    PubMed Central

    Ammayappan, Arun; Vakharia, Vikram N

    2009-01-01

    Background Viral hemorrhagic septicemia virus (VHSV) is a highly contagious viral disease of fresh and saltwater fish worldwide. VHSV caused several large scale fish kills in the Great Lakes area and has been found in 28 different host species. The emergence of VHS in the Great Lakes began with the isolation of VHSV from a diseased muskellunge (Esox masquinongy) caught from Lake St. Clair in 2003. VHSV is a member of the genus Novirhabdovirus, within the family Rhabdoviridae. It has a linear single-stranded, negative-sense RNA genome of approximately 11 kbp, with six genes. VHSV replicates in the cytoplasm and produces six monocistronic mRNAs. The gene order of VHSV is 3'-N-P-M-G-NV-L-5'. This study describes molecular characterization of the Great Lakes VHSV strain (MI03GL), and its phylogenetic relationships with selected European and North American isolates. Results The complete genomic sequences of VHSV-MI03GL strain was determined from cloned cDNA of six overlapping fragments, obtained by RT-PCR amplification of genomic RNA. The complete genome sequence of MI03GL comprises 11,184 nucleotides (GenBank GQ385941) with the gene order of 3'-N-P-M-G-NV-L-5'. These genes are separated by conserved gene junctions, with di-nucleotide gene spacers. The first 4 nucleotides at the termini of the VHSV genome are complementary and identical to other novirhadoviruses genomic termini. Sequence homology and phylogenetic analysis show that the Great Lakes virus is closely related to the Japanese strains JF00Ehi1 (96%) and KRRV9822 (95%). Among other novirhabdoviruses, VHSV shares highest sequence homology (62%) with snakehead rhabdovirus. Conclusion Phylogenetic tree obtained by comparing 48 glycoprotein gene sequences of different VHSV strains demonstrate that the Great Lakes VHSV is closely related to the North American and Japanese genotype IVa, but forms a distinct genotype IVb, which is clearly different from the three European genotypes. Molecular characterization of the

  15. Effect of G gene-deleted recombinant viral hemorrhagic septicemia virus (rVHSV-ΔG) on the replication of wild type VHSV in a fish cell line and in olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Min Sun; Choi, Seung Hyuk; Kim, Ki Hong

    2016-07-01

    In an earlier study, we generated a replicon viral hemorrhagic septicemia virus (VHSV) particle that was lacking the G gene in the genome (rVHSV-ΔG), and proved the potential of it as a protective vaccine through the immunization of olive flounder (Paralichthys olivaceus) fingerlings. Safety is the most important preconsideration for the development of recombinant live vaccines, and a major concern of propagation-incompetent viral particles would be the possible harmful effect to hosts through the interaction with wild-type viruses. Thus, in the present study, we analyzed the replication of rVHSV-ΔG in the presence of wild-type VHSV and the effect of rVHSV-ΔG on the replication of wild-type VHSV in Epithelioma papulosum cyprini (EPC) cells and in olive flounder fingerlings. The replication of wild-type VHSV in EPC cells was severely suppressed when the MOI of rVHSV-ΔG was 0.1 or 0.01, on the other hand, the titers of rVHSV-ΔG were not increased and stayed in a relatively constant according to time lapse. Furthermore, the replication of other novirhabdoviruses, IHNV and HIRRV, was also inhibited by co-infection with high titers of rVHSV-ΔG. There were no big differences in mortalities between groups infected with wild-type VHSV plus rVHSV-ΔG and groups infected with wild-type VHSV alone, when the challenged wild-type VHSV was more than 10(2) PFU/fish. However, a group of fish infected with 10 PFU/fish of wild-type VHSV plus rVHSV-ΔG showed significantly lower and slowly progressing cumulative mortality than a group of fish infected with 10 PFU/fish of wild-type VHSV alone. This result suggests that rVHSV-ΔG has an ability to attenuate the disease progression caused by wild-type VHSV when co-infected with relatively low titers of wild-type VHSV. These results indicate that the propagation-incompetent rVHSV-ΔG would not worsen but attenuate the progression of a disease caused by wild-type VHSV infection. Therefore, rVHSV-ΔG-based vaccines can provide a

  16. QCM DNA biosensor for the diagnosis of a fish pathogenic virus VHSV.

    PubMed

    Hong, Sung-Rok; Jeong, Hyun-Do; Hong, Suhee

    2010-08-15

    Viral haemorrhagic septicaemia (VHS) is one of the most serious viral diseases damaging both fresh and marine fish species. VHS caused by VHSV and diagnosis of VHSV has been dependent on the conventional methods, such as cell culture and RT-PCR, which takes a few days or several hours. This study demonstrates a rapid and sensitive QCM biosensor for diagnosis of VHSV infection in fish. The QCM biosensor was developed to detect a main viral RNA encoding G protein in VHSV using the specific DNA probe. To maximize the sensitivity of the biosensor, we prepared three different DNA probes which modified 3' end of DNA by thiol, amine, or biotin and compared three different immobilisation methods on quartz surface coated with gold: immobilisation of thiol labelled probe DNA on naked gold surface, immobilisation of amino labelled probe DNA on gold surface prepared as carboxyl chip using MPA followed by EDC/NHS activation, and immobilisation of biotin labelled probe DNA on gold surface after immobilising avidin on carboxyl chip prior to biotin. As a result, immobilisation method using avidin-biotin interaction was most efficient to immobilise probe DNA and to detect target DNA. The QCM biosensor system using biotinylated probe DNA was stable enough to withstand 32 times of repeated regenerations and the detection limit was 0.0016muM. Diagnosis using the QCM biosensor system was more sensitive and much faster than a conventional RT-PCR analysis in detecting the viral RNA.

  17. Viral hemorrhagic septicemia virus (VHSV IVb) risk factors and association measures derived by expert panel.

    PubMed

    2010-04-01

    Viral hemorrhagic septicemia virus (VHSV) is an OIE-listed pathogen of fish, recently expanding in known host and geographic range in North America. Through a group process designed for subjective probability assessment, an international panel of fish health experts identified and weighted risk factors perceived important to the emergence and spread of the viral genotype, VHSV IVb, within and from the Great Lakes region of the US and Canada. Identified factors included the presence of known VHSV-susceptible species, water temperatures conducive for disease, hydrologic connectivity and proximity to known VHSV-positive areas, untested shipments of live or frozen fish from known positive regions, insufficient regulatory infrastructure for fish health oversight, and uncontrolled exposure to fomites associated with boat and equipment or fish wastes from known VHSV-positive areas. Results provide qualitative insights for use in VHSV surveillance and risk-management planning, and quantitative estimates of contextual risk for use in a Bayesian model combining multiple evidence streams for joint probability assessment of disease freedom status. Consistency checks suggest that the compiled factors positively reflect expert judgment of watershed risk for acquiring VHSV IVb. External validation is recommended as the availability of empirical data permits.

  18. Viral hemorrhagic septicemia virus (VHSV IVb) risk factors and association measures derived by expert panel

    USGS Publications Warehouse

    ,

    2010-01-01

    Viral hemorrhagic septicemia virus (VHSV) is an OIE-listed pathogen of fish, recently expanding in known host and geographic range in North America. Through a group process designed for subjective probability assessment, an international panel of fish health experts identified and weighted risk factors perceived important to the emergence and spread of the viral genotype, VHSV IVb, within and from the Great Lakes region of the US and Canada. Identified factors included the presence of known VHSV-susceptible species, water temperatures conducive for disease, hydrologic connectivity and proximity to known VHSV-positive areas, untested shipments of live or frozen fish from known positive regions, insufficient regulatory infrastructure for fish health oversight, and uncontrolled exposure to fomites associated with boat and equipment or fish wastes from known VHSV-positive areas. Results provide qualitative insights for use in VHSV surveillance and risk-management planning, and quantitative estimates of contextual risk for use in a Bayesian model combining multiple evidence streams for joint probability assessment of disease freedom status. Consistency checks suggest that the compiled factors positively reflect expert judgment of watershed risk for acquiring VHSV IVb. External validation is recommended as the availability of empirical data permits.

  19. Detection of viral hemorrhagic septicemia virus (VHSV) from Diporeia spp. (Pontoporeiidae, Amphipoda) in the Laurentian Great Lakes, USA.

    PubMed

    Faisal, Mohamed; Winters, Andrew D

    2011-01-06

    The mode of viral hemorrhagic septicemia virus (VHSV) transmission in the Great Lakes basin is largely unknown. In order to assess the potential role of macroinvertebrates in VHSV transmission, Diporeia spp., a group of amphipods that are preyed upon by a number of susceptible Great Lakes fishes, were collected from seven locations in four of the Great Lakes and analyzed for the presence of VHSV. It was demonstrated that VHSV is present in some Diporeia spp. samples collected from lakes Ontario, Huron, and Michigan, but not from Lake Superior. Phylogenetic comparison of partial nucleoprotein (N) gene sequences (737 base pairs) of the five isolates to sequences of 13 other VHSV strains showed the clustering of Diporeia spp. isolates with the VHSV genotype IVb. This study reports the first incidence of a fish-pathogenic rhabdovirus being isolated from Diporeia, or any other crustacean and underscores the role macroinvertebrates may play in VHSV ecology.

  20. Detection of Viral Hemorrhagic Septicemia Virus (VHSV) from Diporeia spp. (Pontoporeiidae, Amphipoda) in the Laurentian Great Lakes, USA

    PubMed Central

    2011-01-01

    The mode of viral hemorrhagic septicemia virus (VHSV) transmission in the Great Lakes basin is largely unknown. In order to assess the potential role of macroinvertebrates in VHSV transmission, Diporeia spp., a group of amphipods that are preyed upon by a number of susceptible Great Lakes fishes, were collected from seven locations in four of the Great Lakes and analyzed for the presence of VHSV. It was demonstrated that VHSV is present in some Diporeia spp. samples collected from lakes Ontario, Huron, and Michigan, but not from Lake Superior. Phylogenetic comparison of partial nucleoprotein (N) gene sequences (737 base pairs) of the five isolates to sequences of 13 other VHSV strains showed the clustering of Diporeia spp. isolates with the VHSV genotype IVb. This study reports the first incidence of a fish-pathogenic rhabdovirus being isolated from Diporeia, or any other crustacean and underscores the role macroinvertebrates may play in VHSV ecology. PMID:21210995

  1. In vivo study of viral haemorrhagic septicaemia virus and infectious pancreatic necrosis virus coexistence in Senegalese sole (Solea senegalensis).

    PubMed

    López-Vázquez, C; Alonso, M C; Dopazo, C P; Bandín, I

    2017-09-01

    The effect of IPNV-VHSV coinfection and superinfection on the mortality caused by both viruses in Senegalese sole has been analysed. No effect was observed after coinfection. However, a clear viral interference was recorded between a primary IPNV and a subsequent VHSV infection, which led to a survival increase in the infected sole of 50% points when compared with fish infected only with VHSV. The significantly higher Mx transcriptional values in the fish pre-exposed to IPNV (at least at first days after superinfection) and the increased daily mortality when low Mx transcriptional levels were recorded suggest that Mx may be involved in the protective effect against VHSV infection. However, in fish subjected to VHSV primary/IPNV secondary infection, no interference was observed. © 2016 John Wiley & Sons Ltd.

  2. Comparative susceptibility among three stocks of yellow perch, Perca flavescens (Mitchill), to viral haemorrhagic septicaemia virus strain IVb from the Great Lakes.

    PubMed

    Olson, W; Emmenegger, E; Glenn, J; Winton, J; Goetz, F

    2013-08-01

    The Great Lakes strain of viral haemorrhagic septicaemia virus IVb (VHSV-IVb) is capable of infecting a wide number of naive species and has been associated with large fish kills in the Midwestern United States since its discovery in 2005. The yellow perch, Perca flavescens (Mitchill), a freshwater species commonly found throughout inland waters of the United States and prized for its high value in sport and commercial fisheries, is a species documented in several fish kills affiliated with VHS. In the present study, differences in survival after infection with VHSV IVb were observed among juvenile fish from three yellow perch broodstocks that were originally derived from distinct wild populations, suggesting innate differences in susceptibility due to genetic variance. While all three stocks were susceptible upon waterborne exposure to VHS virus infection, fish derived from the Midwest (Lake Winnebago, WI) showed significantly lower cumulative % survival compared with two perch stocks derived from the East Coast (Perquimans River, NC and Choptank River, MD) of the United States. However, despite differences in apparent susceptibility, clinical signs did not vary between stocks and included moderate-to-severe haemorrhages at the pelvic and pectoral fin bases and exophthalmia. After the 28-day challenge was complete, VHS virus was analysed in subsets of whole fish that had either survived or succumbed to the infection using both plaque assay and quantitative PCR methodologies. A direct correlation was identified between the two methods, suggesting the potential for both methods to be used to detect virus in a research setting.

  3. Risk of introducing viral hemorrhagic septicemia virus (VHSV) to the Chilean South Pacific via sardine imports from Europe.

    PubMed

    Hervé-Claude, Luis Pablo; Carpenter, Tim E; Hedrick, Ronald P

    2008-01-24

    Chile imports from Spain 100s of metric tons of frozen sardine Sardina pilchardus fished in European oceans, which, with several other clupeids, are presumed susceptible to infection with viral hemorrhagic septicemia virus (VHSV). The frozen sardines are directly introduced into the sea as bait to catch southern hake Merluccius australis in the same areas where wild and pen-raised salmonids are present. A simulation model was therefore developed to evaluate the potential risk of infection of wild Chilean southern hake with VHSV from imported bait. The model indicated that VHSV-susceptible fish species present in Chilean waters, like southern hake, are not at immediate risk of infection. However, sensitivity analyses showed that infectious doses at lower concentrations of VHSV combined with higher VHSV-prevalence import scenarios could likely result in VHSV infections of a moderate number of indigenous southern hake (> or =54 fish yr(-1)).

  4. Development and application of quantitative detection method for viral hemorrhagic septicemia virus (VHSV) genogroup IVa.

    PubMed

    Kim, Jong-Oh; Kim, Wi-Sik; Kim, Si-Woo; Han, Hyun-Ja; Kim, Jin Woo; Park, Myoung Ae; Oh, Myung-Joo

    2014-05-23

    Viral hemorrhagic septicemia virus (VHSV) is a problematic pathogen in olive flounder (Paralichthys olivaceus) aquaculture farms in Korea. Thus, it is necessary to develop a rapid and accurate diagnostic method to detect this virus. We developed a quantitative RT-PCR (qRT-PCR) method based on the nucleocapsid (N) gene sequence of Korean VHSV isolate (Genogroup IVa). The slope and R² values of the primer set developed in this study were -0.2928 (96% efficiency) and 0.9979, respectively. Its comparison with viral infectivity calculated by traditional quantifying method (TCID₅₀) showed a similar pattern of kinetic changes in vitro and in vivo. The qRT-PCR method reduced detection time compared to that of TCID₅₀, making it a very useful tool for VHSV diagnosis.

  5. Development and Application of Quantitative Detection Method for Viral Hemorrhagic Septicemia Virus (VHSV) Genogroup IVa

    PubMed Central

    Kim, Jong-Oh; Kim, Wi-Sik; Kim, Si-Woo; Han, Hyun-Ja; Kim, Jin Woo; Park, Myoung Ae; Oh, Myung-Joo

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a problematic pathogen in olive flounder (Paralichthys olivaceus) aquaculture farms in Korea. Thus, it is necessary to develop a rapid and accurate diagnostic method to detect this virus. We developed a quantitative RT-PCR (qRT-PCR) method based on the nucleocapsid (N) gene sequence of Korean VHSV isolate (Genogroup IVa). The slope and R2 values of the primer set developed in this study were −0.2928 (96% efficiency) and 0.9979, respectively. Its comparison with viral infectivity calculated by traditional quantifying method (TCID50) showed a similar pattern of kinetic changes in vitro and in vivo. The qRT-PCR method reduced detection time compared to that of TCID50, making it a very useful tool for VHSV diagnosis. PMID:24859343

  6. Turbot (Scophthalmus maximus) vs. VHSV (Viral Hemorrhagic Septicemia Virus): A Review.

    PubMed

    Pereiro, Patricia; Figueras, Antonio; Novoa, Beatriz

    2016-01-01

    Turbot (Scophthalmus maximus) is a very valuable fish species both in Europe and China. The culture of this flatfish is well-established but several bacteria, viruses, and parasites can produce mortality or morbidity episodes in turbot farms. Viral Hemorrhagic Septicemia Virus (VHSV) is one of the most threatening pathogens affecting turbot, because neither vaccines nor treatments are commercially available. Although the mortality in the turbot farms is relatively low, when this virus is detected all the stock have to be destroyed. The main goals that need to be improved in order to reduce the incidence of this disease is to know what are the strategies or molecules the host use to fight the virus and, in consequence, try to potentiate this response using different ways. Certain molecules can be selected as potential antiviral treatments because of their high protective effect against VHSV. On the other hand, the use of resistance markers for selective breeding is one of the most attractive approaches. This review englobes all the investigation concerning the immune interaction between turbot and VHSV, which until the last years was very scarce, and the knowledge about VHSV-resistance markers in turbot. Nowadays, the availability of abundant transcriptomic information and the recent sequencing of the turbot genome open the door to a more exhaustive and profuse investigation in these areas.

  7. Turbot (Scophthalmus maximus) vs. VHSV (Viral Hemorrhagic Septicemia Virus): A Review

    PubMed Central

    Pereiro, Patricia; Figueras, Antonio; Novoa, Beatriz

    2016-01-01

    Turbot (Scophthalmus maximus) is a very valuable fish species both in Europe and China. The culture of this flatfish is well-established but several bacteria, viruses, and parasites can produce mortality or morbidity episodes in turbot farms. Viral Hemorrhagic Septicemia Virus (VHSV) is one of the most threatening pathogens affecting turbot, because neither vaccines nor treatments are commercially available. Although the mortality in the turbot farms is relatively low, when this virus is detected all the stock have to be destroyed. The main goals that need to be improved in order to reduce the incidence of this disease is to know what are the strategies or molecules the host use to fight the virus and, in consequence, try to potentiate this response using different ways. Certain molecules can be selected as potential antiviral treatments because of their high protective effect against VHSV. On the other hand, the use of resistance markers for selective breeding is one of the most attractive approaches. This review englobes all the investigation concerning the immune interaction between turbot and VHSV, which until the last years was very scarce, and the knowledge about VHSV-resistance markers in turbot. Nowadays, the availability of abundant transcriptomic information and the recent sequencing of the turbot genome open the door to a more exhaustive and profuse investigation in these areas. PMID:27303308

  8. Generation of Recombinant Viral Hemorrhagic Septicemia Virus (rVHSV) Expressing Two Foreign Proteins and Effect of Lengthened Viral Genome on Viral Growth and In Vivo Virulence.

    PubMed

    Kim, Min Sun; Lee, Su Jin; Kim, Dong Soo; Kim, Ki Hong

    2016-04-01

    In this study, a new recombinant VHSV (rVHSV-Arfp-Bgfp) was generated by insertion of a red fluorescent protein (RFP) gene between N and P genes, a green fluorescent protein (GFP) gene between P and M genes of VHSV genome, the expression of each heterologous gene in infected cells, and effects of the lengthened recombinant VHSV's genome on the replication ability and in vivo virulence to olive flounder (Paralichthys olivaceus) fingerlings were compared with previously generated rVHSVs (rVHSV-wild, rVHSV-Arfp, and rVHSV-Brfp). The expression of RFP and GFP in cells infected with rVHSV-Arfp-Bgfp was verified through fluorescent microscopy and FACS analysis. In the viral growth analysis, rVHSV-Arfp and rVHSV-Brfp showed significantly lower viral titers than rVHSV-wild, and the replication of rVHSV-Arfp-Bgfp was significantly decreased compared to that of even rVHSV-Arfp or rVHSV-Brfp. These results suggest that the genome length is a critical factor for the determination of rVHSVs replication efficiency. In the in vivo virulence experiment, the cumulative mortalities of olive flounder fingerlings infected with each rVHSV were inversely proportional to the length of the viral genome, suggesting that decreased viral growth rate due to the lengthened viral genome is accompanied with the decrease of in vivo virulence of rVHSVs. Recombinant viruses expressing multiple foreign antigens can be used for the development of combined vaccines. However, as the present rVHSV-Arfp-Bgfp still possesses an ability to kill hosts (although very weakened), researches on the producing more attenuated viruses or propagation-deficient replicon particles are needed to solve safety-related problems.

  9. Transcriptomic responses in rainbow trout gills upon infection with viral hemorrhagic septicemia virus (VHSV).

    PubMed

    Aquilino, Carolina; Castro, Rosario; Fischer, Uwe; Tafalla, Carolina

    2014-05-01

    It has been previously demonstrated that even though the fin bases constitute the main portal of entry of viral hemorrhagic septicemia virus (VHSV) in rainbow trout (Oncorhynchus mykiss), an important number of chemokine genes are up-regulated in the gills upon bath exposure to the virus. Because chemokines mediate the recruitment of leukocytes through the action of specific chemokine receptors, in the current study, we have studied the transcription of several immune genes in response to a VHSV bath infection in the gills, focusing both on chemokine receptor genes and on genes characteristic of distinct leukocyte populations such as IgM, IgD, IgT, CD4, CD8, perforin and MHC-II. We have studied the response to the virus in naïve fish as well as in fish that had been previously intramuscularly (i.m.) injected with a VHSV DNA vaccine. Additionally, we have sorted both IgM(+) and CD8(+) cells from the gills of naïve and infected animals to study some of these up-regulated genes in specific leukocyte populations. Our results indicate that despite the low replication level, VHSV provokes an up-regulation of IgM, IgT, CD3 and perforin transcription together with the up-regulation of CCR7, CCR9, CXCR3B and CXCR4 mRNA levels. Interestingly, MHC-II mRNA was up-regulated and CCR7 was down-modulated in IgM(+) cells from infected gills, whereas perforin, CCR7 and CXCR4 mRNA levels were higher in sorted CD8(+) cells from infected animals. Surprisingly, when fish had been previously injected with either the empty plasmid or the VHSV DNA vaccine, these up-regulations in immune gene transcription were no longer observed. Our results point to the gills as an important site for innate and acquired viral defense.

  10. Ontogeny of anti-viral hemorrhagic septicemia virus (VHSV) immunity in developing Japanese flounder.

    PubMed

    Matsuyama, Tomomasa; Nakayasu, Chihaya; Fujiwara, Atsushi; Kurita, Jun; Takano, Tomokazu; Ito, Takafumi; Sano, Motohiko

    2012-07-01

    We examined the ability of developing Japanese flounder (Paralichthys olivaceus) to acquire protective immunity after exposure to viral hemorrhagic septicemia virus (VHSV). Juveniles measuring 9.8 cm average body length were not susceptible to infection with VHSV at 20 °C, while the smaller fish were susceptible. Mortality was not observed after secondary infection at 15 °C in the 9.8 cm cohort that had previously been exposed to the virus at 20 °C, while the smaller fish were susceptible to secondary infection. The expression of interferon (IFN)-related genes was shown to be better developed in larger fish upon virus infection and basal expression levels of the virus recognition proteins were higher in larger fish. Virus-specific antibody was detected in the larger fish, but not in smaller fish. These data indicate that the largest juvenile (9.8 cm) acquired immunity against VHSV infection at the first virus challenge, but smaller fish did not. The anti-viral immune system in the Japanese flounder matures when juveniles reach approximately 10 cm.

  11. Rainbow trout surviving infections of viral haemorrhagic septicemia virus (VHSV) show lasting antibodies to recombinant G protein fragments.

    PubMed

    Encinas, P; Gomez-Casado, E; Fregeneda-Grandes; Olesen, N J; Lorenzen, N; Estepa, A; Coll, J M

    2011-03-01

    Rainbow trout antibodies (Abs) binding to recombinant fragments (frgs) derived from the protein G of the viral haemorrhagic septicemia virus (VHSV)-07.71 strain, could be detected by ELISA (frg-ELISA) in sera from trout surviving laboratory-controlled infections. Abs were detected not only by using sera from trout infected with the homologous VHSV isolate but also with the VHSV-DK-201433 heterologous isolate, which had 13 amino acid changes. Sera from healthy trout and/or from trout surviving infectious haematopoietic necrosis virus (IHNV) infection, were used to calculate cut-off absorbances to differentiate negative from positive sera. Specific anti-VHSV Abs could then be detected by using any of the following frgs: frg11 (56-110), frg15 (65-250), frg16 (252-450) or G21-465. While high correlations were found among the ELISA values obtained with the different frgs, no correlations between any frg-ELISA and complement-dependent 50% plaque neutralization test (PNT) titres could be demonstrated. Between 4 and 10 weeks after VHSV infection, more trout sera were detected as positives by using heterologous frg-ELISA rather than homologous PNT. Furthermore, the percentage of positive sera detected by frg11-ELISA increased with time after infection to reach 100%, while those detected by complement-dependent PNT decreased to 29.4%, thus confirming that the lack of neutralizing Abs does not mean the lack of any anti-VHSV Abs in survivor trout sera. Preliminary results with sera from field samples suggest that further refinements of the frg-ELISA could allow detection of anti-VHSV trout Abs in natural outbreaks caused by different heterologous VHSV isolates. The homologous frg-ELISA method could be useful to follow G immunization attempts during vaccine development and/or to best understand the fish Ab response during VHSV infections. The viral frgs approach might also be used with other fish species and/or viruses.

  12. Comparative susceptibility among three stocks of yellow perch, Perca flavescens (Mitchill), to viral haemorrhagic septicaemia virus strain IVb from the Great Lakes

    USGS Publications Warehouse

    Olson, W.; Emmenegger, E.; Glenn, J.; Winton, J.; Goetz, F.

    2013-01-01

    The Great Lakes strain of viral haemorrhagic septicaemia virus IVb (VHSV-IVb) is capable of infecting a wide number of naive species and has been associated with large fish kills in the Midwestern United States since its discovery in 2005. The yellow perch, Perca flavescens (Mitchill), a freshwater species commonly found throughout inland waters of the United States and prized for its high value in sport and commercial fisheries, is a species documented in several fish kills affiliated with VHS. In the present study, differences in survival after infection with VHSV IVb were observed among juvenile fish from three yellow perch broodstocks that were originally derived from distinct wild populations, suggesting innate differences in susceptibility due to genetic variance. While all three stocks were susceptible upon waterborne exposure to VHS virus infection, fish derived from the Midwest (Lake Winnebago, WI) showed significantly lower cumulative % survival compared with two perch stocks derived from the East Coast (Perquimans River, NC and Choptank River, MD) of the United States. However, despite differences in apparent susceptibility, clinical signs did not vary between stocks and included moderate-to-severe haemorrhages at the pelvic and pectoral fin bases and exophthalmia. After the 28-day challenge was complete, VHS virus was analysed in subsets of whole fish that had either survived or succumbed to the infection using both plaque assay and quantitative PCR methodologies. A direct correlation was identified between the two methods, suggesting the potential for both methods to be used to detect virus in a research setting.

  13. Cowpox virus infection associated with a streptococcal septicaemia in a foal.

    PubMed

    Ellenberger, C; Schüppel, K-F; Möhring, M; Reischauer, A; Alex, M; Czerny, C-P; Fercho, A; Schoon, H-A

    2005-01-01

    Cowpox virus infection associated with a streptococcal septicaemia was diagnosed in a weak German Warmblood filly, born 29 days prematurely, and humanely destroyed on the sixth day of life. At necropsy, ulcerative lesions in the alimentary tract, colitis, polyarthritis and nephritis were observed. Transmission electron microscopical examination of specimens from ulcerative lesions revealed typical orthopox virions. Cowpox virus was unequivocally identified by virological and molecular-biological methods.

  14. Use of a two-step ultrafiltration procedure to concentrate viral hemorrhagic septicemia virus (VHSV) in seawater.

    PubMed

    Kim, Soo-Jin; Kim, Jong-Oh; Kim, Wi-Sik; Oh, Myung-Joo

    2015-11-01

    Viral hemorrhagic septicemia virus (VHSV) has been reported to be stable in both fresh as well as seawater, suggesting that VHSV exists in natural aquatic environments and might have an effect on the wild and cultured fish. However, VHSV is below the detectable limits of laboratory tests in natural seawater. In this study, a two-step ultrafiltration (UF) procedure was used to concentration of VHSV in seawater, providing samples that were tested for infectivity by cell culture and the presence of VHSV by quantitative reverse transcriptase PCR (qRT-PCR) methods. Overall, VHSV was approximately concentrated 100-1000 times in 1, 5 and 10 L, seawater volumes respectively: from 2.81×10(6) to 6.53×10(7)/mL and 10(3.3) to 10(3.8)TCID50/mL prior to the UF procedure, to 3.78×10(8), 1.16 × 10(11), and 9.12 × 10(10)/mL after the procedure. This is the first report of concentrating VHSV using an UF method that was specifically designed for seawater samples. In addition, the two-step UF procedure appears to be compatible with viral cell culture and qRT-PCR methods.

  15. Inter-species transmission of viral hemorrhagic septicemia virus (VHSV) from turbot (Scophthalmus maximus) to rainbow trout (Onchorhynchus mykiss).

    PubMed

    Schönherz, Anna A; Lorenzen, Niels; Einer-Jensen, Katja

    2013-04-01

    Successful viral infection is a complex mechanism, involving many host-pathogen interactions that developed during coevolution of host and pathogen, and often result in host-species specificity. Nevertheless, many viruses are able to infect several host species and sporadically cross species barriers. The viral hemorrhagic septicemia virus (VHSV), a rhabdovirus with high economic impact on the aquaculture industry, has developed an exceptionally wide host range across marine and freshwater environments. Transmission of VHSV between host species therefore represents a potential risk for aquaculture, which currently is not addressed in biosecurity managements. The objective of this study was to investigate the inter-species transmission potential of VHSV and evaluate whether infected marine wild fish pose a potential risk on marine cultured rainbow trout. A cohabitation infection trial with turbot as donor and rainbow trout as recipient host species was conducted. Turbot were intraperitoneally injected with either a marine-adapted (MA) or a trout-adapted (TA) VHSV isolate and subsequently grouped with naïve rainbow trout. Both VHSV isolates were able to replicate and cause mortality in turbot, while only the TA isolate was able to cross the species barrier and infect rainbow trout with fatal outcome. The results demonstrate that a marine fish species can function as reservoir and transmitter of TA VHSV isolates.

  16. DNA Vaccination Partially Protects Muskellunge against Viral Hemorrhagic Septicemia Virus (VHSV-IVb).

    PubMed

    Millard, Elena V; Bourke, Ashley M; LaPatra, Scott E; Brenden, Travis O; Fitzgerald, Scott D; Faisal, Mohamed

    2017-03-01

    A DNA vaccine containing the glycoprotein (G) gene of the North American viral hemorrhagic septicemia virus (VHSV) genotype IVb was developed to evaluate the immune response of fish following vaccination and evaluate its efficacy in protecting a susceptible species, the Muskellunge Esox masquinongy, against VHSV-IVb challenge. Seven weeks (539 degree-days) following vaccination with 10 μg of either pVHSivb-G or a control plasmid, Muskellunge were challenged by immersion with 10(5) plaque-forming units (pfu)/mL of VHSV-IVb. Fish vaccinated with pVHSivb-G had a relative percent survival (RPS) of 45%. Vaccinated fish also had significantly lower mean viral titers in tissues (4.2 × 10(2) pfu/g) and viral prevalence (4%) than fish receiving the plasmid control vaccine (3.3 × 10(5) pfu/g; 82%). Neutralizing antibodies were detected 28 d (308 degree-days) postchallenge (11 weeks postvaccination) in 100% of Muskellunge vaccinated with pVHSivb-G compared with only 12% of plasmid-control-vaccinated Muskellunge, suggesting robust induction of a secondary, adaptive immune response. In addition, pVHSivb-G-vaccinated Rainbow Trout Oncorhynchus mykiss challenged 7 d (100 degree-days) postvaccination with the heterologous novirhabdovirus, infectious hematopoietic necrosis virus (IHNV), experienced an RPS of 61%, compared to control fish, suggesting induction of an early and transient nonspecific antiviral immune response. This study provides an important starting point for VHSV-IVb vaccine development and useful information about the antiviral immune response elicited by DNA vaccination in a nondomesticated fish species. Received May 1, 2016; accepted September 1, 2016.

  17. Effects of NV gene knock-out recombinant viral hemorrhagic septicemia virus (VHSV) on Mx gene expression in Epithelioma papulosum cyprini (EPC) cells and olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Min Sun; Kim, Ki Hong

    2012-03-01

    To determine whether the NV gene of viral hemorrhagic septicemia virus (VHSV) is related to the type I interferon response of hosts, expression of Mx gene in Epithelioma papulosum cyprini (EPC) cells and in olive flounder (Paralichthys olivaceus) in response to infection with either wild-type VHSV or recombinant VHSVs (rVHSV-ΔNV-EGFP and rVHSV-wild) was investigated. A reporter vector was constructed for measuring Mx gene expression using olive flounder Mx promoter, in which the reporter Metridia luciferase was designed to be excreted to culture medium to facilitate measurement. The highest increase of luciferase activity was detected from supernatant of cells infected with rVHSV-ΔNV-EGFP. In contrast cells infected with wild-type VHSV showed a slight increase of the luciferase activity. Interestingly, cells infected with rVHSV-wild that has artificially changed nucleotides just before and after the NV gene ORF, also showed highly increased luciferase activity, but the increased amplitude was lower than that by rVHSV-ΔNV-EGFP. These results strongly suggest that the NV protein of VHSV plays an important role in suppressing interferon response in host cells, which provides a condition for the viruses to efficiently proliferate in host cells. In an in vivo experiment, the Mx gene expression in olive flounder challenged with the rVHSV-ΔNV-EGFP was clearly higher than fish challenged with rVHSV-wild or wild-type VHSV, suggesting that lacking of the NV gene in the genome of rVHSV-ΔNV-EGFP brought to strong interferon response that subsequently inhibit viral replication in fish.

  18. Zebrafish (Danio rerio) as a model for the study of vaccination against viral haemorrhagic septicemia virus (VHSV).

    PubMed

    Novoa, Beatriz; Romero, Alejandro; Mulero, Victoriano; Rodríguez, Iván; Fernández, Ignacio; Figueras, Antonio

    2006-07-26

    The rhabdovirus viral haemorrhagic septicemia virus (VHSV) is the etiological agent of one of the most important salmonid viral diseases. In the present work, the ability of VHSV to infect and replicate in zebrafish at low temperature (15 degrees C) was demonstrated. Zebrafish was also used to determine the effectiveness of the recombinant virus rIHNV-Gvhsv GFP as a live attenuated vaccine against the virulent VHSV strain. Fish intraperitoneally injected with 3 x 10(6) to 3 x 10(5)TCID50/ml of the wild type VHSV showed a 100% of cumulative mortality, meanwhile only 57% of mortality was obtained in bath infections. Infected fish showed external clinical signs and histological observations revealed the appearance of small haemorrhages in the muscle, kidney, liver and dermis. Neither mortalities nor clinical signs were recorded in fish infected with a live attenuated recombinant virus. By RT-PCR technique, VHSV was detected in all the organs as early as 24h, but the recombinant virus was not detected in all the sampled days. VHSV was able to replicate "in vitro" in head kidney cells but the replication capacity of the attenuated viral strain was limited. The recombinant virus rIHNV-Gvhsv GFP was able to protect against VHSV with a survival rate ranging from 20% to 60% depending of the vaccine dose. The increase of TLR3, IFNalphabeta, Mx, IFNgamma and TNFalpha expression at 72h post-infection in the kidney of VHSV-infected fish contrasted with the results obtained with the avirulent virus, which did not induce an increment of this expression in infected fish. Zebrafish is a suitable animal model to study VHSV infection and immune (innate and adaptive) responses and, more importantly, we demonstrate for the first time the usefulness of the zebrafish as a vaccination model to viral diseases. In addition, the high protection obtained with the live attenuated virus demonstrates that the zebrafish is able to mount an efficient antiviral immune response at 15 degrees C.

  19. Molecular Evolution and Phylogeography of Co-circulating IHNV and VHSV in Italy.

    PubMed

    Abbadi, Miriam; Fusaro, Alice; Ceolin, Chiara; Casarotto, Claudia; Quartesan, Rosita; Dalla Pozza, Manuela; Cattoli, Giovanni; Toffan, Anna; Holmes, Edward C; Panzarin, Valentina

    2016-01-01

    Infectious haematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV) are the most important viral pathogens impacting rainbow trout farming. These viruses are persistent in Italy, where they are responsible for severe disease outbreaks (epizootics) that affect the profitability of the trout industry. Despite the importance of IHNV and VHSV, little is known about their evolution at a local scale, although this is likely to be important for virus eradication and control. To address this issue we performed a detailed molecular evolutionary and epidemiological analysis of IHNV and VHSV in trout farms from northern Italy. Full-length glycoprotein gene sequences of a selection of VHSV (n = 108) and IHNV (n = 89) strains were obtained. This revealed that Italian VHSV strains belong to sublineages Ia1 and Ia2 of genotype Ia and are distributed into 7 genetic clusters. In contrast, all Italian IHNV isolates fell within genogroup E, for which only a single genetic cluster was identified. More striking was that IHNV has evolved more rapidly than VHSV (mean rates of 11 and 7.3 × 10(-4) nucleotide substitutions per site, per year, respectively), indicating that these viruses exhibit fundamentally different evolutionary dynamics. The time to the most recent common ancestor of both IHNV and VHSV was consistent with the first reports of these pathogens in Italy. By combining sequence data with epidemiological information it was possible to identify different patterns of virus spread among trout farms, in which adjacent facilities can be infected by either genetically similar or different viruses, and farms located in different water catchments can be infected by identical strains. Overall, these findings highlight the importance of combining molecular and epidemiological information to identify the determinants of IHN and VHS spread, and to provide data that is central to future surveillance strategies and possibly control.

  20. Molecular Evolution and Phylogeography of Co-circulating IHNV and VHSV in Italy

    PubMed Central

    Abbadi, Miriam; Fusaro, Alice; Ceolin, Chiara; Casarotto, Claudia; Quartesan, Rosita; Dalla Pozza, Manuela; Cattoli, Giovanni; Toffan, Anna; Holmes, Edward C.; Panzarin, Valentina

    2016-01-01

    Infectious haematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV) are the most important viral pathogens impacting rainbow trout farming. These viruses are persistent in Italy, where they are responsible for severe disease outbreaks (epizootics) that affect the profitability of the trout industry. Despite the importance of IHNV and VHSV, little is known about their evolution at a local scale, although this is likely to be important for virus eradication and control. To address this issue we performed a detailed molecular evolutionary and epidemiological analysis of IHNV and VHSV in trout farms from northern Italy. Full-length glycoprotein gene sequences of a selection of VHSV (n = 108) and IHNV (n = 89) strains were obtained. This revealed that Italian VHSV strains belong to sublineages Ia1 and Ia2 of genotype Ia and are distributed into 7 genetic clusters. In contrast, all Italian IHNV isolates fell within genogroup E, for which only a single genetic cluster was identified. More striking was that IHNV has evolved more rapidly than VHSV (mean rates of 11 and 7.3 × 10−4 nucleotide substitutions per site, per year, respectively), indicating that these viruses exhibit fundamentally different evolutionary dynamics. The time to the most recent common ancestor of both IHNV and VHSV was consistent with the first reports of these pathogens in Italy. By combining sequence data with epidemiological information it was possible to identify different patterns of virus spread among trout farms, in which adjacent facilities can be infected by either genetically similar or different viruses, and farms located in different water catchments can be infected by identical strains. Overall, these findings highlight the importance of combining molecular and epidemiological information to identify the determinants of IHN and VHS spread, and to provide data that is central to future surveillance strategies and possibly control. PMID:27602026

  1. Generation of a recombinant viral hemorrhagic septicemia virus (VHSV) expressing olive flounder (Paralichthys olivaceus) interferon-γ and its effects on type I interferon response and virulence.

    PubMed

    Kwak, Jun Soung; Kim, Min Sun; Kim, Ki Hong

    2017-09-01

    Rhabdoviruses including viral hemorrhagic septicemia virus (VHSV) are highly susceptible to type I interferon (IFN) responses, and IFN-γ that is belonging to the type II IFN has been known to enhance type I IFN responses in mammals. In this study, we generated a recombinant VHSV that can express olive flounder IFN-γ (rVHSV-A-IFNγ) using reverse genetics technology, and analyzed the effect of rVHSV-A-IFNγ infection on type I IFN response in Epithelioma papulosum cyprini (EPC) cells. Furthermore, the virulence of rVHSV-A-IFNγ was evaluated by infection to olive flounder (Paralichthys olivaceus). Using a recombinant VHSV full genome vector in which the olive flounder IFN-γ ORF was inserted between N and P genes, rVHSV-A-IFNγ was successfully rescued, and the recombinant virus was grown well in EPC cells. On the other hand, the growth of rVHSV-A-IFNγ rescued from EPC cells was severely retarded when infected into hirame natural embryo (HINAE) cells that were originated from olive flounder. These results indicate that the EPC cell's IFN-γ receptor could not bind to olive flounder IFN-γ, but the species-specific binding of IFN-γ in HINAE cells induced antiviral responses. The expression of Mx1 gene in EPC cells infected with rVHSV-A-IFNγ was not greatly different from cells infected with rVHSV-Arfp (a recombinant VHSV harboring red fluorescent protein gene between N and P genes of the genome), however, in HINAE cells, rVHSV-A-IFNγ infection induced distinctively higher Mx1 gene expression compared to other recombinant viruses. These results suggest that olive flounder IFN-γ produced from rVHSV-A-IFNγ effectively enhanced type I IFN response in HINAE cells. In the present study, the lowest mortality of olive flounder fingerlings was recorded in the group of fish challenged with rVHSV-A-IFNγ, suggesting that the recombinant VHSV was attenuated by production of IFN-γ by itself. However, although rVHSV-A-IFNγ induced significantly lower mortality, the

  2. European freshwater VHSV genotype Ia isolates divide into two distinct subpopulations.

    PubMed

    Kahns, S; Skall, H F; Kaas, R S; Korsholm, H; Bang Jensen, B; Jonstrup, S P; Dodge, M J; Einer-Jensen, K; Stone, D; Olesen, N J

    2012-05-15

    Viral haemorrhagic septicaemia (VHS), caused by the novirhabdovirus VHSV, often leads to significant economic losses to European rainbow trout production. The virus isolates are divided into 4 distinct genotypes with additional subgroups including sublineage Ia, isolates of which are the main source of outbreaks in European rainbow trout farming. A significant portion of Danish rainbow trout farms have been considered endemically infected with VHSV since the first disease outbreak was observed in the 1950s. However, following a series of sanitary programs starting in 1965, VHSV has not been detected in Denmark since January 2009. Full-length G-genes of all Danish VHSV isolates that were submitted for diagnostic analyses in the period 2004-2009 were sequenced and analysed. All 58 Danish isolates from rainbow trout grouped with sublineage Ia isolates. Furthermore, VHSV isolates from infected Danish freshwater catchments appear to have evolved into a distinct clade within sublineage Ia, herein designated clade Ia-1, whereas trout isolates originating from other continental European countries cluster in another distinct clade, designated clade Ia-2. In addition, phylogenetic analyses indicate that VHSV Ia-1 strains have caused a few outbreaks in Germany and the UK. It is likely that viruses have been transmitted from infected site(s) out of the Danish environment, although a direct transmission pathway has not been identified. Furthermore, VHSV Ia-2 isolates seem to have been transmitted to Denmark at least once. Interestingly, one viral isolate possibly persisted in a Danish watershed for nearly 4 yr without detection whereas other subclades of VHSV isolates appear to have been eliminated, probably because of implemented eradication procedures.

  3. Changes in microRNAs expression profile of olive flounder (Paralichthys olivaceus) in response to viral hemorrhagic septicemia virus (VHSV) infection.

    PubMed

    Najib, Abdellaoui; Kim, Min Sun; Choi, Seung Hyuk; Kang, Yue Jai; Kim, Ki Hong

    2016-04-01

    To know the effect of viral hemorrhagic septicemia virus (VHSV) infection on the cellular microRNA expression profile in olive flounder (Paralichthys olivaceus), fish were infected with VHSV, and cellular microRNAs expression was analyzed at 0 (control), 6, 12, 24, 48 and 72 h post-infection (h.p.i.) by the high-throughput sequencing. A total of 372 mature miRNAs were identified, and, among them, 63 miRNAs were differentially expressed during VHSV infection. The differentially expressed microRNAs number was greatly increased from 24 h.p.i. compared to the number at 6 and 12 h.p.i., suggesting that the alteration of microRNAs expression by VHSV infection may be related to the progression of VHSV disease. The target prediction analysis, the GO enrichment analysis, and the KEGG pathway analysis of the predicted target genes showed that various biological pathways could be affected by VHSV infection through the down-regulation or up-regulation of host miRNAs. The present results provide a basic information on the microRNAs related to VHSV infection in olive flounder. Considering broad effects of microRNAs on various biological pathways, data in this study can be used to interpret the mechanism of VHSV pathogenesis, which, vice versa, can be used to develop control measures against VHSV.

  4. Variable domain antibodies specific for viral hemorrhagic septicemia virus (VHSV) selected from a randomized IgNAR phage display library.

    PubMed

    Ohtani, Maki; Hikima, Jun-ichi; Jung, Tae-Sung; Kondo, Hidehiro; Hirono, Ikuo; Takeyama, Haruko; Aoki, Takashi

    2013-02-01

    Phage display libraries are used to screen for nucleotide sequences that encode immunoglobulin variable (V) regions that are specific for a target antigen. We previously constructed an immunoglobulin new antigen receptor (IgNAR) phage display library. Here we used this library to obtain an IgNAR V region that is specific for viral hemorrhagic septicemia virus (VHSV). A phage clone (clone 653) was found to be specific for VHSV by the biopanning method. The V region of clone 653 was used to construct a 6 × His tagged recombinant IgNAR-653 V protein (rIgNAR-653) using the Escherichia coli pET system. The rIgNAR-653 protein bound specifically to VHSV, confirming its activity.

  5. Early immune responses in rainbow trout liver upon viral hemorrhagic septicemia virus (VHSV) infection.

    PubMed

    Castro, Rosario; Abós, Beatriz; Pignatelli, Jaime; von Gersdorff Jørgensen, Louise; González Granja, Aitor; Buchmann, Kurt; Tafalla, Carolina

    2014-01-01

    Among the essential metabolic functions of the liver, in mammals, a role as mediator of systemic and local innate immunity has also been reported. Although the presence of an important leukocyte population in mammalian liver is well documented, the characterization of leukocyte populations in the teleost liver has been only scarcely addressed. In the current work, we have confirmed the presence of IgM+, IgD+, IgT+, CD8α+, CD3+ cells, and cells expressing major histocompatibility complex (MHC-II) in rainbow trout (Oncorhynchus mykiss) liver by flow cytometry and/or immunohistochemistry analysis. Additionally, the effect of viral hemorrhagic septicemia virus (VHSV) on the liver immune response was assessed. First, we studied the effect of viral intraperitoneal injection on the transcription of a wide selection of immune genes at days 1, 2 and 5 post-infection. These included a group of leukocyte markers genes, pattern recognition receptors (PRRs), chemokines, chemokine receptor genes, and other genes involved in the early immune response and in acute phase reaction. Our results indicate that T lymphocytes play a key role in the initial response to VHSV in the liver, since CD3, CD8, CD4, perforin, Mx and interferon (IFN) transcription levels were up-regulated in response to VHSV. Consequently, flow cytometry analysis of CD8α+ cells in liver and spleen at day 5 post-infection revealed a decrease in the number of CD8α+ cells in the spleen and an increased population in the liver. No differences were found however in the percentages of B lymphocyte (IgM+ or IgD+) populations. In addition, a strong up-regulation in the transcription levels of several PRRs and chemokines was observed from the second day of infection, indicating an important role of these factors in the response of the liver to viral infections.

  6. Early Immune Responses in Rainbow Trout Liver upon Viral Hemorrhagic Septicemia Virus (VHSV) Infection

    PubMed Central

    Castro, Rosario; Abós, Beatriz; Pignatelli, Jaime; von Gersdorff Jørgensen, Louise; González Granja, Aitor; Buchmann, Kurt; Tafalla, Carolina

    2014-01-01

    Among the essential metabolic functions of the liver, in mammals, a role as mediator of systemic and local innate immunity has also been reported. Although the presence of an important leukocyte population in mammalian liver is well documented, the characterization of leukocyte populations in the teleost liver has been only scarcely addressed. In the current work, we have confirmed the presence of IgM+, IgD+, IgT+, CD8α+, CD3+ cells, and cells expressing major histocompatibility complex (MHC-II) in rainbow trout (Oncorhynchus mykiss) liver by flow cytometry and/or immunohistochemistry analysis. Additionally, the effect of viral hemorrhagic septicemia virus (VHSV) on the liver immune response was assessed. First, we studied the effect of viral intraperitoneal injection on the transcription of a wide selection of immune genes at days 1, 2 and 5 post-infection. These included a group of leukocyte markers genes, pattern recognition receptors (PRRs), chemokines, chemokine receptor genes, and other genes involved in the early immune response and in acute phase reaction. Our results indicate that T lymphocytes play a key role in the initial response to VHSV in the liver, since CD3, CD8, CD4, perforin, Mx and interferon (IFN) transcription levels were up-regulated in response to VHSV. Consequently, flow cytometry analysis of CD8α+ cells in liver and spleen at day 5 post-infection revealed a decrease in the number of CD8α+ cells in the spleen and an increased population in the liver. No differences were found however in the percentages of B lymphocyte (IgM+ or IgD+) populations. In addition, a strong up-regulation in the transcription levels of several PRRs and chemokines was observed from the second day of infection, indicating an important role of these factors in the response of the liver to viral infections. PMID:25338079

  7. Effects of a broad-spectrum caspase inhibitor, Z-VAD(OMe)-FMK, on viral hemorrhagic septicemia virus (VHSV) infection-mediated apoptosis and viral replication.

    PubMed

    Kim, Min Sun; Lee, Ji Ae; Kim, Ki Hong

    2016-04-01

    In the development of inactivated or attenuated viral vaccines for cultured fish, viral titers harvested from the cultured cells would be the most important factor for the determination of vaccine's cost effectiveness. In this study, we hypothesized that the lengthening of cell survival time by the inhibition of apoptosis can lead to an increase of the final titer of viral hemorrhagic septicemia virus (VHSV). To test the hypothesis, we investigated the effects of a broad-spectrum caspase inhibitor, Z-VAD(OMe)-FMK, on VHSV infection-mediated apoptosis in Epithelioma papulosum cyprini (EPC) cells and on the VHSV titers. VHSV infection induced the DNA laddering in EPC cells, and the progression of DNA fragmentation was in proportion to the CPE extension. The progression of DNA fragmentation in EPC cells infected with VHSV was clearly inhibited by exposure to Z-VAD(OMe)-FMK, and the inhibition was intensified according to the increase of the inhibitor concentration. These results confirmed the previous reports that the death of host cells by VHSV infection is through apoptosis. Cells infected with a recombinant VHSV, rVHSV-ΔNV-eGFP, that was generated from our previous study by replacement of the NV gene ORF with the enhanced green fluorescent protein (eGFP) gene ORF, showed earlier and more distinct DNA fragmentations compared to the cells infected with wild-type VHSV, suggesting the inhibitory role of the NV protein in VHSV-mediated apoptosis that was previously reported. The final viral titers in the supernatant isolated from Z-VAD(OMe)-FMK treated cells after showing an extensive CPE were significantly higher than the viral titers from cells infected with virus alone, indicating that the delay of apoptosis by Z-VAD(OMe)-FMK extended the survival time of EPC cells, which lengthen the time for VHSV replication in the cells. In conclusion, Z-VAD(OMe)-FMK-mediated inhibition of apoptosis significantly increased the final titers of both wild-type VHSV and rVHSV

  8. Real-time RT-PCR for detection, identification and absolute quantification of viral haemorrhagic septicaemia virus using different types of standards.

    PubMed

    Lopez-Vazquez, C; Bandín, I; Dopazo, C P

    2015-05-21

    In the present study, 2 systems of real-time RT-PCR-one based on SYBR Green and the other on TaqMan-were designed to detect strains from any genotype of viral haemorrhagic septicaemia virus (VHSV), with high sensitivity and repeatability/reproducibility. In addition, the method was optimized for quantitative purposes (qRT-PCR), and standard curves with different types of reference templates were constructed and compared. Specificity was tested against 26 isolates from 4 genotypes. The sensitivity of the procedures was first tested against cell culture isolation, obtaining a limit of detection (LD) of 100 TCID50 ml-1 (100-fold below the LD using cell culture), at a threshold cycle value (Ct) of 36. Sensitivity was also evaluated using RNA from crude (LD = 1 fg; 160 genome copies) and purified virus (100 ag; 16 copies), plasmid DNA (2 copies) and RNA transcript (15 copies). No differences between both chemistries were observed in sensitivity and dynamic range. To evaluate repeatability and reproducibility, all experiments were performed in triplicate and on 3 different days, by workers with different levels of experience, obtaining Ct values with coefficients of variation always <5. This fact, together with the high efficiency and R2 values of the standard curves, encouraged us to analyse the reliability of the method for viral quantification. The results not only demonstrated that the procedure can be used for detection, identification and quantification of this virus, but also demonstrated a clear correlation between the regression lines obtained with different standards, which will help scientists to compare sensitivity results between different studies.

  9. Effect of chronic exposure to pendimethalin on the susceptibility of rainbow trout, Oncorhynchus mykiss L., to viral hemorrhagic septicemia virus (VHSV).

    PubMed

    Danion, Morgane; Le Floch, Stéphane; Castric, Jeanne; Lamour, François; Cabon, Joëlle; Quentel, Claire

    2012-05-01

    In this study, the in vivo effects of chronic pollution by the active substance (AS) pendimethalin, a dinitroaniline herbicide, on the susceptibility of rainbow trout, Oncorhynchus mykiss L., to an experimental challenge with viral hemorrhagic septicemia virus (VHSV) were assessed. After four weeks of exposure to fresh water (C group) or 500 ng L(-1) of AS (P500 group), the fish were challenged by immersion in water containing 10(4) TCID(50) mL(-1) of VHSV. While exposure to pendimethalin was maintained throughout the experiment, mortalities were recorded during the 40 days post-infection (dpi) and organs were collected from dead fish for virological examination. At the end of the experiment, anti-VHSV antibodies and the classical pathway of complement activity were assessed in trout plasma. Exposure to pendimethalin significantly affected the distribution of cumulative mortality accelerating death in fish infected by VHSV. Pendimethalin appeared to decrease the Mean Time to Death (MTD) after virus treatment from 14.9 days (C-VHSV) to 10.2 days (P500-VHSV). Nevertheless, by the end of the experiment, differences in cumulative mortality were no longer observed between the two groups, which had reached the same stage (50 percent). Furthermore, a higher concentration of the virus was recovered from the pools of organs from the P500-VHSV group than the C-VHSV group. Moreover, at 40 dpi, although no significant difference was observed in the immune response between the two groups, more fish in the P500-VHSV group had set up an immune response by secreting antibodies than in the control viral group (C-VSHV).

  10. Generation of G gene-deleted viral hemorrhagic septicemia virus (VHSV) and evaluation of its vaccine potential in olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Min Sun; Park, Ji Sun; Kim, Ki Hong

    2015-08-01

    Generation of recombinant viruses lacking an essential gene for the production of infective viral particles would be a way to produce safety-enhanced live viral vaccines. The rhabdoviral envelope-spiked glycoprotein (G) plays critical roles in the attachment of viruses on the cell surface receptor and in the transfer of endocytosed viruses to the cytoplasm by fusion to the endosomal membrane. In the present study, we produced a G gene-lacking recombinant viral hemorrhagic septicemia virus (rVHSV-ΔG) that has no ability to form plaques in the cells without a trans-supply of the G protein, which suggests that rVHSV-ΔG is a single cycle virus and progenies of rVHSV-ΔG are not infectious. One of the major advantages of attenuated vaccines is the maintenance of replication ability in the host, which enforces the adaptive immune responses. However, in spite of lacking an ability to produce infective viral particles, immunization with the present rVHSV-ΔG induced significantly higher serum neutralization titers and survival rates against virulent VHSV challenge in olive flounder (Paralichthys olivaceus) fingerlings, indicating that the present rVHSV-ΔG has a high potential as a prophylactic vaccine.

  11. Interchange of L polymerase protein between two strains of viral hemorrhagic septicemia virus (VHSV) genotype IV alters temperature sensitivities in vitro.

    PubMed

    Kim, Sung-Hyun; Yusuff, Shamila; Vakharia, Vikram N; Evensen, Øystein

    2015-01-02

    Viral hemorrhagic septicemia virus (VHSV) has four genotypes (I-IV) and sub-lineages within genotype I and IV. Using a reverse genetics approach, we explored the importance of the L gene for growth characteristics at different temperatures following interchange of the L gene within genotype IV (IVa and IVb) strains. VHSV strains harboring heterologous L gene were recovered and we show that the L gene determines growth characteristics at different temperatures in permissive cell lines.

  12. Transcription of immune genes upon challenge with viral hemorrhagic septicemia virus (VHSV) in DNA vaccinated rainbow trout (Oncorhynchus mykiss).

    PubMed

    Cuesta, A; Tafalla, C

    2009-01-07

    Even though DNA vaccination has proven as one of the most effective methods in controlling fish rhabdoviruses, the immune mechanisms responsible for protection are still unknown. Many studies have focused on studying which cytokines and immune genes are triggered in response to the vaccine at different times post-vaccination. However, to elucidate the mechanism(s) responsible for protection, to our understanding it is also of great relevance to study the immune response to the virus in fish that have been previously vaccinated and compare it to the effects that the virus might have on non-vaccinated fish. This type of study has never been performed to date in fish. Thus, in the current work, we vaccinated rainbow trout (Oncorhynchus mykiss) with a DNA vaccine against viral hemorrhagic septicemia virus (VHSV), and 30 days post-vaccination we challenged the fish with a virulent VHSV. It was then, that we studied the immune response to the virus at very early times post-infection in fish, in order to compare the effects of VHSV on vaccinated or non-vaccinated trout. We studied the levels of expression of interleukin 1beta (IL-1beta), major histocompatibility complex (MHC) class Ialpha and IIalpha genes, immunoglobulin M (IgM), CD8alpha, type I interferon (IFN), Mx, IFN-gamma and natural killer enhancing factor (NKEF) in head kidney, spleen and blood. When we compared the effect that VHSV had on vaccinated fish to the effect that the virus produced in fish vaccinated with the empty plasmid, the genes that were significantly up-regulated were IL-1beta and MHC IIalpha in the spleen at day 1 post-infection, MHC Ialpha in all organs at day 1 post-infection, and IFN and Mx in the spleen and blood at days 1 and 3 post-infection, respectively. Genes that correlate with an increased specific immune response were not significantly increased in response to VHSV in these vaccinated animals. The results suggest that DNA vaccination induces a memory state in fish that, on the

  13. Ultra-deep sequencing of VHSV isolates contributes to understanding the role of viral quasispecies.

    PubMed

    Schönherz, Anna A; Lorenzen, Niels; Guldbrandtsen, Bernt; Buitenhuis, Bart; Einer-Jensen, Katja

    2016-01-08

    The high mutation rate of RNA viruses enables the generation of a genetically diverse viral population, termed a quasispecies, within a single infected host. This high in-host genetic diversity enables an RNA virus to adapt to a diverse array of selective pressures such as host immune response and switching between host species. The negative-sense, single-stranded RNA virus, viral haemorrhagic septicaemia virus (VHSV), was originally considered an epidemic virus of cultured rainbow trout in Europe, but was later proved to be endemic among a range of marine fish species in the Northern hemisphere. To better understand the nature of a virus quasispecies related to the evolutionary potential of VHSV, a deep-sequencing protocol specific to VHSV was established and applied to 4 VHSV isolates, 2 originating from rainbow trout and 2 from Atlantic herring. Each isolate was subjected to Illumina paired end shotgun sequencing after PCR amplification and the 11.1 kb genome was successfully sequenced with an average coverage of 0.5-1.9 × 10(6) sequenced copies. Differences in single nucleotide polymorphism (SNP) frequency were detected both within and between isolates, possibly related to their stage of adaptation to host species and host immune reactions. The N, M, P and Nv genes appeared nearly fixed, while genetic variation in the G and L genes demonstrated presence of diverse genetic populations particularly in two isolates. The results demonstrate that deep sequencing and analysis methodologies can be useful for future in vivo host adaption studies of VHSV.

  14. Isolation and identification of viral hemorrhagic septicemia virus (VHSV) from farmed rainbow trout (Oncorhynchus mykiss) in Iran.

    PubMed

    Ahmadivand, Sohrab; Soltani, Mehdi; Mardani, Karim; Shokrpoor, Sara; Rahmati-Holasoo, Hooman; Mokhtari, Abbas; Hasanzadeh, Reza

    2016-04-01

    Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus that causes one of the most important fish diseases in rainbow trout (Oncorhynchus mykiss) production industry. During the present study from October 2014 to July 2015, the virus causing viral hemorrhagic septicemia (VHS) was isolated and identified in rainbow trout farms from five of sixteen farms experiencing mass mortalities in six provinces of Iran with major trout production. Cumulative mortalities at VHSV-positive farms ranged from 30 to 70%. Clinical signs of disease included exophthalmia, petechial hemorrhages in the mandible and around the eyes, a swollen abdomen and darkening of the integument, widespread petechiae of the musculature and pyloric regions, severe congestion of the kidney, and pale enlarged livers. In addition, histopathologic examinations of tissues showed severe lesions in muscle, kidney and liver, which were compatible with those already described for VHS. Furthermore, homogenates tissues of diseased fish induced cytopathic effects (CPE) in CHSE-214 cells, and confirmatory diagnosis of VHS was made by RT-PCR reactions. To our knowledge, this is the first report of isolation and identification of VHSV from farmed trout in Iran, which may have originated from Europe.

  15. An outbreak of VHSV (viral hemorrhagic septicemia virus) infection in farmed Japanese flounder Paralichthys olivaceus in Japan.

    PubMed

    Isshik, T; Nishizawa, T; Kobayashi, T; Nagano, T; Miyazaki, T

    2001-11-08

    A rhabdoviral disease occurred in farmed populations of market sized Japanese flounder (hirame) Paralichthys olivaceus in the Seto Inland Sea of Japan in 1996. The causative agent was identified as viral hemorrhagic septicemia virus (VHSV) based on morphological, immunological, and genetic analyses. Diseased fish that were artificially injected with a representative virus isolate showed the same pathological signs and high mortality as observed in the natural outbreak. This is the first report of an outbreak of VHSV infection in cultured fish in Japan. Clinical signs of diseased fish included dark body coloration, an expanded abdomen due to ascites, congested liver, splenomegaly, and a swollen kidney. Myocardial necrosis was most prominent and accompanied by inflammatory reactions. Necrotic lesions also occurred in the liver, spleen and hematopoietic tissue, and were accompanied by circulatory disturbances due to cardiac failure. Hemorrhagic lesions did not always appear in the lateral musculature. Transmission electron microscopy revealed many rhabdovirus particles and associated inclusion bodies containing nucleocapsids in the necrotized myocardium. The histopathological findings indicated that the necrotizing myocarditis could be considered a pathognomonic sign of VHSV infection in Japanese flounder.

  16. Effect of truncation of the N-terminal region of the viral hemorrhagic septicemia virus (VHSV) P protein on viral replication.

    PubMed

    Park, Ji Sun; Kim, Min Sun; Choi, Seung Hyuk; Kang, Yue Jai; Kim, Ki Hong

    2015-11-01

    The phosphoprotein (P) of viral hemorrhagic septicemia virus (VHSV) plays an essential role in viral replication by interconnecting the L protein and the N protein-RNA complex. In this study, to investigate the role of the N-terminal region of the P protein in viral replication, we mutated the first or the first and second or the first, second, and third ATG codon into TGA stop codons. The respective mutants were named P1, P2, and P3. Recombinant VHSVs containing each mutated P gene (rVHSV-P1, -P2, and -P3) were successfully generated by supplying the intact P protein in trans. The rVHSV-P2 and -P3 were not generated from cells expressing truncated P proteins (P1, P2 or P3 protein), but the rVHSV-P1 produced infectious viruses, even in cells without any P-protein-expressing plasmids. Nucleotide sequence analysis of the P gene of rVHSV-P1 showed that a mutation had occurred that resulted in the fourth amino acid (isoleucine, ATT) being changed to methionine (ATG) without a frameshift (P0.5), suggesting that strong selection pressure might facilitate mutations that are advantageous or essential for virus replication. Infectious rVHSV-P2 and -P3 were produced in cells expressing the P0.5 protein, suggesting that the first three amino acids of the P protein of VHSV are dispensable for viral replication. Furthermore, although the P1 protein was shorter than the P0.5 protein by only two amino acid residues, no viruses were produced when the P1 protein was supplied indicating that the fourth and the fifth amino acid residues are indispensable for normal P protein functions involved in viral replication.

  17. Detection of VHSV IVb within the gonads of Great Lakes fish using in situ hybridization.

    PubMed

    Al-Hussinee, L; Lumsden, J S

    2011-05-24

    Viral haemorrhagic septicaemia virus (VHSV) genotype IVb was recently detected as the cause of numerous mortality events in Great Lakes fish. In situ hybridization was used to examine the gonads from 13 fish, including freshwater drum Aplodinotus grunniens and muskellunge Esox masquinongy that were infected naturally, as well as rainbow trout Oncorhynchus mykiss and fathead minnows Pimphales promelas, which were experimentally infected. Although the ovaries and testes of fish infected by VHSV IVb had few lesions, viral RNA was present in the ovaries of the rainbow trout and fathead minnow and was abundant in the gonads of muskellunge and in the ovaries of freshwater drum. Viral RNA was present mainly surrounding yolk vacuoles/granules or adjacent to the germinal vesicle, with lesser amounts found within the germinal vesicle, in the mesovarium and/or tunica albuginea and blood vessels of the ovary. Viral RNA was also found in and surrounding primary and secondary spermatocytes of the muskellunge.

  18. Comparison of the efficacy of Poly(I:C) immunization with live vaccine and formalin-killed vaccine against viral hemorrhagic septicemia virus (VHSV) in olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Hyoung Jun; Park, Jeong Su; Choi, Min Chul; Kwon, Se Ryun

    2016-01-01

    Viral hemorrhagic septicemia (VHS) in olive flounder, Paralichthys olivaceus, causes significant economic loss for the flounder aquaculture industry in Korea. In this study, the immunogenicity of Poly(I:C) immunization with a live vaccine against the VHS virus (VHSV) was compared with that of a formalin-treated vaccine in the olive flounder. In vaccine trial I, fish pre-injected with Poly(I:C) were highly protected from VHSV infection 2 d later (survival rate: 96%) and the surviving fish (Poly(I:C)-VHSV group) showed a 100% survival rate against VHSV re-challenge. Mortality in fish pre-injected with diethylpyrocarbonate-treated water followed by injection with formalin-treated VHSV was only 2% (1 of 50 fish), whereas survivors (DEPC-FT VHSV group) showed an 80% survival rate. In vaccine trial II, 100% survival was observed in all Poly(I:C) vaccination groups-Poly(I:C)-VHSV 6, Poly(I:C)-VHSV 5, and Poly(I:C)-VHSV 4. In contrast, the survival rates of the groups administered the formalin-treated VHSV at a dose of 10(6), 10(5), and 10(4) TCID50 100 μL(-1) fish(-1) (DEPC-FT VHSV 6, DEPC-FT VHSV 5, and DEPC-FT VHSV 4) were only 8%, 12%, and 12%, respectively. The differences in the survival rates of the formalin-treated vaccine groups in trial I and trial II were attributed to the difference in the formalin-treatment period: the formalin-treated VHSV administered in trial I was not completely inactivated and worked as a live vaccine, which explains the 80% survival rate against VHSV challenge. Specific antibodies against VHSV were detected in sera from all vaccinated survivors, except the DEPC-VHSV 4 group. Furthermore, the specific antibody titers of fish vaccinated with the live and dead VHSV vaccines were similar, but the protective effects of the live and dead vaccines varied considerably. Our findings show that Poly(I:C) immunization with the live vaccine offers better protection than the formalin-treated vaccine against VHS in olive flounder and revealed that

  19. Virulence of viral hemorrhagic septicemia virus (VHSV) genotypes Ia, IVa, IVb, and IVc in five fish species.

    USGS Publications Warehouse

    Emmenegger, Eveline J.; Moon, Chang Hoon; Hershberger, Paul K.; Kurath, Gael

    2013-01-01

    The susceptibility of yellow perch Perca flavescens, rainbow trout Oncorhynchus mykiss, Chinook salmon O. tshawytscha, koi Cyprinus carpio koi, and Pacific herring Clupea pallasii to 4 strains of viral hemorrhagic septicemia virus (VHSV) was assessed. Fish were challenged via intraperitoneal injection with high (1 × 106 plaque-forming units, PFU) and low (1 × 103 PFU) doses of a European strain (genotype Ia), and North American strains from the West coast (genotype IVa), Great Lakes (genotype IVb), and the East coast (genotype IVc). Pacific herring were exposed to the same VHSV strains, but at a single dose of 5 × 103 PFU ml-1 by immersion in static seawater. Overall, yellow perch were the most susceptible, with cumulative percent mortality (CPM) ranging from 84 to 100%, and 30 to 93% in fish injected with high or low doses of virus, respectively. Rainbow trout and Chinook salmon experienced higher mortalities (47 to 98% CPM) after exposure to strain Ia than to the other virus genotypes. Pacific herring were most susceptible to strain IVa with an average CPM of 80% and moderately susceptible (42 to 52% CPM) to the other genotypes. Koi had very low susceptibility (≤5.0% CPM) to all 4 VHSV strains. Fish tested at 7 d post challenge were positive for all virus strains, with yellow perch having the highest prevalence and concentrations of virus, and koi the lowest. While genotype Ia had higher virulence in salmonid species, there was little difference in virulence or host-specificity between isolates from subtypes IVa, IVb, and IVc.  

  20. Virulence of viral hemorrhagic septicemia virus (VHSV) genotypes Ia, IVa, IVb, and IVc in five fish species.

    PubMed

    Emmenegger, Eveline J; Moon, Chang Hoon; Hershberger, Paul K; Kurath, Gael

    2013-12-12

    The susceptibility of yellow perch Perca flavescens, rainbow trout Oncorhynchus mykiss, Chinook salmon O. tshawytscha, koi Cyprinus carpio koi, and Pacific herring Clupea pallasii to 4 strains of viral hemorrhagic septicemia virus (VHSV) was assessed. Fish were challenged via intraperitoneal injection with high (1 × 106 plaque-forming units, PFU) and low (1 × 103 PFU) doses of a European strain (genotype Ia), and North American strains from the West coast (genotype IVa), Great Lakes (genotype IVb), and the East coast (genotype IVc). Pacific herring were exposed to the same VHSV strains, but at a single dose of 5 × 103 PFU ml-1 by immersion in static seawater. Overall, yellow perch were the most susceptible, with cumulative percent mortality (CPM) ranging from 84 to 100%, and 30 to 93% in fish injected with high or low doses of virus, respectively. Rainbow trout and Chinook salmon experienced higher mortalities (47 to 98% CPM) after exposure to strain Ia than to the other virus genotypes. Pacific herring were most susceptible to strain IVa with an average CPM of 80% and moderately susceptible (42 to 52% CPM) to the other genotypes. Koi had very low susceptibility (≤5.0% CPM) to all 4 VHSV strains. Fish tested at 7 d post challenge were positive for all virus strains, with yellow perch having the highest prevalence and concentrations of virus, and koi the lowest. While genotype Ia had higher virulence in salmonid species, there was little difference in virulence or host-specificity between isolates from subtypes IVa, IVb, and IVc.

  1. Larval Pacific herring, Clupea pallasii (Valenciennes), are highly susceptible to viral haemorrhagic septicaemia and survivors are partially protected after their metamorphosis to juveniles

    USGS Publications Warehouse

    Hershberger, P.K.; Gregg, J.; Pacheco, C.; Winton, J.; Richard, J.; Traxler, G.

    2007-01-01

    Pacific herring were susceptible to waterborne challenge with viral haemorrhagic septicaemia virus (VHSV) throughout their early life history stages, with significantly greater cumulative mortalities occurring among VHSV-exposed groups of 9-, 44-, 54- and 76-day-old larvae than among respective control groups. Similarly, among 89-day-1-year-old and 1+year old post-metamorphosed juveniles, cumulative mortality was significantly greater in VHSV-challenged groups than in respective control groups. Larval exposure to VHSV conferred partial protection to the survivors after their metamorphosis to juveniles as shown by significantly less cumulative mortalities among juvenile groups that survived a VHS epidemic as larvae than among groups that were previously nai??ve to VHSV. Magnitude of the protection, measured as relative per cent survival, was a direct function of larval age at first exposure and was probably a reflection of gradual developmental onset of immunocompetence. These results indicate the potential for easily overlooked VHS epizootics among wild larvae in regions where the virus is endemic and emphasize the importance of early life history stages of marine fish in influencing the ecological disease processes. ?? 2007 The Authors.

  2. Live vaccine of viral hemorrhagic septicemia virus (VHSV) for Japanese flounder at fish rearing temperature of 21°C instead of Poly(I:C) administration.

    PubMed

    Nishizawa, Toyohiko; Takami, Ikuo; Yang, Minji; Oh, Myung-Joo

    2011-10-26

    The process of "Poly(I:C) immunization" involves immunization of fish with a pathogenic live virus, followed by administration of Poly(I:C), which induces a transient, non-specific antiviral state. As a result, fish in an antiviral state survive the initial immunization with live virus. Moreover, these fish are able to mount a specific protective immune response against the injected pathogenic virus. In the present study, we investigated the optimum temperature for Poly(I:C) immunization of Japanese flounder Paralichthys olivaceus with live viral hemorrhagic septicemia virus (VHSV). It was revealed that the optimum temperature was around at 17°C for Poly(I:C) immunization in Japanese flounder. Furthermore, the protection efficacy of Poly(I:C) immunization was significantly decreased by elevation of fish rearing temperature, and no efficacy was observed at a fish rearing temperature of 25°C. Interestingly, no mortality by VHSV infection was observed in fish reared at 21°C and 25°C even when those fish were not administered Poly(I:C). All of the survivors from the first VHSV-challenge at 21°C were strongly protected from re-challenge with VHSV. However, almost all of the survivors (≥82.6%) from the first challenge at 25°C were lost by the second challenge with VHSV. It was thus concluded that by rearing fish at 21°C and challenging with live VHSV, it is possible to induce strong specific immunity in Japanese flounder without Poly(I:C) administration.

  3. Stability and efficacy of the 3'-UTR A4G-G5A variant of viral hemorrhagic septicemia virus (VHSV) as a live attenuated immersion VHSV vaccine in olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Sung-Hyun; Kim, Meesun; Choi, Go-Eun; Lee, Jeong Ho; Kang, Jung-Ha; Evensen, Øystein; Lee, Woo-Jai

    2016-02-17

    Viral hemorrhagic septicemia virus (VHSV) is the causative agent of viral hemorrhagic septicemia in fish, a disease that affects a number of teleost fish species including olive flounder (Paralichthys olivaceus). In this study, we assessed the safety and efficacy of two recombinant attenuated VHSV strains, termed A4G-G5A and ΔNV, with the purpose to select the most suitable vaccine strain. The virus strains were passaged in two commercially available cell lines, EPC and RTG-2, and the strains were also tested for residual virulence in zebrafish (Danio rerio). The A4G-G5A strain showed an attenuated growth profile in both the EPC and RTG-2 cell lines compared to wild-type (WT) VHSV (JF-09, genotype IVa), whereas the growth profile of ΔNV was comparable to the WT strains in RTG-2 cells in contrast to EPC cells. Moreover, ΔNV had higher residual virulence compared to A4G-G5A and was highly pathogenic to zebrafish. The A4G-G5A strain was chosen as vaccine candidate and tested for efficacy in in vivo fish studies in the target species, olive flounder, using an immersion vaccine scheme. Groups of fish were immunized with 10(2.5), 10(3.5), 10(4.5), and 10(5.5) TCID50/ml of A4G-G5A giving 5-13.3 cumulative percent mortality (CPM) post immunization. Immunization was followed by a challenge experiment using VHSV-WT. The relative percent survival (RPS) in immunized groups ranged from 81.6% to 100%, correlating with vaccination dose. This study demonstrates that while strain A4G-G5A has retained some residual virulence it confers high level of protection in immunized olive flounder.

  4. A new StaRT-PCR approach to detect and quantify fish viral hemorrhagic septicemia virus (VHSv): enhanced quality control with internal standards

    USDA-ARS?s Scientific Manuscript database

    Viral Hemorrhagic Septicemia virus (VHSv) causes one of the world’s most important finfish diseases, killing >80 species across Eurasia and North America. A new and especially virulent strain (IVb) emerged in the North American Great Lakes in 2003, threatening fisheries, baitfish, and aquaculture in...

  5. Accurate detection and quantification of the fish viral hemorrhagic septicemia virus (VHSv) with a two-color fluorometric real-time PCR assay

    USDA-ARS?s Scientific Manuscript database

    Viral Hemorrhagic Septicemia virus (VHSv) is one of the world's most serious fish pathogens, infecting > 80 marine, freshwater, and estuarine fish species from Eurasia and North America. A novel and especially virulent strain - IVb - appeared in the Great Lakes in 2003, killed many game fish species...

  6. Evolution and biogeography of an emerging quasispecies: diversity patterns of the fish Viral Hemorrhagic Septicemia virus (VHSv).

    PubMed

    Pierce, Lindsey R; Stepien, Carol A

    2012-05-01

    Viral Hemorrhagic Septicemia virus (VHSv) is an RNA rhabdovirus that causes one of the most important finfish diseases, affecting over 70 marine and freshwater species. It was discovered in European cultured fish in 1938 and since has been described across the Northern Hemisphere. Four strains and several substrains have been hypothesized, whose phylogenetic relationships and evolutionary radiation are evaluated here in light of a quasispecies model, including an in-depth analysis of the novel and especially virulent new substrain (IVb) that first appeared in the North American Laurentian Great Lakes in 2003. We analyze the evolutionary patterns, genetic diversity, and biogeography of VHSv using all available RNA sequences from the glycoprotein (G), nucleoprotein (N), and non-virion (Nv) genes, with Maximum Likelihood and bayesian approaches. Results indicate that the G gene evolves at an estimated rate of μ=2.58×10(-4) nucleotide substitutions per site per year, the N gene at μ=4.26×10(-4), and Nv fastest at μ=1.25×10(-3). Phylogenetic trees from the three genes largely are congruent, distinguishing strains I-IV as reciprocally monophyletic with high bootstrap and posterior probability support. VHSv appears to have originated from a marine ancestor in the North Atlantic Ocean, diverging into two primary clades: strain IV in North America (the Northwestern Atlantic Ocean), and strains I-III in the Northeastern Atlantic region (Europe). Strain II may comprise the basal group of the latter clade and diverged in Baltic Sea estuarine waters; strains I and III appear to be sister groups (according to the G and Nv genes), with the former mostly in European freshwaters and the latter in North Sea marine/estuarine waters. Strain IV is differentiated into three monophyletic substrains, with IVa infecting Northeastern Pacific salmonids and many marine fishes (with 44 unique G gene haplotypes), IVb endemic to the freshwater Great Lakes (11 haplotypes), and a newly

  7. Establishment and characterization of a rainbow trout heart endothelial cell line with susceptibility to viral hemorrhagic septicemia virus (VHSV).

    PubMed

    Luque, Alfonso; González Granja, Aitor; González, Lucia; Tafalla, Carolina

    2014-05-01

    In the current work, we have established and characterized a novel cell line from rainbow trout (Oncorhynchus mykiss). The cell line, designated as RTH (rainbow trout heart), was obtained by immortalizing heart cells with recombinant retroviruses that transduced polyoma middle T antigen. This is the first time such a strategy is used to obtain an immortalized fish cell line. The cells showed an endothelial-like morphology and characteristics, constitutively transcribing collagen, selectin and VCAM (vascular cell adhesion molecule), as well as different chemokines and chemokine receptors, but not cytokeratin. As already described for heart endothelial cells, RTH cells actively phagocytized latex beads. Furthermore, RTH cells showed a high susceptibility to viral hemorrhagic septicemia virus (VHSV). VHSV modulated the transcription of Mx, major histocompatibility complex II (MHC-II), VCAM and many of the chemokine and chemokine receptors expressed in these cells. Therefore, RTH cells constitute an excellent model to study the immune regulation of endothelial cells in fish and their role in leukocyte extravasation.

  8. cDNA microarray analysis of disk abalone genes in gills and hemocytes after viral hemorrhagic septicemia virus (VHSV) challenge.

    PubMed

    De Zoysa, Mahanama; Nikapitiya, Chamilani; Oh, Chulhong; Whang, Ilson; Shin, Hyun-Jin; Lee, Jehee

    2012-06-01

    A disk abalone Haliotis discus discus 4.2 K cDNA microarray was designed by selecting abalone expressed sequence tags (ESTs). Transcriptional profiles in gills and hemocytes were analyzed upon abalone challenged with viral hemorrhagic septicemia virus (VHSV) in order to select candidates for screening of immune response genes. Among the 4188 genes analyzed, 280 (6.6%) transcripts were changed their expression level in gills and hemocytes against VHSV challenge compared to control animals. Total of 88 and 65 genes were up-regulated in gills and hemocytes, respectively. These genes can be grouped under various immune-functional categories such as transcription factors (Krüppell-like factor; ETS-family transcription factor), inflammatory and apoptosis related genes (TNF superfamily members, Fas ligand), IFN regulatory proteins (IFN-44 like, interferon gamma-inducible lysosomal thiol reductase) and detoxification proteins (glutathione peroxidase). In contrast, 25 and 102 genes were shown down-regulation in gills and hemocytes, respectively. Among the differentially expressed transcripts, considerably higher numbers of ESTs were represented as either hypothetical (unknown) proteins or no GenBank match suggesting those may be novel genes associated with internal defense of abalone.

  9. A new StaRT-PCR approach to detect and quantify fish Viral Hemorrhagic Septicemia virus (VHSv): enhanced quality control with internal standards.

    PubMed

    Pierce, Lindsey R; Willey, James C; Crawford, Erin L; Palsule, Vrushalee V; Leaman, Douglas W; Faisal, Mohamed; Kim, Robert K; Shepherd, Brian S; Stanoszek, Lauren M; Stepien, Carol A

    2013-04-01

    Viral Hemorrhagic Septicemia virus (VHSv) causes one of the world's most important finfish diseases, killing >80 species across Eurasia and North America. A new and especially virulent strain (IVb) emerged in the North American Great Lakes in 2003, threatening fisheries, baitfish, and aquaculture industries. Weeks-long and costly cell culture is the OIE and USDA-APHIS approved diagnostic. A new Standardized Reverse Transcriptase Polymerase Chain Reaction (StaRT-PCR) assay that uniquely incorporates internal standards to improve accuracy and prevent false negatives was developed and evaluated for its ability to detect and quantify VHSv. Results from StaRT-PCR, SYBR(®) green real time qRT-PCR, and cell culture were compared, as well as the effects of potential PCR inhibitors (EDTA and high RNA). Findings show that StaRT-PCR is sensitive, detecting a single molecule, with 100% accuracy at six molecules, and had no false negatives. In comparison, false negatives ranged from 14 to 47% in SYBR(®) green real time qRT-PCR tests, and 47-70% with cell culture. StaRT-PCR uniquely controlled for EDTA and RNA interference. Range of VHSv quantitation by StaRT-PCR was 1.0×10(0)-1.2×10(5) VHSv/10(6)actb1 molecules in wild caught fishes and 1.0×10(0)-8.4×10(5) molecules in laboratory challenged specimens. In the latter experiments, muskellunge with skin lesions had significantly more viral molecules (mean=1.9×10(4)) than those without (1.1×10(3)) (p<0.04). VHSv infection was detected earlier in injection than in immersion challenged yellow perch (two versus three days), with molecule numbers in both being comparable and relatively consistent over the remaining course of the experiment. Our results show that the StaRT-PCR test accurately and reliably detects and quantifies VHSv.

  10. A survey of wild marine fish identifies a potential origin of an outbreak of viral haemorrhagic septicaemia in wrasse, Labridae, used as cleaner fish on marine Atlantic salmon, Salmo salar L., farms.

    PubMed

    Wallace, I S; Donald, K; Munro, L A; Murray, W; Pert, C C; Stagg, H; Hall, M; Bain, N

    2015-06-01

    Viral haemorrhagic septicaemia virus (VHSV) was isolated from five species of wrasse (Labridae) used as biological controls for parasitic sea lice predominantly, Lepeophtheirus salmonis (Krøyer, 1837), on marine Atlantic salmon, Salmo salar L., farms in Shetland. As part of the epidemiological investigation, 1400 wild marine fish were caught and screened in pools of 10 for VHSV using virus isolation. Eleven pools (8%) were confirmed VHSV positive from: grey gurnard, Eutrigla gurnardus L.; Atlantic herring, Clupea harengus L.; Norway pout, Trisopterus esmarkii (Nilsson); plaice, Pleuronectes platessa L.; sprat, Sprattus sprattus L. and whiting, Merlangius merlangus L. The isolation of VHSV from grey gurnard is the first documented report in this species. Nucleic acid sequencing of the partial nucleocapsid (N) and glycoprotein (G) genes was carried out for viral characterization. Sequence analysis confirmed that all wild isolates were genotype III the same as the wrasse and there was a close genetic similarity between the isolates from wild fish and wrasse on the farms. Infection from these local wild marine fish is the most likely source of VHSV isolated from wrasse on the fish farms. © 2014 Crown Copyright. Journal of Fish Diseases © 2014 John Wiley & Sons Ltd.

  11. The use of a one-step real-time reverse transcription polymerase chain reaction (rRT-PCR) for the surveillance of viral hemorrhagic septicemia virus (VHSV) in Minnesota.

    PubMed

    Phelps, Nicholas B D; Patnayak, Devi P; Jiang, Yin; Goyal, Sagar M

    2012-12-01

    Viral hemorrhagic septicemia virus (VHSV) is a highly contagious and pathogenic virus of fish. The virus infects more than 70 fish species worldwide, in both fresh and salt water. A new viral strain (VHSV-IVb) has proven both virulent and persistent, spreading throughout the Great Lakes of North America and to inland water bodies in the region. To better understand the geographic distribution of the virus, we used a modified real-time reverse transcription polymerase chain reaction (rRT-PCR) assay for high-throughput testing of fish for VHSV. The assay was shown to be twice as sensitive as the gold standard, virus isolation, and did not cross react with other viruses found in fish. In addition, the diagnostic turnaround time was reduced from 28 to 30 d for virus isolation to 2-4 d for rRT-PCR. To demonstrate the usefulness of the rRT-PCR assay, 115 high-priority water bodies in Minnesota were tested by both methods from April 2010 to June 2011. All survey sites tested negative for VHSV by both methods. The survey results have informed fisheries managers on the absence of VHSV in Minnesota and have better prepared them for the eventual arrival of the disease. In addition, the results demonstrate the value of this rRT-PCR as a surveillance tool to rapidly identify an outbreak so that it can be controlled in a timely manner.

  12. Optimization of a Plaque Neutralization Test (PNT) to Identify the Exposure History of Pacific Herring to Viral Hemorrhagic Septicemia Virus (VHSV).

    PubMed

    Hart, Lucas M; MacKenzie, Ashley; Purcell, Maureen K; Powers, Rachel L; Hershberger, Paul K

    2017-06-01

    Methods for a plaque neutralization test (PNT) were optimized for the detection and quantification of viral hemorrhagic septicemia virus (VHSV) neutralizing activity in the plasma of Pacific Herring Clupea pallasii. The PNT was complement dependent, as neutralizing activity was attenuated by heat inactivation; further, neutralizing activity was mostly restored by the addition of exogenous complement from specific-pathogen-free Pacific Herring. Optimal methods included the overnight incubation of VHSV aliquots in serial dilutions (starting at 1:16) of whole test plasma containing endogenous complement. The resulting viral titers were then enumerated using a viral plaque assay in 96-well microplates. Serum neutralizing activity was virus-specific as plasma from viral hemorrhagic septicemia (VHS) survivors demonstrated only negligible reactivity to infectious hematopoietic necrosis virus, a closely related rhabdovirus. Among Pacific Herring that survived VHSV exposure, neutralizing activity was detected in the plasma as early as 37 d postexposure and peaked at approximately 64 d postexposure. The onset of neutralizing activity was slightly delayed in fish reared at 7.4°C relative to those in warmer temperatures (9.9°C and 13.1°C); however, neutralizing activity persisted for at least 345 d postexposure in all temperature treatments. It is anticipated that this novel ability to assess VHSV neutralizing activity in Pacific Herring will enable retrospective comparisons between prior VHS infections and year-class recruitment failures. Additionally, the optimized PNT could be employed as a forecasting tool capable of identifying the potential for future VHS epizootics in wild Pacific Herring populations. Received November 7, 2016; accepted January 14, 2017.

  13. Optimization of a Plaque Neutralization Test (PNT) to identify the exposure history of Pacific Herring to viral hemorrhagic septicemia virus (VHSV)

    USGS Publications Warehouse

    Hart, Lucas; Mackenzie, Ashley; Purcell, Maureen; Thompson, Rachel L.; Hershberger, Paul

    2017-01-01

    Methods for a plaque neutralization test (PNT) were optimized for the detection and quantification of viral hemorrhagic septicemia virus (VHSV) neutralizing activity in the plasma of Pacific Herring Clupea pallasii. The PNT was complement dependent, as neutralizing activity was attenuated by heat inactivation; further, neutralizing activity was mostly restored by the addition of exogenous complement from specific-pathogen-free Pacific Herring. Optimal methods included the overnight incubation of VHSV aliquots in serial dilutions (starting at 1:16) of whole test plasma containing endogenous complement. The resulting viral titers were then enumerated using a viral plaque assay in 96-well microplates. Serum neutralizing activity was virus-specific as plasma from viral hemorrhagic septicemia (VHS) survivors demonstrated only negligible reactivity to infectious hematopoietic necrosis virus, a closely related rhabdovirus. Among Pacific Herring that survived VHSV exposure, neutralizing activity was detected in the plasma as early as 37 d postexposure and peaked at approximately 64 d postexposure. The onset of neutralizing activity was slightly delayed in fish reared at 7.4°C relative to those in warmer temperatures (9.9°C and 13.1°C); however, neutralizing activity persisted for at least 345 d postexposure in all temperature treatments. It is anticipated that this novel ability to assess VHSV neutralizing activity in Pacific Herring will enable retrospective comparisons between prior VHS infections and year-class recruitment failures. Additionally, the optimized PNT could be employed as a forecasting tool capable of identifying the potential for future VHS epizootics in wild Pacific Herring populations.

  14. Survival of the North American strain of viral hemorrhagic septicemia virus (VHSV) in filtered seawater and seawater containing ovarian fluid, crude oil and serum-enriched culture medium

    USGS Publications Warehouse

    Kocan, R.M.; Hershberger, P.K.; Elder, N.E.

    2001-01-01

     The North American strain of viral hemorrhagic septicemia virus (NA-VHSV) could be recovered for up to 40 h in natural filtered seawater (27 ppt) with a 50% loss of infectivity after approximately 10 h at 15°C. Addition of 10 ppb North Slope crude oil to the seawater had no effect on virus survival. However, when various concentrations of teleost ovarian fluid were added to seawater, virus could be recovered after 72 h at 0.01% ovarian fluid and after 96 h at 1.0%. When cell culture medium supplemented with 10% fetal bovine serum was added to the seawater, 100% of the virus could be recovered for the first 15 d and 60% of the virus remained after 36 d. These findings quantify NA-VHSV infectivity in natural seawater and demonstrate that ovarian fluid, which occurs naturally during spawning events, significantly prolongs the survival and infectivity of the virus. The extended stabilization of virus in culture medium supplemented with serum allows for low titer field samples to be collected and transported in an unfrozen state without significant loss of virus titer.

  15. Persistence of viral RNA in fish infected with VHSV-IVb at 15°C and then moved to warmer temperatures after the onset of disease.

    PubMed

    Goodwin, A E; Merry, G E; Noyes, A D

    2012-07-01

    Smallmouth bass, Micropterus dolomieu Lacepède, bluegill, Lepomis macrochirus Rafinesque (coppernose strain), koi carp, Cyprinus carpio L., and channel catfish Ictalurus punctatus (Rafinesque), were infected by intraperitoneal injection with viral haemorrhagic septicaemia virus genotype IVb (VHSV-IVb) at 15 °C. When clinical signs of disease developed, one-third of the fish was moved to 20°C and one-third to 25°C. Mortality in challenged fish at all three temperatures ranged from 25 to 45% in smallmouth bass and from 70 to 90% in bluegill. No koi carp or channel catfish died during the study. Viral copy numbers detected by quantitative real-time reverse transcriptase PCR (qrt-RTPCR) in fish dying at 20 and 25°C decreased over time. In survivors of the challenge, viral copy numbers were higher in the more susceptible species (smallmouth bass and bluegill) than in the more VHSV-IVb disease-resistant species (koi carp and channel catfish). In fish surviving 28days post-infection, prevalence of infection was 66-100% depending on species and temperature, and VHSV-IVb was detected at 10(3) -10(5) copies μg(-1) host RNA. Our results show that qrt-RTPCR is a useful tool to investigate fish kills even 28days after temperatures are elevated above those known to be permissive for VHSV replication.

  16. Induction of anti-viral genes during acute infection with Viral hemorrhagic septicemia virus (VHSV) genogroup IVa in Pacific herring (Clupea pallasii).

    PubMed

    Hansen, John D; Woodson, James C; Hershberger, Paul K; Grady, Courtney; Gregg, Jacob L; Purcell, Maureen K

    2012-02-01

    Infection with the aquatic rhabdovirus Viral hemorrhagic septicemia virus (VHSV) genogroup IVa results in high mortality in Pacific herring (Clupea pallasii) and is hypothesized to be a potential limiting factor for herring recovery. To investigate anti-viral immunity in the Pacific herring, four immune response genes were identified: the myxovirus resistance (Clpa-Mx), a major histocompatibility complex IB (named Clpa-UAA.001), the inducible immunoproteosome subunit 9 (Clpa-PSMB9) and the neutrophil chemotactic factor (Clpa-LECT2). Reverse transcriptase quantitative PCR (RT-qPCR) assays were developed based on these gene sequences to investigate the host immune response to acute VHSV infection following both injection and immersion challenge. Virus levels were measured by both plaque assay and RT-qPCR and peaked at day 6 during the 10-day exposure period for both groups of fish. The interferon stimulated genes (Clpa-Mx, -UAA.001, and -PSMB9) were significantly up-regulated in response to VHSV infection at both 6 and 10 days post-infection in both spleen and fin. Results from this study indicate that Pacific herring mount a robust, early antiviral response in both fin and spleen tissues. The immunological tools developed in this study will be useful for future studies to investigate antiviral immunity in Pacific herring.

  17. Optimization of fixed-permeabilized cell monolayers for high throughput micro-neutralizing antibody assays: application to the zebrafish/viral hemorrhagic septicemia virus (vhsv) model.

    PubMed

    Chinchilla, Blanca; Encinas, Paloma; Estepa, Amparo; Coll, Julio; Gomez-Casado, Eduardo

    2013-11-01

    A new high throughput centrifugation-free method to estimate viral neutralizing antibody levels in low volumes and large numbers of plasma blood samples is described. Cell monolayers were, (i) plated on poly-d-Lys coated 96-wells, (ii) infected with viruses previously incubated with fish plasma containing antibodies, (iii) fixed with formaldehyde to increase cell recovery and avoid centrifugation steps, (iv) permeabilized with Saponin, (v) immunostained in the presence of Saponin by using a monoclonal antibody (MAb) to viral protein, (vi) digested with trypsin to detach cells from the monolayer, in the absence of Saponin to reduce damage of intracellular MAb-antigen complexes, and (vii) gated by flow cytometry using automatic 96-well batch analysis. The method was applied to the determination of plasma neutralizing antibodies from zebrafish (Danio rerio) surviving infections with viral hemorrhagic septicemia virus (VHSV) (an important rhabdovirus of salmonids). This semi-automatic, rapid and practical assay detected anti-VHSV neutralizing antibodies in the plasma (∼3 μl per fish) of 95.1% of the zebrafish surviving VHSV infections. The fixed-permeabilized monolayer (FIXPERM) micro-neutralization method might help to analyze sera/plasma from small fish under standarized high throughput conditions.

  18. Induction of anti-viral genes during acute infection with Viral hemorrhagic septicemia virus (VHSV) genogroup IVa in Pacific herring (Clupea pallasii)

    USGS Publications Warehouse

    Hansen, John D.; Woodson, James C.; Hershberger, Paul K.; Grady, Courtney; Gregg, Jacob L.; Purcell, Maureen K.

    2012-01-01

    Infection with the aquatic rhabdovirus Viral hemorrhagic septicemia virus (VHSV) genogroup IVa results in high mortality in Pacific herring (Clupea pallasii) and is hypothesized to be a potential limiting factor for herring recovery. To investigate anti-viral immunity in the Pacific herring, four immune response genes were identified: the myxovirus resistance (Clpa-Mx), a major histocompatibility complex IB (named Clpa-UAA.001), the inducible immunoproteosome subunit 9 (Clpa-PSMB9) and the neutrophil chemotactic factor (Clpa-LECT2). Reverse transcriptase quantitative PCR (RT-qPCR) assays were developed based on these gene sequences to investigate the host immune response to acute VHSV infection following both injection and immersion challenge. Virus levels were measured by both plaque assay and RT-qPCR and peaked at day 6 during the 10-day exposure period for both groups of fish. The interferon stimulated genes (Clpa-Mx, −UAA.001, and −PSMB9) were significantly up-regulated in response to VHSV infection at both 6 and 10 days post-infection in both spleen and fin. Results from this study indicate that Pacific herring mount a robust, early antiviral response in both fin and spleen tissues. The immunological tools developed in this study will be useful for future studies to investigate antiviral immunity in Pacific herring.

  19. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viral hemorrhagic septicaemia virus (VHS).

    PubMed

    Soliman, H; El-Matbouli, M

    2006-05-31

    A one step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of viral hemorrhagic septicaemia virus (VHS). A set of six primers were designed, based on the G-protein sequence of the VHS virus serotypes (He, F1, 23.75, Klapmolle and Rindsholm). The assay was optimised to amplify VHS RNA by incubation at 63 degrees C for only 1h, and required only a simple water bath or heating block to provide a constant temperature of 63 degrees C. RT-LAMP amplification products were detected by visual inspection using SYBR Green I stain and had a ladder-like appearance when electrophoresed on an agarose gel. The detection limit of the RT-LAMP assay was found to be similar to the commonly used RT-PCR method: both methods detected VHS RNA at a dilution of 10(6). The assay was evaluated using clinical samples and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for VHS virus.

  20. Transcriptome profiles associated to VHSV infection or DNA vaccination in turbot (Scophthalmus maximus).

    PubMed

    Pereiro, Patricia; Dios, Sonia; Boltaña, Sebastián; Coll, Julio; Estepa, Amparo; Mackenzie, Simon; Novoa, Beatriz; Figueras, Antonio

    2014-01-01

    DNA vaccines encoding the viral G glycoprotein show the most successful protection capability against fish rhabdoviruses. Nowadays, the molecular mechanisms underlying the protective response remain still poorly understood. With the aim of shedding light on the protection conferred by the DNA vaccines based in the G glycoprotein of viral haemorrhagic septicaemia virus (VHSV) in turbot (Scophthalmus maximus) we have used a specific microarray highly enriched in antiviral sequences to carry out the transcriptomic study associated to VHSV DNA vaccination/infection. The differential gene expression pattern in response to empty plasmid (pMCV1.4) and DNA vaccine (pMCV1.4-G860) intramuscular administration with regard to non-stimulated turbot was analyzed in head kidney at 8, 24 and 72 hours post-vaccination. Moreover, the effect of VHSV infection one month after immunization was also analyzed in vaccinated and non-vaccinated fish at the same time points. Genes implicated in the Toll-like receptor signalling pathway, IFN inducible/regulatory proteins, numerous sequences implicated in apoptosis and cytotoxic pathways, MHC class I antigens, as well as complement and coagulation cascades among others were analyzed in the different experimental groups. Fish receiving the pMCV1.4-G860 vaccine showed transcriptomic patterns very different to the ones observed in pMCV1.4-injected turbot after 72 h. On the other hand, VHSV challenge in vaccinated and non-vaccinated turbot induced a highly different response at the transcriptome level, indicating a very relevant role of the acquired immunity in vaccinated fish able to alter the typical innate immune response profile observed in non-vaccinated individuals. This exhaustive transcriptome study will serve as a complete overview for a better understanding of the crosstalk between the innate and adaptive immune response in fish after viral infection/vaccination. Moreover, it provides interesting clues about molecules with a potential

  1. Gilthead seabream (Sparus aurata) Mx gene promoters respond differentially to IPNV and VHSV infections in RTG-2 cells.

    PubMed

    González-Mariscal, J A; Fernández-Trujillo, M A; Alonso, M C; García-Rosado, E; Álvarez, M C; Béjar, J

    2016-03-01

    The understanding of virus-host interactions relies on the knowledge of the regulatory mechanisms of the type I interferon (IFN I)-stimulated genes (ISGs). Among ISGs, those coding Mx proteins play a main role due to their direct antiviral activity. The study of these genes in gilthead seabream is interesting, since this species displays high natural resistance to viral diseases, being asymptomatic carrier of infectious pancreatic necrosis virus (IPNV) and viral haemorrhagic septicaemia virus (VHSV). Gilthead seabream has three Mx genes (Mx1, Mx2, and Mx3), encoding proteins with a wide spectrum of antiviral activity. The structure of the three promoters (pMx1, pMx2 and pMx3) has been previously disclosed, and their response to poly I:C in RTG-2 cells characterized. To further analyze these promoters, their response to two viral infections has been evaluated in the present study. For that purpose, RTG-2 cells transiently transfected with the luciferase gene under the control of each promoter were inoculated with either IPNV or VHSV at two different doses. The highest and lowest fold induction values were recorded for pMx2 and pMx3, respectively. The promoter induction was always stronger after VHSV inoculation than in IPNV-inoculated cells. In addition, the higher dose of VHSV tested induced higher response of the three promoters, whereas in IPNV-infected cells the highest induction was recorded after inoculation with the lower viral dose. To further study the response of the Mx2 promoter, RTG-2 cells stably transfected with the luciferase gene under the control of pMx2 were stimulated with poly I:C and subsequently infected with IPNV or VHSV. Interestingly, IPNV infection inhibited the induction caused by poly I:C, suggesting an antagonistic activity of IPNV on Mx2 transcription. In contrast, VHSV infection did not alter the response triggered by poly I:C. These results highlight the specific regulation that controls the activity of each promoter, and support the

  2. Comparison of quantitative RT-PCR with cell culture to detect viral hemorrhagic septicemia virus (VHSV) IVb infections in the Great Lakes.

    PubMed

    Hope, Kristine M; Casey, Rufina N; Groocock, Geoffrey H; Getchell, Rodman G; Bowser, Paul R; Casey, James W

    2010-03-01

    Viral hemorrhagic septicemia virus (VHSV) is an important pathogen of cultured and wild fish in marine and freshwater environments. A new genotype, VHSV IVb, was isolated from a fish collected from the Great Lakes in 2003. Since the first isolation, VHSV IVb has been confirmed in 28 species, signaling the early invasion and continued spread of this Office International des Epizooties-reportable agent. For surveillance of this virus in both wild and experimental settings, we have developed a rapid and sensitive one-step quantitative real-time polymerase chain reaction (qRT-PCR) assay that amplifies a 100-base-pair conserved segment from both the genomic negative strand and the mRNA positive strand of the nucleoprotein (N) gene of VHSV IVb. This assay is linear over seven orders of magnitude, with an analytical capability of detecting a single copy of viral RNA and reproducibility at 100 copies. The assay is approximately linear with RNA input from 50 to 1000 ng per assay and works equally well with RNA prepared from a column-based or phenol-chloroform-based method. In wild-caught fish, 97% of the cases were found to be more than three orders of magnitude more sensitive using qRT-PCR than using cell culture. Of the 1,428 fish from the Great Lakes region tested in 2006 and 2007, 24% were positive by qRT-PCR whereas only 5% were positive by cell culture. All of the fish that were positive by cell culture were also positive by qRT-PCR. Importantly, qRT-PCR sensitivity is comparable to that of cell culture detection when comparing VHSV viral RNA levels with viral titer stocks, confirming that the high qRT-PCR signals obtained with diagnostic samples are due to the accumulation of N gene mRNA by transcriptional attenuation. The qRT-PCR assay is particularly valuable for rapid and high-throughput prescreening of fish before confirmatory testing by cell culture or sequencing tissue-derived amplicons and especially in detecting infection in fish that do not show clinical

  3. Viral hemorrhagic septicemia virus (VHSV) infection-mediated sequential changes in microRNAs profile of Epithelioma papulosum cyprini (EPC) cells.

    PubMed

    Najib, Abdellaoui; Kim, Min Sun; Kim, Ki Hong

    2017-02-01

    MicroRNAs are small non-coding RNAs and are involved in the regulation of wide biological processes. Viral hemorrhagic septicemia virus (VHSV) is the causative agent of viral hemorrhagic septicemia (VHS) disease causing a heavy loss in aquaculture farms. In this study, we tried to explore the effect of VHSV infection on microRNAs profile of Epithelioma papulosum cyprini (EPC) cells at different points of time (0, 3, 12, 24, and 48 h post infection). A total of 355 conserved microRNAs and 3 novel microRNAs were identified, and among them, 103 microRNAs were differentially expressed. The number of differentially expressed microRNAs was highly increased at 24 h.p.i compared to 3 h.p.i and 12 h.p.i., suggesting that EPC cells might not actively respond to VHSV infection at an early infection period, which can allow viruses to transcript and translate their genes enough to produce viral particles that can infect to another cells. Among the differentially expressed microRNAs, 2 miRNAs (miR-735 and miR-738) that were reported only in fish species were highly upregulated, and based on the target prediction, they could regulate several immune pathways. Furthermore, the present results showed the upregulation of representative immune regulating microRNAs such as miR-146a, miR-155, and miR-99. The target prediction of differentially expressed miRNAs, GO, and KEGG pathways analysis revealed that several biological processes and different pathways were affected by the viral infection. The present dynamical changing patterns of differentially expressed microRNAs in response to the progression of VHSV infection suggest that microRNA profile that was analyzed at one time point cannot provide enough information for the interpretation of the disease mechanism. Considering the wide and complex interactions between microRNAs and genes expression, the present results provide the basis for the understanding of VHSV infection-mediated cellular responses and for future investigations

  4. Accurate detection and quantification of the fish viral hemorrhagic Septicemia virus (VHSv) with a two-color fluorometric real-time PCR assay.

    PubMed

    Pierce, Lindsey R; Willey, James C; Palsule, Vrushalee V; Yeo, Jiyoun; Shepherd, Brian S; Crawford, Erin L; Stepien, Carol A

    2013-01-01

    Viral Hemorrhagic Septicemia virus (VHSv) is one of the world's most serious fish pathogens, infecting >80 marine, freshwater, and estuarine fish species from Eurasia and North America. A novel and especially virulent strain - IVb - appeared in the Great Lakes in 2003, has killed many game fish species in a series of outbreaks in subsequent years, and shut down interstate transport of baitfish. Cell culture is the diagnostic method approved by the USDA-APHIS, which takes a month or longer, lacks sensitivity, and does not quantify the amount of virus. We thus present a novel, easy, rapid, and highly sensitive real-time quantitative reverse transcription PCR (qRT-PCR) assay that incorporates synthetic competitive template internal standards for quality control to circumvent false negative results. Results demonstrate high signal-to-analyte response (slope = 1.00±0.02) and a linear dynamic range that spans seven orders of magnitude (R(2) = 0.99), ranging from 6 to 6,000,000 molecules. Infected fishes are found to harbor levels of virus that range to 1,200,000 VHSv molecules/10(6) actb1 molecules with 1,000 being a rough cut-off for clinical signs of disease. This new assay is rapid, inexpensive, and has significantly greater accuracy than other published qRT-PCR tests and traditional cell culture diagnostics.

  5. Accurate Detection and Quantification of the Fish Viral Hemorrhagic Septicemia virus (VHSv) with a Two-Color Fluorometric Real-Time PCR Assay

    PubMed Central

    Palsule, Vrushalee V.; Yeo, Jiyoun; Shepherd, Brian S.; Crawford, Erin L.; Stepien, Carol A.

    2013-01-01

    Viral Hemorrhagic Septicemia virus (VHSv) is one of the world's most serious fish pathogens, infecting >80 marine, freshwater, and estuarine fish species from Eurasia and North America. A novel and especially virulent strain – IVb – appeared in the Great Lakes in 2003, has killed many game fish species in a series of outbreaks in subsequent years, and shut down interstate transport of baitfish. Cell culture is the diagnostic method approved by the USDA-APHIS, which takes a month or longer, lacks sensitivity, and does not quantify the amount of virus. We thus present a novel, easy, rapid, and highly sensitive real-time quantitative reverse transcription PCR (qRT-PCR) assay that incorporates synthetic competitive template internal standards for quality control to circumvent false negative results. Results demonstrate high signal-to-analyte response (slope = 1.00±0.02) and a linear dynamic range that spans seven orders of magnitude (R2 = 0.99), ranging from 6 to 6,000,000 molecules. Infected fishes are found to harbor levels of virus that range to 1,200,000 VHSv molecules/106 actb1 molecules with 1,000 being a rough cut-off for clinical signs of disease. This new assay is rapid, inexpensive, and has significantly greater accuracy than other published qRT-PCR tests and traditional cell culture diagnostics. PMID:23977162

  6. Efficacy of a glycoprotein DNA vaccine against viral haemorrhagic septicaemia (VHS) in Pacific herring, Clupea pallasii Valenciennes

    USGS Publications Warehouse

    Hart, L.M.; Lorenzen, Niels; LaPatra, S.E.; Grady, C.A.; Roon, S.E.; O’Reilly, J.; Gregg, J.L.; Hershberger, P.K.

    2012-01-01

    Viral haemorrhagic septicaemia virus (VHSV) and its associated disease state, viral haemorrhagic septicaemia (VHS), is hypothesized to be a proximate factor accounting for the decline and failed recovery of Pacific herring populations in Prince William Sound, AK (Marty et al. 1998, 2003, 2010). Survivors of laboratory-induced VHSV epizootics develop resistance to subsequent viral exposure (Kocan et al. 2001; Hershberger et al. 2007, 2010), which is likely the result of immune system recognition of the viral glycoprotein (G) (Lecocq-Xhonneux et al. 1994), a surface antigen that contains neutralizing epitopes (Lorenzen, Olesen & Jorgensen 1990; Jørgensen et al. 1995) and cell attachment domains (Lecocq-Xhonneux et al. 1994; Estepa & Coll 1996). These properties have proven useful in the development of G-gene-based DNA vaccines for VHSV and a related rhabdovirus, infectious haematopoietic necrosis virus (IHNV) (Anderson et al. 1996; Heppell et al. 1998; Corbeil et al. 1999; Einer-Jensen et al. 2009). Rainbow trout fingerlings, Oncorhynchus mykiss (Walbaum), vaccinated with 1 µg of either the VHS or IHN vaccine are protected from VHS when exposed to virus as early as 4 days (44 degree days) post-vaccination (p.v.) (Lorenzen et al. 2002). At later time points (80 days p.v.; 880 degree days), the level of cross-protection against VHS by IHN vaccination is either completely lost (60 days p.v.; 660 degree days) (3 g rainbow trout; 1 µg vaccine dose) (Lorenzen et al. 2002) or present at intermediate levels (6.5 g rainbow trout; 1 µg vaccine dose) (Einer-Jensen et al. 2009). Comparatively, VHS vaccination remains effective as long as 9 months (2520 degree days) p.v. (100 g rainbow trout; 0.5 µg vaccine dose) (McLauchlan et al. 2003). These results suggest that IHN and VHS vaccination activate a rapid transitory innate immune response against VHSV that is followed by long-term adaptive immunity in VHS-vaccinated trout (Lorenzen et al. 2002).

  7. Development of a walleye spleen stromal cell line sensitive to viral hemorrhagic septicemia virus (VHSV IVb) and to protection by synthetic dsRNA.

    PubMed

    Vo, Nguyen T K; Bender, Aaron W; Ammendolia, Dustin A; Lumsden, John S; Dixon, Brian; Bols, Niels C

    2015-07-01

    A cell line, WE-spleen6, has been developed from the stromal layer of primary spleen cell cultures. On conventional plastic, WE-spleen6 cells had a spindle-shaped morphology at low cell density but grew to become epithelial-like at confluency. On the commercial extracellular matrix (ECM), Matrigel, the cells remained spindle-shaped and formed lumen-like structures. WE-spleen6 cells had intermediate filament protein, vimentin and the ECM protein, collagen I, but not smooth muscle α-actin (SMA) and von Willebrand factor (vWF) and lacked alkaline phosphatase and phagocytic activities. WE-spleen6 was more susceptible to infection with VHSV IVb than a fibroblast and epithelial cell lines from the walleye caudal fin, WE-cfin11f and WE-cfin11e, respectively. Viral transcripts and proteins appeared earlier in WE-spleen6 cultures as did cytopathic effect (CPE) and significant virus production. The synthetic double-stranded RNA (dsRNA), polyinosinic: polycytidylic acid (pIC), induced the antiviral protein Mx in both cell lines. Treating WE-spleen6 cultures with pIC prior to infection with VHSV IVb inhibited the early accumulation of viral transcripts and proteins and delayed the appearance of CPE and significant viral production. Of particular note, pIC caused the disappearance of viral P protein 2 days post infection. WE-spleen6 should be useful for investigating the impact of VHSV IVb on hematopoietic organs and the actions of pIC on the rhabdovirus life cycle.

  8. Fugu double U6 promoter-driven long double-stranded RNA inhibits proliferation of viral hemorrhagic septicemia virus (VHSV) in fish cell lines.

    PubMed

    Kim, Min Sun; Jee, Bo Young; Cho, Mi Young; Kim, Jin Woo; Jeong, Hyun Do; Kim, Ki Hong

    2012-06-01

    A long double-stranded RNA (dsRNA)-producing vector driven by fugu double U6 promotors, in which the two promoters were arranged in a head-to-head fashion, was newly constructed. To determine whether the DNA-vector-based long dsRNAs can induce sequence-specific RNA interference (RNAi), Epithelioma papulosum cyprini (EPC) cells and chinook salmon embryonic (CHSE-214) cells were transfected with the long dsRNA vector targeting the G gene of VHSV, and its effect on expression of the G gene and viral proliferation was investigated. The sequence-specific inhibitory effect was further confirmed by analysis of interferon (IFN)-triggered Mx1 gene expression and cross-protection against infectious hematopoietic necrosis virus (IHNV). The fugu double U6 promoter-driven vector successfully produced long dsRNAs in EPC cells, a system that allows continuous production of long dsRNAs in transfected cells. The plasmid-based long dsRNAs targeting the VHSV G gene effectively suppressed G gene expression, but control dsRNAs targeting the EGFP gene did not. Furthermore, there was no significant difference in Mx gene expression between cells transfected with the long dsRNA-producing vector and those transfected with the control empty vector. These results suggest that G gene expression was suppressed not by type-I-IFN-mediated nonspecific inhibition but in a sequence-specific manner. Both EPC and CHSE-214 cells transfected with plasmids producing long dsRNAs targeting the VHSV G gene were protected against VHSV infection but were not protected against IHNV infection, suggesting sequence-specific RNAi-mediated inhibition of viral proliferation. In conclusion, we show, for the first time, long-dsRNA-mediated RNAi in fish cells. The DNA-vector-based long dsRNAs may provide an efficient tool for analysis of gene function in fish cells without preliminary burdensome work for selection of effective siRNA clones, and it may be applied as an antiviral measure in cultured fish.

  9. Montanide IMS 1312 VG adjuvant enhances the efficacy of immersion vaccine of inactivated viral hemorrhagic septicemia virus (VHSV) in olive flounder, Paralichthys olivaceus.

    PubMed

    Hwang, Jee Youn; Kwon, Mun-Gyeong; Kim, Yu Jin; Jung, Sung-Hee; Park, Myoung-Ae; Son, Maeng-Hyun

    2017-01-01

    Vaccination by immersion is suitable for mass vaccination of small size fish. However, no viral vaccine has been developed for immersion applications, because of low efficacy. In this study, we evaluated the efficacy and safety of immersion vaccine against viral hemorrhagic septicemia (VHS) containing Montanide IMS 1312 VG adjuvant in olive flounder (Paralichthys olivaceus). Healthy fish were vaccinated by an immersion method with a heat-inactivated FP-VHS2010-1 strain of VHS virus (VHSV) in combination with Montanide IMS 1312 VG for 5 min at 20 ± 2 °C. The control group was vaccinated with sterile PBS. No toxicity of immersion vaccine with Montanide IMS 1312 VG adjuvant was observed by hematological and histopathological analysis. Immersion vaccine with adjuvant enhanced gene expression of immune-associated genes, i.e., genes encoding interleukin (IL)-1β, IL-6, IL-8, and Toll-like receptor (TLR) 3. Relative percent survival (RPS) of fish was measured on weeks 4 and 8 post vaccination. In fish vaccinated with adjuvant, RPS was significantly higher than that of fish vaccinated without adjuvant. The results of the present study provide evidence that the VHSV immersion vaccine with Montanide IMS 1312 VG induces protective immunity in olive flounder against VHS.

  10. Experimental transmission of VHSV genotype IVb by predation.

    PubMed

    Getchell, Rodman G; Cornwell, Emily R; Groocock, Geoffrey H; Wong, Po Ting; Coffee, Laura L; Wooster, Gregory A; Bowser, Paul R

    2013-12-01

    Preliminary surveillance of wild baitfish during the 2006 viral hemorrhagic septicemia virus genotype IVb (VHSV IVb) outbreaks indicated Emerald Shiners Notropis atherinoides and Bluntnose Minnow Pimephales notatus were infected with high levels of VHSV without showing clinical signs of disease. The movement and use of baitfish was recognized as the most probable vector for the introduction of VHSV to inland waters, such as Conesus Lake and Skaneateles Lake in New York, Budd Lake in Michigan, and Little Lake Butte des Morts and Lake Winnebago in Wisconsin. While numerous government agencies implemented restrictions to stop the movement of potentially infected baitfish into new waters and prevent the spread of VHSV IVb, until now, studies to investigate whether these initial introductions were by an oral route of infection have not occurred. Our studies identified infected Fathead Minnow Pimephales promelas as suitable vectors for transmitting VHSV IVb when fed to Tiger Muskellunge ( ♂ Northern Pike Esox lucius × ♀ Muskellunge Esox masquinongy) during laboratory trials. Six of 16 Tiger Muskellunge were infected with VHSV IVb after consumption of infected Fathead Minnows when assayed with quantitative reverse transcriptase polymerase chain reaction and viral isolation in cell culture. Weekly sampling of water and feces from these Tiger Muskellunge individually reared showed intermittent shedding of VHSV IVb. Those exposed to similarly VHSV IVb-inoculated fathead minnows by cohabitation only became infected in 1 case out of 16. A similar trial of 12 Tiger Muskellunge fed Round Goby Neogobius melanostomus that survived a VHSV IVb immersion challenge did not result in infection. Overall, our findings imply that consumption of infected wild baitfish may be a risk factor for introduction of VHSV.

  11. Enhancement of glycoprotein-based DNA vaccine for viral hemorrhagic septicemia virus (VHSV) via addition of the molecular adjuvant, DDX41.

    PubMed

    Lazarte, Jassy Mary S; Kim, Young Rim; Lee, Jung Seok; Im, Se Pyeong; Kim, Si Won; Jung, Jae Wook; Kim, Jaesung; Lee, Woo Jai; Jung, Tae Sung

    2017-03-01

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 10(6) TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p < 0.05/p < 0.001). After VHSV challenge, the fish group treated with pEF-GD showed higher survival rate than the pEF-G treated group, though difference was not statistically significant in the 15 dpv challenged group however in the 30 dpv challenged group, the difference was statistically significant (p < 0.05). Together, these results clearly demonstrate that DDX41 is an effective adjuvant for the G-based DNA vaccine in olive flounder. Our novel findings could facilitate the development of more effective DNA vaccines for the aquaculture industry.

  12. Transcriptome Profiles Associated to VHSV Infection or DNA Vaccination in Turbot (Scophthalmus maximus)

    PubMed Central

    Pereiro, Patricia; Dios, Sonia; Boltaña, Sebastián; Coll, Julio; Estepa, Amparo; Mackenzie, Simon; Novoa, Beatriz; Figueras, Antonio

    2014-01-01

    DNA vaccines encoding the viral G glycoprotein show the most successful protection capability against fish rhabdoviruses. Nowadays, the molecular mechanisms underlying the protective response remain still poorly understood. With the aim of shedding light on the protection conferred by the DNA vaccines based in the G glycoprotein of viral haemorrhagic septicaemia virus (VHSV) in turbot (Scophthalmus maximus) we have used a specific microarray highly enriched in antiviral sequences to carry out the transcriptomic study associated to VHSV DNA vaccination/infection. The differential gene expression pattern in response to empty plasmid (pMCV1.4) and DNA vaccine (pMCV1.4-G860) intramuscular administration with regard to non-stimulated turbot was analyzed in head kidney at 8, 24 and 72 hours post-vaccination. Moreover, the effect of VHSV infection one month after immunization was also analyzed in vaccinated and non-vaccinated fish at the same time points. Genes implicated in the Toll-like receptor signalling pathway, IFN inducible/regulatory proteins, numerous sequences implicated in apoptosis and cytotoxic pathways, MHC class I antigens, as well as complement and coagulation cascades among others were analyzed in the different experimental groups. Fish receiving the pMCV1.4-G860 vaccine showed transcriptomic patterns very different to the ones observed in pMCV1.4-injected turbot after 72 h. On the other hand, VHSV challenge in vaccinated and non-vaccinated turbot induced a highly different response at the transcriptome level, indicating a very relevant role of the acquired immunity in vaccinated fish able to alter the typical innate immune response profile observed in non-vaccinated individuals. This exhaustive transcriptome study will serve as a complete overview for a better understanding of the crosstalk between the innate and adaptive immune response in fish after viral infection/vaccination. Moreover, it provides interesting clues about molecules with a potential

  13. An isolate and sequence database of infectious haematopoietic necrosis virus (IHNV).

    PubMed

    Jonstrup, S P; Schuetze, H; Kurath, G; Gray, T; Jensen, B Bang; Olesen, N J

    2010-06-01

    In the field of fish diseases, the amount of relevant information available is enormous. Internet-based databases are an excellent tool for keeping track of the available knowledge in the field. Fishpathogens.eu was launched in June 2009 with the aim of collecting, storing and sorting data on fish pathogens. The first pathogen to be included was the rhabdovirus, viral haemorrhagic septicaemia virus (VHSV). Here, we present an extension of the database to also include infectious haematopoietic necrosis virus (IHNV). The database is developed, maintained and managed by the European Community Reference Laboratory for Fish Diseases and collaborators. It is available at http://www.fishpathogens.eu/ihnv.

  14. Recombinant infectious hematopoietic necrosis viruses induce protection for rainbow trout Oncorhynchus mykiss.

    PubMed

    Romero, Alejandro; Figueras, Antonio; Thoulouze, Maria-Isabel; Bremont, Michael; Novoa, Beatriz

    2008-07-07

    Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) are rhabdoviruses that infect salmonids, producing serious economic losses. Two recombinant IHN viruses were generated by reverse genetics. For one (rIHNV GFP) the IHNV NV gene was replaced with the green fluorescent protein (GFP) gene. In the other (rIHNV-Gvhsv GFP) the G gene was also exchanged for that of VHSV. No mortalities, external signs or histological lesions were observed in experimental infections conducted with the recombinant viruses. Neither the rIHNV GFP nor rIHNV-Gvhsv GFP was detected by RT-PCR in any of the examined tissues from experimentally infected fish. In order to assess their potential as vaccines against the wild type viruses, rainbow trout were vaccinated with the recombinant viruses by intraperitoneal injection and challenged 30 d later with virulent IHNV or VHSV. The GFP viruses provided protection against both wild type viruses. None of the recombinant viruses induced antibody production, and the expression of interferon (IFNalpha4) and interferon induced genes such as Mx protein and ISG-15 was not different to that of controls. The rIHNV-Gvhsv GFP did not inhibit cellular apoptosis as it was observed in an IHNV inoculated fish cell line. These studies suggest that the recombinant rIHNV-Gvhsv GFP is a promising candidate as a live recombinant vaccine and also provides a good model to further study viral pathogenicity and the molecular basis of protection against these viral infections.

  15. Serotyping of foot and mouth disease virus and Pasteurella multocida from Indian gaurs (Bos gaurus), concurrently infected with foot and mouth disease and haemorrhagic septicaemia.

    PubMed

    Chandranaik, Basavegowdanadoddi Marinaik; Hegde, Raveendra; Shivashankar, Beechagondahalli Papanna; Giridhar, Papanna; Muniyellappa, Handenahally Kaverappa; Kalge, Rajeshwar; Sumathi, Benamanahalli Raju; Nithinprabhu, Kumble; Chandrashekara, Narasimhaiah; Manjunatha, Venkataramanappa; Jaisingh, Nirupama; Mayanna, Asha; Chandrakala, Gowda Kallenahalli; Kanaka, Sermaraja; Venkatesha, Mudalagiri Dasappagupta

    2015-06-01

    We report the serotyping of foot-and-mouth disease virus (FMDV) and Pasteurella multocida from Indian gaurs which were concurrently infected with foot-and-mouth disease (FMD) and haemorrhagic septicaemia. Bannerghatta biological park (BBP), a national park located in the outskirts of Bengaluru city, Karnataka, India, is bordered by several villages. These villages witnessed massive outbreaks of FMD which spread rapidly to the herbivores at BBP. Post-mortem was conducted on carcasses of two Indian gaurs that died with symptoms of FMD. The salient gross findings included extensive vesicular lesions on the tongue, gums, cheeks, upper palate and hooves. Haemorrhagic tracheitis and ecchymotic haemorrhages on the heart were characteristic. The vesicular lesions of oral cavity were positive for 'O' type of FMD virus by sandwich enzyme-linked immuno sorbent assay (ELISA). The heart blood and spleen samples yielded growth of pure cultures of P. multocida. The isolates were typed as P. multocida type B using KTSP61 and KTT72 primers yielding specific amplicons of 620 bp. The phylogenetic analysis of the isolates was carried by sequencing of 1.4-Kbp nucleotides on the 16S ribosomal RNA (rRNA) gene of the isolates.

  16. Model for ranking freshwater fish farms according to their risk of infection and illustration for viral haemorrhagic septicaemia.

    PubMed

    Oidtmann, Birgit C; Pearce, Fiona M; Thrush, Mark A; Peeler, Edmund J; Ceolin, Chiara; Stärk, Katharina D C; Dalla Pozza, Manuela; Afonso, Ana; Diserens, Nicolas; Reese, R Allan; Cameron, Angus

    2014-08-01

    We developed a model to calculate a quantitative risk score for individual aquaculture sites. The score indicates the risk of the site being infected with a specific fish pathogen (viral haemorrhagic septicaemia virus (VHSV); infectious haematopoietic necrosis virus, Koi herpes virus), and is intended to be used for risk ranking sites to support surveillance for demonstration of zone or member state freedom from these pathogens. The inputs to the model include a range of quantitative and qualitative estimates of risk factors organised into five risk themes (1) Live fish and egg movements; (2) Exposure via water; (3) On-site processing; (4) Short-distance mechanical transmission; (5) Distance-independent mechanical transmission. The calculated risk score for an individual aquaculture site is a value between zero and one and is intended to indicate the risk of a site relative to the risk of other sites (thereby allowing ranking). The model was applied to evaluate 76 rainbow trout farms in 3 countries (42 from England, 32 from Italy and 2 from Switzerland) with the aim to establish their risk of being infected with VHSV. Risk scores for farms in England and Italy showed great variation, clearly enabling ranking. Scores ranged from 0.002 to 0.254 (mean score 0.080) in England and 0.011 to 0.778 (mean of 0.130) for Italy, reflecting the diversity of infection status of farms in these countries. Requirements for broader application of the model are discussed. Cost efficient farm data collection is important to realise the benefits from a risk-based approach. Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

  17. Replacement of the human cytomegalovirus promoter with fish enhancer and core elements to control the expression of the G gene of viral haemorrhagic septicemia virus (VHSV).

    PubMed

    Martinez-Lopez, A; Chinchilla, B; Encinas, P; Gomez-Casado, E; Estepa, A; Coll, J M

    2012-12-15

    This work explores some of the possibilities to replace human cytomegalovirus (CMV) core and/or enhancer promoter control elements to create new expression vectors for use with fish. The work is relevant to fish vaccination, since DNA vaccines use eukaryotic expression plasmids controlled by the human cytomegalovirus (CMV) promoter to be effective against novirhabdoviruses, such as viral haemorrhagic septicemia virus (VHSV), one of the most devastating fish viral European diseases. To reduce possible homologous recombination with fish genome, core and enhancer sequences from fish origin, such as trout interferon-inducible myxovirus protein (Mx), zebrafish retrovirus long terminal repeat (LTR) and carp β-actin (AE6), were combined with those of CMV to design alternative hybrid promoters. The substitution of CMV core and/or enhancer with the corresponding elements of Mx or the LTR core maintained a similar in vitro protein G expression level than that obtained by using the CMV promoter. Vectors using the dsRNA-inducible Mx enhancer followed either by the LTR or the AE6 cores showed the highest in vitro protein G expression levels. Furthermore, synthetic constructs using the Mx enhancer maintained their polyI:C induction capabilities despite the core used. Some of these hybrid promoters might contribute to the development of all-fish-vectors for DNA vaccines while others might be useful for more basic studies.

  18. Resistance to a rhabdovirus (VHSV) in rainbow trout: identification of a major QTL related to innate mechanisms.

    PubMed

    Verrier, Eloi R; Dorson, Michel; Mauger, Stéphane; Torhy, Corinne; Ciobotaru, Céline; Hervet, Caroline; Dechamp, Nicolas; Genet, Carine; Boudinot, Pierre; Quillet, Edwige

    2013-01-01

    Health control is a major issue in animal breeding and a better knowledge of the genetic bases of resistance to diseases is needed in farm animals including fish. The detection of quantitative trait loci (QTL) will help uncovering the genetic architecture of important traits and understanding the mechanisms involved in resistance to pathogens. We report here the detection of QTL for resistance to Viral Haemorrhagic Septicaemia Virus (VHSV), a major threat for European aquaculture industry. Two induced mitogynogenetic doubled haploid F2 rainbow trout (Oncorhynchus mykiss) families were used. These families combined the genome of susceptible and resistant F0 breeders and contained only fully homozygous individuals. For phenotyping, fish survival after an immersion challenge with the virus was recorded, as well as in vitro virus replication on fin explants. A bidirectional selective genotyping strategy identified seven QTL associated to survival. One of those QTL was significant at the genome-wide level and largely explained both survival and viral replication in fin explants in the different families of the design (up to 65% and 49% of phenotypic variance explained respectively). These results evidence the key role of innate defence in resistance to the virus and pave the way for the identification of the gene(s) responsible for resistance. The identification of a major QTL also opens appealing perspectives for selective breeding of fish with improved resistance.

  19. Resistance to a Rhabdovirus (VHSV) in Rainbow Trout: Identification of a Major QTL Related to Innate Mechanisms

    PubMed Central

    Verrier, Eloi R.; Dorson, Michel; Mauger, Stéphane; Torhy, Corinne; Ciobotaru, Céline; Hervet, Caroline; Dechamp, Nicolas; Genet, Carine; Boudinot, Pierre; Quillet, Edwige

    2013-01-01

    Health control is a major issue in animal breeding and a better knowledge of the genetic bases of resistance to diseases is needed in farm animals including fish. The detection of quantitative trait loci (QTL) will help uncovering the genetic architecture of important traits and understanding the mechanisms involved in resistance to pathogens. We report here the detection of QTL for resistance to Viral Haemorrhagic Septicaemia Virus (VHSV), a major threat for European aquaculture industry. Two induced mitogynogenetic doubled haploid F2 rainbow trout (Oncorhynchus mykiss) families were used. These families combined the genome of susceptible and resistant F0 breeders and contained only fully homozygous individuals. For phenotyping, fish survival after an immersion challenge with the virus was recorded, as well as in vitro virus replication on fin explants. A bidirectional selective genotyping strategy identified seven QTL associated to survival. One of those QTL was significant at the genome-wide level and largely explained both survival and viral replication in fin explants in the different families of the design (up to 65% and 49% of phenotypic variance explained respectively). These results evidence the key role of innate defence in resistance to the virus and pave the way for the identification of the gene(s) responsible for resistance. The identification of a major QTL also opens appealing perspectives for selective breeding of fish with improved resistance. PMID:23390526

  20. Genetically similar VHSV isolates are differentially virulent in olive flounder Paralichthys olivaceus.

    PubMed

    Cho, Mi Young; Lee, Unn Hwa; Moon, Chang Hoon; Bang, Jong Deuk; Jee, Bo Young; Cha, Seung Ju; Kim, Jin Woo; Park, Myoung Ae; Do, Jeong Wan; Park, Jeong Woo

    2012-11-08

    Two viral hemorrhagic septicemia virus (VHSV) isolates, VHSV-KR-CJA and VHSV-KR-YGH, were isolated from viral hemorrhagic septicemia disease outbreaks in flounder farms in South Korea. The VHSV-KR-CJA isolate was isolated from a flounder farm with high mortality (80%), while the VHSV-KR-YGH isolate was isolated from a flounder farm with low mortality (15%), suggesting that these isolates differ in virulence. The virulence of these isolates was evaluated in juvenile flounder via intraperitoneal injection. Consistent with their virulence in the field, mortality data revealed that the VHSV-KR-CJA isolate was highly pathogenic (cumulative mortality of 80%), while the VHSV-KR-YGH isolate was less pathogenic in flounder (cumulative mortality of 20%). To characterize the genotypes of these viruses, the full open reading frames (ORFs) encoding nucleoprotein N, phosphoprotein P, matrix protein M, glycoprotein G, nonstructural viral protein NV, and polymerase L of these viruses were sequenced and analyzed. Sequence analysis revealed that both isolates are genetically very similar (identical amino acid sequences for P, M, NV, and L and >99.7 and 99.8% amino acid sequence identity for N and G, respectively). Phylogenetic analysis indicated that both of these viruses belong to the Genotype IVa group, suggesting that they originated from a common ancestral virus. The low pathogenicity VHSV strain may potentially evolve to become a more pathogenic strain through only a few nucleotide substitutions. Further functional analyses of mutations in VHSV genes are necessary to identify factors that determine VHSV pathogenicity in flounder.

  1. Salmonid fish viruses and cell interactions at early steps of the infective cycle.

    PubMed

    de las Heras, A I; Rodríguez Saint-Jean, S; Pérez-Prieto, S I

    2008-07-01

    A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.

  2. Sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral hemorrhagic septicemia virus, a fish rhabdovirus

    USGS Publications Warehouse

    Benmansour, A.; Bascuro, B.; Monnier, A.F.; Vende, P.; Winton, J.R.; de Kinkelin, P.

    1997-01-01

    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 the genus Lyssavirus. Compared with each other, the G genes of VHSV displayed a dissimilar overall genetic diversity which correlated with differences in geographical origin. The multiple sequence alignment of the complete G protein, showed that the divergent positions were not uniformly distributed along the sequence. A central region (amino acid position 245-300) accumulated substitutions and appeared to be highly variable. The genetic heterogeneity within a single isolate was high, with an apparent internal mutation frequency of 1.2 x 10(-3) per nucleotide site, attesting the quasispecies nature of the viral population. The phylogeny separated VHSV strains according to the major geographical area of isolation: genotype I for continental Europe, genotype II for the British Isles, and genotype III for North America. Isolates from continental Europe exhibited the highest genetic variability, with sub-groups correlated partially with the serological classification. Neither neutralizing polyclonal sera, nor monoclonal antibodies, were able to discriminate between the genotypes. The overall structure of the phylogenetic tree suggests that VHSV genetic diversity and evolution fit within the model of random change and positive selection operating on quasispecies.

  3. In vitro search for alternative promoters to the human immediate early cytomegalovirus (IE-cMV) to express the G gene of viral haemorrhagic septicemia virus (VHSV) in fish epithelial cells.

    PubMed

    Ruiz, S; Tafalla, C; Cuesta, A; Estepa, A; Coll, J M

    2008-12-02

    Present DNA vaccines against fish rhabdoviruses require intramuscular injection (fish-to-fish vaccination) of their G-protein gene under the control of the human immediate early cytomegalovirus (IE-CMV) promoter, while immersion delivery (mass DNA vaccination), for instance, by using fish epithelial-specific promoters, would be more practical for aquaculture. To find fish epithelial-specific promoters alternative to the IE-CMV, a comparative study of the effectiveness of different fish promoters constitutively expressing the G gene of the viral haemorrhagic septicemia virus (VHSV) in the epithelial papulosum cyprini (EPC) cell line was performed. The study included MCV1.4 (an alternative IE-CMV promoter version), AE6 (a version of the carp beta-actin promoter), long terminal repeats (LTR) of zebrafish or walleye retroviruses, trout Mx1, carp myosin-heavy-chain and flatfish pleurocidin promoters and salmonid sleeping beauty (SB)/medaka Tol2 transposon repeats. The G-protein expression in transfected EPC cells was studied by estimating the number of cells expressing the G-protein in their membrane and the average expression level per cell. In addition, in an attempt to reduce their sizes, some regions of the MCV1.4 and AE6 promoters were deleted and expression levels compared to those observed for full-length promoters. Since both zebrafish LTR and carp AE6 promoters were the most effective regulatory sequences for expressing the VHSV G-protein in EPC cells, these sequences might be candidates for new DNA vaccine vectors for fish epithelial tissues avoiding the IE-CMV promoter. Furthermore, known transcription factor binding sites (TFBS) common to most of the fish G-expressing promoters, might enable the future design of fully synthetic or hybrid promoters with improved efficacy of VHSV G-protein expression in epithelial fish cells.

  4. Potential Reservoirs and Risk Factors for VHSV IVb in an Enzootic System: Budd Lake, Michigan.

    PubMed

    Throckmorton, Elizabeth; Brenden, Travis; Peters, Amber K; Newcomb, Tammy J; Whelan, Gary E; Faisal, Mohamed

    2017-03-01

    Viral hemorrhagic septicemia virus genotype IVb (VHSV IVb) has caused major, sporadic fish die-offs in the Laurentian Great Lakes region of North America since 2005. Presently, factors affecting VHSV IVb persistence in enzootic systems are not well understood. Even with annual surveillance, the virus can go undetected for several years after an outbreak before again re-emerging, which suggests that the virus is maintained in the system either below detectable levels or in untested reservoirs. The aim of this study was to identify potential reservoirs of VHSV IVb in Budd Lake, Michigan; VHSV IVb was first detected in Budd Lake in 2007 but remained undetected until 2011. Additionally, we explored the susceptibility of naive fish introduced into a water body enzootic for VHSV IVb by stocking age-0 Largemouth Bass Micropterus salmoides at varying densities into enclosures in the lake. The virus was not detected among samples of the fishes Notropis spp. and Lepomis spp., cylindrical papershell mussels Anodontoides ferussacianus, leeches (subclass Hirudinea), sediment, or water. However, the virus was successfully isolated from amphipods (family Hyalellidae) and Largemouth Bass held in the enclosures. Our finding of VHSV IVb in Hyalellidae amphipods in combination with other research that has detected the virus in Diporeia spp., a large benthic amphipod important as a food resource to Great Lake fishes, suggests that benthic macroinvertebrates are a reservoir for VHSV IVb in infected systems. If there are environmental reservoirs for VHSV IVb in infected systems, they are likely unevenly distributed. Findings of this study add to our understanding of the seemingly complex ecology of this deadly and economically detrimental virus. Received February 22, 2016; accepted October 16, 2016.

  5. Role of the IFN I system against the VHSV infection in juvenile Senegalese sole (Solea senegalensis).

    PubMed

    Alvarez-Torres, Daniel; Podadera, Ana M; Bejar, Julia; Bandin, Isabel; Alonso, M Carmen; Garcia-Rosado, Esther

    2016-01-08

    Senegalese sole is susceptible to marine VHSV isolates but is not affected by freshwater isolates, which may indicate differences regarding virus-host immune system interaction. IFN I induces an antiviral state in fish, stimulating the expression of genes encoding antiviral proteins (ISG). In this study, the stimulation of the Senegalese sole IFN I by VHSV infections has been evaluated by the relative quantification of the transcription of several ISG (Mx, Isg15 and Pkr) after inoculation with marine (pathogenic) and freshwater (non-pathogenic) VHSV isolates. Compared to marine VHSV, lower levels of RNA of the freshwater VHSV induced transcription of ISG to similar levels, with the Isg15 showing the highest fold induction. The protective role of the IFN I system was evaluated in poly I:C-inoculated animals subsequently challenged with VHSV isolates. The cumulative mortality caused by the marine isolate in the control group was 68%, whereas in the poly I:C-stimulated group was 5%. The freshwater VHSV isolate did not cause any mortality. Furthermore, viral RNA fold change and viral titers were lower in animals from the poly I:C + VHSV groups than in the controls. The implication of the IFN I system in the protection observed was confirmed by the transcription of the ISG in animals from the poly I:C + VHSV groups. However, the marine VHSV isolate exerts a negative effect on the ISG transcription at 3 and 6 h post-inoculation (hpi), which is not observed for the freshwater isolate. This difference might be partly responsible for the virulence shown by the marine isolate.

  6. Can VHS Virus Bypass the Protective Immunity Induced by DNA Vaccination in Rainbow Trout?

    PubMed

    Sepúlveda, Dagoberto; Lorenzen, Niels

    2016-01-01

    DNA vaccines encoding viral glycoproteins have been very successful for induction of protective immunity against diseases caused by rhabdoviruses in cultured fish species. However, the vaccine concept is based on a single viral gene and since RNA viruses are known to possess high variability and adaptation capacity, this work aimed at evaluating whether viral haemorrhagic septicaemia virus (VHSV), an RNA virus and member of Rhabdoviridae family, was able to evade the protective immune response induced by the DNA vaccination of rainbow trout. The experiments comprised repeated passages of a highly pathogenic VHSV isolate in a fish cell line in the presence of neutralizing fish serum (in vitro approach), and in rainbow trout immunized with the VHS DNA vaccine (in vivo approach). For the in vitro approach, the virus collected from the last passage (passaged virus) was as sensitive as the parental virus to serum neutralization, suggesting that the passaging did not promote the selection of virus populations able to bypass the neutralization by serum antibodies. Also, in the in vivo approach, where virus was passaged several times in vaccinated fish, no increased virulence nor increased persistence in vaccinated fish was observed in comparison with the parental virus. However, some of the vaccinated fish did get infected and could transmit the infection to naïve cohabitant fish. The results demonstrated that the DNA vaccine induced a robust protection, but also that the immunity was non-sterile. It is consequently important not to consider vaccinated fish as virus free in veterinary terms.

  7. Can VHS Virus Bypass the Protective Immunity Induced by DNA Vaccination in Rainbow Trout?

    PubMed Central

    Sepúlveda, Dagoberto; Lorenzen, Niels

    2016-01-01

    DNA vaccines encoding viral glycoproteins have been very successful for induction of protective immunity against diseases caused by rhabdoviruses in cultured fish species. However, the vaccine concept is based on a single viral gene and since RNA viruses are known to possess high variability and adaptation capacity, this work aimed at evaluating whether viral haemorrhagic septicaemia virus (VHSV), an RNA virus and member of Rhabdoviridae family, was able to evade the protective immune response induced by the DNA vaccination of rainbow trout. The experiments comprised repeated passages of a highly pathogenic VHSV isolate in a fish cell line in the presence of neutralizing fish serum (in vitro approach), and in rainbow trout immunized with the VHS DNA vaccine (in vivo approach). For the in vitro approach, the virus collected from the last passage (passaged virus) was as sensitive as the parental virus to serum neutralization, suggesting that the passaging did not promote the selection of virus populations able to bypass the neutralization by serum antibodies. Also, in the in vivo approach, where virus was passaged several times in vaccinated fish, no increased virulence nor increased persistence in vaccinated fish was observed in comparison with the parental virus. However, some of the vaccinated fish did get infected and could transmit the infection to naïve cohabitant fish. The results demonstrated that the DNA vaccine induced a robust protection, but also that the immunity was non-sterile. It is consequently important not to consider vaccinated fish as virus free in veterinary terms. PMID:27054895

  8. Replication and persistence of VHSV IVb in freshwater turtles.

    PubMed

    Goodwin, Andrew E; Merry, Gwenn E

    2011-05-09

    With the emergence of viral hemorrhagic septicemia virus (VHSV) strain IVb in the Great Lakes of North America, hatchery managers have become concerned that this important pathogen could be transmitted by animals other than fish. Turtles are likely candidates because they are poikilotherms that feed on dead fish, but there are very few reports of rhabdovirus infections in reptiles and no reports of the fish rhabdoviruses in animals other than teleosts. We injected common snapping turtles Chelydra serpentine and red-eared sliders Trachemys scripta elegans intraperitoneally with 10(4) median tissue culture infectious dose (TCID50) of VHSV-IVb and 21 d later were able to detect the virus by quantitative real-time reverse transcriptase PCR (qrt-RTPCR) in pools of kidney, liver, and spleen. In a second experiment, snapping turtles, red-eared sliders, yellow-bellied sliders T. scripta scripta, and northern map turtles Grapetemys geographica at 14 degrees C were allowed to feed on tissues from bluegill dying of VHSV IVb disease. Turtle kidney, spleen, and brain pools were not positive by qrt-RTPCR on Day 3 post feeding, but were positive on Days 10 and 20. Map turtles on Day 20 post-feeding were positive by both qrt-RTPCR and by cell culture. Our work shows that turtles that consume infected fish are a possible vector for VHSV IVb, and that the fish rhabdoviruses may have a broader host range than previously suspected.

  9. Septicaemia in the Neutropenic Patient

    PubMed Central

    Atkinson, K.; Kay, H. E. M.; McElwain, T. J.

    1974-01-01

    A total of 29 cases of septicaemia proved by blood culture in 22 severely neutropenic patients with acute leukaemia or aplastic anaemia have been studied. The recovery rate was 75% in the Gram-positive septicaemias and 60% in the Gram-negative septicaemias in which treatment response could be evaluated. Neutropenia predisposed to septicaemia and its degree seemed to be important. The underlying state of the bone marrow was an important prognostic factor; the neutrophil count at the time of diagnosis and the infecting organism were less important. Gentamicin was the single most useful antibiotic, and the infection was controlled largely with gentamicin and one other antibiotic, most often carbenicillin. Possibly a similar result could have been obtained with gentamicin alone, but since the bacterial flora in a given environment is changeable empirical antibiotic regimens should remain flexible. PMID:4846132

  10. Vaccine Adjuvants in Fish Vaccines Make a Difference: Comparing Three Adjuvants (Montanide ISA763A Oil, CpG/Poly I:C Combo and VHSV Glycoprotein) Alone or in Combination Formulated with an Inactivated Whole Salmonid Alphavirus Antigen.

    PubMed

    Thim, Hanna L; Villoing, Stéphane; McLoughlin, Marian; Christie, Karen Elina; Grove, Søren; Frost, Petter; Jørgensen, Jorunn B

    2014-03-25

    Most commercial vaccines offered to the aquaculture industry include inactivated antigens (Ag) formulated in oil adjuvants. Safety concerns are related to the use of oil adjuvants in multivalent vaccines for fish, since adverse side effects (e.g., adhesions) can appear. Therefore, there is a request for vaccine formulations for which protection will be maintained or improved, while the risk of side effects is reduced. Here, by using an inactivated salmonid alphavirus (SAV) as the test Ag, the combined use of two Toll-like receptor (TLR) ligand adjuvants, CpG oligonucleotides (ODNs) and poly I:C, as well as a genetic adjuvant consisting of a DNA plasmid vector expressing the viral haemorrhagic septicaemia virus (VHSV) glycoprotein (G) was explored. VHSV-G DNA vaccine was intramuscularly injected in combination with intraperitoneal injection of either SAV Ag alone or combined with the oil adjuvant, Montanide ISA763, or the CpG/polyI:C combo. Adjuvant formulations were evaluated for their ability to boost immune responses and induce protection against SAV in Atlantic salmon, following cohabitation challenge. It was observed that CpG/polyI:C-based formulations generated the highest neutralizing antibody titres (nAbs) before challenge, which endured post challenge. nAb responses for VHSV G-DNA- and oil-adjuvanted formulations were marginal compared to the CpG/poly I:C treatment. Interestingly, heat-inactivated sera showed reduced nAb titres compared to their non-heated counterparts, which suggests a role of complement-mediated neutralization against SAV. Consistently elevated levels of innate antiviral immune genes in the CpG/polyI:C injected groups suggested a role of IFN-mediated responses. Co-delivery of the VHSV-G DNA construct with either CpG/polyI:C or oil-adjuvanted SAV vaccine generated higher CD4 responses in head kidney at 48 h compared to injection of this vector or SAV Ag alone. The results demonstrate that a combination of pattern recognizing receptor (PRR

  11. Vaccine Adjuvants in Fish Vaccines Make a Difference: Comparing Three Adjuvants (Montanide ISA763A Oil, CpG/Poly I:C Combo and VHSV Glycoprotein) Alone or in Combination Formulated with an Inactivated Whole Salmonid Alphavirus Antigen

    PubMed Central

    Thim, Hanna L.; Villoing, Stéphane; McLoughlin, Marian; Christie, Karen Elina; Grove, Søren; Frost, Petter; Jørgensen, Jorunn B.

    2014-01-01

    Most commercial vaccines offered to the aquaculture industry include inactivated antigens (Ag) formulated in oil adjuvants. Safety concerns are related to the use of oil adjuvants in multivalent vaccines for fish, since adverse side effects (e.g., adhesions) can appear. Therefore, there is a request for vaccine formulations for which protection will be maintained or improved, while the risk of side effects is reduced. Here, by using an inactivated salmonid alphavirus (SAV) as the test Ag, the combined use of two Toll-like receptor (TLR) ligand adjuvants, CpG oligonucleotides (ODNs) and poly I:C, as well as a genetic adjuvant consisting of a DNA plasmid vector expressing the viral haemorrhagic septicaemia virus (VHSV) glycoprotein (G) was explored. VHSV-G DNA vaccine was intramuscularly injected in combination with intraperitoneal injection of either SAV Ag alone or combined with the oil adjuvant, Montanide ISA763, or the CpG/polyI:C combo. Adjuvant formulations were evaluated for their ability to boost immune responses and induce protection against SAV in Atlantic salmon, following cohabitation challenge. It was observed that CpG/polyI:C-based formulations generated the highest neutralizing antibody titres (nAbs) before challenge, which endured post challenge. nAb responses for VHSV G-DNA- and oil-adjuvanted formulations were marginal compared to the CpG/poly I:C treatment. Interestingly, heat-inactivated sera showed reduced nAb titres compared to their non-heated counterparts, which suggests a role of complement-mediated neutralization against SAV. Consistently elevated levels of innate antiviral immune genes in the CpG/polyI:C injected groups suggested a role of IFN-mediated responses. Co-delivery of the VHSV-G DNA construct with either CpG/polyI:C or oil-adjuvanted SAV vaccine generated higher CD4 responses in head kidney at 48 h compared to injection of this vector or SAV Ag alone. The results demonstrate that a combination of pattern recognizing receptor (PRR

  12. Interaction of the attenuated recombinant rIHNV-Gvhsv GFP virus with macrophages from rainbow trout (Oncorhynchus mykiss).

    PubMed

    Romero, Alejandro; Dios, Sonia; Bremont, Michel; Figueras, Antonio; Novoa, Beatriz

    2011-03-15

    One of the most important threats to the salmonid aquaculture industry is infection caused by novirhabdoviruses such as infectious haematopoietic necrosis virus (IHNV) or viral haemorrhagic septicaemia virus (VHSV). Using reverse genetics, an avirulent recombinant rIHNV-Gvhsv GFP strain was generated, which was able to replicate as effectively as wild type IHNV in a fish cell line and in macrophages. Although this recombinant virus induced protective responses against IHNV and VHSV, the response did not involve the production of antibodies or modulate the expression of some antiviral genes. To determine the immune mechanisms underlying the protection conferred by the rIHNV-Gvhsv GFP virus, different immune parameters (NO production, respiratory burst activity and the induction of apoptosis) were assessed in the macrophage population. The results obtained in the present work may indicate that the Nv protein could be important in the modulation of NO and ROS production. rIHNV-Gvhsv GFP did not appear to have a clear effect on nitric oxide production or apoptosis. However, an increased respiratory burst activity (with levels induced by the recombinant virus significantly higher than the levels induced by the wild type virus), suggests a stimulation of the macrophage population, which could be related to the protection against virulent viruses.

  13. Low prevalence of VHSV detected in round goby collected in offshore regions of Lake Ontario

    USGS Publications Warehouse

    Cornwell, Emily R.; Getchell, Rodman G.; Groocock, Geoffrey H.; Walsh, Maureen G.; Bowser, Paul R.

    2012-01-01

    Since the first reports of mortalities due to viral hemorrhagic septicemia virus (VHSV) type IVb in the Laurentian Great Lakes basin during 2005 (Lake St. Clair, USA and Bay of Quinte, Lake Ontario, Canada), many groups have conducted surveillance efforts for the virus, primarily in nearshore areas. The round goby (Neogobius melanostomus) has been identified as a key species to target for surveillance, because they have a very high probability of infection at a given site. Our objective in this study was to document and quantify VHSV in round gobies in offshore waters of Lake Ontario using molecular techniques. We collected 139 round gobies from depths ranging from 55 to 150 m using bottom trawls during the early spring of 2011 and detected VHSV in 4 individuals (1/26 fish at 95 m, 2/12 fish at 105 m, and 1/24 fish at 135 m). These results expand the known depth range of VHSV in the Great Lakes. They also have implications on the management of the spread of VHSV within infected bodies of water related to the mixing of populations of fish that would remain distinct in their breeding habitats, but then have the opportunity to mix in their overwintering habitats, as well as to increase overlap of predator and prey species in overwintering habitats.

  14. Burkholderia pseudomallei septicaemia - a case report.

    PubMed

    Dias, M; Antony, B; Aithala, S; Hanumanthappa, B; Pinto, H; Rekha, B

    2004-01-01

    Burkholderia pseudomallei, a natural saprophyte widely distributed in soil, stagnant waters of endemic areas, is said to infect humans through breaks in the skin or through inhalation causing protean clinical manifestations including fatal septicaemia. A case of septicaemia in a elderly female diabetic due to B. pseudomallei following a history of fall is being reported with complete details.

  15. Comparative immunogenecity of foot and mouth disease virus antigens in FMD-haemorrhagic septicaemia combined vaccine and FMD vaccine alone in buffalo calves.

    PubMed

    Chhabra, Rajesh; Sharma, R; Kakker, N K

    2004-03-01

    Humoral immune response was evaluated by monitoring the serum antibody titres and virus specific IgM titres against Foot and Mouth Disease (FMD) virus antigens in serum samples obtained from different groups of calves inoculated with combined vaccine or FMD vaccine alone, on 0, 7, 14, 21, 28, 42 and 56 days post-vaccination (DPV). The cellular immune response was monitored by MTT based lymphoproliferation in peripheral blood mononuclear cell cultures. Higher liquid phase blocking (LPB) ELISA antibody titres were observed in calves receiving combined vaccine as compared to calves immunized with FMD vaccine alone with the peak titres in both the groups obtained on 21 days post-vaccination. However, the virus specific IgM titres were significantly higher in group of calves inoculated with combined vaccine than FMD vaccine alone. The lymphoproliferative responses against FMDV types O, A22 and Asia 1 in the groups receiving combined vaccine and FMD vaccine alone started increasing gradually after day 14 and reached peak levels on 28 DPV followed by a gradual decline subsequently. The group receiving combined vaccine showed higher proliferative responses on in vitro stimulation with FMD virus type O, whereas, with FMD virus type Asia 1, the responses were significantly higher on 14 and 21 DPV as compared to the group immunized with FMD vaccine alone. However, in the group receiving combined vaccine, the responses on in vitro stimulation with FMD virus type A22 were significantly higher than FMD vaccine alone group on all DPV except on 42 DPV.

  16. VHSV G glycoprotein major determinants implicated in triggering the host type I IFN antiviral response as DNA vaccine molecular adjuvants.

    PubMed

    Martinez-Lopez, A; Garcia-Valtanen, P; Ortega-Villaizan, M; Chico, V; Gomez-Casado, E; Coll, J M; Estepa, A

    2014-10-14

    We have recently identified the two major determinants of the glycoprotein G of the viral hemorrhagic septicaemia rhabdovirus (gpGVHSV), peptides p31 and p33 implicated in triggering the host type I IFN antiviral response associated to these rhabdoviral antigens. With the aim to investigate the properties of these viral glycoprotein regions as DNA molecular adjuvants, their corresponding cDNA sequences were cloned into a plasmid (pMCV1.4) flanked by the signal peptide and transmembrane sequences of gpGVHSV. In addition, a plasmid construct encoding both sequences p31 and p33 (pMCV1.4-p31+p33) was also designed. In vitro transitory cell transfection assays showed that these VHSV gpG regions were able to induce the expression of type I IFN stimulated genes as well as to confer resistance to the infection with a different fish rhabdovirus, the spring viremia of carp virus (SVCV). In vivo, zebrafish intramuscular injection of only 1μg of the construct pMCV1.4-p31+p33 conferred fish protection against SVCV lethal challenge up to 45 days post-immunization. Moreover, pMCV1.4-p31+p33 construct was assayed for molecular adjuvantcity's for a DNA vaccine against SVCV based in the surface antigen of this virus (pAE6-GSVCV). The results showed that the co-injection of the SVCV DNA vaccine and the molecular adjuvant allowed (i) a ten-fold reduction in the dose of pAE6-Gsvcv without compromising its efficacy (ii) an increase in the duration of protection, and (iii) an increase in the survival rate. To our knowledge, this is the first report in which specific IFN-inducing regions from a viral gpG are used to design more-efficient and cost-effective viral vaccines, as well as to improve our knowledge on how to stimulate the innate immune system.

  17. A reverse genetics system for the Great Lakes strain of viral hemorrhagic septicemia virus: the NV gene is required for pathogenicity

    USGS Publications Warehouse

    Ammayappan, Arun; Kurath, Gael; Thompson, Tarin M.; Vakharia, Vikram N.

    2011-01-01

    Viral hemorrhagic septicemia virus (VHSV), belonging to the genus Novirhabdovirus in the family of Rhabdoviridae, causes a highly contagious disease of fresh and saltwater fish worldwide. Recently, a novel genotype of VHSV, designated IVb, has invaded the Great Lakes in North America, causing large-scale epidemics in wild fish. An efficient reverse genetics system was developed to generate a recombinant VHSV of genotype IVb from cloned cDNA. The recombinant VHSV (rVHSV) was comparable to the parental wild-type strain both in vitro and in vivo, causing high mortality in yellow perch (Perca flavescens). A modified recombinant VHSV was generated in which the NV gene was substituted with an enhanced green fluorescent protein gene (rVHSV-ΔNV-EGFP), and another recombinant was made by inserting the EGFP gene into the full-length viral clone between the P and M genes (rVHSV-EGFP). The in vitro replication kinetics of rVHSV-EGFP was similar to rVHSV; however, the rVHSV-ΔNV-EGFP grew 2 logs lower. In yellow perch challenges, wtVHSV and rVHSV induced 82-100% cumulative per cent mortality (CPM), respectively, whereas rVHSV-EGFP produced 62% CPM and rVHSV-ΔNV-EGFP caused only 15% CPM. No reversion of mutation was detected in the recovered viruses and the recombinant viruses stably maintained the foreign gene after several passages. These results indicate that the NV gene of VHSV is not essential for viral replication in vitro and in vivo, but it plays an important role in viral replication efficiency and pathogenicity. This system will facilitate studies of VHSV replication, virulence, and production of viral vectored vaccines.

  18. Antiviral activity produced by an IPNV-carrier EPC cell culture confers resistance to VHSV infection.

    PubMed

    Jurado, María Teresa; García-Valtanen, Pablo; Estepa, Amparo; Perez, Luis

    2013-10-25

    Infectious pancreatic necrosis virus (IPNV), a fish birnavirus, can establish a persistent infection on epithelioma papulosum cyprinid (EPC) cells producing a carrier state where a small fraction of IPNV-infected cells is maintained in the culture after continuous subculture. The EPC(IPNV) cells are resistant to challenge with IPNV as well as to challenge with viral hemorrhagic septicemia virus (VHSV), a rhabdovirus. In this work, the antiviral effect of the IPNV carrier culture conditioned medium (EPC(IPNV)-CM) was tested and analyzed in detail. EPC cells treated with the carrier culture supernatant become protected against VHSV challenge. Size-fractionation by filtration and acid and heat treatment showed that the IPNV persistently infected cells release an acid-resistant soluble factor in the molecular weight fraction bellow 50 kDa. The capacity of the EPC(IPNV)-CM to induce cytokine genes in EPC cells was also determined by real-time RT-PCR. We found that there is a positive correlation between up-regulation of mx gene expression in EPC cells treated with EPC(IPNV)-CM and protection against VHSV challenge. Our findings indicate that the control of IPNV multiplication in the carrier culture as well as the interference with rhabdovirus replication are connected to the production and release of an antiviral (interferon-like) factor to the medium.

  19. About viral hemorrhagic septicemia (VHS) virus. Potential threat of Great Lakes VHS virus in Western United States

    USGS Publications Warehouse

    Bartholomew, Jerri L; Kurath, Gael; Emmenegger, Evi

    2011-01-01

    Viral hemorrhagic septicemia (VHS) is a disease caused by a virus (VHSV). There are different strains of the virus that can infect marine and freshwater fish species, and the different strains may affect species differently. VHSV has recently invaded the Great Lakes, resulting in many large-scale fish die-offs and new regulatory restrictions for aquaculture throughout the region.

  20. Immunization of olive flounder (Paralichthys olivaceus) with an auxotrophic Edwardsiella tarda mutant harboring the VHSV DNA vaccine.

    PubMed

    Choi, Seung Hyuk; Kim, Min Sun; Kim, Ki Hong

    2012-09-01

    The aims of the present study were to find more powerful promoter for DNA vaccines in olive flounder (Paralichthys olivaceus) and to evaluate the availability of the auxotrophic Edwardsiella tarda mutant (Δalr Δasd E. tarda) as a delivery vehicle for DNA vaccine against VHSV in olive flounder. The marine medaka (Oryzias dancena) β-actin promoter was clearly stronger than cytomegalovirus (CMV) promoter when the vectors were transfected to Epithelioma papulosum cyprini (EPC) cells or injected into the muscle of olive flounder, suggesting that marine medaka β-actin promoter would be more appropriate promoter for DNA vaccines in olive flounder than CMV promoter. Olive flounder immunized with the Δalr Δasd E. tarda harboring viral hemorrhagic septicemia virus (VHSV) DNA vaccine vector driven by the marine medaka β-actin promoter showed significantly higher serum neutralization titer and higher survival rates against challenge with VHSV than fish immunized with the bacteria carrying VHSV DNA vaccine vector driven by CMV promoter. These results indicate that auxotrophic E. tarda mutant harboring marine medaka β-actin promoter-driven DNA vaccine vectors would be a potential system for prophylactics of infectious diseases in olive flounder.

  1. Citrobacter koseri septicaemia in a holstein calf.

    PubMed

    Komine, M; Massa, A; Moon, L; Mullaney, T

    2014-11-01

    A 4-day-old male Holstein calf with dull mentation, nystagmus and blindness was humanely destroyed and subject to necropsy examination. Gross lesions included severe suppurative meningitis characterized by diffuse cloudy thickening of the meninges, bilateral hypopyon and fibrinosuppurative polyarthritis affecting the hocks. Citrobacter koseri was isolated from the meninges, ocular fluid, synovial fluid, spleen and small intestine. Microscopically, there was neutrophilic and histiocytic meningitis with intralesional bacilli, endophthalmitis, neutrophilic splenitis and multiple renal microabscesses. Failure of passive transfer of colostrum was confirmed. This appears to be the first characterization of septicaemia in a calf caused by C. koseri, with lesions comparable with those described in human neonates.

  2. Capnocytophaga canimorsus: infection, septicaemia, recovery and reconstruction.

    PubMed

    Low, Stephanie Chiang-Mei; Greenwood, John Edward

    2008-07-01

    A case is presented of a life-threatening septicaemia and associated peripheral necrosing microembolic phenomenon, resulting from a dog lick to an insignificant burn wound. The isolated bacterium was Capnocytophaga canimorsus, a slow-growing Gram-negative bacillus commonly found in dog saliva. Any clinician seeing patients with a history of dog bite/saliva contact and progressive illness should consider this bacterium as a possible offender and take special care to elicit an accurate history, specifically including questions regarding animal contact.

  3. Atlantic salmon endothelial cells from the heart were more susceptible than fibroblasts from the bulbus arteriosus to four RNA viruses but protected from two viruses by dsRNA pretreatment.

    PubMed

    Pham, Phuc H; Tong, Winnie W L; Misk, Ehab; Jones, Ginny; Lumsden, John S; Bols, Niels C

    2017-09-04

    Heart diseases caused by viruses are major causes of Atlantic salmon aquaculture loss. Two Atlantic salmon cardiovascular cell lines, an endothelial cell line (ASHe) from the heart and a fibroblast cell line (BAASf) from the bulbus arteriosus, were evaluated for their response to four fish viruses, CSV, IPNV, VHSV IVa and VHSV IVb, and the innate immune agonist, double-stranded RNA mimic poly IC. All four viruses caused cytopathic effects in ASHe and BAASf. However, ASHe was more susceptible to all four viruses than BAASf. When comparing between the viruses, ASHe cells were found to be moderately susceptible to CSV and VHSV IVb, but highly susceptible to IPNV and VHSV IVa induced cell death. All four viruses were capable of propagating in the ASHe cell line, leading to increases in virus titre over time. In BAASf, CSV and IPNV produced more than one log increase in titre from initial infection, but VHSV IVb and IVa did not. When looking at the antiviral response of both cell lines, Mx proteins were induced in ASHe and BAASf by poly IC. All four viruses induced Mx proteins in BAASf, while only CSV and VHSV IVb induced Mx proteins in ASHe. IPNV and VHSV IVa suppressed Mx proteins expression in ASHe. Pretreatment of ASHe with poly IC to allow for Mx proteins accumulation protected the culture from subsequent infections with IPNV and VHSV IVa, resulting in delayed cell death, reduced virus titres and reduced viral proteins expression. These data suggest that endothelial cells potentially can serve as points of infections for viruses in the heart and that two of the four viruses, IPNV and VHSV IVa, have mechanisms to avoid or downregulate antiviral responses in ASHe cells. Furthermore, the high susceptibility of the ASHe cell line to IPNV and VHSV IVa can make it a useful tool for studying antiviral compounds against these viruses and for general detection of fish viruses. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Development of an aquatic pathogen database (AquaPathogen X) and its utilization in tracking emerging fish virus pathogens in North America

    USGS Publications Warehouse

    Emmenegger, E.J.; Kentop, E.; Thompson, T.M.; Pittam, S.; Ryan, A.; Keon, D.; Carlino, J.A.; Ranson, J.; Life, R.B.; Troyer, R.M.; Garver, K.A.; Kurath, G.

    2011-01-01

    The AquaPathogen X database is a template for recording information on individual isolates of aquatic pathogens and is freely available for download (http://wfrc.usgs.gov). This database can accommodate the nucleotide sequence data generated in molecular epidemiological studies along with the myriad of abiotic and biotic traits associated with isolates of various pathogens (e.g. viruses, parasites and bacteria) from multiple aquatic animal host species (e.g. fish, shellfish and shrimp). The cataloguing of isolates from different aquatic pathogens simultaneously is a unique feature to the AquaPathogen X database, which can be used in surveillance of emerging aquatic animal diseases and elucidation of key risk factors associated with pathogen incursions into new water systems. An application of the template database that stores the epidemiological profiles of fish virus isolates, called Fish ViroTrak, was also developed. Exported records for two aquatic rhabdovirus species emerging in North America were used in the implementation of two separate web-accessible databases: the Molecular Epidemiology of Aquatic Pathogens infectious haematopoietic necrosis virus (MEAP-IHNV) database (http://gis.nacse.org/ihnv/) released in 2006 and the MEAP- viral haemorrhagic septicaemia virus (http://gis.nacse.org/vhsv/) database released in 2010.

  5. Development of an aquatic pathogen database (AquaPathogen X) and its utilization in tracking emerging fish virus pathogens in North America.

    PubMed

    Emmenegger, E J; Kentop, E; Thompson, T M; Pittam, S; Ryan, A; Keon, D; Carlino, J A; Ranson, J; Life, R B; Troyer, R M; Garver, K A; Kurath, G

    2011-08-01

    The AquaPathogen X database is a template for recording information on individual isolates of aquatic pathogens and is freely available for download (http://wfrc.usgs.gov). This database can accommodate the nucleotide sequence data generated in molecular epidemiological studies along with the myriad of abiotic and biotic traits associated with isolates of various pathogens (e.g. viruses, parasites and bacteria) from multiple aquatic animal host species (e.g. fish, shellfish and shrimp). The cataloguing of isolates from different aquatic pathogens simultaneously is a unique feature to the AquaPathogen X database, which can be used in surveillance of emerging aquatic animal diseases and elucidation of key risk factors associated with pathogen incursions into new water systems. An application of the template database that stores the epidemiological profiles of fish virus isolates, called Fish ViroTrak, was also developed. Exported records for two aquatic rhabdovirus species emerging in North America were used in the implementation of two separate web-accessible databases: the Molecular Epidemiology of Aquatic Pathogens infectious haematopoietic necrosis virus (MEAP-IHNV) database (http://gis.nacse.org/ihnv/) released in 2006 and the MEAP- viral haemorrhagic septicaemia virus (http://gis.nacse.org/vhsv/) database released in 2010. © 2011 Blackwell Publishing Ltd.

  6. Emerging vesiculo-type virus infections of freshwater fishes in Europe.

    PubMed

    Betts, A M; Stone, D M; Way, K; Torhy, C; Chilmonczyk, S; Benmansour, A; de Kinkelin, P

    2003-12-29

    Rhabdoviruses were isolated from perch Perca fluviatilis and largemouth bass Micropterus salmoides exhibiting clinical signs of disease. Preliminary studies indicated that these viruses could be neutralised by antisera to perch rhabdovirus (Dorson et al. 1984) and may be similar to those previously isolated from grayling Thymallus thymallus and pike-perch Stizostedion stizostedion. The relationship between these viruses and the previously characterised fish rhabdoviruses, pike fry rhabdovirus (PFRV), spring viraemia of carp virus (SVCV) and lake trout rhabdovirus, was investigated. Viruses were propagated in bluegill fry (BF-2) cells and were characterised using electron microscopy, serum neutralisation tests, immunofluorescence tests, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and nucleotide sequence analysis. The bullet-shaped viral particles appeared to be compact, with spikes visible at the surface, a morphology similar to that of the vesiculovirus group of rhabdoviruses. Serum neutralisation tests showed that the viruses were antigenically closely related to the previously characterised perch rhabdovirus, but were not significantly neutralised by antisera to PFRV, SVCV or viral haemorrhagic septicaemia virus (VHSV). In immunofluorescence tests with perch rhabdovirus antisera, strong specific fluorescence was observed in cell cultures infected with the new rhabdovirus isolates, but no fluorescence was observed with antisera to PFRV, SVCV or VHSV. SDS-PAGE analysis revealed a polypeptide profile typical of vesiculoviruses, but the novel virus isolates had different relative mobilities of their P and M proteins compared to PFRV and SVCV. Nucleotide sequence analysis was carried out using reverse transcriptase-polymerase chain reaction (RT-PCR) and DNA sequencing of a 439 base-pair region of the viral L gene. The novel rhabdovirus isolates had <76% nucleotide sequence identity to PFRV, SVCV and lake trout rhabdovirus and >95% identity to

  7. Development and evaluation of a blocking enzyme-linked immunosorbent assay and virus neutralization assay to detect antibodies to viral hemorrhagic septicemia virus

    USGS Publications Warehouse

    Wilson, Anna; Goldberg, Tony; Marcquenski, Susan; Olson, Wendy; Goetz, Frederick; Hershberger, Paul; Hart, Lucas M.; Toohey-Kurth, Kathy

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a target of surveillance by many state and federal agencies in the United States. Currently, the detection of VHSV relies on virus isolation, which is lethal to fish and indicates only the current infection status. A serological method is required to ascertain prior exposure. Here, we report two serologic tests for VHSV that are nonlethal, rapid, and species independent, a virus neutralization (VN) assay and a blocking enzyme-linked immunosorbent assay (ELISA). The results show that the VN assay had a specificity of 100% and sensitivity of 42.9%; the anti-nucleocapsid-blocking ELISA detected nonneutralizing VHSV antibodies at a specificity of 88.2% and a sensitivity of 96.4%. The VN assay and ELISA are valuable tools for assessing exposure to VHSV.

  8. Viral hemorrhagic septicemia virus in North America

    USGS Publications Warehouse

    Meyers, Theodore R; Winton, James R.

    1995-01-01

    The first detections of viral hemorrhagic septicemia virus (VHSV) in North America were in Washington State from adult coho (Oncorhynchus kisutch) and chinook (O. tshawytscha) salmon in 1988. Subsequently, VHSV was isolated from adult coho salmon returning to hatcheries in the Pacific Northwest in 1989, 1991 and 1994. These isolates represented a strain of VHSV that was genetically different from European VHSV as determined by DNA sequence analysis and T1 ribonuclease fingerprinting. The North American strain of VHSV was also isolated from skin lesions of Pacific cod (Gadus macrocephalus) taken from Prince William Sound (PWS), Alaska in 1990, 1991 and 1993. In 1993 and 1994, the virus was isolated from Pacific herring (Clupea harengus pallasi) in Alaskan waters of PWS, Kodiak Island, Auke Bay and Port Frederick. During 1993 and 1994 the herring fishery in PWS failed from a probable complex of environmental stressors but VHSV isolates were associated with hemorrhages of the skin and fins in fish that returned to spawn. Also in 1993 and 1994, VHSV was isolated from apparently healthy stocks of herring in British Columbia, Canada and Puget Sound, Washington. Thus, the North American strain of VHSV is enzootic in the Northeastern Pacific Ocean among Pacific herring stocks with Pacific cod serving as a secondary reservoir. Although the North American strain of the virus appears to be moderately pathogenic for herring, causing occasional self-limiting epizootics, it was shown to be relatively avirulent for several species of salmonids. Pacific herring are common prey for cod and salmon and were most probably the source of the VHSV isolates from the adult salmon returning to spawn in rivers or at hatcheries in Washington State. Compelling circumstances involving the European isolates of VHSV suggest that this strain of the virus also is enzootic among marine fish in the Atlantic Oean. The highly pathogenic nature of the European strain of VHSV for salmonid fish may be the

  9. Detection of viral hemorrhagic septicemia virus

    USGS Publications Warehouse

    Winton, James; Kurath, Gael; Batts, William

    2007-01-01

    Viral hemorrhagic septicemia virus (VHSV) is considered to be one of the most important viral pathogens of finfish and is listed as reportable by many nations and international organizations (Office International des Epizooties 2006). Prior to 1988, VHSV was thought to be limited to Europe (Wolf 1988; Smail 1999). Subsequently, it was shown that the virus is endemic among many marine and anadromous fish species in both the Pacific and Atlantic Oceans (Meyers and Winton 1995; Skall et al. 2005). Genetic analysis reveals that isolates of VHSV can be divided into four genotypes that generally correlate with geographic location with the North American isolates generally falling into VHSV Genotype IV (Snow et al. 2004). In 2005-2006, reports from the Great Lakes region indicated that wild fish had experienced disease or, in some cases, very large die-offs from VHSV (Elsayed et al. 2006, Lumsden et al. 2007). The new strain from the Great Lakes, now identified as VHSV Genotype IVb, appears most closely related to isolates of VHSV from mortalities that occurred during 2000-2004 in rivers and near-shore areas of New Brunswick and Nova Scotia, Canada (Gagne et al. 2007). The type IVb isolate found in the Great Lakes region is the only strain outside of Europe that has been associated with significant mortality in freshwater species.

  10. Nonvirion Protein of Novirhabdovirus Suppresses Apoptosis at the Early Stage of Virus Infection ▿

    PubMed Central

    Ammayappan, Arun; Vakharia, Vikram N.

    2011-01-01

    Viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) are members of the genus Novirhabdovirus within the Rhabdoviridae family, which can cause severe hemorrhagic disease in fresh- and saltwater fish worldwide. These viruses carry an additional nonvirion (NV) gene, which codes for the nonstructural NV protein that has been implicated to play a role in viral pathogenesis. To determine the precise biological function of this NV gene and its gene product, we generated NV-deficient and NV knockout recombinant VHSVs, using reverse genetics. Comparisons of the replication kinetics and markers for virus-induced apoptosis indicated that the NV-deficient and NV knockout mutant viruses induce apoptosis earlier in cell culture than the wild-type recombinant VHSV. These results suggest that the NV protein has an antiapoptotic function at the early stage of virus infection. Furthermore, we created a chimeric VHSV, in which the NV gene of VHSV was replaced by the IHNV NV gene, which was capable of suppressing apoptosis in cell culture. These results show that the NV protein of other members of Novirhabdovirus can restore the NV protein function. In this study, we also investigated the kinetics of VHSV replication during a single round of viral replication and examined the mechanism of VHSV-induced apoptosis. Our results show that VHSV infection induced caspases 3, 8 and 9 in cell culture. PMID:21653667

  11. Assessment of the inhibitory effect of ribavirin on the rainbow trout rhabdovirus VHSV by real-time reverse-transcription PCR.

    PubMed

    Marroquí, Laura; Estepa, Amparo; Perez, Luis

    2007-05-16

    Viral hemorrhagic septicemia virus (VHSV) is one of the most ubiquitous viruses in salmonid aquaculture in Europe. This infectious disease results in significant losses in the farming industry and therefore effective therapeutic agents are needed to control outbreaks caused by this pathogen. Thus, accurate methods to test new antiviral compounds need to be developed. Our goal was to establish a model system for testing novel antivirals with potential applications to aquaculture. In a previous study, a TaqMan real-time RT-PCR assay was designed to detect and quantitate VHSV in rainbow trout tissues [Chico, V., Gomez, N., Estepa, A., Perez, L., 2006. Rapid detection and quantitation of viral hemorrhagic septicemia virus in experimentally challenged rainbow trout by real-time RT-PCR. J. Virol. Methods 132, 154-159]. In this report, we applied the real-time RT-PCR assay to the evaluation of the inhibitory effect of ribavirin, a well-known broad spectrum antiviral drug, in a cell culture system. When added from the beginning of the infection, ribavirin caused a dose-dependent reduction of VHSV RNA accumulation. Real-time RT-PCR measurements showed 99.8% inhibition at 25 microg/ml ribavirin, with an IC50 of 0.43 microg/ml. Ribavirin maintained its inhibitory activity against VHSV when added at 6 h post-infection. Quantitation of N protein messenger RNA and plus-stranded RNA showed a substantial decrease of viral transcription in ribavirin-treated cells. Partial reversion of the effect of ribavirin by addition of GTP was observed, confirming that ribavirin targets the synthesis of guanidine nucleotides in the cells. This is the first report of a real-time PCR-based assay for addressing the efficacy and mechanism of action of an antiviral agent for rainbow trout.

  12. Screening for Viral Hemorrhagic Septicemia Virus in Marine Fish along the Norwegian Coastal Line

    PubMed Central

    Sandlund, Nina; Gjerset, Britt; Bergh, Øivind; Modahl, Ingebjørg; Olesen, Niels Jørgen; Johansen, Renate

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) infects a wide range of marine fish species. To study the occurrence of VHSV in wild marine fish populations in Norwegian coastal waters and fjord systems a total of 1927 fish from 39 different species were sampled through 5 research cruises conducted in 2009 to 2011. In total, VHSV was detected by rRT-PCR in twelve samples originating from Atlantic herring (Clupea harengus), haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and silvery pout (Gadiculus argenteus). All fish tested positive in gills while four herring and one silvery pout also tested positive in internal organs. Successful virus isolation in cell culture was only obtained from one pooled Atlantic herring sample which shows that today's PCR methodology have a much higher sensitivity than cell culture for detection of VHSV. Sequencing revealed that the positive samples belonged to VHSV genotype Ib and phylogenetic analysis shows that the isolate from Atlantic herring and silvery pout are closely related. All positive fish were sampled in the same area in the northern county of Finnmark. This is the first detection of VHSV in Atlantic herring this far north, and to our knowledge the first detection of VHSV in silvery pout. However, low prevalence of VHSV genotype Ib in Atlantic herring and other wild marine fish are well known in other parts of Europe. Earlier there have been a few reports of disease outbreaks in farmed rainbow trout with VHSV of genotype Ib, and our results show that there is a possibility of transfer of VHSV from wild to farmed fish along the Norwegian coast line. The impact of VHSV on wild fish is not well documented. PMID:25248078

  13. Screening for viral hemorrhagic septicemia virus in marine fish along the Norwegian coastal line.

    PubMed

    Sandlund, Nina; Gjerset, Britt; Bergh, Øivind; Modahl, Ingebjørg; Olesen, Niels Jørgen; Johansen, Renate

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) infects a wide range of marine fish species. To study the occurrence of VHSV in wild marine fish populations in Norwegian coastal waters and fjord systems a total of 1927 fish from 39 different species were sampled through 5 research cruises conducted in 2009 to 2011. In total, VHSV was detected by rRT-PCR in twelve samples originating from Atlantic herring (Clupea harengus), haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and silvery pout (Gadiculus argenteus). All fish tested positive in gills while four herring and one silvery pout also tested positive in internal organs. Successful virus isolation in cell culture was only obtained from one pooled Atlantic herring sample which shows that today's PCR methodology have a much higher sensitivity than cell culture for detection of VHSV. Sequencing revealed that the positive samples belonged to VHSV genotype Ib and phylogenetic analysis shows that the isolate from Atlantic herring and silvery pout are closely related. All positive fish were sampled in the same area in the northern county of Finnmark. This is the first detection of VHSV in Atlantic herring this far north, and to our knowledge the first detection of VHSV in silvery pout. However, low prevalence of VHSV genotype Ib in Atlantic herring and other wild marine fish are well known in other parts of Europe. Earlier there have been a few reports of disease outbreaks in farmed rainbow trout with VHSV of genotype Ib, and our results show that there is a possibility of transfer of VHSV from wild to farmed fish along the Norwegian coast line. The impact of VHSV on wild fish is not well documented.

  14. Treatment of endotoxaemia and septicaemia in the equine patient.

    PubMed

    Werners, A H

    2017-01-01

    Endotoxins, constituents of the cell wall of gram-positive and gram-negative bacteria, regularly result in severe illness and death in horses. In endotoxaemia, these constituents are present in the systemic circulation; in septicaemia, whole microbes invade normally sterile parts of the body. Interaction of these endotoxins with pathogen recognition receptors leads to an inflammatory response that cannot always be sufficiently contained and hence needs direct treatment. Over the last decennia, our understanding of the pathophysiology of endotoxaemia and septicaemia has significantly increased. Based on improved understanding of the interaction between receptors and endotoxins as well as the subsequent downstream signalling pathways, new therapeutic targets have been identified in laboratory animal species and humans. Important species differences in the recognition of endotoxins and pathogens by their receptors as well as the inflammatory response to receptor activation hamper extrapolation of this information to the horse (and other species). Historically, horses with endotoxaemia and septicaemia have been treated mainly symptomatically and supportively. Based on the identified therapeutic targets, this review describes the current knowledge of the treatment for endotoxaemia and septicaemia in the horse with reference to the findings in other animal species and humans. © 2016 John Wiley & Sons Ltd.

  15. Re-emergence of bovine haemorrhagic septicaemia in Hungary.

    PubMed

    Magyar, Tibor; Ujvári, Barbara; Szeredi, Levente; Virsinger, Norbert; Albert, Ervin; Német, Zoltán; Csuka, Edit; Biksi, Imre

    2017-03-01

    This paper reports an outbreak of haemorrhagic septicaemia caused by Pasteurella multocida B:2 in beef calves, a disease that has not been described in the Hungarian literature since 1943, and has not been reported to the World Organisation For Animal Health (OIE) since 1970. Acute haemorrhagic septicaemia was confirmed in beef calves on one small farm, and was suspected on two further nearby holdings with concomitant unexplained losses. The source of the infection could not be determined. Apart from a short duration of depression and loss of appetite, the affected calves developed characteristic distal limb oedema. Gross findings in two calves submitted for laboratory examinations included subcutaneous oedema and haemorrhages on serous membranes, and in one case severe pharyngeal lymph node enlargement was observed. Histological examinations revealed lesions characteristic of septicaemia. Moderate to large amounts of Pasteurella antigens were detected in all organs tested by immunohistochemistry. Two isolates of P. multocida (Pm240, Pm241) were cultured from these cases and examined in detail. These were identified as P. multocida ssp. multocida biovar 3. Both were toxA negative and belonged to serotype B:2. Multilocus sequence typing was used to assign these to a new sequence type (ST64) that is closely related to other haemorrhagic septicaemia causing strains of P. multocida regardless of the host.

  16. Genetic diversification of an emerging pathogen: A decade of mutation by the fish Viral Hemorrhagic Septicemia (VHS) virus in the Laurentian Great Lakes

    USDA-ARS?s Scientific Manuscript database

    Viral Hemorrhagic Septicemia virus (VHSv) is an RNA rhabdovirus, which causes one of the world's most serious fish diseases, infecting >80 freshwater and marine species across the Northern Hemisphere. A new, novel, and especially virulent substrain - VHSv-IVb - first appeared in the Laurentian Gre...

  17. Distribution of an Invasive Aquatic Pathogen (Viral Hemorrhagic Septicemia Virus) in the Great Lakes and Its Relationship to Shipping

    PubMed Central

    Bain, Mark B.; Cornwell, Emily R.; Hope, Kristine M.; Eckerlin, Geofrey E.; Casey, Rufina N.; Groocock, Geoffrey H.; Getchell, Rodman G.; Bowser, Paul R.; Winton, James R.; Batts, William N.; Cangelosi, Allegra; Casey, James W.

    2010-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus found in fish from oceans of the northern hemisphere and freshwaters of Europe. It has caused extensive losses of cultured and wild fish and has become established in the North American Great Lakes. Large die-offs of wild fish in the Great Lakes due to VHSV have alarmed the public and provoked government attention on the introduction and spread of aquatic animal pathogens in freshwaters. We investigated the relations between VHSV dispersion and shipping and boating activity in the Great Lakes by sampling fish and water at sites that were commercial shipping harbors, recreational boating centers, and open shorelines. Fish and water samples were individually analyzed for VHSV using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and cell culture assays. Of 1,221 fish of 17 species, 55 were VHSV positive with highly varied qRT-PCR titers (1 to 5,950,000 N gene copies). The detections of VHSV in fish and water samples were closely associated and the virus was detected in 21 of 30 sites sampled. The occurrence of VHSV was not related to type of site or shipping related invasion hotspots. Our results indicate that VHSV is widely dispersed in the Great Lakes and is both an enzootic and epizootic pathogen. We demonstrate that pathogen distribution information could be developed quickly and is clearly needed for aquatic ecosystem conservation, management of affected populations, and informed regulation of the worldwide trade of aquatic organisms. PMID:20405014

  18. Distribution of an invasive aquatic pathogen (viral hemorrhagic septicemia virus) in the Great Lakes and its relationship to shipping

    USGS Publications Warehouse

    Bain, Mark B.; Cornwell, Emily R.; Hope, Kristine M.; Eckerlin, Geofrey E.; Casey, Rufina N.; Groocock, Geoffrey H.; Getchell, Rodman G.; Bowser, Paul R.; Winton, James R.; Batts, William N.; Cangelosi, Allegra; Casey, James W.

    2010-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus found in fish from oceans of the northern hemisphere and freshwaters of Europe. It has caused extensive losses of cultured and wild fish and has become established in the North American Great Lakes. Large die-offs of wild fish in the Great Lakes due to VHSV have alarmed the public and provoked government attention on the introduction and spread of aquatic animal pathogens in freshwaters. We investigated the relations between VHSV dispersion and shipping and boating activity in the Great Lakes by sampling fish and water at sites that were commercial shipping harbors, recreational boating centers, and open shorelines. Fish and water samples were individually analyzed for VHSV using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and cell culture assays. Of 1,221 fish of 17 species, 55 were VHSV positive with highly varied qRT-PCR titers (1 to 5,950,000 N gene copies). The detections of VHSV in fish and water samples were closely associated and the virus was detected in 21 of 30 sites sampled. The occurrence of VHSV was not related to type of site or shipping related invasion hotspots. Our results indicate that VHSV is widely dispersed in the Great Lakes and is both an enzootic and epizootic pathogen. We demonstrate that pathogen distribution information could be developed quickly and is clearly needed for aquatic ecosystem conservation, management of affected populations, and informed regulation of the worldwide trade of aquatic organisms.

  19. Distribution of an invasive aquatic pathogen (viral hemorrhagic septicemia virus) in the Great Lakes and its relationship to shipping.

    PubMed

    Bain, Mark B; Cornwell, Emily R; Hope, Kristine M; Eckerlin, Geofrey E; Casey, Rufina N; Groocock, Geoffrey H; Getchell, Rodman G; Bowser, Paul R; Winton, James R; Batts, William N; Cangelosi, Allegra; Casey, James W

    2010-04-13

    Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus found in fish from oceans of the northern hemisphere and freshwaters of Europe. It has caused extensive losses of cultured and wild fish and has become established in the North American Great Lakes. Large die-offs of wild fish in the Great Lakes due to VHSV have alarmed the public and provoked government attention on the introduction and spread of aquatic animal pathogens in freshwaters. We investigated the relations between VHSV dispersion and shipping and boating activity in the Great Lakes by sampling fish and water at sites that were commercial shipping harbors, recreational boating centers, and open shorelines. Fish and water samples were individually analyzed for VHSV using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and cell culture assays. Of 1,221 fish of 17 species, 55 were VHSV positive with highly varied qRT-PCR titers (1 to 5,950,000 N gene copies). The detections of VHSV in fish and water samples were closely associated and the virus was detected in 21 of 30 sites sampled. The occurrence of VHSV was not related to type of site or shipping related invasion hotspots. Our results indicate that VHSV is widely dispersed in the Great Lakes and is both an enzootic and epizootic pathogen. We demonstrate that pathogen distribution information could be developed quickly and is clearly needed for aquatic ecosystem conservation, management of affected populations, and informed regulation of the worldwide trade of aquatic organisms.

  20. Protective effect of a recombinant VHSV-G vaccine using poly(I:C) loaded nanoparticles as an adjuvant in zebrafish (Danio rerio) infection model.

    PubMed

    Kavaliauskis, Arturas; Arnemo, Marianne; Speth, Martin; Lagos, Leidy; Rishovd, Anne-Lise; Estepa, Amparo; Griffiths, Gareth; Gjøen, Tor

    2016-08-01

    There is a constant need to increase the efficiency of vaccines in the aquaculture industry. Although several nano-based vaccine formulations have been reported, to the best of our knowledge so far only one of them have been implemented in the industry. Here we report on chitosan-poly(I:C) nanoparticles (NPs) that could be used as a non-specific adjuvant in antiviral vaccines in aquaculture. We have characterized the physical parameters of the NPs, studied the in vivo and in vitro bio-distribution of fluorescent NPs and verified NP uptake by zebrafish leucocytes. We used the zebrafish model to test the protective efficiency of the recombinant glycoprotein G (rgpG) of VHSV compared to inactivated whole virus (iV) against VHSV using NPs as an adjuvant in both formulations. In parallel we tested free poly(I:C) and rgpG (pICrgpG), and free chitosan and rgpG (CSrgpG) vaccine formulations. While the iV group (with NP adjuvant) provided the highest overall survival, all vaccine formulations with poly(I:C) provided a significant protection against VHSV; possibly through an early induction of an anti-viral state. Our results suggest that chitosan-poly(I:C) NPs are a promising adjuvant candidate for future vaccine formulations.

  1. Development and validation of a reverse transcription quantitative PCR for universal detection of viral hemorrhagic septicemia virus.

    PubMed

    Garver, Kyle A; Hawley, Laura M; McClure, Carol A; Schroeder, Tamara; Aldous, Sandra; Doig, Fiona; Snow, Michael; Edes, Sandra; Baynes, Catherine; Richard, Jon

    2011-06-16

    Viral hemorrhagic septicemia virus (VHSV) infects over 70 fish species inhabiting marine, brackish or freshwater environments throughout the Northern Hemisphere. Over its geographic range, 4 VHSV genotypes and multiple subtypes exist. Here, we describe the development and validation of a rapid, sensitive and specific real-time reverse transcription quantitative PCR assay (RT-qPCR) that amplifies sequence from representative isolates of all VHSV genotypes (I, II, III and IV). The pan-specific VHSV RT-qPCR assay reliably detects 100 copies of VHSV nucleoprotein RNA without cross-reacting with infectious hematopoietic necrosis virus, spring viremia of carp virus or aquatic birnavirus. Test performance characteristics evaluated on experimentally infected Atlantic salmon Salmo salar L. revealed a diagnostic sensitivity (DSe) > or = 93% and specificity (DSp) = 100%. The repeatability and reproducibility of the procedure was exceptionally high, with 93% agreement among test results within and between 2 laboratories. Furthermore, proficiency testing demonstrated the VHSV RT-qPCR assay to be easily transferred to and performed by a total of 9 technicians representing 4 laboratories in 2 countries. The assay performed equivalent to the traditional detection method of virus isolation via cell culture with the advantage of faster turnaround times and high throughput capacity, further suggesting the suitability of the use of this VHSV RT-qPCR in a diagnostic setting.

  2. Dual DNA vaccination of rainbow trout (Oncorhynchus mykiss) against two different rhabdoviruses, VHSV and IHNV, induces specific divalent protection.

    PubMed

    Einer-Jensen, Katja; Delgado, Lourdes; Lorenzen, Ellen; Bovo, Giuseppe; Evensen, Øystein; Lapatra, Scott; Lorenzen, Niels

    2009-02-18

    DNA vaccines encoding the glycoprotein genes of the salmonid rhabdoviruses VHSV and IHNV are very efficient in eliciting protective immune responses against their respective diseases in rainbow trout (Oncorhynchus mykiss). The early anti-viral response (EAVR) provides protection by 4 days post vaccination and is non-specific and transient while the specific anti-viral response (SAVR) is long lasting and highly specific. Since both VHSV and IHNV are endemic in rainbow trout in several geographical regions of Europe and Atlantic salmon (Salmo salar) on the Pacific coast of North America, co-vaccination against the two diseases would be a preferable option. In the present study we demonstrated that a single injection of mixed DNA vaccines induced long-lasting protection against both individual and a simultaneous virus challenge 80 days post vaccination. Transfected muscle cells at the injection site expressed both G proteins. This study confirms the applied potential of using a combined DNA vaccination for protection of fish against two different rhabdoviral diseases.

  3. Comparative susceptibility of representative Great Lakes fish species to the North American viral hemorrhagic septicemia virus Sublineage IVb.

    PubMed

    Kim, Robert; Faisal, Mohamed

    2010-07-26

    The present study compared the susceptibility of representative Laurentian Great Lakes fish species to the emerging viral hemorrhagic septicemia virus (VHSV) Genotype IVb. The median lethal dose of infection by intraperitoneal injection (IP-LD50) was obtained from fish that were experimentally infected with the MI03 index strain of VHSV-IVb. Fish were injected at doses ranging from 7 x 10(7) to 7 x 10(-2) plaque-forming units (pfu) and maintained at 12 +/- 1 degrees C. The infection trials identified species of high, medium, and low susceptibility based on the IP-LD50 values. Pathogenicity of VHSV-IVb was highest in largemouth bass Micropterus salmoides, which resulted in an IP-LD50 of 1.5 x 10(2) pfu, while also demonstrating the clinical diathesis of VHSV-infected fish. The virus was moderately pathogenic in yellow perch Perca flavescens (IP-LD50 of 2.5 x 10(5) pfu), but also showed the classical signs of VHSV infection. Salmonids were the least susceptible to VHSV-IVb with IP-LD50 values of no less than 1.4 x 106 pfu; however, in fish that succumbed to infection, characteristic VHSV lesions were observed. Histopathologic alterations were most profound in gill, skin, muscle, gonads, and liver of largemouth bass and yellow perch, while in salmonids, hemorrhages in the swimbladder and/or degenerative changes in the liver were the most common lesions noticed. VHSV was isolated from infected fish, and its identity was confirmed by the reverse transcriptase polymerase chain reaction. These results highlight the variations among fish species susceptibility to this emerging strain of VHSV and offer insights into the potential impact of VHSV-IVb on the Laurentian Great Lakes fish community.

  4. Haemorrhagic smolt syndrome (HSS) in Norway: pathology and associated virus-like particles.

    PubMed

    Nylund, A; Plarre, H; Hodneland, K; Devold, M; Aspehaug, V; Aarseth, M; Koren, C; Watanabe, K

    2003-03-17

    Atlantic salmon Salmo salar pre-smolt, smolt and post-smolt, with clinical signs of haemorrhagic smolt syndrome (HSS) have been found in several locations along the Norwegian coast (Rogaland to Troms). Affected fish had pale gills and bleeding at the fin bases, but seemed to be in good physical condition with no obvious weight loss. The internal organs and body cavity showed distinct bleedings. Petechiae were found on the gastrointestinal tract, swim bladder and peritoneum, visceral adipose tissue, heart and somatic musculature. The liver was bright yellow and sometimes mottled with petechiae and ecchymoses. Acitic fluid was found in the visceral cavity and fluid was also present in the pericardial cavity. Histological examination revealed haemorrhage in most organs. The glomeruli were degenerated and the renal tubules were filled with erythrocytes. The aims of this study were to describe the pathology and discover, if possible, the aetiology of the HSS. Tissues were collected for light and transmission electron microscopy (TEM), immunofluorescence (IFAT), reverse transcription (RT)-PCR diagnostics (screening for infectious salmon anaemia virus [ISAV], viral haemorrhagic septicaemia virus [VHSV], salmon pancreas disease virus [SPDV], sleeping disease virus [SDV] and infectious haematopoetic necrosis virus [IHNV]), and tissue homogenates (heart, liver, kidney and spleen) were sterile-filtered and inoculated into cell cultures. Homogenates made from several tissues were also injected intraperitoneally into salmon and rainbow trout Oncorhynchus mykiss. The diagnostic tests revealed no consistent findings of any pathogens, with the exception of TEM which showed 2 types of virus-like particles: Type I was 50 to 60 nm in diameter and Type II about 50 nm in diameter. These virus-like particles were found in salmon from all farms affected by HSS and screened by TEM. Several different cells, blood vessel endothelial cells, endocardial cells, heart myofibres, and leukocytes

  5. Complement depletion aggravates Staphylococcus aureus septicaemia and septic arthritis

    PubMed Central

    Sakiniene, E; Bremell, T; Tarkowski, A

    1999-01-01

    The aim of the study was to assess the role of the complement system in Staphylococcus aureus arthritis and septicaemia. The murine model of haematogenously acquired septic arthritis was used, injecting intravenously toxic shock syndrome toxin-1 (TSST-1), producing S. aureus LS-1. Complement was depleted using cobra venom factor (CVF). Evaluation of arthritis was performed clinically and histopathologically. In addition, the effect of complement depletion on the phagocytic activity of leucocytes was assessed in vivo and in vitro. Six days after inoculation of S. aureus the prevalence of arthritis in decomplemented mice was three-fold higher than that in controls (91% versus 25%). The clinical severity of arthritis at the end of the experiment, expressed as arthritic index, was 7.3 and 1.9, respectively. These findings were confirmed by histological index of synovitis as well as of cartilage and/or bone destruction being significantly higher in decomplemented mice than in controls (9.8 ± 1.7 versus 4.9 ± 1.2, P < 0.05; and 7.9 ± 1.7 versus 3.0 ± 0.9, P < 0.05, respectively). Also, the septicaemia-induced mortality was clearly higher in decomplemented mice compared with the controls. CVF treatment significantly reduced in vivo polymorphonuclear cell-dependent inflammation induced by subcutaneous injection of olive oil and mirroring the capacity of polymorphonuclear cells (PMNC) to migrate and/or extravasate. Besides, the decomplementation procedure significantly impaired phagocytic activity of peripheral blood leucocytes in vitro, since the number of phagocytes being able to ingest bacteria decreased by 50% when the cells were maintained in decomplemented serum compared with those in intact serum. The conclusion is that complement depletion aggravates the clinical course of S. aureus arthritis and septicaemia, possibly by a combination of decreased migration/extravasation of PMNC and an impairment of phagocytosis. PMID:9933426

  6. Bacteroides fragilis septicaemia and meningitis in early infancy.

    PubMed Central

    Cooke, R W

    1975-01-01

    A case of recurrent Bacteroides fragilis septicaemia leading eventually to meningitis in a 6-week-old infant is reported. Perforation of the small bowel at the constriction ring of a strangulated inguinal hernia caused a faecal peritonitis and was the primary source of infection. Erythromycin, to which the isolate was fully sensitive in vitro, was only temporarily an effective treatment; the infection was finally eradicated with chloramphenicol, and the baby made a full recovery. Bacteroides infections in infancy and childhood are briefly reviewed. PMID:1147659

  7. RNA-Seq transcriptome analysis of the olive flounder (Paralichthys olivaceus) kidney response to vaccination with heat-inactivated viral hemorrhagic septicemia virus.

    PubMed

    Hwang, Jee Youn; Kwon, Mun Gyeoung; Jung, Sung-Hee; Park, Myoung Ae; Kim, Dong-Wook; Cho, Wang Sik; Park, Jeong Woo; Son, Maeng-Hyun

    2017-03-01

    Viral hemorrhagic septicemia (VHS) is a highly contagious disease of cultured flounder caused by VHS virus (VHSV). To develop effective VHSV vaccines, it is essential to understand the molecular mechanisms underlying the host's protective response against VHSV. The purpose of this study is to clarify which genes are involved in the protective response of olive flounder after VHSV vaccination. We first injected olive flounder intraperitoneally with 10(7) TCID50 heat-inactivated VHSV vaccine and evaluated the vaccine efficacy at 20 °C. Fish vaccinated with heat-inactivated VHSV were significantly protected compared to non-vaccinated fish, with a relative percentage survival of 83%. To analyze the vaccination-induced changes in the expression profiles of genes, kidneys were collected from control and vaccinated fish at days 1, 3, and 7 after vaccination and global gene expression profiling was carried out by RNA sequencing. The analysis revealed that 15,001 genes were differentially expressed by at least 2-fold between vaccinated fish and non-vaccinated controls. Of these, 58 genes clustered into the acute phase response, Toll-like receptor, interferon-inducible/regulatory proteins, and apoptosis pathways. These data provided insights into the molecular mechanisms underlying the protective immune response of olive flounder against heat-inactivated VHSV vaccine and might aid future studies to develop a highly immunogenic vaccine against VHSV in flounder.

  8. Spread of the Emerging Viral Hemorrhagic Septicemia Virus Strain, Genotype IVb, in Michigan, USA

    PubMed Central

    Faisal, Mohamed; Shavalier, Megan; Kim, Robert K.; Millard, Elena V.; Gunn, Michelle R.; Winters, Andrew D.; Schulz, Carolyn A.; Eissa, Alaa; Thomas, Michael V.; Wolgamood, Martha; Whelan, Gary E.; Winton, James

    2012-01-01

    In 2003, viral hemorrhagic septicemia virus (VHSV) emerged in the Laurentian Great Lakes causing serious losses in a number of ecologically and recreationally important fish species. Within six years, despite concerted managerial preventive measures, the virus spread into the five Great Lakes and to a number of inland waterbodies. In response to this emerging threat, cooperative efforts between the Michigan Department of Natural Resources (MI DNR), the Michigan State University Aquatic Animal Health Laboratory (MSU-AAHL), and the United States Department of Agriculture-Animal and Plant Health Inspection Services (USDA-APHIS) were focused on performing a series of general and VHSV-targeted surveillances to determine the extent of virus trafficking in the State of Michigan. Herein we describe six years (2005–2010) of testing, covering hundreds of sites throughout Michigan’s Upper and Lower Peninsulas. A total of 96,228 fish representing 73 species were checked for lesions suggestive of VHSV and their internal organs tested for the presence of VHSV using susceptible cell lines. Of the 1,823 cases tested, 30 cases from 19 fish species tested positive for VHSV by tissue culture and were confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Gene sequence analyses of all VHSV isolates retrieved in Michigan demonstrated that they belong to the emerging sublineage “b” of the North American VHSV genotype IV. These findings underscore the complexity of VHSV ecology in the Great Lakes basin and the critical need for rigorous legislation and regulatory guidelines in order to reduce the virus spread within and outside of the Laurentian Great Lakes watershed. PMID:22754647

  9. Comparative studies for serodiagnosis of haemorrhagic septicaemia in cattle sera

    PubMed Central

    El-Jakee, Jakeen K.; Ali, Samah Said; El-Shafii, Soumaya Ahmed; Hessain, Ashgan M.; Al-Arfaj, Abdullah A.; Mohamed, Moussa I.

    2015-01-01

    Haemorrhagic septicaemia caused by Pasteurella multocida is a major epizootic disease in cattle and buffaloes in developing countries with high morbidity and mortality rate. In the present study, a total of 88 P. multocida isolates were isolated from 256 nasopharyngeal swabs and lung tissues samples (34.4%) during the period from January, 2013 to March, 2014 from different governorates located in Egypt. Dead calves showed the highest percentage of P. multocida isolation followed by the emergency slaughtered calves, diseased calves then apparently healthy ones. These isolates were confirmed as P. multocida microscopically, biochemically by traditional tests and by API 20E commercial kit then by PCR. The percentages of positive serum samples using somatic antigen and micro-agglutination test at 1/1280 diluted serum were 10%, 54.49% and 0% in apparently healthy, diseased and emergency slaughtered samples, respectively whereas, the percentages using capsular antigen and indirect haemagglutination test were 40%, 60.89% and 60% in apparently healthy, diseased and emergency slaughtered samples, respectively. The ELISA showed the highest sensitivity for diagnosing P. multocida in apparently healthy, diseased and emergency slaughtered animals with percentages of 42%; 92.9% and 80%, respectively. The obtained results revealed that the ELISA using capsular antigen of P. multocida is a more sensitive and specific serological test for diagnosis of haemorrhagic septicaemia. PMID:26858538

  10. A Recombinant Novirhabdovirus Presenting at the Surface the E Glycoprotein from West Nile Virus (WNV) Is Immunogenic and Provides Partial Protection against Lethal WNV Challenge in BALB/c Mice

    PubMed Central

    Nzonza, Angella; Lecollinet, Sylvie; Chat, Sophie; Lowenski, Steeve; Mérour, Emilie; Biacchesi, Stéphane; Brémont, Michel

    2014-01-01

    West Nile Virus (WNV) is a zoonotic mosquito-transmitted flavivirus that can infect and cause disease in mammals including humans. Our study aimed at developing a WNV vectored vaccine based on a fish Novirhabdovirus, the Viral Hemorrhagic Septicemia virus (VHSV). VHSV replicates at temperatures lower than 20°C and is naturally inactivated at higher temperatures. A reverse genetics system has recently been developed in our laboratory for VHSV allowing the addition of genes in the viral genome and the recovery of the respective recombinant viruses (rVHSV). In this study, we have generated rVHSV vectors bearing the complete WNV envelope gene (EWNV) (rVHSV-EWNV) or fragments encoding E subdomains (either domain III alone or domain III fused to domain II) (rVHSV-DIIIWNV and rVHSV-DII-DIIIWNV, respectively) in the VHSV genome between the N and P cistrons. With the objective to enhance the targeting of the EWNV protein or EWNV-derived domains to the surface of VHSV virions, Novirhadovirus G-derived signal peptide and transmembrane domain (SPG and TMG) were fused to EWNV at its amino and carboxy termini, respectively. By Western-blot analysis, electron microscopy observations or inoculation experiments in mice, we demonstrated that both the EWNV and the DIIIWNV could be expressed at the viral surface of rVHSV upon addition of SPG. Every constructs expressing EWNV fused to SPG protected 40 to 50% of BALB/cJ mice against WNV lethal challenge and specifically rVHSV-SPGEWNV induced a neutralizing antibody response that correlated with protection. Surprisingly, rVHSV expressing EWNV-derived domain III or II and III were unable to protect mice against WNV challenge, although these domains were highly incorporated in the virion and expressed at the viral surface. In this study we demonstrated that a heterologous glycoprotein and non membrane-anchored protein, can be efficiently expressed at the surface of rVHSV making this approach attractive to develop new vaccines against

  11. An active DNA vaccine against infectious pancreatic necrosis virus (IPNV) with a different mode of action than fish rhabdovirus DNA vaccines.

    PubMed

    Cuesta, A; Chaves-Pozo, E; de Las Heras, A I; Saint-Jean, S Rodríguez; Pérez-Prieto, S; Tafalla, C

    2010-04-26

    Although there are some commercial vaccines available against infectious pancreatic necrosis virus (IPNV), the disease still continues to be a major problem for aquaculture development worldwide. In the current work, we constructed a DNA vaccine against IPNV (pIPNV-PP) by cloning the long open reading frame of the polyprotein encoded by the viral RNA segment A. In vitro, the vaccine is properly translated giving the functional IPNV polyprotein since preVP2, VP2 and VP3 proteins were detected because of the VP4-protease cleavage. EPC cells transfected with the vaccine plasmid expressed the viral proteins and induced the expression of type I interferon (IFN)-induced Mx genes. Furthermore, IPNV synthesized proteins seemed to assemble in virus-like particles as evidenced by electron microscopy. In vivo, rainbow trout specimens were intramuscularly injected with the vaccine and expression of immune-relevant genes, the presence of neutralizing antibodies and effect on viral load was determined. The pIPNV-PP vaccine was expressed at the injection site and up-regulated MHC Ialpha, MHC IIalpha, type-I interferon (IFN), Mx, CD4 and CD8alpha gene expression in the muscle, head kidney or spleen, although to a much lower extent than the up-regulations observed in response to an effective DNA vaccine against viral hemorrhagic septicaemia virus (VHSV). However, the IPNV vaccine was also very effective in terms of acquired immunity since it elicited neutralizing antibodies (in 6 out of 8 trout fingerlings) and decreased 665-fold the viral load after IPNV infection. The effectiveness of this new IPNV DNA vaccine and its possible mechanism of action are discussed and compared to other viral vaccines.

  12. An unusual culprit: Klebsiella pneumoniae causing septicaemia outbreaks in neonatal pigs?

    PubMed

    Bowring, Bethany G; Fahy, V Anthony; Morris, Andrew; Collins, Alison M

    2017-05-01

    This study investigated the cause of recent outbreaks of septicaemia in neonatal pigs in Australia (Victoria and Queensland). Septicaemia in neonatal pigs is commonly caused by enterotoxigenic E. coli, extraintestinal pathogenic E. coli and beta-haemolytic streptococci. Infrequent causes of septicaemia are Actinobacillus suis and Citrobacter freundii. Therefore, it was quite unexpected when Klebsiella pneumoniae was isolated in predominant growth from multiple organs of pigs dying of septicaemia from six days of age. Two cases in Victoria were associated with multilocus sequence type 25 (ST25), whereas the cases on a single farm in Queensland were associated with two different sequence types (ST278 and ST1978). Similar outbreaks of septicaemia have also occurred in England, but all were associated with K. pneumoniae ST25. The K. pneumoniae sequence types did not cluster phylogenetically into groups of isolates from the same geographical location or into groups which caused either septicaemia or pneumonia. Antibiotic resistance also varied between isolates and showed neomycin resistance in Queensland. These results suggest that several sequence types of K. pneumoniae are involved in causing outbreaks of septicaemia in neonatal pigs, in addition to its previously recognised role in pneumonia and mastitis. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Autophagy-inducing peptides from mammalian VSV and fish VHSV rhabdoviral G glycoproteins (G) as models for the development of new therapeutic molecules.

    PubMed

    García-Valtanen, Pablo; Ortega-Villaizán, María Del Mar; Martínez-López, Alicia; Medina-Gali, Regla; Pérez, Luis; Mackenzie, Simon; Figueras, Antonio; Coll, Julio M; Estepa, Amparo

    2014-09-01

    It has not been elucidated whether or not autophagy is induced by rhabdoviral G glycoproteins (G) in vertebrate organisms for which rhabdovirus infection is lethal. Our work provides the first evidence that both mammalian (vesicular stomatitis virus, VSV) and fish (viral hemorrhagic septicemia virus, VHSV, and spring viremia carp virus, SVCV) rhabdoviral Gs induce an autophagic antiviral program in vertebrate cell lines. The transcriptomic profiles obtained from zebrafish genetically immunized with either Gsvcv or Gvhsv suggest that autophagy is induced shortly after immunization and therefore, it may be an important component of the strong antiviral immune responses elicited by these viral proteins. Pepscan mapping of autophagy-inducing linear determinants of Gvhsv and Gvsv showed that peptides located in their fusion domains induce autophagy. Altogether these results suggest that strategies aimed at modulating autophagy could be used for the prevention and treatment of rhabdoviral infections such as rabies, which causes thousands of human deaths every year.

  14. Autophagy-inducing peptides from mammalian VSV and fish VHSV rhabdoviral G glycoproteins (G) as models for the development of new therapeutic molecules

    PubMed Central

    García-Valtanen, Pablo; Ortega-Villaizán, María del Mar; Martínez-López, Alicia; Medina-Gali, Regla; Pérez, Luis; Mackenzie, Simon; Figueras, Antonio; Coll, Julio M; Estepa, Amparo

    2014-01-01

    It has not been elucidated whether or not autophagy is induced by rhabdoviral G glycoproteins (G) in vertebrate organisms for which rhabdovirus infection is lethal. Our work provides the first evidence that both mammalian (vesicular stomatitis virus, VSV) and fish (viral hemorrhagic septicemia virus, VHSV, and spring viremia carp virus, SVCV) rhabdoviral Gs induce an autophagic antiviral program in vertebrate cell lines. The transcriptomic profiles obtained from zebrafish genetically immunized with either Gsvcv or Gvhsv suggest that autophagy is induced shortly after immunization and therefore, it may be an important component of the strong antiviral immune responses elicited by these viral proteins. Pepscan mapping of autophagy-inducing linear determinants of Gvhsv and Gvsv showed that peptides located in their fusion domains induce autophagy. Altogether these results suggest that strategies aimed at modulating autophagy could be used for the prevention and treatment of rhabdoviral infections such as rabies, which causes thousands of human deaths every year. PMID:25046110

  15. Effect of viral hemorrhagic septicemia virus on Pacific herring in Prince William Sound, Alaska, from 1989 to 2005.

    PubMed

    Elston, Ralph A; Meyers, Theodore R

    2009-02-25

    We critically review the role of viral hemorrhagic septicemia virus (VHSV) in the 1992-1993 collapse of the Prince William Sound (PWS) herring fishery. VHSV was detected in samples of moribund Pacific herring from PWS in spring 1993 when about 63% of the expected fish failed to appear. A low prevalence and severity of VHSV were observed in adult pre-spawning PWS herring in most of the years from 1994 to 2002. The North American strain of VHSV became established about 500 yr ago in many northeast Pacific marine fish species, including herring. In Alaska, the typical annual prevalence of VHSV in pre-spawning herring ranges from 0 to 17%. New threshold analysis of a 9 yr study indicates that only about half of the virus-infected adult fish in PWS were clinically affected; ulcers formerly attributed to VHS have been overestimated by a factor of about 3. We conclude that VHSV was not a primary causative factor in the PWS herring population collapse or in its failure to recover. Because older age classes of herring were not disproportionately missing in 1993, the protozoan Ichthyophonus hoferi was also not a likely cause of losses. The 'Exxon Valdez' oil spill occurred in PWS, Alaska, U.S.A., in 1989. Evidence for interaction of oil and VHSV expression is also evaluated. A study exposing herring to varying concentrations of weathered crude oil showed increasing prevalences of VHSV correlated with oil concentration; however, repeated experiments with juvenile and adult fish failed to corroborate these results or link oil to VHSV infection in herring.

  16. In vivo and in vitro phenotypic differences between Great Lakes VHSV genotype IVb isolates with sequence types vcG001 and vcG002.

    PubMed

    Imanse, Sierra M; Cornwell, Emily R; Getchell, Rodman G; Kurath, Gael; Bowser, Paul R

    2014-12-01

    Viral hemorrhagic septicemia virus (VHSV) is an aquatic rhabdovirus first recognized in farmed rainbow trout in Denmark. In the past decade, a new genotype of this virus, IVb was discovered in the Laurentian Great Lakes basin and has caused several massive die-offs in some of the 28 species of susceptible North American freshwater fishes. Since its colonization of the Great Lakes, several closely related sequence types within genotype IVb have been reported, the two most common of which are vcG001 and vcG002. These sequence types have different spatial distributions in the Great Lakes. The aim of this study was to determine whether the genotypic differences between representative vcG001 (isolate MI03) and vcG002 (isolate 2010-030 #91) isolates correspond to phenotypic differences in terms of virulence using both an in vitro and in vivo approach. In vitro infection of epithelioma papulosum cyprini (EPC), bluegill fry (BF-2), and Chinook salmon embryo (CHSE) cells demonstrated some differences in onset and rate of growth in EPC and BF-2 cells, without any difference in the quantity of RNA produced. In vivo infection of round gobies (Neogobius melanostomus) via immersion exposure to different concentrations of vcG001 or vcG002 caused a significantly greater mortality in round gobies exposed to 10(2) plaque forming units ml(-1) of vcG001. These experiments suggest that there are phenotypic differences between Great Lakes isolates of VHSV genotype IVb.

  17. In vivo and in vitro phenotypic differences between Great Lakes VHSV genotype IVb isolates with sequence types vcG001 and vcG002

    USGS Publications Warehouse

    Imanse, Sierra M.; Cornwell, Emily R.; Getchell, Rodman G.; Kurath, Gael; Bowser, Paul R.

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) is an aquatic rhabdovirus first recognized in farmed rainbow trout in Denmark. In the past decade, a new genotype of this virus, IVb was discovered in the Laurentian Great Lakes basin and has caused several massive die-offs in some of the 28 species of susceptible North American freshwater fishes. Since its colonization of the Great Lakes, several closely related sequence types within genotype IVb have been reported, the two most common of which are vcG001 and vcG002. These sequence types have different spatial distributions in the Great Lakes. The aim of this study was to determine whether the genotypic differences between representative vcG001 (isolate MI03) and vcG002 (isolate 2010-030 #91) isolates correspond to phenotypic differences in terms of virulence using both in vitro and in vivo approaches. In vitro infection of epithelioma papulosum cyprini (EPC), bluegill fry (BF-2), and Chinook salmon embryo (CHSE) cells demonstrated some differences in onset and rate of growth in EPC and BF-2 cells, without any difference in the quantity of RNA produced. In vivo infection of round gobies (Neogobius melanostomus) via immersion exposure to different concentrations of vcG001 or vcG002 caused a significantly greater mortality in round gobies exposed to 102 plaque forming units ml− 1 of vcG001. These experiments suggest that there are phenotypic differences between Great Lakes isolates of VHSV genotype IVb.

  18. In vivo and in vitro phenotypic differences between Great Lakes VHSV genotype IVb isolates with sequence types vcG001 and vcG002

    PubMed Central

    Imanse, Sierra M.; Cornwell, Emily R.; Getchell, Rodman G.; Kurath, Gael; Bowser, Paul R.

    2014-01-01

    Viral hemorrhagic septicemia virus (VHSV) is an aquatic rhabdovirus first recognized in farmed rainbow trout in Denmark. In the past decade, a new genotype of this virus, IVb was discovered in the Laurentian Great Lakes basin and has caused several massive die-offs in some of the 28 species of susceptible North American freshwater fishes. Since its colonization of the Great Lakes, several closely related sequence types within genotype IVb have been reported, the two most common of which are vcG001 and vcG002. These sequence types have different spatial distributions in the Great Lakes. The aim of this study was to determine whether the genotypic differences between representative vcG001 (isolate MI03) and vcG002 (isolate 2010-030 #91) isolates correspond to phenotypic differences in terms of virulence using both an in vitro and in vivo approach. In vitro infection of epithelioma papulosum cyprini (EPC), bluegill fry (BF-2), and Chinook salmon embryo (CHSE) cells demonstrated some differences in onset and rate of growth in EPC and BF-2 cells, without any difference in the quantity of RNA produced. In vivo infection of round gobies (Neogobius melanostomus) via immersion exposure to different concentrations of vcG001 or vcG002 caused a significantly greater mortality in round gobies exposed to 102 plaque forming units ml−1 of vcG001. These experiments suggest that there are phenotypic differences between Great Lakes isolates of VHSV genotype IVb. PMID:25722533

  19. Specific Regulation of the Chemokine Response to Viral Hemorrhagic Septicemia Virus at the Entry Site▿

    PubMed Central

    Montero, Jana; Garcia, Jessica; Ordas, M. Camino; Casanova, Isabel; Gonzalez, Antonia; Villena, Alberto; Coll, Julio; Tafalla, Carolina

    2011-01-01

    The fin bases constitute the main portal of rhabdovirus entry into rainbow trout (Oncorhynchus mykiss), and replication in this first site strongly conditions the outcome of the infection. In this context, we studied the chemokine response elicited in this area in response to viral hemorrhagic septicemia virus (VHSV), a rhabdovirus. Among all the rainbow trout chemokine genes studied, only the transcription levels of CK10 and CK12 were significantly upregulated in response to VHSV. As the virus had previously been shown to elicit a much stronger chemokine response in internal organs, we compared the effect of VHSV on the gills, another mucosal site which does not constitute the main site of viral entry or rhabdoviral replication. In this case, a significantly stronger chemokine response was triggered, with CK1, CK3, CK9, and CK11 being upregulated in response to VHSV and CK10 and CK12 being down-modulated by the virus. We then conducted further experiments to understand how these different chemokine responses of mucosal tissues could correlate with their capacity to support VHSV replication. No viral replication was detected in the gills, while at the fin bases, only the skin and the muscle were actively supporting viral replication. Within the skin, viral replication took place in the dermis, while viral replication was blocked within epidermal cells at some point before protein translation. The different susceptibilities of the different skin layers to VHSV correlated with the effect that VHSV has on their capacity to secrete chemotactic factors. Altogether, these results suggest a VHSV interference mechanism on the early chemokine response at its active replication sites within mucosal tissues, a possible key process that may facilitate viral entry. PMID:21325404

  20. Vaccines for piscirickettsiosis (salmonid rickettsial septicaemia, SRS): the Chile perspective.

    PubMed

    Maisey, Kevin; Montero, Ruth; Christodoulides, Myron

    2017-03-01

    Piscirickettsia salmonis (P. salmonis) is the aetiological bacterium of the contagious disease piscirickettsiosis or salmonid rickettsial septicaemia (SRS) and causes significant economic losses to aquaculture production in Chile. Current strategies to control infection are i) indiscriminate antibiotic use and ii) vaccination with predominantly P. salmonis bacterin vaccines that do not provide acceptable levels of protection against piscirickettsiosis. Areas covered: This review covers the basic biology of P. salmonis, clinical piscirickettsiosis and disease control, the development of current P. salmonis vaccines, innate and adaptive immunity and a 5-year plan to develop new piscirickettsiosis vaccines. Expert commentary: Fundamental knowledge is lacking on the complexities of P. salmonis-host interactions, relating to bacterial virulence and host innate and adaptive immune responses, which needs to be addressed. The development of new P. salmonis vaccines needs the application of comprehensive 'omics' technologies to identify candidate vaccine antigens capable of stimulating long-lasting protective immune responses.

  1. Staphylococcal septicaemia, endocarditis, and osteomyelitis in dialysis and renal transplant patients.

    PubMed Central

    Nicholls, A.; Edward, N.; Catto, G. R.

    1980-01-01

    Septicaemia is a common and potentially lethal hazard of haemodialysis and renal transplantation; it is usually caused by Staphylococcus pyogenes. In 6 patients with S. pyogenes septicaemia, fatal endocarditis and spinal osteomyelitis have each occurred once, and 3 patients have had recurrent episodes of septicaemia. The management of septicaemia in these patients must include a search for metastatic infection, and prolonged therapy with 2 antistaphylococcal agents is necessary to ensure eradication of infection. Access site infection in dialysis patients must be treated vigorously, and recognized as potentially hazardous by patients. The risk of sepsis in dialysis and transplant patients cannot be excluded, but devastating consequences may be avoided by simple measures. Images Fig. 1 Fig. 2 Fig. 3 PMID:7008049

  2. Increased risk of stroke after septicaemia: a population-based longitudinal study in Taiwan.

    PubMed

    Lee, Jiunn-Tay; Chung, Wen Ting; Lin, Jin-Ding; Peng, Giia-Sheun; Muo, Chih-Hsin; Lin, Che-Chen; Wen, Chi-Pang; Wang, I-Kuan; Tseng, Chun-Hung; Kao, Chia-Hung; Hsu, Chung Y

    2014-01-01

    Inflammation and infection have been noted to increase stroke risk. However, the association between septicaemia and increased risk of stroke remains unclear. This population-based cohort study, using a National Health Insurance database, aimed to investigate whether patients with septicaemia are predisposed to increased stroke risk. The study included all patients hospitalised for septicaemia for the first time between 2000 and 2003 without prior stroke. Patients were followed until the end of 2010 to evaluate incidence of stroke. An age-, gender- and co-morbidities-matched cohort without prior stroke served as the control. Cox's proportional hazards regressions were used to assess differences in stroke risk between groups. Based on hazard ratios (HRs), patients with septicaemia had greater stroke risk, especially in the younger age groups (age <45: HR = 4.16, 95% CI: 2.39-7.24, p<0.001; age 45-64: HR = 1.76, 95% CI: 1.41-2.19, p<0.001; age ≥ 65: HR = 1.05, 95% CI: 0.91-1.22, p>0.05). Haemorrhagic stroke was the dominant type (ischaemic stroke: HR = 1.20, 95% CI: 1.06-1.37, p<0.01; haemorrhagic stroke: HR = 1.82, 95% CI: 1.35-2.46, p<0.001) and patients without co-morbidities were at slightly higher risk (without co-morbidities: HR = 1.49, 95% CI: 1.02-2.17, p<0.05; with co-morbidities: HR = 1.24, 95% CI: 1.10-1.41, p<0.001). The impact of septicaemia on stroke risk was highest within 6 months of the event and gradually declined over time. Our results suggest that septicaemia is associated with an increase in stroke risk, which is greatest in haemorrhagic stroke. Closer attention to patients with history of septicaemia may be warranted for stroke preventive measures, especially for younger patients without co-morbidities.

  3. In vitro neutralization of viral hemorrhagic septicemia virus by plasma from immunized zebrafish.

    PubMed

    Chinchilla, Blanca; Gomez-Casado, Eduardo; Encinas, Paloma; Falco, Alberto; Estepa, Amparo; Coll, Julio

    2013-03-01

    We studied humoral long-term adaptive viral neutralization responses in zebrafish (Danio rerio), an increasingly useful vertebrate model for viral diseases actually limited by the absence of standardized anti-zebrafish immunoglobulin M (IgM) antibodies. We established an alternative method, similar to those used in other fish, to achieve a first estimation of zebrafish anti-viral antibody-like responses. We used the viral hemorrhagic septicemia virus (VHSV) model because, although protection after this non-natural infection was demonstrated in cold-acclimatized zebrafish, little is known about their induced anti-VHSV antibody-like responses. Therefore, we first optimized a micro-neutralization method based on immunostaining VHSV-infected fish cell monolayers to detect zebrafish neutralizing activity in plasma samples in one day. We then used the method to measure the specific anti-VHSV neutralization in plasma obtained from individual zebrafish under various VHSV challenges or immunization protocols. The neutralizing activity was inhibited by protein A-sepharose and rabbit anti-zebrafish IgM antibodies, suggesting the implication of IgM zebrafish antibodies in such responses. To our knowledge, this is the first report to demonstrate detectable and significant VHSV neutralization titers in zebrafish surviving VHSV infections. This micro-method might be useful, not only for the follow-up of infection/vaccine development in the zebrafish/VHSV model in particular, but also for similar work involving other in vitro neutralizable zebrafish pathogens. This technique might also further the development of alternative ELISA assay methods to measure specific immunoglobulins in zebrafish.

  4. Complete Protection against Influenza Virus H1N1 Strain A/PR/8/34 Challenge in Mice Immunized with Non-Adjuvanted Novirhabdovirus Vaccines

    PubMed Central

    Rouxel, Ronan N.; Mérour, Emilie; Biacchesi, Stéphane; Brémont, Michel

    2016-01-01

    Novirhabdoviruses like Viral Hemorrhagic Septicemia Virus (VHSV) and Infectious Hematopoietic Necrosis Virus (IHNV) are fish-infecting Rhabdoviruses belonging to the Mononegavirales order. By reverse genetics, we previously showed that a recombinant VHSV expressing the West Nile Virus (WNV) E glycoprotein could serve as a vaccine platform against WNV. In the current study, we aimed to evaluate the potential of the Novirhabdovirus platform as a vaccine against influenza virus. Recombinant Novirhabdoviruses, rVHSV-HA and rIHNV-HA, expressing at the viral surface the hemagglutinin HA ectodomain were generated and used to immunized mice. We showed that mice immunized with either, rVHSV-HA or rIHNV-HA, elicited a strong neutralizing antibody response against influenza virus. A complete protection was conferred to the immunized mice when challenged with a lethal dose of influenza H1N1 A/PR/8/34 virus. Furthermore we showed that although acting as inert antigen in mice, since naturally inactivated over 20°C, mice immunized with rVHSV-HA or rIHNV-HA in the absence of adjuvant were also completely protected from a lethal challenge. Novirhabdoviruses platform are of particular interest as vaccines for mammals since they are cost effective to produce, relatively easy to generate and very effective to protect immunized animals. PMID:27711176

  5. [Septicaemia in piglets associated with a positive finding of a methicillin-resistant S. aureus strain].

    PubMed

    Graage, R; Ganter, M; Verspohl, J; Strommenger, B; Waldmann, K-H; Baumgärtner, W; Hennig-Pauka, I

    2014-01-01

    Staphylococcus (S.) aureus has been associated with septicaemia, mastitis, vaginitis, metritis, osteomyelitis, and endocarditis. This case report demonstrates S. aureus-induced septicaemia in suckling pig- lets. Three days after birth, littermates displayed severe ecchymosis and cyanosis, with a 50% mortality rate. The surviving littermates were cross-fostered, but died 1 day later. Other piglets, which were allowed to suck at the sow, developed similar clinical signs. Haematological findings were anaemia, thrombocytopenia, and leukopenia; therefore, neither isohaemolytic anaemia nor septicaemia could be excluded as differential diagnoses. At necropsy, petechial bleeding on inner organ surfaces and free blood in body cavities were found. Bacteriological examination of the sow's milk and of the spleen of one piglet detected a methicillin-resistent S. aureus strain (MRSA CC398), which was in all likelihood the cause of the disease. Potential differential diagnoses are discussed.

  6. Predisposing factors to neonatal septicaemia at ahmadu bello university teaching hospital, zaria Nigeria.

    PubMed

    Onalo, R; Ogala, W N; Ogunrinde, G O; Olayinka, A T; Adama, S A; Ega, B A

    2011-03-01

    The study aimed at identifying the risk factors for neonatal septicaemia in Zaria. Consecutive newborns admitted into the Special Care Baby Unit of Ahmadu Bello University Teaching Hospital, Zaria with the presumptive test diagnosis of neonatal septicaemia between 25 May, 2004 and 31 May, 2005 were studied. History of events in the antenatal and neonatal periods was obtained and physical examination was done. Blood, urine, cerebrospinal fluid and swabs of body discharges were taken for culture and sensitivity studies. The data were analysed using Epi Info version 6 software. Associations were tested using Chi square, with Yates correction, or Fisher's exact tests where appropriate, while statistical significance was set at p < 0.05. A total of 211 neonates, consisting of 69 in-born and 142 out-born infants were studied. There were 122 (57.8%) males and 89 (42.2%) females; giving a male: female ratio of 1.4:1. Seventy-five (35.5%) of the newborns had bacteriologically proven septicaemia consisting of 54 (38.0%) of the outborn and 21 (30.4%) of the inborn babies (p = 0.3535); 42 males and 33 females (p = 0.8011). The male: female ratio of newborns with culture-proven septicaemia was 1.2:1. The predisposing factors that were associated with culture-proven septicaemia were lack of antenatal care (p = 0.0234), prolonged rupture of membranes (p = 0.0085), prolonged labour (p = 0.0032), preterm delivery (p = 0.0125) and perinatal asphyxia (p = 0.0078). The risk factors in neonates with septicaemia in this study emphasise the need for timely improvement in the implementation of existing public health strategies.

  7. Conjugate vaccines for preventing meningococcal C meningitis and septicaemia.

    PubMed

    Conterno, L O; Silva Filho, C R; Rüggeberg, J U; Heath, P T

    2006-07-19

    Meningococcal polysaccharide (MPLS) vaccines protect against Serogroup C disease, but do not produce an immune response in infants less than two years of age. This limitation can be overcome by linking C polysaccharide to carrier proteins ('conjugating'), to create meningococcal serogroup C conjugate (MCC) vaccines. In the absence of trial data, the immune response to vaccination has been considered to be a reasonable surrogate for vaccine protection. To assess the immunogenicity, safety and efficacy of MCC vaccines for preventing meningitis and septicaemia. We searched the Cochrane Central Register Controlled Trials (CENTRAL) (The Cochrane Library Issue 3, 2005); MEDLINE (1966 to September, Week 1 2005); and EMBASE (1990 to June 2005) and references of studies. Randomised controlled trials (RCTs) and controlled clinical trials (CCTs) in humans comparing MCC vaccines against a control vaccine or none. In the absence of any trials on vaccine efficacy, population-based observational studies about effectiveness were included. Two authors independently screened the results of the literature searches, selected eligible studies, extracted the data and evaluated the quality of them. The studies showed that MCC vaccine was highly immunogenic in infants after two and three doses, in toddlers after one and two doses and in older age groups after one dose. In general higher titres were generated after MCC than after MPLS vaccines. Immunological hypo-responsiveness seen after repeated doses of MPLS vaccine may be overcome with MCC. Observational studies have documented a significant decline in meningococcal C disease in countries where MCC vaccines have been widely used. The timing of the vaccinations schedules, the specific conjugate used, and the vaccines given concomitantly or combined, may be important. The MCC vaccine appears to be safe, immunogenic and able to induce immunological memory in all age groups. Observational studies strongly suggest that MCC is clinically

  8. WITHDRAWN: Conjugate vaccines for preventing meningococcal C meningitis and septicaemia.

    PubMed

    Conterno, Lucieni O; da Silva Filho, Carlos Rodrigues; Ruggeberg, Jens U; Heath, Paul T

    2011-07-06

    Meningococcal polysaccharide (MPLS) vaccines protect against Serogroup C disease, but do not produce an immune response in infants less than two years of age. This limitation can be overcome by linking C polysaccharide to carrier proteins ('conjugating'), to create meningococcal serogroup C conjugate (MCC) vaccines. In the absence of trial data, the immune response to vaccination has been considered to be a reasonable surrogate for vaccine protection. To assess the immunogenicity, safety and efficacy of MCC vaccines for preventing meningitis and septicaemia. We searched the Cochrane Central Register Controlled Trials (CENTRAL) (The Cochrane Library 2005, issue 3); MEDLINE (1966 to September, Week 1 2005); and EMBASE (1990 to June 2005) and references of studies. Randomised controlled trials (RCTs) and controlled clinical trials (CCTs) in humans comparing MCC vaccines against a control vaccine or none. In the absence of any trials on vaccine efficacy, population-based observational studies about effectiveness were included. Two authors independently screened the results of the literature searches, selected eligible studies, extracted the data and evaluated the quality of them. The studies showed that MCC vaccine was highly immunogenic in infants after two and three doses, in toddlers after one and two doses and in older age groups after one dose. In general higher titres were generated after MCC than after MPLS vaccines. Immunological hypo-responsiveness seen after repeated doses of MPLS vaccine may be overcome with MCC. Observational studies have documented a significant decline in meningococcal C disease in countries where MCC vaccines have been widely used. The timing of the vaccinations schedules, the specific conjugate used, and the vaccines given concomitantly or combined, may be important. The MCC vaccine appears to be safe, immunogenic and able to induce immunological memory in all age groups. Observational studies strongly suggest that MCC is clinically

  9. Detection of viral hemorrhagic septicemia virus by quantitative reverse transcription polymerase chain reaction from two fish species at two sites in Lake Superior.

    PubMed

    Cornwell, Emily R; Eckerlin, Geofrey E; Getchell, Rodman G; Groocock, Geoffrey H; Thompson, Tarin M; Batts, William N; Casey, Rufina N; Kurath, Gael; Winton, James R; Bowser, Paul R; Bain, Mark B; Casey, James W

    2011-12-01

    Viral hemorrhagic septicemia virus (VHSV) was first detected in the Laurentian Great Lakes in 2005 during a mortality event in the Bay of Quinte, Lake Ontario. Subsequent analysis of archived samples determined that the first known isolation of VHSV in the Laurentian Great Lakes was from a muskellunge Esox masquinongy collected in Lake St. Clair in 2003. By the end of 2008, mortality events and viral isolations had occurred in all of the Laurentian Great Lakes except Lake Superior. In 2009, a focused disease surveillance program was designed to determine whether VHSV was also present in Lake Superior. In this survey, 874 fish from 7 sites along the U.S. shoreline of Lake Superior were collected during June 2009. Collections were focused on nearshore species known to be susceptible to VHSV. All fish were dissected individually by using aseptic techniques and were tested for the presence of VHSV genetic material by use of a quantitative reverse transcription (qRT) polymerase chain reaction (PCR) targeting the viral nucleoprotein gene. Seventeen fish from two host species at two different sites tested positive at low levels for VHSV. All attempts to isolate virus in cell culture were unsuccessful. However, the presence of viral RNA was confirmed independently in five fish by using a nested PCR that targeted the glycoprotein (G) gene. Partial G gene sequences obtained from three fish were identical to the corresponding sequence from the original 2003 VHSV isolate (MI03) from muskellunge. These detections represent the earliest evidence for the presence of VHSV in Lake Superior and illustrate the utility of the highly sensitive qRT-PCR assay for disease surveillance in aquatic animals.

  10. Detection of Viral Hemorrhagic Septicemia Virus by Quantitative Reverse Transcription Polymerase Chain Reaction from Two Fish Species at Two Sites in Lake Superior

    USGS Publications Warehouse

    Cornwell, Emily R.; Eckerlin, Geofrey E.; Getchell, Rodman G.; Groocock, Geoffrey H.; Thompson, Tarin M.; Batts, William N.; Casey, Rufina N.; Kurath, Gael; Winton, James R.; Bowser, Paul R.; Bain, Mark B.; Casey, James W.

    2011-01-01

    Viral hemorrhagic septicemia virus (VHSV) was first detected in the Laurentian Great Lakes in 2005 during a mortality event in the Bay of Quinte, Lake Ontario. Subsequent analysis of archived samples determined that the first known isolation of VHSV in the Laurentian Great Lakes was from a muskellunge Esox masquinongy collected in Lake St. Clair in 2003. By the end of 2008, mortality events and viral isolations had occurred in all of the Laurentian Great Lakes except Lake Superior. In 2009, a focused disease surveillance program was designed to determine whether VHSV was also present in Lake Superior. In this survey, 874 fish from 7 sites along the U.S. shoreline of Lake Superior were collected during June 2009. Collections were focused on nearshore species known to be susceptible to VHSV. All fish were dissected individually by using aseptic techniques and were tested for the presence of VHSV genetic material by use of a quantitative reverse transcription (qRT) polymerase chain reaction (PCR) targeting the viral nucleoprotein gene. Seventeen fish from two host species at two different sites tested positive at low levels for VHSV. All attempts to isolate virus in cell culture were unsuccessful. However, the presence of viral RNA was confirmed independently in five fish by using a nested PCR that targeted the glycoprotein (G) gene. Partial G gene sequences obtained from three fish were identical to the corresponding sequence from the original 2003 VHSV isolate (MI03) from muskellunge. These detections represent the earliest evidence for the presence of VHSV in Lake Superior and illustrate the utility of the highly sensitive qRT-PCR assay for disease surveillance in aquatic animals.

  11. Epizootiology of viral hemorrhagic septicemia virus in Pacific herring from the spawn-on-kelp fishery in Prince William Sound, Alaska, USA

    USGS Publications Warehouse

    Hershberger, P.K.; Kocan, R.M.; Elder, N.E.; Meyers, T.R.; Winton, J.R.

    1999-01-01

    Both the prevalence and tissue titer of viral hemorrhagic septicemia virus (VHSV) increased in Pacific herring Clupea pallasi following their introduction into net pens (pounds) used in the closed pound spawn-on-kelp (SOK) fishery in Prince William Sound, Alaska. VHSV was also found in water samples from inside and outside the SOK pounds after herring had been confined for several days; however, water samples taken near wild free-ranging, spawning herring either failed to test positive or tested weakly positive for virus. Little or no virus was found in tissue samples from free-ranging, spawning herring captured from the vicinity of the pounds, nor did the prevalence of VHSV increase following spawning as it did in impounded herring. The data indicated that increased prevalences of VHSV were correlated with confinement of herring for the closed pound SOK fishery and that infection was spread within the pounds through waterborne exposure to virus particles originating from impounded fish. In addition, pounds containing predominantly young fish had higher prevalences of VHSV, suggesting that older fish may be partially immune, perhaps as a result of previous infection with the virus. Operation of SOK pounds during spawning seasons in which young herring predominate may amplify the disease and possibly exacerbate the population fluctuations observed in wild herring stocks.

  12. Episootiology of viral hemorrhagic septicemia virus in Pacific herring from the spawn-on-kelp fishery in Prince William Sound, Alaska, USA

    USGS Publications Warehouse

    Hershberger, P.K.; Kocan, R.M.; Elder, N.E.; Meyers, T.R.; Winton, J.R.

    1999-01-01

    Both the prevalence and tissue titer of viral hemorrhagic septicemia virus (VHSV) increased in Pacific herring Clupea pallasi following their introduction into net pens (pounds) used in the closed pound spawn-on-kelp (SOK) fishery in Prince William Sound, Alaska. VHSV was also found in water samples from inside and outside the SOK pounds after herring had been confined for several days; however, water samples taken near wild free-ranging, spawning herring either failed to test positive or tested weakly positive for virus. Little or no virus was found in tissue samples from free-ranging, spawning herring captured from the vicinity of the pounds, nor did the prevalence of VHSV increase following spawning as it did in impounded herring. The data indicated that increased prevalences of VHSV were correlated with confinement of herring for the closed pound SOK fishery and that infection was spread within the pounds through waterborne exposure to virus particles originating from impounded fish. In addition, pounds containing predominantly young fish had higher prevalences of VHSV, suggesting that older fish may be partially immune, perhaps as a result of previous infection with the virus. Operation of SOK pounds during spawning seasons in which young herring predominate may amplify the disease and possibly exacerbate the population fluctuations observed in wild herring stocks.

  13. Establishment and partial characterization of a cell line from burbot Lota lota maculosa: susceptibility to IHNV, IPNV and VHSV.

    PubMed

    Polinski, Mark P; Drennan, John D; Batts, William N; Ireland, Susan C; Cain, Kenneth D

    2010-05-18

    This study describes the development and partial characterization of a continuous fibroblastic-like cell line (BEF-1) developed from late stage embryos of North American burbot Lota lota maculosa. This cell line has been maintained for over 5 yr and 100 passages in vitro. Cells were cultured using Eagle's minimum essential medium with Earle's salts (MEM) supplemented with GlutaMAX, and 10% fetal bovine serum (FBS), pH 7.4. The addition of penicillin-streptomycin-neomycin (PSN) antibiotic mixture (0.05, 0.05, 0.1 mg m(-1), respectively) did not negatively influence cell replication; however, the antimycotic FungizoneTM (2.5 microg m(-1), amphotericin B) caused cell rounding and resulted in a severe decrease in cell proliferation. Optimal incubation temperature has been observed between 15 and 23 degrees C, and at these temperatures cultures are routinely passed using standard trypsinization methods every 5 to 7 d at a split ratio of 1:3 or 1:4. The cell line was susceptible to isolates of the M and U North American genotypes of infectious hematopoietic necrosis virus (IHNV), and to isolates of genotypes I, IVa, and IVb of viral hemorrhagic septicemia virus (VHSV). In contrast, the cell line was refractory to infection by 2 North American isolates of infectious pancreatic necrosis virus (IPNV) from serotypes A1 and A9. This cell line provides a new laboratory tool, will allow further investigation into viral diseases of burbot and possibly other species, and is the first immortalized cell line reported from a species in the Gadidae (cod) family.

  14. Establishment and partial characterization of a cell line from burbot Lota lota maculosa: susceptibility to IHNV, IPNV and VHSV.

    USGS Publications Warehouse

    Batts, William N.; Polinski, Mark P.; Drennan, John D.; Ireland, Susan C.; Cain, Kenneth D.

    2010-01-01

    This study describes the development and partial characterization of a continuous fibroblastic-like cell line (BEF-1) developed from late stage embryos of North American burbot Lota lota maculosa. This cell line has been maintained for over 5 yr and 100 passages in vitro. Cells were cultured using Eagle’s minimum essential medium with Earle’s salts (MEM) supplemented with GlutaMAX™, and 10% fetal bovine serum (FBS), pH 7.4. The addition of penicillin-streptomycin-neomycin (PSN) antibiotic mixture (0.05, 0.05, 0.1 mg ml–1, respectively) did not negatively influence cell replication; however, the antimycotic Fungizone™ (2.5 µg ml–1, amphotericin B) caused cell rounding and resulted in a severe decrease in cell proliferation. Optimal incubation temperature has been observed between 15 and 23°C, and at these temperatures cultures are routinely passed using standard trypsinization methods every 5 to 7 d at a split ratio of 1:3 or 1:4. The cell line was susceptible to isolates of the M and U North American genotypes of infectious hematopoietic necrosis virus (IHNV), and to isolates of genotypes I, IVa, and IVb of viral hemorrhagic septicemia virus (VHSV). In contrast, the cell line was refractory to infection by 2 North American isolates of infectious pancreatic necrosis virus (IPNV) from serotypes A1 and A9. This cell line provides a new laboratory tool, will allow further investigation into viral diseases of burbot and possibly other species, and is the first immortalized cell line reported from a species in the Gadidae (cod) family.

  15. Rainbow trout Oncorhynchus mykiss ladderlectin, but not intelectin, binds viral hemorrhagic septicemia virus IVb.

    PubMed

    Reid, A; Young, K M; Lumsden, J S

    2011-06-16

    The innate immune system of fish is critical for rapid detection and immediate response to infection, as well as to orchestrate the adaptive branch of the immune system. Rainbow trout Oncorhynchus mykiss ladderlectin and intelectin are plasma pattern recognition receptors (PRR) for bacterial and fungal pathogens of rainbow trout, but their role as PRRs for virus is unknown. Viral hemorrhagic septicemia virus (VHSV) IVb is a recently described fish pathogen in the Great Lakes, and rainbow trout can be experimentally infected. Using an indirect enzyme-linked immunosorbent assay, rainbow trout plasma ladderlectin significantly (p < 0.05) bound purified VHSV while intelectin did not. In addition, plasma ladderlectin but not intelectin was eluted from a VHSV-conjugated Toyopearl column using EDTA. Protein identification was confirmed with polyclonal antiserum used with slot immunoblot, 1-dimensional sodium dodecyl sulphate polyacrylamide electrophoresis, and Western immunoblot.

  16. Septicaemia due to Neisseria lactamica--initial confusion with Neisseria meningitidis.

    PubMed

    Brown, N M; Ragge, N K; Speller, D C

    1987-11-01

    Neisseria lactamica, isolated from a baby with septicaemia, was at first thought to be Neisseria meningitidis, possibly acquired in hospital. Extensive investigation of contacts was made until the O-nitrophenyl-D-galactopyranoside reaction proved positive. Distinction between the two species, easily made in this way, is important both in individual patients and in population surveys.

  17. Aetiology, risk factors and immediate outcome of bacteriologically confirmed neonatal septicaemia in Mulago hospital, Uganda.

    PubMed

    Mugalu, J; Nakakeeto, M K; Kiguli, S; Kaddu-Mulindwa, Deo H

    2006-06-01

    Neonatal septicaemia remains a major cause of morbidity and mortality. The aetiology, risk factors and outcome of this problem need to understood. To determine the aetiology, risk factors and immediate outcome of bacteriologically confirmed neonatal septicaemia in Mulago hospital. Blood cultures were aseptically obtained from neonates presenting with clinical sepsis by WHO criteria to Mulago during a five month period between July and November 2002. Blood was placed in Brain Heart Infusion media and incubated within 30 minutes. Subcultures were plated daily up to 7 days on blood, chocolate and MacConkey agar and incubated in aerobic and 5% carbon dioxide conditions. Pure colonies were identified by Gram stain and biochemical tests and antibiotic sensitivities were obtained. Gram positive organisms were predominant (69.2%) followed by E. coli (17%) and Group B Streptococci (GBS) (7%). Staphylococcus aureus and E. coli dominated isolates in early and late onset sepsis. S. aureus was more sensitive to gentamicin than to cloxacillin. The sensitivity of E. coli to ceftriaxone was 94.1%. Factors significantly associated with neonatal septicaemia were male sex, history of convulsions, hypoglycaemia, lack of antenatal care, late onset sepsis and umbilical pus discharge. Mortality in sepsis cases was 18.1%, and 84% of deaths occurred in the first 2 days of admission. Hypoglycaemia was significantly associated with death (p < 0.01). S. aureus predominates the aetiology of neonatal septicaemia followed by E.coli. Most deaths occur in the first 48 hours of admission and hypoglycaemia is significantly associated with death.

  18. Neonatal Klebsiella Septicaemia in Ibadan: Implications for Neonatal Care in Developing Countries.

    ERIC Educational Resources Information Center

    Omokhodion, S. I.; And Others

    1993-01-01

    The antecedent events, clinical features, prevalence, and complications of neonatal Klebsiella septicaemia in 73 infants admitted to a special care baby unit in Nigeria are retrospectively reviewed and compared with those of 72 infants who had no risk factors for sepsis admitted to the same unit during the same period. A nosocomial acquisition of…

  19. Neonatal Klebsiella Septicaemia in Ibadan: Implications for Neonatal Care in Developing Countries.

    ERIC Educational Resources Information Center

    Omokhodion, S. I.; And Others

    1993-01-01

    The antecedent events, clinical features, prevalence, and complications of neonatal Klebsiella septicaemia in 73 infants admitted to a special care baby unit in Nigeria are retrospectively reviewed and compared with those of 72 infants who had no risk factors for sepsis admitted to the same unit during the same period. A nosocomial acquisition of…

  20. A survey of fish viruses isolated from wild marine fishes from the coastal waters of southern Korea.

    PubMed

    Kim, Wi-Sik; Choi, Shin-Young; Kim, Do-Hyung; Oh, Myung-Joo

    2013-11-01

    A survey was conducted to investigate viral infection in 253 wild marine fishes harvested in the southern coastal area of Korea from 2010 to 2012. The fish that were captured by local anglers were randomly bought and sampled for virus examination. The samples were tested for presence of virus by virus isolation with FHM, FSP, and BF-2 cells and molecular methods (polymerase chain reaction and sequencing). Of the 253 fish sampled, 9 fish were infected with virus. Aquabirnaviruses (ABVs), Viral hemorrhagic septicemia virus (VHSV), and Red seabream iridovirus (RSIV) were detected in 7, 1, and 1 fish, respectively. Molecular phylogenies demonstrated the detected viruses (ABV, VHSV, and RSIV) were more closely related to viruses reported of the same type from Korea and Japan than from other countries, suggesting these viruses may be indigenous to Korean and Japanese coastal waters.

  1. In vitro infection of salmonid epidermal tissues by infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus

    USGS Publications Warehouse

    Yamamoto, T.; Batts, W.N.; Winton, J.R.

    1992-01-01

    The ability of two rhabdoviruses, infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV), to infect fish skin was investigated by in vitro infection of excised tissues. Virus replication was determined by plaque assay of homogenized tissue extracts, and the virus antigen was detected by immunohistology of tissue sections. Gill, fin, and ventral abdominal skin tissues of rainbow trout Oncorhynchus mykiss that had been infected in vitro with a virulent strain of IHNV (193–110) produced substantial increases in virus titer within 24 h. Titers continued to increase up until day 3 of incubation; by this time, virus had increased 1,000-fold or more. This increase in IHNV titer occurred in epidermal tissues of fingerlings and of older fish. In another experiment, IHNV replicated in excised rainbow trout tissues whether the fish had been subject to prior infection with a virulent strain of IHNV (Western Regional Aquaculture Consortium isolate) or whether the fish had been infected previously with an attenuated strain of the virus (Nan Scott Lake, with 100 passes in culture). A virulent strain of VHSV (23/75) replicated effectively in excised gill tissues and epidermal tissues of rainbow trout and chinook salmon O. tshawytscha; however, the avirulent North American strain of VHSV (Makah) replicated poorly or not at all.

  2. Host and geographic range extensions of the North American strain of viral hemorrhagic septicemia virus

    USGS Publications Warehouse

    Hedrick, R.P.; Batts, W.N.; Yun, S.; Traxler, G.S.; Kaufman, J.; Winton, J.R.

    2003-01-01

    Viral hemorrhagic septicemia virus (VHSV) was isolated from populations of Pacific sardine Sardinops sagaxfrom the coastal waters of Vancouver Island, British Columbia, Canada, and central and southern California, USA. The virus was also isolated from Pacific mackerel Scomber japonicus in southern California, from eulachon or smeltThaleichthys pacificus, and surf smelt Hypomesus pretiosus pretiosus from Oregon, USA. Mortality and skin lesions typical of viral hemorrhagic septicemia in other marine fish species were observed among sardine in Canada and in a few surf smelt from Oregon, but the remaining isolates of VHSV were obtained from healthy appearing fish. The prevalence of VHSV among groups of apparently healthy sardine, mackerel and smelt ranged from 4 to 8% in California and Oregon. A greater prevalence of infection (58%) occurred in groups of sardine sampled in Canada that sustained a naturally occurring epidemic during 1998-99. A captive group of surf smelt in Oregon exhibited an 81% prevalence of infection with clinical signs in only a few fish. The new isolates were confirmed as North American VHSV and were closely related based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene. The VHSV isolates from sardine in Canada and California were the most closely related, differing from isolates obtained from other marine fish species and salmonids in British Columbia, Canada, Alaska and Washington, USA. These new virus isolations extend both the known hosts (sardine, mackerel and 2 species of smelt) and geographic range (Oregon and California, USA) of VHSV.

  3. Detection of viruses in farmed rainbow trout (Oncorhynchus mykiss) in Korea by RT-LAMP assay.

    PubMed

    Suebsing, Rungkarn; Kim, Jeong-Ho; Kim, Seok Ryel; Park, Myung-Ae; Oh, Myung-Joo

    2011-10-01

    The viral diseases have been the serious problem in salmonid farming, and rainbow trout is not an exception. In this study, routine surveys were conducted for detecting of viruses in farmed rainbow trout (Oncorhynchus mykiss) in Korea during 2009-2010. Head kidneys from individual fish were employed for virus detection by using a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV) were the target viruses in this study. 53.5% (46/86) were found to be IPNV-positive, while IHNV and VHSV showed RT-LAMP negative during examination for 2 years. Ten IPNV-positive samples were randomly selected for viral isolation and the cells showing CPEs were subjected to RT-LAMP, RT-PCR, and direct sequencing. Phylogenetic analysis showed that the rainbow trout isolate has high similarity homologies with the VR-299 strain, as previously described.

  4. Vibrio vulnificus septicaemia in Japan: an estimated number of infections and physicians' knowledge of the syndrome.

    PubMed Central

    Osaka, K.; Komatsuzaki, M.; Takahashi, H.; Sakano, S.; Okabe, N.

    2004-01-01

    Questionnaire surveys were implemented to study the incidence and physicians' knowledge of Vibrio vulnificus infections in Japan. Registered emergency physicians were selected by stratified random sampling for a questionnaire survey. A total of 235 out of 386 physicians (61%) responded to the questionnaire and 12 V. vulnificus septicaemia cases were reported from 10 respondents. The annual estimated number of V. vulnificus septicaemia was calculated as 425 (95 % CI 238-752). The study also revealed that only 15.7 % (95 % CI 11.3-21.0) of responding physicians had a basic knowledge of V. vulnificus infection. Education for both physicians and people in the high-risk group for developing the infection (e.g. immunocompromised, chronic liver disease) will be necessary for the prevention, early diagnosis and appropriate treatment of the disease. PMID:15473164

  5. Pyogranulomatous myocarditis due to Staphylococcus aureus septicaemia in two harbour porpoises (Phocoena phocoena).

    PubMed

    Siebert, U; Müller, G; Desportes, G; Weiss, R; Hansen, K; Baumgärtner, W

    2002-03-02

    Staphylococcus aureus septicaemia was diagnosed in a dead, stranded harbour porpoise from the German Baltic Sea and in a live harbour porpoise by-caught in inner Danish waters and taken into captivity. Lesions included pyogranulomatous myocarditis, necrotising suppurative bronchopneumonia, pyelonephritis, osteomyelitis and leptomeningitis, and abscesses in lymph nodes and skeletal muscles. The captive animal had fibrinous suppurative epicarditis and pyogranulomatous myocarditis with abscesses. In both animals the organism was suspected to have entered through skin lesions or via the respiratory tract.

  6. In vitro and in vivo study of fosfomycin in methicillin-resistant Staphylococcus aureus septicaemia.

    PubMed Central

    Lau, W. Y.; Teoh-Chan, C. H.; Fan, S. T.; Lau, K. F.

    1986-01-01

    Five hundred strains of methicillin-resistant Staphylococcus aureus were tested against various anti-staphylococcal agents. Vancomycin, fusidic acid and fosfomycin were found to be the most effective. Only 1 strain out of 500 was resistant to fosfomycin. Three patients with methicillin-resistant Staphylococcus aureus septicaemia were successfully treated by fosfomycin. We conclude that fosfomycin could be the drug of choice for methicillin-resistant Staphylococcus aureus infection. PMID:3637200

  7. Molecular epidemiology of viral hemorrhagic septicemia virus in the Great Lakes region

    USGS Publications Warehouse

    Winton, James; Kurath, Gael; Batts, William

    2008-01-01

    Viral hemorrhagic septicemia virus (VHSV) is considered by many nations and international organizations to be one of the most important viral pathogens of finfish (Office International des Epizooties 2007). For several decades following its initial characterization in the 1950s, VHSV was thought to be limited to Europe where it was regarded as an endemic pathogen of freshwater fish that was especially problematic for farmed rainbow trout, an introduced species (Wolf 1988; Smail 1999). Subsequently, it was shown that VHSV was present among many species of marine and anadromous fishes in both the Pacific and Atlantic Oceans where it has been associated with substantial mortality among both wild and cultured fish (Meyers and Winton 1995; Skall et al. 2005).

  8. Temporal Variation in Viral Hemorrhagic Septicemia Virus Antibodies in Freshwater Drum (Aplodinotus grunniens) Indicates Cyclic Transmission in Lake Winnebago, Wisconsin.

    PubMed

    Wilson-Rothering, Anna; Marcquenski, Susan; Koenigs, Ryan; Bruch, Ronald; Kamke, Kendall; Isermann, Daniel; Thurman, Andrew; Toohey-Kurth, Kathy; Goldberg, Tony

    2015-09-01

    Viral hemorrhagic septicemia virus (VHSV) is an emerging pathogen that causes mass mortality in multiple fish species. In 2007, the Great Lakes freshwater strain, type IVb, caused a large die-off of freshwater drum (Aplodinotus grunniens) in Lake Winnebago, Wisconsin, USA. To evaluate the persistence and transmission of VHSV, freshwater drum from Lake Winnebago were tested for antibodies to the virus using recently developed virus neutralization (VN) and enzyme-linked immunosorbent (ELISA) assays. Samples were also tested by real-time reverse transcription-PCR (rRT-PCR) to detect viral RNA. Of 548 serum samples tested, 44 (8.03%) were positive by VN (titers ranging from 1:16 to 1:1,024) and 45 (8.21%) were positive by ELISA, including 7 fish positive by both assays. Antibody prevalence increased with age and was higher in one northwestern area of Lake Winnebago than in other areas. Of 3,864 tissues sampled from 551 fish, 1 spleen and 1 kidney sample from a single adult female fish collected in the spring of 2012 tested positive for VHSV by rRT-PCR, and serum from the same fish tested positive by VN and ELISA. These results suggest that VHSV persists and viral transmission may be active in Lake Winnebago even in years following outbreaks and that wild fish may survive VHSV infection and maintain detectable antibody titers while harboring viral RNA. Influxes of immunologically naive juvenile fish through recruitment may reduce herd immunity, allow VHSV to persist, and drive superannual cycles of transmission that may sporadically manifest as fish kills. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Temporal variation in viral hemorrhagic septicemia virus antibodies in freshwater drum (Aplodinotus grunniens) indicates cyclic transmission in Lake Winnebago, Wisconsin

    USGS Publications Warehouse

    Wilson-Rothering, Anna; Marcquenski, Susan; Koenigs, Ryan; Bruch, Ronald; Kamke, Kendall; Isermann, Daniel A.; Thurman, Andrew; Toohey-Kurth, Kathy; Goldberg, Tony

    2015-01-01

    Viral hemorrhagic septicemia virus (VHSV) is an emerging pathogen that causes mass mortality in multiple fish species. In 2007, the Great Lakes freshwater strain, type IVb, caused a large die-off of freshwater drum (Aplodinotus grunniens) in Lake Winnebago, Wisconsin, USA. To evaluate the persistence and transmission of VHSV, freshwater drum from Lake Winnebago were tested for antibodies to the virus using recently developed virus neutralization (VN) and enzyme-linked immunosorbent (ELISA) assays. Samples were also tested by real-time reverse transcription-PCR (rRT-PCR) to detect viral RNA. Of 548 serum samples tested, 44 (8.03%) were positive by VN (titers ranging from 1:16 to 1:1,024) and 45 (8.21%) were positive by ELISA, including 7 fish positive by both assays. Antibody prevalence increased with age and was higher in one northwestern area of Lake Winnebago than in other areas. Of 3,864 tissues sampled from 551 fish, 1 spleen and 1 kidney sample from a single adult female fish collected in the spring of 2012 tested positive for VHSV by rRT-PCR, and serum from the same fish tested positive by VN and ELISA. These results suggest that VHSV persists and viral transmission may be active in Lake Winnebago even in years following outbreaks and that wild fish may survive VHSV infection and maintain detectable antibody titers while harboring viral RNA. Influxes of immunologically naive juvenile fish through recruitment may reduce herd immunity, allow VHSV to persist, and drive superannual cycles of transmission that may sporadically manifest as fish kills.

  10. Temporal Variation in Viral Hemorrhagic Septicemia Virus Antibodies in Freshwater Drum (Aplodinotus grunniens) Indicates Cyclic Transmission in Lake Winnebago, Wisconsin

    PubMed Central

    Wilson-Rothering, Anna; Marcquenski, Susan; Koenigs, Ryan; Bruch, Ronald; Kamke, Kendall; Isermann, Daniel; Thurman, Andrew; Toohey-Kurth, Kathy

    2015-01-01

    Viral hemorrhagic septicemia virus (VHSV) is an emerging pathogen that causes mass mortality in multiple fish species. In 2007, the Great Lakes freshwater strain, type IVb, caused a large die-off of freshwater drum (Aplodinotus grunniens) in Lake Winnebago, Wisconsin, USA. To evaluate the persistence and transmission of VHSV, freshwater drum from Lake Winnebago were tested for antibodies to the virus using recently developed virus neutralization (VN) and enzyme-linked immunosorbent (ELISA) assays. Samples were also tested by real-time reverse transcription-PCR (rRT-PCR) to detect viral RNA. Of 548 serum samples tested, 44 (8.03%) were positive by VN (titers ranging from 1:16 to 1:1,024) and 45 (8.21%) were positive by ELISA, including 7 fish positive by both assays. Antibody prevalence increased with age and was higher in one northwestern area of Lake Winnebago than in other areas. Of 3,864 tissues sampled from 551 fish, 1 spleen and 1 kidney sample from a single adult female fish collected in the spring of 2012 tested positive for VHSV by rRT-PCR, and serum from the same fish tested positive by VN and ELISA. These results suggest that VHSV persists and viral transmission may be active in Lake Winnebago even in years following outbreaks and that wild fish may survive VHSV infection and maintain detectable antibody titers while harboring viral RNA. Influxes of immunologically naive juvenile fish through recruitment may reduce herd immunity, allow VHSV to persist, and drive superannual cycles of transmission that may sporadically manifest as fish kills. PMID:26135873

  11. Bacterial isolates in blood cultures of children with suspected septicaemia in Kano: a two-year study.

    PubMed

    Nwadioha, S I; Kashibu, E; Alao, O O; Aliyu, I

    2011-06-01

    Septicaemia is a common condition in children with a resultant high morbidity and mortality. The gold standard for diagnosis of septicaemia is the isolation of bacterial agents from blood cultures. The study was done to determine the common aetiology of septicaemia in children and their antibiotic susceptibility pattern in Kano, Nigeria. A retrospective study with a review of blood culture reports of paediatric patients aged 0-15 years, suspected of septicaemia, from October 2006 to October 2008 in the Medical Microbiology department of Aminu Kano Teaching Hospital Kano. Kano. Nigeria was carried out. Out of a total of 3840 blood culture samples, only 18.2% (n=700) was culture positive. Gram-negative and gram - positive bacteria constituted 69.3% (n=2661) and 30.7% (n=1179) respectively. The most prevalent bacterial isolates were Escherichia coli with 44.3% (n=310/700) and Staphylococcus aureus 30.7% (n=215/700). Escherichia coli were sensitive to ceftriaxone, ciprofloxacin, gentamicin and clavulinate - amoxyl. The commonest bacterial isolate from blood culture of children with suspected septicaemia in Kano is Escherichia coli. The most sensitive and preferable among the tested antibiotics is ceftriaxone. Rational use of antibiotics with regular antibiotic susceptibility surveillance studies is recommended to maintain high antibiotic therapeutic profile.

  12. Prediction of G gene epitopes of viral hemorrhagic septicemia virus and eukaryotic expression of major antigen determinant sequence.

    PubMed

    Sun, T; Yin, W-L; Fang, B-H; Wang, Q; Liang, C-Z; Yue, Z-Q

    2017-08-15

    This study aims to express fish Viral hemorrhagic septicemia virus (VHSV) G main antigen domain by using Bac-to-bac expression system. Using bioinformatics tools, B cell epitope of VHSV G gene was predicted, and G main antigen domain was optimized. GM gene was inserted into pFastBac1 vector, then transferred recombinant plasmid into DH10Bac to get recombinant rBacmid-GM. Obtained shuttle plasmid rBacmid-GM was transfected into sf9 cells. GM expression was examined using by PCR and western-blot. Results indicated that G main antigen domain gene of VHSV was successfully cloned and sequenced which contains 1209 bp. PCR proved that shuttle plasmid rBacmid-GM was constructed correctly. SDS-PAGE electrophoresis analysis detected a band of protein about 45kD in expression product of G gene. Obtained recombinant G protein reacted with VHSV-positive serum that was substantiated by western-blot analysis. In conclusion, the main antigen domain of VHSV G was successfully expressed in the Bac-to-Bac baculovirus system.

  13. The management of the orthopaedic sequelae of meningococcal septicaemia: patients treated to skeletal maturity.

    PubMed

    Park, D H; Bradish, C F

    2011-07-01

    Septicaemia resulting from meningococcal infection is a devastating illness affecting children. Those who survive can develop late orthopaedic sequelae from growth plate arrests, with resultant complex deformities. Our aim in this study was to review the case histories of a series of patients with late orthopaedic sequelae, all treated by the senior author (CFB). We also describe a treatment strategy to address the multiple deformities that may occur in these patients. Between 1997 and 2009, ten patients (seven girls and three boys) were treated for late orthopaedic sequelae following meningococcal septicaemia. All had involvement of the lower limbs, and one also had involvement of the upper limbs. Each patient had a median of three operations (one to nine). Methods of treatment included a combination of angular deformity correction, limb lengthening and epiphysiodesis. All patients were skeletally mature at the final follow-up. One patient with bilateral below-knee amputations had satisfactory correction of her right amputation stump deformity, and has complete ablation of both her proximal tibial growth plates. In eight patients length discrepancy in the lower limb was corrected to within 1 cm, with normalisation of the mechanical axis of the lower limb. Meningococcal septicaemia can lead to late orthopaedic sequelae due to growth plate arrests. Central growth plate arrests lead to limb-length discrepancy and the need for lengthening procedures, and peripheral growth plate arrests lead to angular deformities requiring corrective osteotomies and ablation of the damaged physis. In addition, limb amputations may be necessary and there may be altered growth of the stump requiring further surgery. Long-term follow-up of these patients is essential to recognise and treat any recurrence of deformity.

  14. The Macrophage Scavenger Receptor A Is Host-Protective in Experimental Meningococcal Septicaemia

    PubMed Central

    Makepeace, Katherine; Moxon, E. Richard; Gordon, Siamon

    2009-01-01

    Macrophage Scavenger Receptor A (SR-A) is a major non-opsonic receptor for Neisseria meningitidis on mononuclear phagocytes in vitro, and the surface proteins NMB0278, NMB0667, and NMB1220 have been identified as ligands for SR-A. In this study we ascertain the in vivo role of SR-A in the recognition of N. meningitidis MC58 (serogroup B) in a murine model of meningococcal septicaemia. We infected wild-type and SR-A−/− animals intraperitoneally with N. meningitidis MC58 and monitored their health over a period of 50 hours. We also determined the levels of bacteraemia in the blood and spleen, and measured levels of the pro-inflammatory cytokine interleukin-6 (IL-6). The health of SR-A−/− animals deteriorated more rapidly, and they showed a 33% reduction in survival compared to wild-type animals. SR-A−/− animals consistently exhibited higher levels of bacteraemia and increased levels of IL-6, compared to wild-type animals. Subsequently, we constructed a bacterial mutant (MC58-278-1220) lacking two of the SR-A ligands, NMB0278 and NMB1220. Mutation of NMB0667 proved to be lethal. When mice were infected with the mutant bacteria MC58-278-1220, no significant differences could be observed in the health, survival, bacteraemia, and cytokine production between wild-type and SR-A−/− animals. Overall, mutant bacteria appeared to cause less severe symptoms of septicaemia, and a competitive index assay showed that higher levels of wild-type bacteria were recovered when animals were infected with a 1∶1 ratio of wild-type MC58 and mutant MC58-278-1220 bacteria. These data represent the first report of the protective role of SR-A, a macrophage-restricted, non-opsonic receptor, in meningococcal septicaemia in vivo, and the importance of the recognition of bacterial protein ligands, rather than lipopolysaccharide. PMID:19214213

  15. [Fulminant pneumococcal septicaemia in a patient with systemic lupus erythematosus and functional asplenia].

    PubMed

    Troldborg, Anne; Deleuran, Bent; Storgaard, Merete

    2014-06-30

    Autosplenectomy is a rare phenomenon in patients with systemic lupus erythematosus (SLE). We present a case of pneumococcal septicaemia in an SLE patient. A CT-scan of the abdomen showed complete autosplenectomy, but a CT scan from years prior showed a normal size spleen. SLE patients have an increased risk of invasive pneumococcal disease and asplenia increases this risk. In Denmark there are no guidelines for vaccination of SLE patients. On the basis of this case report and the guidelines from European League Against Rheumatism we propose that the 23-polyvalent pneumococcal vaccine should be part of the standard regime in the management of SLE.

  16. Wide range of susceptibility to rhabdoviruses in homozygous clones of rainbow trout.

    PubMed

    Quillet, Edwige; Dorson, Michel; Le Guillou, Sandrine; Benmansour, Abdenour; Boudinot, Pierre

    2007-05-01

    Inbred lines differentially susceptible to diseases are a powerful tool to get insights into the mechanisms of genetic resistance to pathogens. In fish, chromosome manipulation techniques allow a quick production of such homozygous lines. Using gynogenesis, we produced nine homozygous clones of rainbow trout from a domestic population (INRA Sy strain). We examined the variability between clones for resistance to two rhabdoviruses, the viral haemorrhagic septicaemia virus (VHSV) and the infectious haematopoietic necrosis virus (IHNV). Intraperitoneal injections and waterborne infections were performed in parallel for both viruses. No survival was recorded after intraperitoneal injection of VHSV or IHNV, indicating that fish from all clones were fully susceptible to both viruses by this route of infection. In contrast, the different clones showed a wide range of survival frequency after waterborne infection. The resistance levels to VHSV ranged from 0 to 99% and resistance was not abrogated when resistant and sensitive animals were mixed and subjected to waterborne infection. VHSV was recovered from 10% of resistant fish after waterborne infection, confirming that virus replication was possible in this context but effective only in a low proportion of the population. The different clones also exhibited a wide range of survival (0-68%) after a waterborne infection with IHNV. Although VHSV-resistant clones were not fully resistant to IHNV, the susceptibility to IHNV and VHSV tended to be correlated, suggesting that non-specific mechanisms common to both viruses were involved.

  17. Genetic comparison of viral hemorrhagic septicemia virus isolates from North America and Europe

    USGS Publications Warehouse

    Oshima, K.H.; Higman, K.H.; Arakawa, C.K.; de Kinkelin, P.; Jorgensen, P.E.V.; Meyers, T.R.; Winton, J.R.

    1993-01-01

    Viral hemorrhagic septicemia virus (VHSV) is the cdusative agent of a serious rhabdoviral d~sease of rainbow trout Oncorhynchus myklss in Europe The first isolation of the vlrus in North Amenca occurred In the fall of 1988 when it was recovered from adult chinook 0 tshawytscha and coho 0 klsutch salmon returning to 2 hatcher~es in the state of Washington, USA The following year, VHSV was isolated from adult coho salmon at 2 other hatcher~es in northwestern Washington In 1990 and 1991, VHSV was recovered from Pacific cod Gadus macrocephalus caught in Pnnce Willlam Sound, Alaska Genetic vanation among the 4 isolates from salmon and the 1990 ~solate from Pacific cod was determ~ned uslng T1 nbonuclease finqerprlnt~ng In addition, 4 d~verse isolates from Europe were lncluded for companson The North Amencan isolates of VHSV formed a slngle fingerprint group In which the 4 isolates from salmonids were h~ghly similar to each other and the isolate from Pacific cod was related but less s~milar The 4 European ~solates which included an isolate from Atlantic cod G morhua, formed a second fingerpnnt group The genetic d~vers~ty among the isolates within each fingerpnnt group was estimated to be less than 5 % whlle the North Amencan and European strains of the virus were judged to differ by more than 5% The results indicate that the North Amerlcan isolates of VHSV are not of European ongln and that the virus may be enzootic wlthin the manne environment.

  18. Haemophilia-associated Yersinia pseudotuberculosis serotype O:1 septicaemia: the role of iron.

    PubMed

    Mischnik, Alexander; Dahme, Tillman; Bekeredjian, Raffi; Zimmermann, Stefan

    2012-01-01

    Septicaemia and septic arthritis due to Yersinia pseudotuberculosis are rare diseases with high mortality rates. Reactive arthritis caused by Yersinia infection is a well-known complication but septic arthritis is found at a much lower frequency. It has already been established that there is a relationship between yersiniosis and iron but there are currently no data about yersiniosis and haematological disorders such as haemophilia. We report a case of septic arthritis due to Y. pseudotuberculosis as an early manifestation of Yersinia septicaemia in a patient with severe haemophilia A. The patient had no history of immunosuppression and presented with a repeat case of haemarthrosis with a fever of unknown origin. Furthermore, he suffered from acute-on-chronic renal failure and non-ST segment elevation myocardial infarction. Arthrocentesis and blood culture tested positive for Y. pseudotuberculosis. Iron deposits at localized sites in the synovium in patients with haemophilia have been described, and as Yersinia infections are common in patients with secondary iron overload, we felt that a review of the literature was in order. Severe Yersinia infection is often associated with iron overload, a condition that might occur as a side effect in the treatment of haemophilia. Iron overload, which plays an important role in the pathogenesis of haemophilic arthropathy, may have increased the virulence of Y. pseudotuberculosis in the present case.

  19. Identification and characterization of pathogen to bacterial septicaemia in cultured turbot, Scophthalmus maximus

    NASA Astrophysics Data System (ADS)

    Qin, Guomin; Zhang, Xiaojun; Chen, Cuizhen; Fang, Hai; Zhan, Wenbin

    2007-10-01

    Bacteria-infected turbots Scophthalmus maximus with septicaemia were examined between 2001 and 2004 in aspects of the conditions of disease occurrence, clinical syndromes and pathological changes. The phenotypic information of pathogenic bacteria was studied, including morphology, physiological and biochemical characteristics, and the mol% G+C of the DNA. In addition, representative strains (S010623-1, LH031120-1) were selected for molecular identification by partial 16S rRNA gene sequencing. The results show that the isolates (LH031120-1 to LH031120-6, HT040308-1 to HT040308-6, HT040620-1 to HT040620-6) from three farms were identified as Edwardsiella tarda. The isolates (S010610-1 to S010610-10, S010623-1 to S010623-20) from one farm were identified as Listonella anguillarum. We conducted studies on the pathogenicity of isolates by artificial infection, and revealed all infected groups in morbidity and mortality. The septicaemia infected turbot showed a syndrome similar to that of the naturally infected fish. Antibiotic sensitivity showed that of 37 antimicrobial agents, E. tarda was sensitive to 27 agents, and L. anguillarum was sensitive to 21 agents.

  20. Infectious hematopoietic necrosis (IHN) and viral hemorrhagic septicemia (VHS): Detection of the trout antibodies to the causative viruses by means of plaque neutralization, immunofluorescence, and enzyme-linked immunosorbent assay

    USGS Publications Warehouse

    Vestergard Jorgensen, P. E.; Olesen, N.J.; Lorenzen, N.; Winton, J.R.; Ristow, S.S.

    1991-01-01

    Sera collected from cultured rainbow trout Oncorhynchus mykiss surviving outbreaks of infectious hematopoietic necrosis (IHN) or viral hemorrhagic septicemia (VHS) were examined for the presence of antibodies to both of the causative viruses, infectious hematopoietic necrosis virus (IHNV) and Egtved virus (viral hemorrhagic septicemia virus: VHSV). Sera were screened with three serological tests: 50% plaque neutralization test (PNT), immunofluorescence (IF), and enzyme-linked immunosorbent assay (ELISA). In sera from 20 rainbow trout surviving IHN, antibodies to IHNV were detected in 9 fish by PNT, in 12 fish by IF, and in 9 fish by ELISA. In these sera, antibodies cross-reacting with VHSV were rare (detected in 0 fish by PNT, in 1 by IF, and in 1 by ELISA). In sera from 20 rainbow trout surviving VHS, antibodies to VHSV were detected in 9 fish by PNT, in 16 fish by IF, and in 18 fish by ELISA. A considerable percentage of the VHS-survivor sera contained antibodies that cross-reacted with IHNV, as detected by ELISA (16 fish) and 1F (7 fish) but not by PNT (0 fish). The three serological tests appear to be useful tools for IHNV and VHSV epidemiology; however, the presence of cross-reacting antibodies in some sera suggests caution when farms require specific pathogen-free certification for one of the viruses in the presence of the other.

  1. Bacteriological spectrum and antimicrobial susceptibility pattern of neonatal septicaemia in a tertiary care hospital of North India.

    PubMed

    Lamba, Mamta; Sharma, Rajni; Sharma, Deepak; Choudhary, Mukesh; Maheshwari, Rakesh Kumar

    2016-12-01

    Neonatal septicaemia is a clinical entity that is characterised by systemic signs and symptoms of infection and accompanied by bacteraemia in first 4 weeks of life and is one of the four leading causes of neonatal mortality and morbidity in India. To determine the bacterial spectrum and antimicrobial susceptibility pattern of neonatal septicaemia in a tertiary care hospital of North India. In this prospective observational study, 850 blood samples were collected and processed from clinically suspected neonates according to standard laboratory protocol. Antimicrobial susceptibility of the isolates was done by Kirby Bauer disc diffusion method according to Clinical and Laboratory Standard Institution (CLSI) recommendations. Blood culture reports were positive in 322 (37.8%) cases. Early onset sepsis (EOS) was present in 61.41% and late onset sepsis (LOS) in 38.59% of cases. Gram-negative septicaemia (60.67%) was encountered more than Gram-positive (32.01%). Coagulase negative Staphylococci (17.43%) was the predominant isolate followed by, Klebsiella spp in 16.11% cases. Best overall sensitivity among Gram-negative isolates was to Colistin (89.94%), Imipenem (86.43%) and Meropenam (77.88%). Gram-positive isolates had good (97.15%) sensitivity to linezolid, (95.23%) vancomycin and (88.57%) Teicoplanin. Gram-negative organisms are the leading cause of neonatal septicaemia with Klebsiella spp being commonest. Coagulase negative Staphylococci is the predominant isolate among Gram-positive organisms. Most of the isolates are resistant to common antibiotics.

  2. Antiviral effects of extracts from Celosia cristata and Raphanus sativus roots against viral hemorrhagic septicemia virus.

    PubMed

    Park, Yeon Jung; Moon, Chiung; Kang, Jung-Ha; Choi, Tae-Jin

    2017-06-01

    The antiviral activity of an extract mixture from Celosia cristata and Raphanus sativus was tested against viral hemorrhagic septicemia virus (VHSV). Pretreatment of EPC cells with this extract up to 72 h before VHSV infection markedly reduced the virus titer, but it had no effect when added after virus inoculation. In olive flounder that received 5 µg of extract per fish, Mx expression peaked at 48 h after treatment. In contrast, ISG15 and TLR2 expression peaked at 72 h, and that of TLR7 peaked at 48 h, followed by a slight decrease at 72 h, indicating that the antiviral activity was mediated by induction of gene expression involved in the innate immune response.

  3. Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes

    PubMed Central

    Moustafa, Ahmed M.; Seemann, Torsten; Gladman, Simon; Adler, Ben; Harper, Marina; Boyce, John D.; Bennett, Mark D.

    2015-01-01

    Pasteurella multocida is the primary causative agent of a range of economically important diseases in animals, including haemorrhagic septicaemia (HS), a rapidly fatal disease of ungulates. There is limited information available on the diversity of P. multocida strains that cause HS. Therefore, we determined draft genome sequences of ten disease-causing isolates and two vaccine strains and compared these genomes using a range of bioinformatic analyses. The draft genomes of the 12 HS strains were between 2,298,035 and 2,410,300 bp in length. Comparison of these genomes with the North American HS strain, M1404, and other available P. multocida genomes (Pm70, 3480, 36950 and HN06) identified a core set of 1,824 genes. A set of 96 genes was present in all HS isolates and vaccine strains examined in this study, but absent from Pm70, 3480, 36950 and HN06. Moreover, 59 genes were shared only by the Asian B:2 strains. In two Pakistani isolates, genes with high similarity to genes in the integrative and conjugative element, ICEPmu1 from strain 36950 were identified along with a range of other antimicrobial resistance genes. Phylogenetic analysis indicated that the HS strains formed clades based on their country of isolation. Future analysis of the 96 genes unique to the HS isolates will aid the identification of HS-specific virulence attributes and facilitate the development of disease-specific diagnostic tests. PMID:26151935

  4. Evaluation of the Efficacy of Iodophor Disinfection of Walleye and Northern Pike Eggs to Eliminate Viral Hemorrhagic Septicemia Virus

    USGS Publications Warehouse

    Tuttle-Lau, M.T.; Phillips, K.A.; Gaikowski, M.P.

    2009-01-01

    Viral hemorrhagic septicemia virus (VHSv) is a serious fish pathogen that has been responsible for large-scale fish kills in the Great Lakes since 2005. It causes high mortality and resulting outbreaks have severe economic consequences for aquaculture. Iodophor disinfection of salmonid eggs is a standard hatchery practice to reduce the risk of pathogen transfer during gamete collection ('spawning') operations and is thus a leading candidate for reducing VHSv transmission during and after spawning of nonsalmonid fishes. However, before it is incorporated by hatcheries during nonsalmonid fish spawning efforts, its safety and effectiveness needs to be evaluated. The USGS Fact Sheet 2009-3107, 'Evaluation of the Efficacy of Iodophor Disinfection of Walleye and Northern Pike Eggs to Eliminate Viral Hemorrhagic Septicemia Virus' presents the results of a study to assess the effectiveness of iodophor disinfection for eliminating VHSv (strain IVb) from fertilized eggs of walleye and northern pike intentionally challenged with VHSv following egg fertilization. Walleye and northern pike egg survival (hatch) following iodophor egg disinfection also was assessed.

  5. Pepscan mapping of viral hemorrhagic septicemia virus glycoprotein G major lineal determinants implicated in triggering host cell antiviral responses mediated by type I interferon.

    PubMed

    Chico, V; Martinez-Lopez, A; Ortega-Villaizan, M; Falco, A; Perez, L; Coll, J M; Estepa, A

    2010-07-01

    Surface glycoproteins of enveloped virus are potent elicitors of type I interferon (IFN)-mediated antiviral responses in a way that may be independent of the well-studied genome-mediated route. However, the viral glycoprotein determinants responsible for initiating the IFN response remain unidentified. In this study, we have used a collection of 60 synthetic 20-mer overlapping peptides (pepscan) spanning the full length of glycoprotein G (gpG) of viral hemorrhagic septicemia virus (VHSV) to investigate what regions of this protein are implicated in triggering the type I IFN-associated immune responses. Briefly, two regions with ability to increase severalfold the basal expression level of the IFN-stimulated mx gene and to restrict the spread of virus among responder cells were mapped to amino acid residues 280 to 310 and 340 to 370 of the gpG protein of VHSV. In addition, the results obtained suggest that an interaction between VHSV gpG and integrins might trigger the host IFN-mediated antiviral response after VHSV infection. Since it is known that type I IFN plays an important role in determining/modulating the protective-antigen-specific immune responses, the identification of viral glycoprotein determinants directly implicated in the type I IFN induction might be of special interest for designing new adjuvants and/or more-efficient and cost-effective viral vaccines as well as for improving our knowledge on how to stimulate the innate immune system.

  6. Pepscan Mapping of Viral Hemorrhagic Septicemia Virus Glycoprotein G Major Lineal Determinants Implicated in Triggering Host Cell Antiviral Responses Mediated by Type I Interferon▿

    PubMed Central

    Chico, V.; Martinez-Lopez, A.; Ortega-Villaizan, M.; Falco, A.; Perez, L.; Coll, J. M.; Estepa, A.

    2010-01-01

    Surface glycoproteins of enveloped virus are potent elicitors of type I interferon (IFN)-mediated antiviral responses in a way that may be independent of the well-studied genome-mediated route. However, the viral glycoprotein determinants responsible for initiating the IFN response remain unidentified. In this study, we have used a collection of 60 synthetic 20-mer overlapping peptides (pepscan) spanning the full length of glycoprotein G (gpG) of viral hemorrhagic septicemia virus (VHSV) to investigate what regions of this protein are implicated in triggering the type I IFN-associated immune responses. Briefly, two regions with ability to increase severalfold the basal expression level of the IFN-stimulated mx gene and to restrict the spread of virus among responder cells were mapped to amino acid residues 280 to 310 and 340 to 370 of the gpG protein of VHSV. In addition, the results obtained suggest that an interaction between VHSV gpG and integrins might trigger the host IFN-mediated antiviral response after VHSV infection. Since it is known that type I IFN plays an important role in determining/modulating the protective-antigen-specific immune responses, the identification of viral glycoprotein determinants directly implicated in the type I IFN induction might be of special interest for designing new adjuvants and/or more-efficient and cost-effective viral vaccines as well as for improving our knowledge on how to stimulate the innate immune system. PMID:20463070

  7. Walleye Sander vitreus (Mitchill) are relatively resistant to experimental infection with VHSV IVb and extant walleye strains vary in susceptibility.

    PubMed

    Grice, J; Reid, A; Peterson, A; Blackburn, K; Tubbs, L; Lord, S; Huber, P; Horricks, R; Dixon, B; Bols, N C; Lumsden, J S

    2014-09-15

    Compared to fathead minnow, walleye demonstrate low susceptibility to experimental infection with VHSV IVb, regardless of route of exposure or water temperature at time of infection. In triplicate and duplicate groups, walleye were intraperitoneally (i.p.) injected (10(2) -10(8)  pfu/fish) or waterborne-exposed (w; 1.4 × 10(7)  pfu mL(-1) ) with VHSV IVb. High cumulative mortality (64-100%) and severe gross lesions associated with VHSV IVb infection were evident only in fish i.p. injected with 10(8)  pfu at 12 °C. These fish had multifocal necrosis of several tissues including the gill and heart. There was no difference in mortality between walleye infected (w or i.p.) at 12 °C (spring stocking) compared with a declining temperature profile from 18 to 12 °C (fall stocking). There were significant differences (P < 0.05) in mortality between four extant walleye strains following i.p. infection, indicating that the choice of walleye strain for stocking might be an important consideration. Viral antigen was found in both i.p. and w-exposed walleye using immunohistochemistry, mostly within the gill and skin of w-exposed fish and most prominently in dermal fibrocytes. VHSV IVb was detected in multiple tissues from 6 to 21 days post-infection using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR).

  8. Edwardsiella Septicaemias

    USDA-ARS?s Scientific Manuscript database

    The genus Edwardsiella includes two species of bacteria that cause major diseases in fish: Edwardsiella tarda (1965) infects fish and other animals and Edwardsiella ictaluri (Hawke 1979; Hawke et al. 1981) infects primarily fish. A third species, Edwardsiella hoshinae (Grimont et al. 1980), infects ...

  9. DNA analysis of nosocomial infection by Enterobacter aerogenes in three cases of septicaemia in Japan.

    PubMed

    Goshi, S; Taneike, I; Nakagawa, S; Kojio, S; Tamura, Y; Ohara, T; Ozaki, K; Tsukada, H; Aoki, Y; Asakura, H; Gejyo, F; Itoh, M; Yamamoto, T

    2002-07-01

    Ceftazidime-resistant Enterobacter aerogenes was isolated from blood cultures of three patients with fever. DNA analysis using pulsed-field gel electrophoresis and ribosomal RNA gene restriction digest pattern analysis revealed that the strains were clonally similar to each other with a 79.3-96.0% homology. The same strain of E. aerogenes was isolated from a three-way stopcock connected to the indwelling catheter in one of the patients at a concentration of 45 cfu/mL. A similar strain was also isolated from the urine of one other patient on the same floor. The data suggest that E. aerogenes caused septicaemia via low bacterial contamination of a three-way stopcock in a peripheral drip intravenous infusion system in at least one patient, and that the outbreak of E. aerogenes infections was due to clonally-related strains.

  10. Bilateral choroidal detachment with exudative retinal detachment in a patient with septicaemia.

    PubMed

    Rani, Padmaja Kumari; Ambiya, Vikas; Senthil, Sirisha; Jalali, Subhadra

    2016-12-20

    We present a case of bilateral choroidal and exudative retinal detachment with hypotony in a patient with septicaemia. Our patient after initial hospitalisation was treated by the local ophthalmologist as a case of sterile pan uveitis with systemic and topical steroids that did not have any effect. Our patient responded well to timely surgical intervention of choroidal drainage and vitrectomy with silicone oil tamponade followed by steroids and antibiotics. Cataract progression in both eyes occurred due to hypotony, inflammation, use of steroids and the vitreous surgery. It was successfully managed with timely surgical removal of cataract and intraocular lens implantation in both eyes facilitating early visual rehabilitation. Topical medications included frequent topical steroids and antibiotics as per our published protocols. 2016 BMJ Publishing Group Ltd.

  11. Distal tibial physeal arrest after meningococcal septicaemia: management and outcome in seven ankles.

    PubMed

    Monsell, F P; Barnes, J R; Kirubanandan, R; McBride, A M B

    2011-06-01

    Survivors of infantile meningococcal septicaemia often develop progressive skeletal deformity as a result of physeal damage at many sites, particularly in the lower limb. Distal tibial physeal arrest typically occurs with sparing of the distal fibular physis leading to a rapidly progressive varus deformity. There have been reports of isolated cases of this deformity, but to our knowledge there have been no papers which specifically describe the development of the deformity and the options for treatment. Surgery to correct this deformity is complex because of the patient's age, previous scarring and the multiplanar nature of the deformity. The surgical goal is to restore leg-length equality and the mechanical axis at the end of growth. Surgery should be planned and staged throughout growth in order to achieve the best functional results. We report our experience in six patients (seven ankles) with this deformity, who were managed by corrective osteotomy using a programmable circular fixator.

  12. Association of Streptococcus pluranimalium with valvular endocarditis and septicaemia in adult broiler parents.

    PubMed

    Hedegaard, L; Christensen, H; Chadfield, M S; Christensen, J P; Bisgaard, M

    2009-04-01

    The genus Streptococcus consists of more than 60 species, but only Streptococcus equi subspecies zooepidemicus, Streptococcus gallolyticus ssp. gallolyticus, Streptococcus gallinaceus, Streptococcus dysgalactiae, Streptococcus mutans and Streptococcus suis have been isolated from poultry. During investigations of the aetiology of increased mortality in broiler parent stock at the end of production, pure cultures of streptococcal-like organisms that could not be classified among these six species were obtained from 24 cases of septicaemia or valvular endocarditis and septicaemia. Phenotypic characterization using the API20 STREP kit identified the isolates as Aerococcus viridans (10), Aerococcus urinae (2), Leuconostoc species (4), Streptococcus salivarius (2), Streptococcus bovis II 3 (1), Enterococcus avium (3), Enterococcus faecium (1) or Gemella morbillorum (1). However, this identification was misleading as subsequent genetic investigations using pulse field gel electrophoresis and sequencing of 16S rRNA genes showed that 19 isolates were classified as Streptococcus pluranimalium, while the remaining isolates were E. avium (3), E. faecium (1) or Lactobacillus species (1). Misidentification by API20 STREP was related to the database provided by the manufacturer, as the phenotypic characteristics could identify these organisms as S. pluranimalium. The isolates of S. pluranimalium belonged to at least three different clones as determined by pulsed field gel electrophoresis of SmaI-digested genomic DNA. The capacity that these isolates had to colonize the valvular endothelium was suggested by the occurrence of valvular endocarditis in 12 of 19 cases. Demonstration of the same clone in all four houses on a farm suggested the pathogenic potential of this organism.

  13. Costs of surviving meningococcal disease in Spain: evaluation for two cases of severe meningitis and septicaemia.

    PubMed

    Darbà, Josep; Kaskens, Lisette; Hark, Mareile; Wright, Claire

    2014-09-03

    The aim of this study was to count the lifelong rehabilitation costs associated with surviving meningococcal disease with major sequelae from the perspective of the Spanish National Healthcare System (NHS) and the national government. Two severe scenarios describing meningococcal disease were developed, one case that represented meningococcal septicaemia and another case for meningococcal meningitis. The scenarios were developed based on a literature review on severe sequelae of meningococcal disease, and discussions with paediatricians who have been responsible for the treatment of children with this disease in Spain. Second, a detailed list of all health, educational and social care resources used by survivors during their acute illness and during the rest of their lives and by family members was obtained by interviewing survivors and their families. Professionals in health and social care were also interviewed to complete the list of resources and ensure the scenario's were accurate. The costs attributed to these resources were obtained from tariff lists, catalogues and published information by the national authorities. All costs were based on a life expectancy of a survivor of 70 years and expressed in EUR 2012. In this study it was estimated that the lifelong discounted rehabilitation costs associated with the treatment of long-term sequelae due to meningococcal disease are approximately €1180,000-€1400,000. Medical care and social care were the main cost drivers for both septicaemia and meningitis. Annual costs showed to be the largest in the first year after diagnosis of the disease for both cases, due to high hospital admission and medical care costs during this period and decreased significantly over the years. This study shows that the lifelong rehabilitation costs associated with the survival of meningococcal disease with severe sequelae place an important burden on the NHS budget and governmental resources in Spain. Copyright © 2014 Elsevier Ltd

  14. Pharmacokinetics and clinical efficacy of cefotaxime for the treatment of septicaemia in dogs.

    PubMed

    Sumano, H; Gutierrez, Lilia; Ocampo, L

    2004-01-01

    Considering the already known pharmacological features of cefotaxime, a study with two approaches of pharmacokinetics and clinical efficacy in septicaemic dogs was carried out. Pharmacokinetic variables were defined for doses of 10 mg/kg, and 20 mg/kg, utilising a quantitative bacteriological analysis. Values for half-life (T1/2 beta) at 10 mg/kg were 0.8, 1.48 and 1.52 h for the i.v., s.c. and i.m. routes, respectively. Corresponding values for the 20 mg/kg dose for the same routes were 0.8, 1.49 and 1.53 h, respectively. Relatively fast clearance (ranging from 0.58 to 0.64 L/kg/h) allowed a maximum dose interval of 12 h. The above-stated doses of cefotaxime were administered i.v. to 40 cases of septicaemia, clinically divided into 20 moderately severe cases treated with 10 mg/kg i.v., of cefotaxime bid, and 20 severe ones, treated with 20 mg/kg i.v. of cefotaxime bid. Injections continued until a previously defined criterion of 'clinically recovered' was obtained. Thereafter, a follow-up treatment was established using the same dose and dose-interval but through the s.c. route. Due to the apparent volumes of distribution obtained (ranging from 0.48 to 0.51 L/kg), considering the overall clinical efficacy obtained (90% for the 10 mg/kg dose and 75% for the 20 mg/kg dose), and due to the rapid improvement observed after a few doses of the drug (1.8 to 2.5 doses to 'clinical improvement'), it is safe to postulate such doses of cefotaxime as excellent choices for the treatment of septicaemia in dogs.

  15. Streptococcus viridans septicaemia: a comparison study in patients admitted to the departments of infectious diseases and haematology in a university hospital.

    PubMed

    Westling, Katarina; Ljungman, Per; Thalme, Anders; Julander, Inger

    2002-01-01

    Infective endocarditis caused by viridans streptococci is a well-described disease. Streptococcus viridans is also an important etiologic agent causing septicaemia in neutropenic patients with haematological diseases. In this study we retrospectively reviewed charts from 111 patients with 121 episodes of viridans streptococci septicaemia during the period 1992-97 for clinical data, presence of endocarditis, subtype and outcome. Forty-seven episodes of S. viridans septicaemia were documented in 45 non-neutropenic patients treated at the Department of Infectious Diseases (Group A). Thirty of these episodes were defined as definite and 9 as possible infective endocarditis, using Duke's critera. Seventy-four episodes of S. viridans septicaemia were identified in 66 patients treated at the Department of Haematology (Group B), only 1 of which fulfilled the criteria for possible infective endocarditis. S. sanguis was the most common subtype (18/47; 38%) in Group A and S. mitis was the major subtype (51/74; 69%) in Group B.

  16. A study on blood culture positivity and C-reactive protein variability in neonatal septicaemia at neonatal intensive care unit of a tertiary care hospital.

    PubMed

    Bhattacharyya, Kumkum; Bandyopadhyay, Maitreyi; Karmakar, Balai Chandra; Bhattacharya, Sujata; Banerjee, Parthajit; Chatterjee, Sumanta; Dutta, Riju

    2012-12-01

    Neonatal septicaemia is an important cause of neonatal morbidity and mortality. Blood culture is the gold standard for diagnosis of neonatal septicaemia. Several laboratory investigations are available to detect neonatal sepsis, one important is 'sepsis screen' which includes C-reactive protein (CRP), micro -ESR, total WBC count including immature to mature (VT) ratio and absolute neutrophil count. Blood culture was done among 210 samples from neonatal intensive care unit (NICU), RG Kar Medical College with suspected septicaemia along with CRP estimation. Among all the parameters, clinical correlation of CRP is significant; 65.2% of patients has blood culture positive. CRP positivity varied in different organisms. CRP is a non-specific acute phase reactant and rises significantly after 12 hours onwards. It can be used as an important parameter in infant at risk of septicaemia (significant > 6 mg/dl) and early institution of antimicrobials therapy. It has got prognostic value.

  17. Corexit 9500 inactivates two enveloped viruses of aquatic animals but enhances the infectivity of a nonenveloped fish virus.

    PubMed

    Pham, P H; Huang, Y J; Chen, C; Bols, N C

    2014-02-01

    The effects of Corexit 9500, a dispersant used to clean up oil spills, on invertebrates, lower vertebrates, birds, and human health have been examined, but there is a significant lack of study of the effect of this dispersant on aquatic viruses. In this study, the effects of Corexit 9500 on four aquatic viruses of differing structural composition were examined. Corexit 9500 reduced the titer of the enveloped viral hemorrhagic septicemia virus (VHSV) at all concentrations (10% to 0.001%) examined. The titer of frog virus 3 (FV3), a virus with both enveloped and nonenveloped virions, was reduced only at the high Corexit 9500 concentrations (10% to 0.1%). Corexit 9500 was unable to reduce the titer of nonenveloped infectious pancreatic necrosis virus (IPNV) but enhanced the titer of chum salmon reovirus (CSV) by 2 to 4 logs. With the ability to inactivate enveloped viruses and possibly enhance some nonenveloped viruses, Corexit 9500 has the potential to alter the aquatic virosphere.

  18. Septicaemia in a population-based HIV clinical cohort in rural Uganda, 1996-2007: incidence, aetiology, antimicrobial drug resistance and impact of antiretroviral therapy.

    PubMed

    Mayanja, B N; Todd, J; Hughes, P; Van der Paal, L; Mugisha, J O; Atuhumuza, E; Tabuga, P; Maher, D; Grosskurth, H

    2010-06-01

    To describe the incidence and aetiology of septicaemia, and antimicrobial drug resistance in HIV-infected and uninfected individuals, and the impact of antiretroviral therapy (ART) on septicaemia. Between 1996 and 2007, we followed up a rural population-based cohort of HIV-infected and uninfected participants. The aetiology and incidence of septicaemia, and antimicrobial drug resistances were determined. ART became available in 2004, and its impact on the incidence of septicaemia was examined. The overall septicaemia incidence (per 1000 pyrs) was 32.4 (95% CI 26.2-40.6) but was only 2.6 (95% CI 1.3-6.2) in HIV-negative patients and 67.1 (95% CI 53.4-85.4) in HIV-positive patients not on ART. Among those on ART, the overall incidence was 71.5 (95% CI 47.1-114.3), although it was 121.4 (95%CI 77.9-200.4) in the first year on ART and 37.4 (95%CI 18.9-85.2) in the subsequent period. Septicaemia incidence was significantly associated with lower CD4 counts. The commonest isolates were Streptococcus pneumoniae (SPN, n = 68) and Non-typhi salmonellae (NTS, n = 42). Most SPN isolates were susceptible to ceftriaxone and erythromycin, while resistance to cotrimoxazole and penicillin was common. All NTS isolates were susceptible to ciprofloxacin, but resistance to cotrimoxazole and chloramphenicol was common. Septicaemia incidence was higher in HIV-infected than in HIV-uninfected participants, and it remained high for some time among those who started ART. Starting ART earlier at higher CD4 counts is likely to lead to lower septicaemia incidence. Both SPN and NTS, the commonest isolates, were resistant to most commonly available antimicrobials. Blood culture laboratory surveillance systems to monitor antibiotic susceptibility and inform treatment guidelines are needed in Africa.

  19. Chronic and persistent viral hemorrhagic septicemia virus infections in Pacific herring

    USGS Publications Warehouse

    Hershberger, P.K.; Gregg, J.L.; Grady, C.A.; Taylor, L.; Winton, J.R.

    2010-01-01

    Chronic viral hemorrhagic septicemia virus (VHSV) infections were established in a laboratory stock of Pacific herring Clupea pallasii held in a large-volume tank supplied with pathogenfree seawater at temperatures ranging from 6.8 to 11.6??C. The infections were characterized by viral persistence for extended periods and near-background levels of host mortality. Infectious virus was recovered from mortalities occurring up to 167 d post-exposure and was detected in normal-appearing herring for as long as 224 d following initial challenge. Geometric mean viral titers were generally as high as or higher in brain tissues than in pools of kidney and spleen tissues, with overall prevalence of infection being higher in the brain. Upon re-exposure to VHSV in a standard laboratory challenge, negligible mortality occurred among groups of herring that were either chronically infected or fully recovered, indicating that survival from chronic manifestations conferred protection against future disease. However, some survivors of chronic VHS infections were capable of replicating virus upon re-exposure. Demonstration of a chronic manifestation of VHSV infection among Pacific herring maintained at ambient seawater temperatures provides insights into the mechanisms by which the virus is maintained among populations of endemic hosts. ?? 2010 Inter-Research.

  20. Chronic and persistent viral hemorrhagic septicemia virus infections in Pacific herring

    USGS Publications Warehouse

    Hershberger, Paul K.; Gregg, Jacob L.; Winton, James R.; Grady, Cortney A.; Taylor, L.

    2010-01-01

    Chronic viral hemorrhagic septicemia virus (VHSV) infections were established in a laboratory stock of Pacific herring Clupea pallasii held in a large-volume tank supplied with pathogen-free seawater at temperatures ranging from 6.8 to 11.6°C. The infections were characterized by viral persistence for extended periods and near-background levels of host mortality. Infectious virus was recovered from mortalities occurring up to 167 d post-exposure and was detected in normal-appearing herring for as long as 224 d following initial challenge. Geometric mean viral titers were generally as high as or higher in brain tissues than in pools of kidney and spleen tissues, with overall prevalence of infection being higher in the brain. Upon re-exposure to VHSV in a standard laboratory challenge, negligible mortality occurred among groups of herring that were either chronically infected or fully recovered, indicating that survival from chronic manifestations conferred protection against future disease. However, some survivors of chronic VHS infections were capable of replicating virus upon re-exposure. Demonstration of a chronic manifestation of VHSV infection among Pacific herring maintained at ambient seawater temperatures provides insights into the mechanisms by which the virus is maintained among populations of endemic hosts.

  1. Identification of viral hemorrhagic septicemia virus isolated from Pacific cod Gadus macrocephalus in Prince William Sound Alaska, USA

    USGS Publications Warehouse

    Meyers, T.R.; Sullivan, J.; Emmenegger, E.; Follett, J.; Short, S.; Batts, W.N.; Winton, J.R.

    1992-01-01

    Ulcerative slun tissues from 2 Pacific cod Gadus rnacrocephalus caught in Prince William Sound, Alaska, USA, were examined for virus by Fish Pathology staff within the F.R.E.D. Division of the Alaska Department of Fish and Game. Six days after inoculation of Epitheliorna papulosum cyprini (EPC) cells at 14"C, diffuse rounding and lifting of cells from the monolayers suggestive of cytopathlc effect became visible in the lower sample dilutions. Ultrastructural examinations of affected EPC cells showed rhabdovirus particles within cytoplasmic vacuoles and on the cell surface membranes. Virus isolates from both cod were subsequently confirmed as viral hemorrhagic septicemia virus (VHSV) by serum neutralizabon and immunoblot assay. This is the first VHSV isolated from Pacific cod, which represents a new host species for the virus. Histologically, cod skin ulcers appeared to be caused by a foreign-body-type inflammatory response to foci of protozoa resembling X cells that also had plasmodial stages. Whether the rhabdovirus was incidental to the slun lesion or played a role in its etiology remains to be determined. The possible relationship between thls virus and the recent occurrences of VHSV in anadromous salmoruds from Washington State, USA, is discussed.

  2. Association of viral hemorrhagic septicemia virus with epizootic hemorrhages of the skin in Pacific herring Clupea harengus pallasi from Prince William Sound and Kodiak Island, Alaska, USA

    USGS Publications Warehouse

    Meyers, T.R.; Short, S.; Lipson, K.; Batts, W.N.; Winton, J.R.; Wilcock, J.; Brown, E.

    1994-01-01

    Only one-third of the Pacific hernng Clupea harengus pallasi expected to spawn in Prince William Sound (PWS), Alaska, USA, in sprlng 1993 were observed. Of these herring, 15 to 43 '% had external ulcers or subdermal hemorrhages of the skln and fins. A rhabdovirus identified as the North American strain of viral hemorrhagic septicemia virus (VHSV) was isolated from affected herring and 1 Pacific cod Gadus macrocephalus with skin lesions from PWS, and from herring with similar lesions collected near Kodiak Island. No other pathogens were detected in the herring examined. Although VHSV may have been responsible for the skin lesions, there was no confirmed mass herring mortality observed in PWS; hence the actual cause of the reduced herring numbers is still unknown. The same strain of VHSV was subsequently isolated from captive juvenile herring collected from Auke Bay, Alaska, near Juneau, from herring in British Columbia, Canada, and from Puget Sound, Washington, USA. These findings suggest the virus is an opportunistic pathogen that is widely indigenous to Pacific herring populations in the Pacific Northwest and that herring are a significant marine reservoir for North American VHSV.

  3. Expression of NLRC4 in children with septicaemia and mechanisms of NLRC4 in in vitro cytokine secretion.

    PubMed

    Zhu, Guoji; Chen, Jing; Tian, Jianmei; Ge, Lingqing; Xing, Aixia; Tang, Guangming

    2016-07-01

    Septicaemia, a systemic bacterial infection, frequently leads to morbidity and mortality in children. The NOD-like receptor (NLR) family, CARD domain containing 4 (NLRC4) is involved in the control of infections. The aim of the present study was to detect the expression of NLRC4 in the blood samples of children with septicaemia, in addition to investigating the importance of NLRC4 in cytokine production, and the signaling pathways that regulate NLRC4 expression in lipopolysaccharide (LPS)-stimulated macrophages. It was determined that when compared with the control, the mRNA and protein expression levels of NLRC4 were significantly increased in the blood samples of children with septicaemia, as demonstrated by the reverse transcription‑quantitative polymerase chain reaction and western blot analysis. The results from the western blotting indicated that treatment with LPS induced NLRC4 expression in a time‑ and dose‑dependent manner in RAW264.7 cells. A knockdown of NLRC4 by siRNA transfection enhanced the effect of LPS on interleukin (IL)‑1β and IL‑18 production, as determined by enzyme‑linked immunosorbent assay. Inhibitors of extracellular regulated protein kinases, c‑Jun N‑terminal kinases and p38 were used in the present study to block the mitogen‑activated protein kinase (MAPK) signaling pathway, and it was determined that LPS‑induced NLRC4 expression was reversed by the suppression of the MAPK signaling pathway. To the best of our knowledge, this is the first report regarding the expression of NLRC4 in children with septicaemia. Furthermore, a novel molecular mechanism for NLRC4 regulation in LPS‑induced RAW264.7 macrophage cells has been elucidated. The data in the present study supports the hypothesis that LPS activates the MAPK pathway in macrophages, thus resulting in the upregulation of NLRC4; however, NLRC4 inhibits IL‑1β and IL‑18 production, contributing to the anti-inflammatory response.

  4. Histological, serological and virulence studies on rainbow trout experimentally infected with recombinant infectious hematopoietic necrosis viruses.

    PubMed

    Romero, Alejandro; Figueras, Antonio; Tafalla, Carolina; Thoulouze, Maria-Isabel; Bremont, Michel; Novoa, Beatriz

    2005-12-30

    Several recombinant infectious hematopoietic necrosis viruses (IHNV) were produced by reverse genetics and their pathogenicity in trout was evaluated and compared to that of the wild type (wt) viruses IHNV and viral haemorrhagic septicemia virus (VHSV). Recombinant IHNVs used in this study were: rIHNV, identical to the wtIHNV; rIHNV-Gvhsv, a recombinant virus expressing the VHSV G gene instead of the IHNV G gene; rIHNV-Gmut, which possesses 2 targeted mutations in the glycoprotein; and rIHNVmut-Gmut, which is similar to the rIHNV-Gmut, but exhibits additional mutations along the genome. Results obtained in experimental infections showed that the rIHNV and rIHNV-Gmut were the most virulent recombinant viruses. Severity of the lesions induced by the different recombinant viruses was in agreement with mortality data. The kidney and the liver were the organs most affected by the most pathogenic viruses, and the lesions observed resembled those produced by wtIHNV. The introduction of mutations did not alter the tissue tropism of the virus. The recombinant viruses were able to replicate in fish, as shown by immunoperoxidase assay and RT-PCR. Antibodies against IHNV were detected in the fish inoculated with IHNV, rIHNV, rIHNV-Gmut and rIHNVmut-Gmut, and antibodies against VHSV were also found in fish infected with rIHNV-Gvhsv. Finally, antibody production was highest in fish infected with the rIHNVmut-Gmut even though this virus was the least virulent.

  5. Immunohistochemistry and pathology of multiple Great Lakes fish from mortality events associated with viral hemorrhagic septicemia virus type IVb.

    PubMed

    Al-Hussinee, L; Lord, S; Stevenson, R M W; Casey, R N; Groocock, G H; Britt, K L; Kohler, K H; Wooster, G A; Getchell, R G; Bowser, P R; Lumsden, J S

    2011-01-21

    A novel viral hemorrhagic septicemia virus (VHSV) (genotype IVb) has been isolated from mortality events in a range of wild freshwater fish from the Great Lakes since 2005. In 2005 and 2006, numerous new freshwater host species (approximately 90 fish from 12 different species) were confirmed to have VHSV by cell culture and reverse transcriptase polymerase chain reaction. A prominent feature observed in infected fish were the petechial and ecchymotic haemorrhages on the body surface and in visceral organs, as well as serosanguinous ascites; however, many fish had few and subtle, gross lesions. Histologically, virtually all fish had a vasculitis and multifocal necrosis of numerous tissues. Excellent correlation was found between the presence of VHSV IVb antigen detected by immunohistochemistry and the pathological changes noted by light microscopy. Intact and degenerate leukocytes, including cells resembling lymphocytes and macrophages, also had cytoplasmic viral antigen. By contrast, renal tubules and gonadal tissues (ovary and testis), were strongly immunopositive for VHSV IVb, but no lesions were noted.

  6. New sites of localisation of Pasteurella multocida B:2 in buffalo surviving experimental haemorrhagic septicaemia

    PubMed Central

    2014-01-01

    Background Haemorrhagic septicaemia (HS) is an acute septicaemic disease of buffalo and cattle caused by Pasteurella multocida B:2 and E:2. Field outbreaks of HS are known to result in localisation of bacteria in the tonsils of surviving buffalo, confirming that animals can become carriers and the role of respiratory tract in the transmission of the disease. This report describes additional sites of localisation of P. multocida B:2 in surviving buffalo following experimental induction of HS. Results Following P. multocida B:2 infection, all calves in group 1 and one calf in group 2 that was allowed to commingle with infected calves from group 1 were euthanised within 48 h. Pasteurella multocida B:2 was detected from the nasal and rectal swab samples on days 5 and 6 from the remaining calves in group 2. The first injection of dexamethasone into the carrier animals resulted in reemergence in samples from the nose, rectum and vagina. However, subsequent dexamethasone injections failed to re-activate P. multocida B:2. When surviving carrier calves in group 2 were euthanised at the end of the experiment, P. multocida B:2 was detected in the lungs and various organs of the respiratory, gastrointestinal and urinary tracts. Conclusions Commingling naive buffalo calves with calves acutely infected with P. multocida B:2 resulted in carriers among surviving buffalo. Pasteurella was found in various organs of the respiratory, gastrointestinal and urinary tracts, suggesting their role in the pathogenesis of HS. PMID:24721163

  7. Correlation between electrophoretic types B1 and B2 of carboxylesterase B and host-dependent factors in Escherichia coli septicaemia.

    PubMed Central

    Picard, B.; Goullet, P.

    1988-01-01

    Electrophoretic types B1 and B2 of carboxylesterase B produced by strains of Escherichia coli isolated from 100 septicaemia cases were correlated with alpha-haemolysin and mannose resistant haemagglutinin (MRHA) production and with clinical data including eventual underlying diseases, origin of septicaemia and evolution. Electrophoretic type B2 was phenotypically linked with alpha-haemolysin and MRHA production. The proportion of type B2 isolates varied significantly with occurrence of an underlying illness (45% for patients without an underlying disease and 22% for compromised patients) and with the site of origin of the septicaemia (40% for those of urinary origin and 18% for infection of digestive origin). In the former infections, type B2 isolates were obtained in the majority from male patients while type B1 isolates predominated in women. The septicaemias associated with type B1 were characterized by a lower proportion of isolates producing alpha-haemolysin and MRHA and by a greater frequency of septic shock and death than those associated with type B2. These facts emphasize the importance of host-dependent factors in E. coli septicaemia. PMID:3276540

  8. Viruses

    USDA-ARS?s Scientific Manuscript database

    Lytic bacteriophages, viruses which infect and lyse bacterial cells, can provide a natural method to reduce bacterial pathogens on produce commodities. The use of multi-phage cocktails is most likely to be effective against bacterial pathogens on produce commodities, and minimize the development of...

  9. Suppression subtractive hybridization coupled with microarray analysis to examine differential expression of genes in Japanese flounder Paralichthys olivaceus leucocytes during Edwardsiella tarda and viral hemorrhagic septicemia virus infection.

    PubMed

    Matsuyama, Tomomasa; Fujiwara, Atushi; Takano, Tomokazu; Nakayasu, Chihaya

    2011-10-01

    Transcriptional changes in the peripheral blood leucocytes (PBL) of Japanese flounder Paralichthys olivaceus challenged by Edwardsiella tarda and viral hemorrhagic septicemia virus (VHSV) were investigated using suppression subtractive hybridization (SSH) coupled with cDNA microarray analysis. First, we constructed an SSH cDNA library using mRNA samples isolated from PBL of P. olivaceus that had been experimentally infected with E. tarda. We then examined the transcriptional changes occurring in the PBL due to E. tarda and VHSV infection using a cDNA microarray produced using clones produced from the SSH library. A total of 565 and 180 cDNA sequences corresponding to mRNA species that are either up- or down-regulated by E. tarda infection were isolated by SSH. While host gene expression responses in response to E. tarda and VHSV infection share several response elements, distinct patterns of gene expression were also observed. Specifically, E. tarda infection enhanced the expression of cell adhesion molecules while VHSV enhanced the expression of interferon and proteasome-related genes. In challenge trials of the two infectious agents, expression profiles of chemokines were also observed to differ. The results indicated that distinguishing between viral and bacterial infection is possible based on the RNA expression profiles of PBL from infected fish.

  10. Trends in mortality from septicaemia and pneumonia with economic development: an age-period-cohort analysis.

    PubMed

    Wong, Irene O L; Cowling, Benjamin J; Leung, Gabriel M; Schooling, C Mary

    2012-01-01

    Hong Kong population has experienced drastic changes in its economic development in the 1940s. Taking advantage of Hong Kong's unique demographic and socioeconomic history, characterized by massive, punctuated migration waves from Southern China, and recent, rapid transition from a pre-industrialized society to the first ethnic Chinese community reaching "first world" status over the last 60 years (i.e., in two or three generations), we examined the longitudinal trends in infection related mortality including septicemia compared to trends in non-bacterial pneumonia to generate hypotheses for further testing in other recently transitioned economies and to provide generalized aetiological insights on how economic transition affects infection-related mortality. We used deaths from septicemia and pneumonia not specified as bacterial, and population figures in Hong Kong from 1976-2005. We fitted age-period-cohort models to decompose septicemia and non-bacterial pneumonia mortality rates into age, period and cohort effects. Septicaemia-related deaths increased exponentially with age, with a downturn by period. The birth cohort curves had downward inflections in both sexes in the 1940s, with a steeper deceleration for women. Non-bacterial pneumonia-related deaths also increased exponentially with age, but the birth cohort patterns showed no downturns for those born in the 1940s. The observed changes appeared to suggest that better early life conditions may enable better development of adaptive immunity, thus enhancing immunity against bacterial infections, with greater benefits for women than men. Given the interaction between the immune system and the gonadotropic axis, these observations are compatible with the hypothesis that upregulation of the gonadotropic axis underlies some of the changes in disease patterns with economic development.

  11. Trends in Mortality from Septicaemia and Pneumonia with Economic Development: An Age-Period-Cohort Analysis

    PubMed Central

    Wong, Irene O. L.; Cowling, Benjamin J.; Leung, Gabriel M.; Schooling, C. Mary

    2012-01-01

    Background Hong Kong population has experienced drastic changes in its economic development in the 1940s. Taking advantage of Hong Kong’s unique demographic and socioeconomic history, characterized by massive, punctuated migration waves from Southern China, and recent, rapid transition from a pre-industrialized society to the first ethnic Chinese community reaching “first world” status over the last 60 years (i.e., in two or three generations), we examined the longitudinal trends in infection related mortality including septicemia compared to trends in non-bacterial pneumonia to generate hypotheses for further testing in other recently transitioned economies and to provide generalized aetiological insights on how economic transition affects infection-related mortality. Methods We used deaths from septicemia and pneumonia not specified as bacterial, and population figures in Hong Kong from 1976–2005. We fitted age-period-cohort models to decompose septicemia and non-bacterial pneumonia mortality rates into age, period and cohort effects. Results Septicaemia-related deaths increased exponentially with age, with a downturn by period. The birth cohort curves had downward inflections in both sexes in the 1940s, with a steeper deceleration for women. Non-bacterial pneumonia-related deaths also increased exponentially with age, but the birth cohort patterns showed no downturns for those born in the 1940s. Conclusion The observed changes appeared to suggest that better early life conditions may enable better development of adaptive immunity, thus enhancing immunity against bacterial infections, with greater benefits for women than men. Given the interaction between the immune system and the gonadotropic axis, these observations are compatible with the hypothesis that upregulation of the gonadotropic axis underlies some of the changes in disease patterns with economic development. PMID:22720008

  12. Angular deformity of the ankle with sparing of the distal fibula following meningococcal septicaemia: a case series involving 14 ankles in ten children.

    PubMed

    Monsell, F P; McBride, A R T; Barnes, J R; Kirubanandan, R

    2011-08-01

    Progressive angular deformity of an extremity due to differential physeal arrest is the most common late orthopaedic sequela following meningococcal septicaemia in childhood. A total of ten patients (14 ankles) with distal tibial physeal arrest as a consequence of meningococcal septicaemia have been reviewed. Radiological analysis of their ankles has demonstrated a distinct pattern of deformity. In 13 of 14 cases the distal fibular physis was unaffected and continued distal fibular growth contributed to a varus deformity. We recommend that surgical management should take account of this consistent finding during the correction of these deformities.

  13. Molecular typing of haemorrhagic septicaemia-associated Pasteurella multocida isolates from Pakistan and Thailand using multilocus sequence typing and pulsed-field gel electrophoresis.

    PubMed

    Moustafa, Ahmed M; Bennett, Mark D; Edwards, John; Azim, Kamran; Mesaik, Muhammed A; Choudhary, M Iqbal; Pathanasophon, Pornpen; Worarach, Apasara; Ali, Qurban; Abubakar, Muhammad; Anjum, Rehana

    2013-12-01

    A comparative genetic study of 23 field isolates and vaccine strains of Pasteurella multocida associated with haemorrhagic septicaemia cases from Pakistan and Thailand was done using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The MLST sequence type (ST) for all 20 of the 23 isolates tested was 122. The PFGE results showed one band difference between the Pakistani and the Thai isolates. Sequence type 122 is the dominant associated profile with haemorrhagic septicaemia (HS) cases in South Asia. The study supports the concept of using PFGE for short-term epidemiology and MLST for long-term epidemiology. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Experimental infection studies demonstrating Atlantic salmon as a host and reservoir of viral hemorrhagic septicemia virus type IVa with insights into pathology and host immunity

    USGS Publications Warehouse

    Lovy, Jan; Piesik, P.; Hershberger, P.K.; Garver, K.A.

    2013-01-01

    In British Columbia, Canada (BC), aquaculture of finfish in ocean netpens has the potential for pathogen transmission between wild and farmed species due to the sharing of an aquatic environment. Viral hemorrhagic septicemia virus (VHSV) is enzootic in BC and causes serious disease in wild Pacific herring, Clupea pallasii, which often enter and remain in Atlantic salmon, Salmo salar, netpens. Isolation of VHSV from farmed Atlantic salmon has been previously documented, but the effects on the health of farmed salmon and the wild fish sharing the environment are unknown. To determine their susceptibility, Atlantic salmon were exposed to a pool of 9 isolates of VHSV obtained from farmed Atlantic salmon in BC by IP-injection or by waterborne exposure and cohabitation with diseased Pacific herring. Disease intensity was quantified by recording mortality, clinical signs, histopathological changes, cellular sites of viral replication, expression of interferon-related genes, and viral tissue titers. Disease ensued in Atlantic salmon after both VHSV exposure methods. Fish demonstrated gross disease signs including darkening of the dorsal skin, bilateral exophthalmia, light cutaneous hemorrhage, and lethargy. The virus replicated within endothelial cells causing endothelial cell necrosis and extensive hemorrhage in anterior kidney. Infected fish demonstrated a type I interferon response as seen by up-regulation of genes for IFNα, Mx, and ISG15. In a separate trial infected salmon transmitted the virus to sympatric Pacific herring. The results demonstrate that farmed Atlantic salmon can develop clinical VHS and virus can persist in the tissues for at least 10 weeks. Avoiding VHS epizootics in Atlantic salmon farms would limit the potential of VHS in farmed Atlantic salmon, the possibility for further host adaptation in this species, and virus spillback to sympatric wild fishes.

  15. Experimental infection studies demonstrating Atlantic salmon as a host and reservoir of viral hemorrhagic septicemia virus type IVa with insights into pathology and host immunity.

    PubMed

    Lovy, J; Piesik, P; Hershberger, P K; Garver, K A

    2013-09-27

    In British Columbia, Canada (BC), aquaculture of finfish in ocean netpens has the potential for pathogen transmission between wild and farmed species due to the sharing of an aquatic environment. Viral hemorrhagic septicemia virus (VHSV) is enzootic in BC and causes serious disease in wild Pacific herring, Clupea pallasii, which often enter and remain in Atlantic salmon, Salmo salar, netpens. Isolation of VHSV from farmed Atlantic salmon has been previously documented, but the effects on the health of farmed salmon and the wild fish sharing the environment are unknown. To determine their susceptibility, Atlantic salmon were exposed to a pool of 9 isolates of VHSV obtained from farmed Atlantic salmon in BC by IP-injection or by waterborne exposure and cohabitation with diseased Pacific herring. Disease intensity was quantified by recording mortality, clinical signs, histopathological changes, cellular sites of viral replication, expression of interferon-related genes, and viral tissue titers. Disease ensued in Atlantic salmon after both VHSV exposure methods. Fish demonstrated gross disease signs including darkening of the dorsal skin, bilateral exophthalmia, light cutaneous hemorrhage, and lethargy. The virus replicated within endothelial cells causing endothelial cell necrosis and extensive hemorrhage in anterior kidney. Infected fish demonstrated a type I interferon response as seen by up-regulation of genes for IFNα, Mx, and ISG15. In a separate trial infected salmon transmitted the virus to sympatric Pacific herring. The results demonstrate that farmed Atlantic salmon can develop clinical VHS and virus can persist in the tissues for at least 10 weeks. Avoiding VHS epizootics in Atlantic salmon farms would limit the potential of VHS in farmed Atlantic salmon, the possibility for further host adaptation in this species, and virus spillback to sympatric wild fishes.

  16. The health, social and educational needs of children who have survived meningitis and septicaemia: the parents' perspective.

    PubMed

    Clark, Laura J; Glennie, Linda; Audrey, Suzanne; Hickman, Matthew; Trotter, Caroline L

    2013-10-10

    Survivors of bacterial meningitis and septicaemia can experience a range of after-effects. There is little published research on the needs and provision of aftercare for children surviving bacterial meningitis and septicaemia. Mixed methods study employing a survey and follow-up interviews with a sample of survey participants recruited from Meningitis Research Foundation's member database and social media. Of 194 eligible survey respondents, 77% reported at least moderate short-term after-effects, and 57% a need for aftercare or support. Most parents reported that their child received a hearing test (98%) and follow-up appointment with a paediatrician (66%). Psychosocial after-effects were most common and the greatest need was for educational support. About half of participants felt their children's needs for aftercare were met. We conducted interviews with 18 parents. Findings suggest access could be limited by: parents' inability to navigate systems in place, child's age, and delayed identification of sequelae. Parents felt a comprehensive explanation of possible after-effects on discharge from hospital was required, and found uncertain prognoses difficult. Good communication between professionals enabled a service tailored to the child's needs. Our study supports the NICE and SIGN guidelines and highlights areas for improvement in the aftercare of these children.

  17. Utility of qualitative C- reactive protein assay and white blood cells counts in the diagnosis of neonatal septicaemia at Bugando Medical Centre, Tanzania.

    PubMed

    Chacha, Flora; Mirambo, Mariam M; Mushi, Martha F; Kayange, Neema; Zuechner, Antke; Kidenya, Benson R; Mshana, Stephen E

    2014-10-03

    Neonatal septicaemia diagnosis based on clinical features alone is non-specific leading to the initiation of unnecessary antibiotic treatment posing a danger of increased antibiotic resistance. In the present study the utility of serial qualitative C-reactive protein (CRP) assay and white blood cells count (WBC) in the diagnosis of neonatal septicaemia was investigated using blood culture as gold standard. A total of 305 neonates admitted at Bugando Medical Centre (BMC) neonatal units between September 2013 and April 2014 were enrolled. Demographic and clinical data were collected using standardized data collection tool. Blood specimens were collected for blood culture, WBC count and qualitative CRP assay. Of 305 neonates; 224 (73.4%) were ≤ 72 hrs of age and 91(29.8%) had low birth weight. The positive CRP assay was observed in 67 (22.0%), 80 (26.2%) and 88 (28.9%) of neonates on day 1, 2 and 3 respectively; with any CRP positive occurred in 104 (34.1%) of neonates. The sensitivities of CRP assay in the diagnosis of septicaemia using culture as gold standard on day 1, 2, 3 and any positive were 40.4%, 53.2%, 54.8% and 62.9% respectively. While specificities were 82.7%, 80.7%, 77.8% and 73.3% respectively. Higher sensitivity of 75% was observed when CRP was used to diagnose gram negative septicaemia compared to 50% that was observed in the diagnosis of gram positive septicaemia. WBC count of ≥13 × 109 /L had sensitivity and specificity of 64.5% and 66.7% respectively with area under the curve of 0.694. When the any positive CRP and WBC of ≥13 × 109 /L were used the sensitivity increased to 90.3% with specificity of 50%. Neonates with septicaemia due to gram negative bacteria were significantly found to have higher rates of positive CRP than neonates with gram positive septicaemia and with negative culture (p < 0.001, OR 8.2, 95 CI; 2.9-26). In place where blood culture is limited neonates having clinical features of neonatal sepsis with positive qualitative

  18. Heterogeneity in levels of serum neutralizing antibodies against viral hemorrhagic septicemia virus genotype IVB among fish species in Lake St. Clair, Michigan, USA.

    PubMed

    Millard, Elena V; Faisal, Mohamed

    2012-04-01

    The presence of neutralizing antibodies against viral hemorrhagic septicemia virus (VHSV-IVb) was investigated in sera of 13 fish species collected from Lake St. Clair, Michigan, USA, a VHSV-endemic water body. We tested 297 sera collected May 2004-June of 2010, using a complement-dependent 50% plaque neutralization test (50% PNT). Neutralizing antibodies were detected in 23% (67/297) of the samples. The highest overall antibody prevalence (85%, 34/40) and mean positive antibody titer (12,113 ± 11,699 SD) were detected in muskellunge (Esox masquinongy). Antibodies were also detected in 50% (15/30) of sampled northern pike (E. lucius), 25% (15/61) of freshwater drum (Aplodinotus grunniens), and 7% (3/41) of smallmouth bass (Micropterus dolomieu). All sera from channel catfish (Ictalurus punctatus), lake sturgeon (Acipenser fulvescens), quillback (Carpiodes cyprinus), rock bass (Ambloplites rupestris), shorthead redhorse (Moxostoma macrolepidotum), silver redhorse (M. anisurum), walleye (Sander vitreus), white perch (Morone americana), and yellow perch (Perca flavescens) were negative. Antibodies in one or more fish species were detected in all sampling years (2004, 2006, 2007, 2009, and 2010), whereas in parallel sampling periods, VHS virus was detected only in 2006 and 2009. Our results suggest the continued presence of VHSV-IVb in the Lake St. Clair ecosystem, and underscore the importance of assessing immune responses of fish populations to determine prior virus exposure.

  19. Emergence of Viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity.

    PubMed

    Thompson, Tarin M; Batts, William N; Faisal, Mohamed; Bowser, Paul; Casey, James W; Phillips, Kenneth; Garver, Kyle A; Winton, James; Kurath, Gael

    2011-08-29

    Viral hemorrhagic septicemia virus (VHSV) is a fish rhabdovirus that causes disease in a broad range of marine and freshwater hosts. The known geographic range includes the Northern Atlantic and Pacific Oceans, and recently it has invaded the Great Lakes region of North America. The goal of this work was to characterize genetic diversity of Great Lakes VHSV isolates at the early stage of this viral emergence by comparing a partial glycoprotein (G) gene sequence (669 nt) of 108 isolates collected from 2003 to 2009 from 31 species and at 37 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVb within the major VHSV genetic group IV. Among these 108 isolates, genetic diversity was low, with a maximum of 1.05% within the 669 nt region. There were 11 unique sequences, designated vcG001 to vcG011. Two dominant sequence types, vcG001 and vcG002, accounted for 90% (97 of 108) of the isolates. The vcG001 isolates were most widespread. We saw no apparent association of sequence type with host or year of isolation, but we did note a spatial pattern, in which vcG002 isolates were more prevalent in the easternmost sub-regions, including inland New York state and the St. Lawrence Seaway. Different sequence types were found among isolates from single disease outbreaks, and mixtures of types were evident within 2 isolates from individual fish. Overall, the genetic diversity of VHSV in the Great Lakes region was found to be extremely low, consistent with an introduction of a new virus into a geographic region with previously naive host populations.

  20. Emergence of viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity

    USGS Publications Warehouse

    Thompson, T.M.; Batts, W.N.; Faisal, M.; Bowser, P.; Casey, J.W.; Phillips, K.; Garver, K.A.; Winton, J.; Kurath, G.

    2011-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a fish rhabdovirus that causes disease in a broad range of marine and freshwater hosts. The known geographic range includes the Northern Atlantic and Pacific Oceans, and recently it has invaded the Great Lakes region of North Ame­rica. The goal of this work was to characterize genetic diversity of Great Lakes VHSV isolates at the early stage of this viral emergence by comparing a partial glycoprotein (G) gene sequence (669 nt) of 108 isolates collected from 2003 to 2009 from 31 species and at 37 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVb within the major VHSV genetic group IV. Among these 108 isolates, genetic diversity was low, with a maximum of 1.05% within the 669 nt region. There were 11 unique sequences, designated vcG001 to vcG011. Two dominant sequence types, vcG001 and vcG002, accounted for 90% (97 of 108) of the isolates. The vcG001 isolates were most widespread. We saw no apparent association of sequence type with host or year of isolation, but we did note a spatial pattern, in which vcG002 isolates were more prevalent in the easternmost sub-regions, including inland New York state and the St. Lawrence Seaway. Different sequence types were found among isolates from single disease outbreaks, and mixtures of types were evident within 2 isolates from ­individual fish. Overall, the genetic diversity of VHSV in the Great Lakes region was found to be extremely low, consistent with an introduction of a new virus into a geographic region with ­previously naïve host populations.

  1. The BAFF / APRIL axis plays an important role in virus-induced peritoneal responses in rainbow trout.

    PubMed

    Soleto, Irene; Abós, Beatriz; Castro, Rosario; González, Lucia; Tafalla, Carolina; Granja, Aitor G

    2017-05-01

    IgM(+) B cells have been recently demonstrated to be key regulators of peritoneal inflammation in teleost, as a large number of them occupy the peritoneal cavity after 48 h of antigenic stimulation. Despite this, the number of studies addressing the mechanism through which this cell population expands and differentiates in response to stimuli has been scarcely addressed. Because the BAFF/APRIL axis is known to play a major role in B cell survival and differentiation in mammals, we hypothesized that it could be affected in the peritoneal cavity in response to an inflammatory stimulus. To verify this hypothesis, we studied how BAFF, APRIL and the fish-specific related cytokine BALM as well as their putative receptors are regulated in rainbow trout after intraperitoneal (i.p.) injection of viral hemorrhagic septicemia virus (VHSV). When the transcriptional analysis was performed in total cells from the peritoneum, we observed that VHSV provoked an up-regulation of both BAFF and BAFF receptor (BAFF-R) mRNA levels. However, when we examined how isolated peritoneal IgM(+) B cells were transcriptionally affected by VHSV i.p. injection, we found that APRIL, BALM and the transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) were also up-regulated in response to the virus. IgM(-) cells, on the other hand, only up-regulated BALM transcription in response to VHSV. Finally, to gain further insight on the role that these cytokines play in the peritoneum, we have studied their effect on the survival of peritoneal IgM(+) B cells. This work demonstrates a key role for the BAFF/APRIL axis in the peritoneal inflammatory response and contributes to further understanding how IgM(+) B cells are regulated at this specific peripheral site. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Companion Animals as a Source of Viruses for Human Beings and Food Production Animals.

    PubMed

    Reperant, L A; Brown, I H; Haenen, O L; de Jong, M D; Osterhaus, A D M E; Papa, A; Rimstad, E; Valarcher, J-F; Kuiken, T

    2016-07-01

    Companion animals comprise a wide variety of species, including dogs, cats, horses, ferrets, guinea pigs, reptiles, birds and ornamental fish, as well as food production animal species, such as domestic pigs, kept as companion animals. Despite their prominent place in human society, little is known about the role of companion animals as sources of viruses for people and food production animals. Therefore, we reviewed the literature for accounts of infections of companion animals by zoonotic viruses and viruses of food production animals, and prioritized these viruses in terms of human health and economic importance. In total, 138 virus species reportedly capable of infecting companion animals were of concern for human and food production animal health: 59 of these viruses were infectious for human beings, 135 were infectious for food production mammals and birds, and 22 were infectious for food production fishes. Viruses of highest concern for human health included hantaviruses, Tahyna virus, rabies virus, West Nile virus, tick-borne encephalitis virus, Crimean-Congo haemorrhagic fever virus, Aichi virus, European bat lyssavirus, hepatitis E virus, cowpox virus, G5 rotavirus, influenza A virus and lymphocytic choriomeningitis virus. Viruses of highest concern for food production mammals and birds included bluetongue virus, African swine fever virus, foot-and-mouth disease virus, lumpy skin disease virus, Rift Valley fever virus, porcine circovirus, classical swine fever virus, equine herpesvirus 9, peste des petits ruminants virus and equine infectious anaemia virus. Viruses of highest concern for food production fishes included cyprinid herpesvirus 3 (koi herpesvirus), viral haemorrhagic septicaemia virus and infectious pancreatic necrosis virus. Of particular concern as sources of zoonotic or food production animal viruses were domestic carnivores, rodents and food production animals kept as companion animals. The current list of viruses provides an objective

  3. Detection of Infectious Pancreatic Necrosis Virus from the Leeches Hemiclepsis marginata and Hirudo medicinalis.

    PubMed

    Salimi, Behnam; Abdi, Kazem

    2016-12-01

    Leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV), and also may contain blood protozoa. In the present study, leeches were collected from water bodies located in Kurdistan province, Iran. The specimens were tested for IHNV, VHSV, and infectious pancreatic necrosis virus (IPNV) using the PCR method. The results showed that two different species of leeches, Hemiclepsis marginata and Hirudo medicinalis, were infected by IPNV among the seven species studied. The infected leeches were found in areas that were polluted with untreated sewage coming from upstream fish farms culturing Rainbow Trout Oncorhynchus mykiss. In addition, the fish at fish farms in the vicinity had been infected with IPNV 9 months previously. Our results showed that the virus causing infectious pancreatic necrosis is present in the leeches H. marginata and H. medicinalis, suggesting that leeches are a potential source of IPNV in fish farms. Received October 14, 2015; accepted June 1, 2016.

  4. A DNA vaccine encoding the viral hemorrhagic septicemia virus genotype IVb glycoprotein confers protection in muskellunge (Esox masquinongy), rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta), and lake trout (Salvelinus namaycush).

    PubMed

    Standish, Isaac F; Millard, Elena V; Brenden, Travis O; Faisal, Mohamed

    2016-12-02

    The viral hemorrhagic septicemia virus (VHSV) is one of the most serious fish pathogens. In 2003, a novel sublineage (genotype IVb) of this deadly virus emerged in the Great Lakes basin causing serious fish kills. We have previously demonstrated that a DNA plasmid (pcDNA), containing a cytomegalovirus (CMV) promoter and the viral hemorrhagic septicemia virus (VHSV) genotype IVb glycoprotein (G) gene insert (designated pVHSivb-G) confers moderate protection in muskellunge (Esox masquinongy), a highly susceptible species upon challenge. In order to achieve optimal protection, we investigated a number of factors including the incubation time [i.e. the number of degree days (° days)] before challenge, and viral challenge dose and route. Additionally, we tested if pVHSivb-G provides protection against VHSV-IVb to less susceptible salmonids such as rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta) and lake trout (Salvelinus namaycush). An increase in the period lapsed between vaccination and challenge to 1880° days resulted in 95% relative percent protection (RPS) in muskellunge following a single administration of the pVHSivb-G plasmid and viral challenge. An RPS of 100% for muskellunge was achieved with a longer incubation period (2400° days) and in conjunction with a booster dose of the plasmid. The pVHSivb-G vaccine also elicited significant protection in all three salmonid species, reaching 100% RPS in lake trout following an incubation period of 1001° days prior to viral challenge. Vaccination with pVHSivb-G was also associated with the development of significant levels of circulating VHSV-binding antibodies in muskellunge as measured by indirect ELISA, which reached peak levels 6-7 weeks post-vaccination. Viral shedding in vaccinated survivors was minimal and of transient nature. The study shows that the pVHSivb-G plasmid can elicit a protective response against the wild virus strain in a range of species important in recreational and commercial

  5. Expression kinetics of key genes in the early innate immune response to Great Lakes viral hemorrhagic septicemia virus IVb infection in yellow perch (Perca flavescens)

    USGS Publications Warehouse

    Olson, Wendy; Emmenegger, Eveline; Glenn, Jolene; Simchick, Crystal; Winton, Jim; Goetz, Frederick

    2013-01-01

    The recently discovered strain of viral hemorrhagic septicemia virus, VHSV-IVb, represents an example of the introduction of an extremely pathogenic rhabdovirus capable of infecting a wide variety of new fish species in a new host-environment. The goal of the present study was to delineate the expression kinetics of key genes in the innate immune response relative to the very early stages of VHSV-IVb infection using the yellow perch (Perca flavescens) as a model. Administration of VHSV-IVb by IP-injection into juvenile yellow perch resulted in 84% cumulative mortality, indicating their high susceptibility to this disease. In fish sampled in the very early stages of infection, a significant up-regulation of Mx gene expression in the liver, as well as IL-1β and SAA activation in the head kidney, spleen, and liver was directly correlated to viral load. The potential down-regulation of Mx in the hematopoietic tissues, head kidney and spleen, may represent a strategy utilized by the virus to increase replication.

  6. Surveillance of viruses in wild fish populations in areas around the Gulf of Cadiz (South Atlantic Iberian Peninsula).

    PubMed

    Moreno, Patricia; Olveira, José G; Labella, Alejandro; Cutrín, Juan Manuel; Baro, Jorge C; Borrego, Juan Jose; Dopazo, Carlos P

    2014-10-01

    This report describes a viral epidemiological study of wild fish around the Gulf of Cadiz (southwestern Iberian Peninsula) and is focused on infectious pancreatic necrosis virus (IPNV), viral hemorrhagic septicemia virus (VHSV), and viral nervous necrosis virus (VNNV). One fish species (Chelon labrosus) was sampled inside the gulf, at the mouth of the San Pedro River. Another 29 were sampled, in three oceanographic campaigns, at sites around the Bay of Cadiz. The fish were processed individually and subjected to isolation in cell culture and molecular diagnosis. VHSV was not isolated from any species. Thirteen IPNV-type isolates were obtained from barracuda (Sphyraena sphyraena), axillary seabream (Pagellus acarne), common two-banded seabream (Diplodus vulgaris), common pandora (P. erythrinus), Senegal seabream (D. bellottii), and surmullet (Mullus surmuletus). Six VNNV isolates were obtained from axillary seabream, common pandora, black seabream (Spondyliosoma cantharus), red mullet (Mullet barbatus), Lusitanian toadfish (Halobatrachus didactylus), and tub gurnard (Chelidonichtys lucerna). In the river mouth, viruses were detected only after reamplification, obtaining prevalence percentages of IPNV and VNNV (44.4 and 63.0%, respectively) much higher than those observed in the oceanographic campaigns (25.7 and 19.6%, respectively). The opposite results were obtained in the case of VHSV after reamplification: 11.1% in the river mouth and 43.6% in the oceanic locations. Analyzing the results with respect to the proximity of the sampling sites to the coast, an anthropogenic influence on wild fish is suggested and discussed. The type of viruses and the presence of natural reassortants are also discussed.

  7. Surveillance of Viruses in Wild Fish Populations in Areas around the Gulf of Cadiz (South Atlantic Iberian Peninsula)

    PubMed Central

    Moreno, Patricia; Olveira, José G.; Labella, Alejandro; Cutrín, Juan Manuel; Baro, Jorge C.; Borrego, Juan Jose

    2014-01-01

    This report describes a viral epidemiological study of wild fish around the Gulf of Cadiz (southwestern Iberian Peninsula) and is focused on infectious pancreatic necrosis virus (IPNV), viral hemorrhagic septicemia virus (VHSV), and viral nervous necrosis virus (VNNV). One fish species (Chelon labrosus) was sampled inside the gulf, at the mouth of the San Pedro River. Another 29 were sampled, in three oceanographic campaigns, at sites around the Bay of Cadiz. The fish were processed individually and subjected to isolation in cell culture and molecular diagnosis. VHSV was not isolated from any species. Thirteen IPNV-type isolates were obtained from barracuda (Sphyraena sphyraena), axillary seabream (Pagellus acarne), common two-banded seabream (Diplodus vulgaris), common pandora (P. erythrinus), Senegal seabream (D. bellottii), and surmullet (Mullus surmuletus). Six VNNV isolates were obtained from axillary seabream, common pandora, black seabream (Spondyliosoma cantharus), red mullet (Mullet barbatus), Lusitanian toadfish (Halobatrachus didactylus), and tub gurnard (Chelidonichtys lucerna). In the river mouth, viruses were detected only after reamplification, obtaining prevalence percentages of IPNV and VNNV (44.4 and 63.0%, respectively) much higher than those observed in the oceanographic campaigns (25.7 and 19.6%, respectively). The opposite results were obtained in the case of VHSV after reamplification: 11.1% in the river mouth and 43.6% in the oceanic locations. Analyzing the results with respect to the proximity of the sampling sites to the coast, an anthropogenic influence on wild fish is suggested and discussed. The type of viruses and the presence of natural reassortants are also discussed. PMID:25128341

  8. Characterization of a rhabdovirus isolated from carpione Salmo trutta carpio in Italy

    USGS Publications Warehouse

    Bovo, G.; Olesen, N.J.; Jorgensen, P.E.V.; Ahne, W.; Winton, J.R.

    1995-01-01

    A virus, strain 583, was isolated from carpione Salmo trutta carpio fry exhibiting high mortality. The virus was not neutralized by rabbit antisera against the fish rhabdoviruses viral haemorrhagic septicaemia virus (VHSV), infectious hematopoietic necrosis virus, eel rhabdovirus European X, spring viraemia of carp virus or pike fry rhabdovirus, or against the birnavirus infectious pancreatic necrosis virus. The virus replicated in several fish cell lines incubated at 20 to 25*C and grew optimally in the bluegill fry (BF-2) and fathead minnow (FHM) cell lines. Electron microscopy of infected BF-2 cell cultures revealed the presence of typical rhabdovirus particles, and immunofluorescent staining was observed using various polyclonal and monoclonal antibodies (MAbs) against Egtved virus, the causative agent of viral haemorrhagic septicaemia. The staining by a MAb against the nucleoprotein (N) of VHSV was particularly strong, a MAb against the glycoprotein (G) gave a moderate reaction, whereas a second MAb against the G protein and MAbs against the matrix proteins, M_(1) and M_(2), of VHSV did not react. Fluorescence titres using 3 rabbit antisera against whole Egtved virus varied between negative and moderately positive. Western blotting using polyclonal and monoclonal sera confirmed that both the N and G proteins of the carpione virus shared some epitopes with those of VHSV, but the M_(1) and M_(2) proteins did not. SDS-PAGE showed the structural proteins of the carpione virus produced a pattern typical of members of the Lyssavirus genus of the Rhabdoviridae and the molecular weights were very similar to those of VHSV, except for the M_(2) protein which was somewhat smaller. Infection trials showed the carpione virus induced high mortalities in carpione fry but not in rainbow trout Oncorhynchus mykiss fry. The carpione virus was clearly distinguishable from Egtved virus despite limited serological cross reaction. Since it was also easily distinguishable by

  9. Phylogeny of the Viral Hemorrhagic Septicemia Virus in European Aquaculture

    PubMed Central

    Cieslak, Michael; Mikkelsen, Susie S.; Skall, Helle F.; Baud, Marine; Diserens, Nicolas; Engelsma, Marc Y.; Haenen, Olga L. M.; Mousakhani, Shirin; Panzarin, Valentina; Wahli, Thomas; Olesen, Niels J.; Schütze, Heike

    2016-01-01

    One of the most valuable aquaculture fish in Europe is the rainbow trout, Oncorhynchus mykiss, but the profitability of trout production is threatened by a highly lethal infectious disease, viral hemorrhagic septicemia (VHS), caused by the VHS virus (VHSV). For the past few decades, the subgenogroup Ia of VHSV has been the main cause of VHS outbreaks in European freshwater-farmed rainbow trout. Little is currently known, however, about the phylogenetic radiation of this Ia lineage into subordinate Ia clades and their subsequent geographical spread routes. We investigated this topic using the largest Ia-isolate dataset ever compiled, comprising 651 complete G gene sequences: 209 GenBank Ia isolates and 442 Ia isolates from this study. The sequences come from 11 European countries and cover the period 1971–2015. Based on this dataset, we documented the extensive spread of the Ia population and the strong mixing of Ia isolates, assumed to be the result of the Europe-wide trout trade. For example, the Ia lineage underwent a radiation into nine Ia clades, most of which are difficult to allocate to a specific geographic distribution. Furthermore, we found indications for two rapid, large-scale population growth events, and identified three polytomies among the Ia clades, both of which possibly indicate a rapid radiation. However, only about 4% of Ia haplotypes (out of 398) occur in more than one European country. This apparently conflicting finding regarding the Europe-wide spread and mixing of Ia isolates can be explained by the high mutation rate of VHSV. Accordingly, the mean period of occurrence of a single Ia haplotype was less than a full year, and we found a substitution rate of up to 7.813 × 10−4 nucleotides per site per year. Finally, we documented significant differences between Germany and Denmark regarding their VHS epidemiology, apparently due to those countries’ individual handling of VHS. PMID:27760205

  10. Comparison of genome size and synthesis of structural proteins of Hirame Rhabdovirus, infectious hematopoietic necrosis virus, and viral hemorrhagic Septicemia virus

    USGS Publications Warehouse

    Nishizawa, Toyohiko; Yoshimizu, Mamoru; Winton, James R.; Kimura, Takahisa

    1991-01-01

    Genomic RNA was extracted from purified virions of hirame rhabdovirus (HRV), infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV). The full-length RNA was analyzed using formaldehyde agarose gel electrophoresis followed by ethidium bromide staining. Compared with an internal RNA size standard, all three viral genomic RNAs appeared to have identical relative mobilities and were estimated to be approximately 10.7 kilobases in length or about 3.7 megadaltons in molecular mass. Structural protein synthesis of HRV, IHNV, and VHSV was studied using cell cultures treated with actinomycin D. At 2 h intervals, proteins were labeled with 35S-methionine, extracted, and analyzed by SDS-polyacrylamide gel electrophoresis and autoradiography. The five structural proteins of each of the three viruses appeared in the following order : nucleoprotein (N), matrix protein 1 (M1), matrix protein 2 (M2), glycoprotein (G), and polymerase (L) reflecting both the approximate relative abundance of each protein within infected cells and the gene order within the viral genome.

  11. Non-O1/non-O139 Vibrio cholerae septicaemia in a Saudi man: a case report

    PubMed Central

    El-Hossary, Dalia; Jiman-Fatani, Asif; Al-Ghamdi, Rahaf

    2017-01-01

    Background. The non-O1/non-O139 serogroups of Vibrio cholerae occur in diverse natural niches, and usually cause mild and self-limiting gastrointestinal illness. However, they have well-documented potential to cause invasive and extra-intestinal infections among immunocompromised patients. Furthermore, their ability to grow in low-salinity surface water, and the existence of asymptomatic human carriers, suggest novel acquisition routes for this unusual infection, even in people without obvious risk factors. Case presentation. A 62-year-old man presented with epigastric pain, vomiting and fever. The patient had a history of diabetes and cholecystectomy, although our initial examination did not reveal any significant findings that might indicate V. cholerae infection. However, blood cultures subsequently revealed the presence of V. cholerae, which was positively identified using both conventional and modern non-conventional technologies. The identity of the V. cholerae isolate was confirmed using Vitek MS (matrix–assisted laser desorption ionization-time of flight MS) and the FilmArray system, in addition to its initial identification using the Vitek 2 system. The septicaemia was successfully treated using a 14 day course of ciprofloxacin. Conclusion. The present case highlights the need to remain highly suspicious of non-O1/non-O139 V. cholerae infections in patients with known risk factors, as well as in healthy individuals with epidemiological exposure and compatible clinical symptoms. Special care should be taken to avoid false-positive results from confirmatory laboratory tests, as the organism can grow in fresh water, and the results should be verified using multiple methods. PMID:28348803

  12. Non-O1/non-O139 Vibrio cholerae septicaemia in a Saudi man: a case report.

    PubMed

    Kaki, Reham; El-Hossary, Dalia; Jiman-Fatani, Asif; Al-Ghamdi, Rahaf

    2017-02-01

    Background. The non-O1/non-O139 serogroups of Vibrio cholerae occur in diverse natural niches, and usually cause mild and self-limiting gastrointestinal illness. However, they have well-documented potential to cause invasive and extra-intestinal infections among immunocompromised patients. Furthermore, their ability to grow in low-salinity surface water, and the existence of asymptomatic human carriers, suggest novel acquisition routes for this unusual infection, even in people without obvious risk factors. Case presentation. A 62-year-old man presented with epigastric pain, vomiting and fever. The patient had a history of diabetes and cholecystectomy, although our initial examination did not reveal any significant findings that might indicate V. cholerae infection. However, blood cultures subsequently revealed the presence of V. cholerae, which was positively identified using both conventional and modern non-conventional technologies. The identity of the V. cholerae isolate was confirmed using Vitek MS (matrix-assisted laser desorption ionization-time of flight MS) and the FilmArray system, in addition to its initial identification using the Vitek 2 system. The septicaemia was successfully treated using a 14 day course of ciprofloxacin. Conclusion. The present case highlights the need to remain highly suspicious of non-O1/non-O139 V. cholerae infections in patients with known risk factors, as well as in healthy individuals with epidemiological exposure and compatible clinical symptoms. Special care should be taken to avoid false-positive results from confirmatory laboratory tests, as the organism can grow in fresh water, and the results should be verified using multiple methods.

  13. Monitoring of viruses in chum salmon (Oncorhynchus keta) migrating to Korea.

    PubMed

    Jeon, C-H; Kim, S-R; Kim, W-S; Lee, C-H; Seong, K-B; Lee, C-S; Oh, M-J; Kim, J-H

    2011-06-01

    It is important to investigate the prevalence of salmonid pathogens because they can affect the amount of release of salmonid fry and the migration rate of adult salmonids. In this study, routine surveys were conducted for investigating virus distribution in migrating chum salmon spawners (Oncorhynchus keta) and their offsprings at the Namdae River, Yangyang, Korea, during 2006-2008. Anterior kidneys were removed from chum salmon spawner individuals, homogenized with minimal essential medium, and centrifuged to make supernatants for conducting RT-PCR. Five offspring were pooled to for conducting RT-PCR. Infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) were the target viruses for monitoring. In 2006, only spawners were investigated, and 27.5% of fish (22/80) were found to be IHNV-positive by nested PCR. In 2007, 65.6% of pooled fry (21/32) were IHNV-positive, and 9.4% (3/32) were IPNV-positive by one-step PCR. When nested PCR was conducted, 84.4% (27/32) were IHNV-positive, and 28.1% (9/32) were IPNV-positive. However, only 1.3% of spawners (1/80) were IHNV-positive by nested PCR. In 2008, 25% (8/32) of pooled fry were IHNV-positive by one-step PCR, but 59.4% (19/32) were IHNV-positive and 12.5% (4/32) were IPNV-positive by nested PCR. All of the samples tested were VHSV-negative. Although all viruses detected in this study were from chum salmon, phylogenetic analysis showed that they possibly originated from rainbow trout or clustered with the rainbow trout isolates. More extensive long-term studies are needed to clarify the origins of these viruses and their potential effects on chum salmon migration in Korea.

  14. A survey of the frequency of aminoglycoside antibiotic-resistant genotypes and phenotypes in Escherichia coli in broilers with septicaemia in Hebei, China.

    PubMed

    Zhang, F Y; Huo, S Y; Li, Y R; Xie, R; Wu, X J; Chen, L G; Gao, Y H

    2014-01-01

    1. The aim of this study was to investigate the occurrence of aminoglycoside resistance and the prevalence of 6 important modifying enzyme genes, i.e. (strA, strB, aph(3')-IIa, aac(3)-IIa, aac(6')-Ib and ant(3")-Ia), in Escherichia coli strains in broilers with septicaemia in Hebei, China. 2. A total of 111 clinical isolates of E. coli were collected from 46 large-scale farms. Antimicrobial susceptibility tests, using the Kirby-Bauer disc diffusion method, were performed on all 111 isolates. In addition, all were screened for the presence of modifying enzyme genes using the polymerase chain reaction (PCR). 3. The results show that the rates of resistance were as follows: streptomycin: 97.3%, kanamycin: 97.0%, gentamicin: 95.5%, neomycin: 50.5%, amikacin: 46.0%, spectinomycin: 22.5%. Of the genes examined, strB (73.9%) was the most frequently identified gene in the phenotypic resistant isolates, followed in order by: ant(3")-Ia, aac(3)-IIa, aac(6')-Ib, aph(3')-IIa and strA. 4. It is concluded that aminoglycoside resistance in E. coli from broilers with septicaemia remains a serious problem in Hebei, China. This emphasises the need to ban the non-therapeutic use of antibiotics, discourage their misuse and to be continually vigilant by providing appropriate scientific and technological support for the poultry industry.

  15. A five-year experience of carbapenem resistance in Enterobacteriaceae causing neonatal septicaemia: predominance of NDM-1.

    PubMed

    Datta, Saswati; Roy, Subhasree; Chatterjee, Somdatta; Saha, Anindya; Sen, Barsha; Pal, Titir; Som, Tapas; Basu, Sulagna

    2014-01-01

    Treatment of neonatal sepsis has become a challenge with the emergence of carbapenemase-producing bacteria. This study documents the trend of carbapenem susceptibility in Enterobacteriaceae that caused septicaemia in neonates over a five year period (2007-2011) and the molecular characterisation of Enterobacteriaceae resistant to carbapenems and cephalosporins. Hundred and five Enterobacteriaceae including Escherichia coli (n = 27), Klebsiella pneumoniae (n = 68) and Enterobacter spp. (n = 10) were isolated from blood of septicaemic neonates followed by antibiotic susceptibility tests, determination of MIC values, phenotypic and genotypic detection of β-lactamases. Carbapenem was the most active antimicrobial tested after tigecycline. CTX-M type was the most prevalent ESBL throughout the period (82%). New Delhi Metallo-β-lactamase-1 (NDM-1), which is a recent addition to the carbapenemase list, was the only carbapenemase identified in our setting. Fourteen percent of the isolates possessed blaNDM-1. Carbapenem non-susceptibility was first observed in 2007 and it was due to loss of Omp F/Ompk36 in combination with the presence of ESBLs/AmpCs. NDM-1 first emerged in E. coli during 2008; later in 2010, the resistance was detected in K. pneumoniae and E. cloacae isolates. NDM-1-producing isolates were resistant to other broad-spectrum antibiotics and possessed ESBLs, AmpCs, 16S-rRNA methylases, AAC(6')-Ib-cr, bleomycin resistant gene and class 1 integron. Pulsed field gel electrophoresis of the NDM-1-producing isolates indicated that the isolates were clonally diverse. The study also showed that there was a significantly higher incidence of sepsis caused by NDM-1-harbouring isolates in the male sex, in neonates with low birth weight and neonates born at an extramural centre. However, sepsis with NDM-1-harbouring isolates did not result in a higher mortality rate. The study is the first to review the carbapenem resistance patterns in neonatal sepsis over

  16. Child Mortality after Discharge from a Health Facility following Suspected Pneumonia, Meningitis or Septicaemia in Rural Gambia: A Cohort Study.

    PubMed

    Chhibber, Aakash Varun; Hill, Philip C; Jafali, James; Jasseh, Momodou; Hossain, Mohammad Ilias; Ndiaye, Malick; Pathirana, Jayani C; Greenwood, Brian; Mackenzie, Grant A

    2015-01-01

    To measure mortality and its risk factors among children discharged from a health centre in rural Gambia. We conducted a cohort study between 12 May 2008 and 11 May 2012. Children aged 2-59 months, admitted with suspected pneumonia, sepsis, or meningitis after presenting to primary and secondary care facilities, were followed for 180 days after discharge. We developed models associating post-discharge mortality with clinical syndrome on admission and clinical risk factors. One hundred and five of 3755 (2.8%) children died, 80% within 3 months of discharge. Among children aged 2-11 and 12-59 months, there were 30 and 29 deaths per 1000 children per 180 days respectively, compared to 11 and 5 respectively in the resident population. Children with suspected pneumonia unaccompanied by clinically severe malnutrition (CSM) had the lowest risk of post-discharge mortality. Mortality increased in children with suspected meningitis or septicaemia without CSM (hazard ratio [HR] 2.6 and 2.2 respectively). The risk of mortality greatly increased with CSM on admission: CSM with suspected pneumonia (HR 8.1; 95% confidence interval (CI) 4.4 to 15), suspected sepsis (HR 18.4; 95% CI 11.3 to 30), or suspected meningitis (HR 13.7; 95% CI 4.2 to 45). Independent associations with mortality were: mid-upper arm circumference (MUAC) of 11.5-13.0 cm compared to >13.0 cm (HR 7.2; 95% CI 3.0 to 17.0), MUAC 10.5-11.4 cm (HR 24; 95% CI 9.4 to 62), and MUAC <10.5 cm (HR 44; 95% CI 18 to 108), neck stiffness (HR 10.4; 95% CI 3.1 to 34.8), non-medical discharge (HR 4.7; 95% CI 2.0 to 10.9), dry season discharge (HR 2.0; 95% CI 1.2 to 3.3), while greater haemoglobin (HR 0.82; 0.73 to 0.91), axillary temperature (HR 0.71; 95% CI 0.58 to 0.87), and oxygen saturation (HR 0.96; 95% CI 0.93 to 0.99) were associated with reduced mortality. Gambian children experience increased mortality after discharge from primary and secondary care. Interventions should target both moderately and severely

  17. Child Mortality after Discharge from a Health Facility following Suspected Pneumonia, Meningitis or Septicaemia in Rural Gambia: A Cohort Study

    PubMed Central

    Chhibber, Aakash Varun; Hill, Philip C.; Jafali, James; Jasseh, Momodou; Hossain, Mohammad Ilias; Ndiaye, Malick; Pathirana, Jayani C.; Greenwood, Brian; Mackenzie, Grant A.

    2015-01-01

    Objective To measure mortality and its risk factors among children discharged from a health centre in rural Gambia. Methods We conducted a cohort study between 12 May 2008 and 11 May 2012. Children aged 2–59 months, admitted with suspected pneumonia, sepsis, or meningitis after presenting to primary and secondary care facilities, were followed for 180 days after discharge. We developed models associating post-discharge mortality with clinical syndrome on admission and clinical risk factors. Findings One hundred and five of 3755 (2.8%) children died, 80% within 3 months of discharge. Among children aged 2–11 and 12–59 months, there were 30 and 29 deaths per 1000 children per 180 days respectively, compared to 11 and 5 respectively in the resident population. Children with suspected pneumonia unaccompanied by clinically severe malnutrition (CSM) had the lowest risk of post-discharge mortality. Mortality increased in children with suspected meningitis or septicaemia without CSM (hazard ratio [HR] 2.6 and 2.2 respectively). The risk of mortality greatly increased with CSM on admission: CSM with suspected pneumonia (HR 8.1; 95% confidence interval (CI) 4.4 to 15), suspected sepsis (HR 18.4; 95% CI 11.3 to 30), or suspected meningitis (HR 13.7; 95% CI 4.2 to 45). Independent associations with mortality were: mid-upper arm circumference (MUAC) of 11.5–13.0 cm compared to >13.0 cm (HR 7.2; 95% CI 3.0 to 17.0), MUAC 10.5–11.4 cm (HR 24; 95% CI 9.4 to 62), and MUAC <10.5 cm (HR 44; 95% CI 18 to 108), neck stiffness (HR 10.4; 95% CI 3.1 to 34.8), non-medical discharge (HR 4.7; 95% CI 2.0 to 10.9), dry season discharge (HR 2.0; 95% CI 1.2 to 3.3), while greater haemoglobin (HR 0.82; 0.73 to 0.91), axillary temperature (HR 0.71; 95% CI 0.58 to 0.87), and oxygen saturation (HR 0.96; 95% CI 0.93 to 0.99) were associated with reduced mortality. Conclusion Gambian children experience increased mortality after discharge from primary and secondary care. Interventions should

  18. [Haemorrhagic septicaemia in a pig caused by extraintestinal pathogenic Escherichia coli (ExPEC) as a differential diagnosis in classical swine fever--case report and review of the literature].

    PubMed

    Reiner, Gerald; von Berg, Stephan; Hillen, Sonja; Clemens, Nina; Huisinger, Maike; Burkhardt, Eberhard; Weiss, Reinhardt; Reinacher, Manfred

    2010-01-01

    Domestic pig herds in some regions of Germany are permanently threatened by Classical Swine Fever. In the case of suspicion, a series of infectious and non infectious causes has to be excluded. The present paper describes a case of Escherichia coli septicaemia, with clinical and pathological symptoms that could not be differentiated from European or African Swine Fever. The E. coli strain could not be classified by standard serotyping. Virulence factors common for ETEC (enterotoxic E. coli) or EDEC (edema-disease E. coli) were not detected. Instead, we found P-fimbriae and aerobactin, thus characterising this strain as an extraintestinal pathogenic strain. Such strains have sporadicly been reported as the cause of septicaemia in piglets or weaners, but the present case is the first report of an E. coli-associated septicaemia in an adult pig. This case shows that extraintestinal pathogenic E. coli can be the cause of severe septicaemia and haemorrhagia. They thus have to be considered as a further differential diagnosis in swine fever.

  19. Ebola Virus and Marburg Virus

    MedlinePlus

    Diseases and Conditions Ebola virus and Marburg virus By Mayo Clinic Staff Ebola virus and Marburg virus are related viruses that cause hemorrhagic ... Africa, where sporadic outbreaks have occurred for decades. Ebola virus and Marburg virus live in animal hosts, ...

  20. Characterization of an infectious pancreatic necrosis (IPN) virus carrier cell culture with resistance to superinfection with heterologous viruses.

    PubMed

    García, Inmaculada; Galiana, Antonio; Falcó, Alberto; Estepa, Amparo; Perez, Luis

    2011-04-21

    A state of persistence of a non susceptible fish cell line with infectious pancreatic necrosis virus (IPNV) was established in vitro by experimental infection. The persistently infected culture showed sustained production of infectious virus and could be continuously passaged for months. A distinct feature of this culture is that only a very small fraction of the cells harbours virus replication, in contrast to other reported IPNV-persistently infected cells from salmonid fish, where nearly all the cells express viral antigens. In spite of the small number of detectable IPNV-infected cells, the carrier culture shows resistance to superinfection with homologous as well as heterologous viruses. Temperature shift-up experiments indicate that viral interference is due to continuous replication of IPNV in the culture. Quantitation of Mx gene expression suggested that the interference phenomenon could be mediated by the activation of the interferon (IFN) system. However, conditioned medium from the IPNV-infected cell cultures only marginally protected other cells against VHSV infection, indicating that other type I IFN-independent mechanism may be underlying the resistance of the persistently infected culture to infection with heterologous viruses. Our study defines a novel in vitro model of IPNV persistence and contributes to the understanding of the widespread distribution of aquabirnaviruses in marine and fresh water environments by establishing a carrier state in non susceptible fish species. Copyright © 2010 Elsevier B.V. All rights reserved.

  1. Influence of age and foaling on plasma protein electrophoresis and serum amyloid A and their possible role as markers of equine neonatal septicaemia.

    PubMed

    Paltrinieri, Saverio; Giordano, Alessia; Villani, Marta; Manfrin, Margherita; Panzani, Sara; Veronesi, Maria Cristina

    2008-06-01

    A field study was undertaken to investigate plasma protein electrophoresis (PPE) and serum amyloid A (SAA) concentrations at 1, 3 and 7 days of age in foals born by oxytocin-induced (group A, n =17) or spontaneous (group B, n =12) parturition. The putative diagnostic role of PPE and SAA in four septicaemic foals was also examined. At birth, beta-gamma-globulins were higher in group B, and then increased in both groups, probably due to colostrum intake. At day 3, no significant differences between the groups were detectable. In both groups, SAA values increased from day 0 to day 3, then decreased at day 7. Septicaemic foals had decreased concentrations of many electrophoretic fractions (probably reflecting decreased colostrum intake and circulating bacterial proteases) and an increased SAA concentration. It was concluded that both PPE and SAA concentrations vary with age and type of foaling and may be useful in the early diagnosis of neonatal septicaemia.

  2. Viral hemorrhagic septicemia virus, Ichthyophonus hoferi, and other causes of morbidity in Pacific herring Clupea pallasi spawning in Prince William Sound, Alaska, USA.

    PubMed

    Marty, G D; Freiberg, E F; Meyers, T R; Wilcock, J; Farver, T B; Hinton, D E

    1998-02-26

    Pacific herring Clupea pallasi populations in Prince William Sound, Alaska, USA, declined from an estimated 9.8 x 10(7) kg in 1992 to 1.5 x 10(7) kg in 1994. To determine the role of disease in population decline, 233 Pacific herring from Prince William Sound were subjected to complete necropsy during April 1994. The North American strain of viral hemorrhagic septicemia virus (VHSV) was isolated from 11 of 233 fish (4.7%). VHSV was significantly related to myocardial mineralization, hepatocellular necrosis, submucosal gastritis, and meningoencephalitis. Ichthyophonus hoferi infected 62 of 212 (29%) fish. I. hoferi infections were associated with severe, disseminated, granulomatous inflammation and with increased levels of plasma creatine phosphokinase (CPK) and aspartate aminotransferase (AST). I. hoferi prevalence in 1994 was more than double that of most previous years (1989 to 1993). Plasma chemistry values significantly greater (p < 0.01) in males than females included albumin, total protein, cholesterol, chloride, glucose, and potassium; only alkaline phosphatase was significantly greater in females. Hypoalbuminemia was relatively common in postspawning females; other risk factors included VHSV and moderate or severe focal skin reddening. Pacific herring had more than 10 species of parasites, but they were not associated with significant lesions. Two of the parasites have not previously been described: a renal intraductal myxosporean (11% prevalence) and an intestinal coccidian (91% prevalence). Transmission electron microscopy of a solitary mesenteric lesion revealed viral particles consistent with lymphocystis virus. No fish had viral erythrocytic necrosis (VEN). Prevalence of external gross lesions and major parasites was not related to fish age, and fish that were year-lings at the time of the 1989 'Exxon Valdez' oil spill (1988 year class) had no evidence of increased disease prevalence.

  3. The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: analysis of relationships with other rhabdoviruses.

    PubMed

    Björklund, H V; Higman, K H; Kurath, G

    1996-06-01

    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 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2-33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

  4. The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: Analysis of relationships with other rhabdoviruses

    USGS Publications Warehouse

    Bjorklund, H.V.; Higman, K.H.; Kurath, G.

    1996-01-01

    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 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2–33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

  5. The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: Analysis of relationships with other rhabdoviruses

    USGS Publications Warehouse

    Bjorklund, H.V.; Higman, K.H.; Kurath, G.

    1996-01-01

    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 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2-33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

  6. Development of a multiplex assay to measure the effects of shipping and storage conditions on the quality of RNA used in molecular assays for detection of viral haemorrhagic septicemia virus.

    PubMed

    Siah, A; Duesund, H; Frisch, K; Nylund, A; McKenzie, P; Saksida, S

    2014-09-01

    Abstract In routine diagnostics, real-time reverse transcriptase quantitative PCR (RT-qPCR) has become a powerful method for fish health screening. Collection, transportation, and storage conditions of specimens could dramatically affect their integrity and could consequently affect RT-qPCR test results. In this study, to assess the expression profile of elongation factor 1 alpha (ELF-1α) gene, head kidney (HK) tissues from Atlantic Salmon Salmo salar were exposed at room temperature, 4°C, -20°C, and -80°C as well as in 70% ethanol for 6, 12, 24, 48, and 72 h. Data showed a significant increase of RT-qPCR cycle threshold (Ct) values for ELF-1α ranging from 14.7 to 26.5 cycles for tissues exposed to room temperature. In order to mimic the sample transportation conditions, different temperatures of storage were used and tissue quality was evaluated using ELF-1α gene expression. Data showed that Ct values for ELF-1α increased significantly when the tissues were transported on ice for 2 h, stored at -20°C, thawed on ice for 6 h, and stored again at -80°C. The HK tissues collected from Atlantic Salmon challenged with viral hemorrhagic septicemia virus (VHSV) through intraperitoneal injection were exposed at room temperature for 0, 6, 12, 24, 48, 72, and 96 h. Data showed a good correlation of values for ELF-1α and VHSV Ct although the ELF-1α mRNA of the host degraded faster than the RNA of VHSV. Based on these data, HK tissues could be transported on ice or ice packs without the quality of the tissue being affected when stored at -80°C upon arrival at the laboratory. In addition, 70% ethanol could be used as a preservative for long-distance transportation. For an efficient diagnostic test, a duplex VHSV-ELF-1α was developed and optimized. Data showed that the sensitivity of the duplex assay for VHSV was similar to the singleplex. Received November 25, 2013; accepted February 14, 2014.

  7. Isolates of viral hemorrhagic septicemia virus from North America and Europe can be detected and distinguished by DNA probes

    USGS Publications Warehouse

    Batts, W.N.; Arakawa, C.K.; Bernard, J.; Winton, J.R.

    1993-01-01

    Biotinylated DNA probes were constructed to hybndize with speclfic sequences within the messenger RNA (mRNA) of the nucleoprotein (N) gene of vlral hemorrhagic septicemia virus (VHSV) reference strains from Europe (07-71) and North Arnenca (Makah) Probes were synthesized that were complementary to (1) a 29-nucleotide sequence near the center of the N gene conlmon to both the 07-71 and Makah reference strains of the virus (2) a unique 28- nucleotide sequence that followed the open readng frame of the Makah N gene mRNA most of which was absent In the 07-71 strain, and (3) a 22-nucleobde sequence wthin the 07-71 N gene that had 6 nllsmatches \

  8. Heartland Virus

    MedlinePlus

    ... Vector-Borne Diseases (DVBD) NCEZID Share Compartir Heartland virus On this Page What is Heartland virus? How ... Do I Need to Know? What is Heartland virus? Heartland virus belongs to a family of viruses ...

  9. Overview of recent DNA vaccine development for fish

    USGS Publications Warehouse

    Kurath, G.; ,

    2005-01-01

    Since the first description of DNA vaccines for fish in 1996, numerous studies of genetic immunisation against the rhabdovirus pathogens infectious haematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV) have established their potential as both highly efficacious biologicals and useful basic research tools. Single small doses of rhabdovirus DNA constructs provide extremely strong protection against severe viral challenge under a variety of conditions. DNA vaccines for several other important fish viruses, bacteria, and parasites are under investigation, but they have not yet shown high efficacy. Therefore, current research is focussed on mechanistic studies to understand the basis of protection, and on improvement of the nucleic acid vaccine applications against a wider range of fish pathogens.

  10. Effect of alum co-adjuvantation of oil adjuvant vaccine on emulsion stability and immune responses against haemorhagic septicaemia in mice.

    PubMed

    Kumar, Sujeet; Chaturvedi, Vinod Kumar; Kumar, Bablu; Kumar, Pankaj; Somarajan, Sudha Rani; Mishra, Anil Kumar; Sharma, Bhaskar

    2015-04-01

    Haemorrhagic septicaemia (HS), caused by Pasteurella multocida, is the most important bacterial disease of cattle and buffaloes in India. Oil adjuvant vaccine (OAV) is the most potent vaccine available for the control of HS. The study aims to evaluate the effect of alum co-adjuvantation of OAV on emulsion stability and immune response. Two different oil adjuvant vaccines viz., standard oil adjuvant vaccine (OAV) and alum precipitated oil adjuvant vaccine (A-OAV) were prepared with Pasteurella multocida antigen. Emulsion stability was tested by centrifugation, storage at 37 °C for 3 months and microscopy. Immune responses were evaluated by ELISA antibody titer, CD4, CD8 T cell populations and survival post challenge by P. multocida in mice. The separation of aqueous and oil phase of emulsion by centrifugation and storage test were 0 and 6.76% in A-OAV as compared to 11.00 and 26.39% in OAV, respectively. The mean droplet size was significantly smaller (p<0.01) in A-OAV as compared to OAV. The A-OAV recorded higher ELISA antibody titer (p<0.05) up to 21st days post vaccination, and higher CD4 (p>0.05) and CD8 T cell (p<0.05) populations compared to OAV. The A-OAV group conferred 100% protection after challenge with both 100 LD50 and 1000 LD50 as compared to 100 and 60% respective protection by OAV group. The results indicates that A-OAV had better emulsion stability, produces higher level of CD4, CD8 T cells and antibody titer with better protection compared to oil adjuvant vaccine.

  11. Effect of alum co-adjuvantation of oil adjuvant vaccine on emulsion stability and immune responses against haemorhagic septicaemia in mice

    PubMed Central

    Kumar, Sujeet; Chaturvedi, Vinod Kumar; Kumar, Bablu; Kumar, Pankaj; Somarajan, Sudha Rani; Mishra, Anil Kumar; Sharma, Bhaskar

    2015-01-01

    Background and Objectives: Haemorrhagic septicaemia (HS), caused by Pasteurella multocida, is the most important bacterial disease of cattle and buffaloes in India. Oil adjuvant vaccine (OAV) is the most potent vaccine available for the control of HS. The study aims to evaluate the effect of alum co-adjuvantation of OAV on emulsion stability and immune response. Materials and Methods: Two different oil adjuvant vaccines viz., standard oil adjuvant vaccine (OAV) and alum precipitated oil adjuvant vaccine (A–OAV) were prepared with Pasteurella multocida antigen. Emulsion stability was tested by centrifugation, storage at 37 °C for 3 months and microscopy. Immune responses were evaluated by ELISA antibody titer, CD4, CD8 T cell populations and survival post challenge by P. multocida in mice. Results: The separation of aqueous and oil phase of emulsion by centrifugation and storage test were 0 and 6.76% in A-OAV as compared to 11.00 and 26.39% in OAV, respectively. The mean droplet size was significantly smaller (p<0.01) in A–OAV as compared to OAV. The A–OAV recorded higher ELISA antibody titer (p<0.05) up to 21st days post vaccination, and higher CD4 (p>0.05) and CD8 T cell (p<0.05) populations compared to OAV. The A–OAV group conferred 100% protection after challenge with both 100 LD50 and 1000 LD50 as compared to 100 and 60% respective protection by OAV group. Conclusion: The results indicates that A–OAV had better emulsion stability, produces higher level of CD4, CD8 T cells and antibody titer with better protection compared to oil adjuvant vaccine. PMID:26622968

  12. Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells.

    PubMed

    Miller, T A; Rapp, J; Wastlhuber, U; Hoffmann, R W; Enzmann, P J

    1998-09-11

    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 permitted us to read the agarose gels within 7 to 9 h after samples, cultured cells and whole fish arrived, which is of great importance when there is reason to believe that VHSV or IHNV may be present. This is also the first report of a large-scale field trial comparing the RT-PCR assay in trout from 30 German fish farms (a total of 330 rainbow trout) with the usual virus isolation and identification method in order to evaluate the efficiency of the RT-PCR assay for general use in fish health management programs. RT-PCR followed by semi-nested PCR using RNA directly extracted from fish tissue turned out to be the most sensitive method. It recognized 9 fish farms as VHS-positive and 7 as IHN-positive. This is 3 VHS- and 4 IHN-farms more than detected by the traditional virus isolation method. By directly examining the tissue by means of a PCR test it was possible to detect viral RNA in acutely and subacutely to chronically diseased fish as well as in asymptomatic VHS/IHN-carrier fish. Therefore, this effective and powerful assay for detecting VHSV and IHNV by means of PCR has great advantages compared with the presently used procedures.

  13. Increasing versatility of the DNA vaccines through modification of the subcellular location of plasmid-encoded antigen expression in the in vivo transfected cells.

    PubMed

    Martinez-Lopez, Alicia; García-Valtanen, Pablo; Ortega-Villaizan, María Del Mar; Chico, Verónica; Medina-Gali, Regla María; Perez, Luis; Coll, Julio; Estepa, Amparo

    2013-01-01

    The route of administration of DNA vaccines can play a key role in the magnitude and quality of the immune response triggered after their administration. DNA vaccines containing the gene of the membrane-anchored glycoprotein (gpG) of the fish rhabdoviruses infectious haematopoietic necrosis virus (IHNV) or viral haematopoietic septicaemia virus (VHSV), perhaps the most effective DNA vaccines generated so far, confer maximum protection when injected intramuscularly in contrast to their low efficacy when injected intraperitoneally. In this work, taking as a model the DNA vaccine against VHSV, we focused on developing a more versatile DNA vaccine capable of inducing protective immunity regardless of the administration route used. For that, we designed two alternative constructs to gpG₁₋₅₀₇ (the wild type membrane-anchored gpG of VHSV) encoding either a soluble (gpG₁₋₄₆₂) or a secreted soluble (gpG(LmPle20-462)) form of the VHSV-gpG. In vivo immunisation/challenge assays showed that only gpG(LmPle20-462) (the secreted soluble form) conferred protective immunity against VHSV lethal challenge via both intramuscular and intraperitoneal injection, being this the first description of a fish viral DNA vaccine that confers protection when administered intraperitoneally. Moreover, this new DNA vaccine construct also conferred protection when administered in the presence of an oil adjuvant suggesting that DNA vaccines against rhabdoviruses could be included in the formulation of current multicomponent-intaperitoneally injectable fish vaccines formulated with an oil adjuvant. On the other hand, a strong recruitment of membrane immunoglobulin expressing B cells, mainly membrane IgT, as well as t-bet expressing T cells, at early times post-immunisation, was specifically observed in the fish immunised with the secreted soluble form of the VHSV-gpG protein; this may indicate that the subcellular location of plasmid-encoded antigen expression in the in vivo

  14. The North American strain of viral hemorrhagic septicemia virus is highly pathogenic for laboratory-reared Pacific herring (Clupea pallasi)

    USGS Publications Warehouse

    Kocan, R.; Bradley, M.; Elder, N.; Meyers, T.; Batts, W.; Winton, J.

    1997-01-01

    Specific-pathogen-free Pacific herring Clupea pallasi were reared in the laboratory from eggs and then challenged at 5, 9, and 13 months of age by waterborne exposure to low (101.5–2.5 plaque-forming units [PFU] per milliliter), medium (103.5–4.5 PFU/mL), or high (105.5–6.5 PFU/mL) levels of a North American isolate of viral hemorrhagic septicemia virus (VHSV). The fish were extremely susceptible to the virus, showing clinical disease, mortality approaching 100%, and only a limited increase in resistance with age. Mortality began 4–6 d after exposure and peaked at approximately day 7 in fish exposed to high levels of virus. Whereas the mean time to death showed a significant dose response (P < 0.001), the percent mortality and virus titers in dead fish were generally high in all groups regardless of initial challenge dose. External signs of disease were usually limited to 1–2-mm hemorrhagic areas on the lower jaw and isthmus and around the eye, but 2 of 130 infected fish exhibited extensive cutaneous hemorrhaging. Histopathologic examination of tissues from moribund fish sampled at 2–8 d after exposure revealed multifocal coagulative necrosis of hepatocytes, diffuse necrosis of interstitial hematopoietic tissues in the kidney, diffuse necrosis of the spleen, epidermis, and subcutis, and occasional necrosis of pancreatic acinar cells. Virus titers in tissues of experimentally infected herring were first detected 48 h after exposure and peaked 6-8 d after exposure at 107.7 PFU/g. Fish began shedding virus at 48 h after exposure with titers in the flow-through aquaria reaching 102.5 PFU/mL at 4–5 d after exposure, just before peak mortality. When the water flow was turned off for 3 h, titers in the water rose to 103.5 PFU/mL, and the amount of virus shed by infected fish (on average, greater than 106.5 PFU/h per fish) appeared sufficient to sustain a natural epizootic among schooling herring. Taken together, these data suggest that VHSV could be a

  15. A retrospective study on the epidemiology of anthrax, foot and mouth disease, haemorrhagic septicaemia, peste des petits ruminants and rabies in Bangladesh, 2010-2012.

    PubMed

    Mondal, Shankar P; Yamage, Mat

    2014-01-01

    Anthrax, foot and mouth disease (FMD), haemorrhagic septicaemia (HS), peste des petits ruminants (PPR) and rabies are considered to be endemic in Bangladesh. This retrospective study was conducted to understand the geographic and seasonal distribution of these major infectious diseases in livestock based on data collected through passive surveillance from 1 January 2010 to 31 December 2012. Data analysis for this period revealed 5,937 cases of anthrax, 300,333 of FMD, 13,436 of HS, 247,783 of PPR and 14,085 cases of dog bite/rabies. While diseases were reported in almost every district of the country, the highest frequency of occurrence corresponded to the susceptible livestock population in the respective districts. There was no significant difference in the disease occurrences between districts bordering India/Myanmar and non-border districts (p>0.05). Significantly higher (p<0.01) numbers of anthrax (84.5%), FMD (88.3%), HS (84.9%) and dog bite/rabies (64.3%) cases were reported in cattle than any other species. PPR cases were reported mostly (94.8%) in goats with only isolated cases (5.2%) in sheep. The diseases occur throughout the year with peak numbers reported during June through September and lowest during December through April, with significant differences (p<0.01) between the months. The annual usages of vaccines for anthrax, FMD, HS and PPR were only 7.31%, 0.61%, 0.84% and 11.59% of the susceptible livestock population, respectively. Prophylactic vaccination against rabies was 21.16% of cases. There were significant differences (p<0.01) in the administration of anthrax, FMD and HS vaccines between border and non-border districts, but not PPR or rabies vaccines. We recommend that surveillance and reporting of these diseases need to be improved throughout the country. Furthermore, all suspected clinical cases should be confirmed by laboratory examination. The findings of this study can be used in the formulation of more effective disease management and

  16. A Retrospective Study on the Epidemiology of Anthrax, Foot and Mouth Disease, Haemorrhagic Septicaemia, Peste des Petits Ruminants and Rabies in Bangladesh, 2010-2012

    PubMed Central

    Mondal, Shankar P.; Yamage, Mat

    2014-01-01

    Anthrax, foot and mouth disease (FMD), haemorrhagic septicaemia (HS), peste des petits ruminants (PPR) and rabies are considered to be endemic in Bangladesh. This retrospective study was conducted to understand the geographic and seasonal distribution of these major infectious diseases in livestock based on data collected through passive surveillance from 1 January 2010 to 31 December 2012. Data analysis for this period revealed 5,937 cases of anthrax, 300,333 of FMD, 13,436 of HS, 247,783 of PPR and 14,085 cases of dog bite/rabies. While diseases were reported in almost every district of the country, the highest frequency of occurrence corresponded to the susceptible livestock population in the respective districts. There was no significant difference in the disease occurrences between districts bordering India/Myanmar and non-border districts (p>0.05). Significantly higher (p<0.01) numbers of anthrax (84.5%), FMD (88.3%), HS (84.9%) and dog bite/rabies (64.3%) cases were reported in cattle than any other species. PPR cases were reported mostly (94.8%) in goats with only isolated cases (5.2%) in sheep. The diseases occur throughout the year with peak numbers reported during June through September and lowest during December through April, with significant differences (p<0.01) between the months. The annual usages of vaccines for anthrax, FMD, HS and PPR were only 7.31%, 0.61%, 0.84% and 11.59% of the susceptible livestock population, respectively. Prophylactic vaccination against rabies was 21.16% of cases. There were significant differences (p<0.01) in the administration of anthrax, FMD and HS vaccines between border and non-border districts, but not PPR or rabies vaccines. We recommend that surveillance and reporting of these diseases need to be improved throughout the country. Furthermore, all suspected clinical cases should be confirmed by laboratory examination. The findings of this study can be used in the formulation of more effective disease management and

  17. ECHO virus

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/001340.htm ECHO virus To use the sharing features on this page, please enable JavaScript. Enteric cytopathic human orphan (ECHO) viruses are a group of viruses that can lead ...

  18. Virus Carcinogenesis

    DTIC Science & Technology

    1961-01-01

    viruses are capable of inducing cancer, it is obvious that virus carcinogenesis cannot be considered in an isolated fashion, without some reference to...intradermal inoculations of vaccinia virus . One of the viruses most widely investigated with respect to quantitative dose- response relationships is the...than the rule. Figure 6 shows the type of deviation most commonly observed with viruses of infectious diseases. VIRUS CARCINOGENESIS 131 It is a

  19. Foodborne viruses

    USDA-ARS?s Scientific Manuscript database

    Testing for human pathogenic viruses in foods represents a formidable task requiring the extraction, concentration, and assay of a host of viruses from a wide range of food matrices. The enteric viruses, particularly genogroup I and II (GI and GII) noroviruses and hepatitis A virus, are the princip...

  20. Inventing Viruses.

    PubMed

    Summers, William C

    2014-11-01

    In the nineteenth century, "virus" commonly meant an agent (usually unknown) that caused disease in inoculation experiments. By the 1890s, however, some disease-causing agents were found to pass through filters that retained the common bacteria. Such an agent was called "filterable virus," the best known being the virus that caused tobacco mosaic disease. By the 1920s there were many examples of filterable viruses, but no clear understanding of their nature. However, by the 1930s, the term "filterable virus" was being abandoned in favor of simply "virus," meaning an agent other than bacteria. Visualization of viruses by the electron microscope in the late 1930s finally settled their particulate nature. This article describes the ever-changing concept of "virus" and how virologists talked about viruses. These changes reflected their invention and reinvention of the concept of a virus as it was revised in light of new knowledge, new scientific values and interests, and new hegemonic technologies.

  1. An unusual cause of septicaemia.

    PubMed

    Nadarajah, Channa Vasanth; Saif, Imran; Asghar, Syed Ali

    2011-01-01

    Splenic infarction occurs when occlusion of splenic vasculature leads to ischemia, and subsequent tissue necrosis. It is a rare condition. Most patients have an underlying haematological or malignant process or a potential source of embolism. This article describes a patient who presented with unexplained sepsis to the acute medical unit; investigation revealed a splenic abscess and primary hyperparathyroidism, but no evidence of an underlying cause.

  2. Hospital admission rates for meningitis and septicaemia caused by Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae in children in England over five decades: a population-based observational study.

    PubMed

    Martin, Natalie G; Sadarangani, Manish; Pollard, Andrew J; Goldacre, Michael J

    2014-05-01

    Infection with Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae causes substantial mortality and long-term morbidity in children. We know of no study to assess the long-term trends in hospital admission rates for meningitis and septicaemia caused by these pathogens in children in England. We aimed to do such a study using routinely reported data in England. In this population-based observational study, we used datasets that include routinely collected administrative statistics for hospital care: the Hospital In-Patient Enquiry (data for England from 1968 to 1985), the Hospital Episode Statistics dataset (data for England from 1989 onwards), and the Oxford record linkage study (data for Oxfordshire and surrounding areas from 1963 to 2011). We analysed annual age-specific and age-standardised admission rates in children younger than 15 years with H influenzae, meningococcal and pneumococcal meningitis, and septicaemia. We saw a reduction in hospital admission rates for childhood invasive bacterial disease after the introduction of conjugate vaccines against H influenzae, N meningitidis, and S pneumoniae in England. Annual incidence of H influenzae meningitis per 100,000 children decreased from 6·72 admissions (95% CI 6·18-7·26) in 1992 to 0·39 admissions (0·26-0·52) in 1994, after the introduction of routine H influenzae type b vaccination. We saw a small rise in admissions in the early 2000s, peaking at 1·24 admissions per 100,000 children (0·99-1·48) in 2003, which decreased to 0·28 per 100,000 children (0·17-0·39) by 2008 after the introduction of catch-up (2003) and routine (2006) booster programmes for young children. Meningococcal disease increased during the 1990s, reaching a peak in 1999, with 34·54 admissions (33·30-35·78) per 100,000 children. Hospital admissions decreased after the meningococcal serogroup C vaccine was introduced in 1999 and was 12·40 admissions (11·68-13·12) per 100,000 in 2011. Admissions for

  3. 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).

    PubMed

    Johnson, M C; Maxwell, J M; Loh, P C; Leong, J A

    1999-11-01

    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 report of SVCV isolation outside of Europe and Asia, and it is also the first report of SVCV infecting a non-vertebrate species. The G gene from SHRV was most closely related to the G genes from the three Novirhabdoviruses, viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), and hirame rhabdovirus (HIRRV), with 47, 37, and 36% amino acid identity, respectively. In addition, a phylogenetic analysis using the amino acid sequence from rhabdovirus G genes indicated that SHRV should be classified within the Novirhabdovirus genus. Finally, the SHRV-G gene was successfully expressed in mammalian cells under the control of the cytomegalovirus (CMV) promoter, establishing that it can potentially be used in the production of pseudotyped retroviruses designed to infect fish.

  4. Ammocoetes of Pacific Lamprey are not susceptible to common fish rhabdoviruses of the U.S. Pacific northwest.

    PubMed

    Kurath, G; Jolley, J C; Thompson, T M; Thompson, D; Whitesel, T A; Gutenberger, S; Winton, J R

    2013-12-01

    Pacific Lampreys Entosphenus tridentatus have experienced severe population declines in recent years and efforts to develop captive rearing programs are under consideration. However, there is limited knowledge of their life history, ecology, and potential to harbor or transmit pathogens that may cause infectious disease. As a measure of the possible risks associated with introducing wild lampreys into existing fish culture facilities, larval lampreys (ammocoetes) were tested for susceptibility to infection and mortality caused by experimental exposures to the fish rhabdovirus pathogens: infectious hematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV). Two IHNV isolates, representing the U and M genogroups, and one VHSV isolate from the IVa genotype were each delivered to groups of ammocoetes by immersion at moderate and high viral doses, and by intraperitoneal injection. Ammocoetes were then held in triplicate tanks with no substrate or sediment. During 41 d of observation postchallenge there was low or no mortality in all groups, and no virus was detected in the small number of fish that died. Ammocoetes sampled for incidence of infection at 6 and 12 d after immersion challenges also had no detectable virus, and no virus was detected in surviving fish from any group. A small number of ammocoetes sampled 6 d after the injection challenge had detectable virus, but at levels below the original quantity of virus injected. Overall there was no evidence of infection, replication, or persistence of any of the viruses in any of the treatment groups. Our results suggest that Pacific Lampreys are highly unlikely to serve as hosts that maintain or transmit these viruses.

  5. Ammocoetes of Pacific lamprey are not susceptible to common fish rhabdoviruses of the U.S. Pacific Northwest

    USGS Publications Warehouse

    Kurath, Gael; Jolley, C J.; Thompson, Tarin M.; Thompson, D.; Whitesel, A.T.; Gutenberger, S.; Winton, James R.

    2013-01-01

    Pacific Lampreys Entosphenus tridentatus have experienced severe population declines in recent years and efforts to develop captive rearing programs are under consideration. However, there is limited knowledge of their life history, ecology, and potential to harbor or transmit pathogens that may cause infectious disease. As a measure of the possible risks associated with introducing wild lampreys into existing fish culture facilities, larval lampreys (ammocoetes) were tested for susceptibility to infection and mortality caused by experimental exposures to the fish rhabdovirus pathogens: infectious hematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV). Two IHNV isolates, representing the U and M genogroups, and one VHSV isolate from the IVa genotype were each delivered to groups of ammocoetes by immersion at moderate and high viral doses, and by intraperitoneal injection. Ammocoetes were then held in triplicate tanks with no substrate or sediment. During 41 d of observation postchallenge there was low or no mortality in all groups, and no virus was detected in the small number of fish that died. Ammocoetes sampled for incidence of infection at 6 and 12 d after immersion challenges also had no detectable virus, and no virus was detected in surviving fish from any group. A small number of ammocoetes sampled 6 d after the injection challenge had detectable virus, but at levels below the original quantity of virus injected. Overall there was no evidence of infection, replication, or persistence of any of the viruses in any of the treatment groups. Our results suggest that Pacific Lampreys are highly unlikely to serve as hosts that maintain or transmit these viruses.

  6. Chikungunya Virus

    MedlinePlus

    ... is key! Prevent Infection. Use mosquito repellent. Chikungunya Virus Distribution Chikungunya in the U.S. What's New Surveillance ... Clinical Challenge For Travelers CDC Travelers' Health Chikungunya Virus Home Prevention Transmission Symptoms & Treatment Geographic Distribution Chikungunya ...

  7. Zika Virus

    MedlinePlus

    Zika is a virus that is spread mostly by mosquitoes. A pregnant mother can pass it to ... through blood transfusions. There have been outbreaks of Zika virus in the United States, Africa, Southeast Asia, ...

  8. Hepadna viruses

    SciTech Connect

    Robinson, W.; Koike, K.; Will, H.

    1987-01-01

    This book examines the molecular biology, disease pathogenesis, epidemiology, and clinical features of hepadna and other viruses with hepatic tropism and outlines future directions and approaches for their management. The volume's six sections provide a review of the various features, mechanisms, and functions of these viruses, ranging from hepadna virus replication and regulation of gene expression to the structure and function of hepadna-virus gene products.

  9. Virus maturation.

    PubMed

    Delgui, Laura R; Rodríguez, José F

    2013-01-01

    The formation of infectious virus particles is a highly complex process involving a series of sophisticated molecular events. In most cases, the assembly of virus structural elements results in the formation of immature virus particles unable to initiate a productive infection. Accordingly, for most viruses the final stage of the assembly pathway entails a set of structural transitions and/or biochemical modifications that transform inert precursor particles into fully infectious agents. In this chapter, we review the most relevant maturation mechanisms involved in the generation of infectious virions for a wide variety of viruses.

  10. Emergence of a new rhabdovirus associated with mass mortalities in eelpout (Zoarces viviparous) in the Baltic Sea.

    PubMed

    Axén, C; Hakhverdyan, M; Boutrup, T S; Blomkvist, E; Ljunghager, F; Alfjorden, A; Hagström, Å; Olesen, N J; Juremalm, M; Leijon, M; Valarcher, J-F

    2017-02-01

    We report the first description of a new Rhabdoviridae tentatively named eelpout rhabdovirus (EpRV genus Perhabdovirus). This virus was associated with mass mortalities in eelpout (Zoarces viviparous, Linnaeus) along the Swedish Baltic Sea coast line in 2014. Diseased fish showed signs of central nervous system infection, and brain lesions were confirmed by histology. A cytopathogenic effect was observed in cell culture, but ELISAs for the epizootic piscine viral haemorrhagic septicaemia virus (VHSV), infectious pancreas necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and spring viraemia of carp virus (SVCV) were negative. Further investigations by chloroform inactivation, indirect fluorescence antibody test and electron microscopy indicated the presence of a rhabdovirus. By deep sequencing of original tissue suspension and infected cell culture supernatant, the full viral genome was assembled and we confirmed the presence of a rhabdovirus with 59.5% nucleotide similarity to the closest relative Siniperca chuatsi rhabdovirus. The full-genome sequence of this new virus, eelpout rhabdovirus (EpRV), has been deposited in GenBank under accession number KR612230. An RT-PCR based on the L-gene sequence confirmed the presence of EpRV in sick/dead eelpout, but the virus was not found in control fish. Additional investigations to characterize the pathogenicity of EpRV are planned.

  11. Mycoplasma viruses.

    PubMed

    Maniloff, J

    1988-01-01

    Unlike bacterial viruses that infect cells bounded by a cell wall, mycoplasma viruses have evolved to enter and propagate in mycoplasma cells bounded only by a single lipid-protein cell membrane. In addition, mycoplasmas have the smallest amount of genetic information of any known cells, so their complexity is constrained by a limited genetic coding capacity. As a consequence of these host cell differences, mycoplasma viruses have been found to have a variety of structures and replication strategies which are different from those of the bacterial viruses. This article is a critical review of mycoplasma viruses infecting the genera Acholeplasma, Spiroplasma, and Mycoplasma; included are data on classification, morphology and structure, biological and physical properties, chemical composition, and productive and lysogenic replication cycles.

  12. Computer viruses

    NASA Technical Reports Server (NTRS)

    Denning, Peter J.

    1988-01-01

    The worm, Trojan horse, bacterium, and virus are destructive programs that attack information stored in a computer's memory. Virus programs, which propagate by incorporating copies of themselves into other programs, are a growing menace in the late-1980s world of unprotected, networked workstations and personal computers. Limited immunity is offered by memory protection hardware, digitally authenticated object programs,and antibody programs that kill specific viruses. Additional immunity can be gained from the practice of digital hygiene, primarily the refusal to use software from untrusted sources. Full immunity requires attention in a social dimension, the accountability of programmers.

  13. Lassa virus.

    PubMed

    Günther, Stephan; Lenz, Oliver

    2004-01-01

    Lassa virus is a RNA virus belonging to the family of Arenaviridae. It was discovered as the causative agent of a hemorrhagic fever--Lassa fever--about 30 years ago. Lassa fever is endemic in West Africa and is estimated to affect some 100,000 people annually. Great progress in the understanding of the life cycle of arenaviruses, including Lassa virus, has been made in recent years. New insights have been gained in the pathogenesis and molecular epidemiology of Lassa fever, and state-of the-art technologies for diagnosing this life-threatening disease have been developed. The intention of this review is to summarize in particular the recent literature on Lassa virus and Lassa fever. Several aspects ranging from basic research up to clinical practice and laboratory diagnosis are discussed and linked together.

  14. Zika Virus

    MedlinePlus

    ... Search Form Controls Cancel Submit Search the CDC Zika Virus Note: Javascript is disabled or is not ... Recommend on Facebook Tweet Share Compartir Men and Zika Zika Cases in Texas What’s your Zika IQ? ...

  15. Quantitative expression profiling of immune response genes in rainbow trout following infectious haematopoietic necrosis virus (IHNV) infection or DNA vaccination.

    PubMed

    Purcell, Maureen K; Kurath, Gael; Garver, Kyle A; Herwig, Russell P; Winton, James R

    2004-11-01

    Infectious haematopoietic necrosis virus (IHNV) is a well-studied virus of salmonid fishes. A highly efficacious DNA vaccine has been developed against this virus and studies have demonstrated that this vaccine induces both an early and transient non-specific anti-viral phase as well as long-term specific protection. The mechanisms of the early anti-viral phase are not known, but previous studies noted changes in Mx gene expression, suggesting a role for type I interferon. This study used quantitative real-time reverse transcriptase PCR methodology to compare expression changes over time of a number of cytokine or cytokine-related genes in the spleen of rainbow trout following injection with poly I:C, live IHNV, the IHNV DNA vaccine or a control plasmid encoding the non-antigenic luciferase gene. The target genes included Mx-1, viral haemorrhagic septicaemia virus induced gene 8 (Vig-8), TNF-alpha1, TNF-alpha2, IL-1beta1, IL-8, TGF-beta1 and Hsp70. Poly I:C stimulation induced several genes but the strongest and significant response was observed in the Mx-1 and Vig-8 genes. The live IHN virus induced a significant response in all genes examined except TGF-beta1. The control plasmid construct and the IHNV DNA vaccine marginally induced a number of genes, but the main difference between these two groups was a statistically significant induction of the Mx-1 and Vig-8 genes by the IHNV vaccine only. The gene expression profiles elicited by the live virus and the IHNV DNA vaccine differed in a number of aspects but this study confirms the clear role for a type I interferon-like response in early anti-viral defence.

  16. Quantitative expression profiling of immune response genes in rainbow trout following infectious haematopoietic necrosis virus (IHNV) infection or DNA vaccination

    USGS Publications Warehouse

    Purcell, Maureen K.; Kurath, Gael; Garver, Kyle A.; Herwig, Russell P.; Winton, James R.

    2004-01-01

    Infectious haematopoietic necrosis virus (IHNV) is a well-studied virus of salmonid fishes. A highly efficacious DNA vaccine has been developed against this virus and studies have demonstrated that this vaccine induces both an early and transient non-specific anti-viral phase as well as long-term specific protection. The mechanisms of the early anti-viral phase are not known, but previous studies noted changes in Mx gene expression, suggesting a role for type I interferon. This study used quantitative real-time reverse transcriptase PCR methodology to compare expression changes over time of a number of cytokine or cytokine-related genes in the spleen of rainbow trout following injection with poly I:C, live IHNV, the IHNV DNA vaccine or a control plasmid encoding the non-antigenic luciferase gene. The target genes included Mx-1, viral haemorrhagic septicaemia virus induced gene 8 (Vig-8), TNF-α1, TNF-α2, IL-1β1, IL-8, TGF-β1 and Hsp70. Poly I:C stimulation induced several genes but the strongest and significant response was observed in the Mx-1 and Vig-8 genes. The live IHN virus induced a significant response in all genes examined except TGF-β1. The control plasmid construct and the IHNV DNA vaccine marginally induced a number of genes, but the main difference between these two groups was a statistically significant induction of the Mx-1 and Vig-8 genes by the IHNV vaccine only. The gene expression profiles elicited by the live virus and the IHNV DNA vaccine differed in a number of aspects but this study confirms the clear role for a type I interferon-like response in early anti-viral defence.

  17. Virophages or satellite viruses?

    PubMed

    Krupovic, Mart; Cvirkaite-Krupovic, Virginija

    2011-11-01

    It has been argued that the smaller viruses associated with giant DNA viruses are a new biological entity. However, Mart Krupovic and Virginija Cvirkaite-Krupovic argue here that these smaller viruses should be classified with the satellite viruses.

  18. Ebola Virus and Marburg Virus

    MedlinePlus

    ... chimps and fruit bats in Africa. Transmission from animals to humans Experts suspect that both viruses are transmitted to humans through an infected animal's bodily fluids. Examples include: Blood. Butchering or eating ...

  19. Computer Viruses. Technology Update.

    ERIC Educational Resources Information Center

    Ponder, Tim, Comp.; Ropog, Marty, Comp.; Keating, Joseph, Comp.

    This document provides general information on computer viruses, how to help protect a computer network from them, measures to take if a computer becomes infected. Highlights include the origins of computer viruses; virus contraction; a description of some common virus types (File Virus, Boot Sector/Partition Table Viruses, Trojan Horses, and…

  20. Foodborne viruses.

    PubMed

    Koopmans, Marion; von Bonsdorff, Carl Henrik; Vinjé, Jan; de Medici, Dario; Monroe, Steve

    2002-06-01

    Foodborne and waterborne viral infections are increasingly recognized as causes of illness in humans. This increase is partly explained by changes in food processing and consumption patterns that lead to the worldwide availability of high-risk food. As a result, vast outbreaks may occur due to contamination of food by a single foodhandler or at a single source. Although there are numerous fecal-orally transmitted viruses, most reports of foodborne transmission describe infections with Norwalk-like caliciviruses (NLV) and hepatitis A virus (HAV), suggesting that these viruses are associated with the greatest risk of foodborne transmission. NLV and HAV can be transmitted from person to person, or indirectly via food, water, or fomites contaminated with virus-containing feces or vomit. People can be infected without showing symptoms. The high frequency of secondary cases of NLV illness and - to a lesser extent - of hepatitis A following a foodborne outbreak results in amplification of the problem. The burden of illness is highest in the elderly, and therefore is likely to increase due to the aging population. For HAV, the burden of illness may increase following hygienic control measures, due to a decreasing population of naturally immune individuals and a concurrent increase in the population at risk. Recent advances in the research of NLV and HAV have led to the development of molecular methods which can be used for molecular tracing of virus strains. These methods can be and have been used for the detection of common source outbreaks. While traditionally certain foods have been implicated in virus outbreaks, it is clear that almost any food item can be involved, provided it has been handled by an infected person. There are no established methods for detection of viruses in foods other than shellfish. Little information is available on disinfection and preventive measures specifically for these viruses. Studies addressing this issue are hampered by the lack of

  1. Zika Virus.

    PubMed

    FitzSimmons, Jack; Shah, Shailen

    2016-06-29

    To the Editor: Petersen et al. (April 21 issue)(1) provide a detailed review of Zika virus. We have some concern regarding diagnostic criteria for microcephaly in fetuses and newborns exposed to the virus. According to the Centers for Disease Control and Prevention (CDC) recommendation that microcephaly should be defined as an occipitofrontal circumference below the third percentile, nearly 3% of newborns would be categorized as having microcephaly. In Brazil, where there are 3 million live births per year, the application of this definition would result in nearly 90,000 infants being labeled as having microcephaly - a far greater number than . . .

  2. [Influenza virus].

    PubMed

    Juozapaitis, Mindaugas; Antoniukas, Linas

    2007-01-01

    Every year, especially during the cold season, many people catch an acute respiratory disease, namely flu. It is easy to catch this disease; therefore, it spreads very rapidly and often becomes an epidemic or a global pandemic. Airway inflammation and other body ailments, which form in a very short period, torment the patient several weeks. After that, the symptoms of the disease usually disappear as quickly as they emerged. The great epidemics of flu have rather unique characteristics; therefore, it is possible to identify descriptions of such epidemics in historic sources. Already in the 4th century bc, Hippocrates himself wrote about one of them. It is known now that flu epidemics emerge rather frequently, but there are no regular intervals between those events. The epidemics can differ in their consequences, but usually they cause an increased mortality of elderly people. The great flu epidemics of the last century took millions of human lives. In 1918-19, during "The Spanish" pandemic of flu, there were around 40-50 millions of deaths all over the world; "Pandemic of Asia" in 1957 took up to one million lives, etc. Influenza virus can cause various disorders of the respiratory system: from mild inflammations of upper airways to acute pneumonia that finally results in the patient's death. Scientist Richard E. Shope, who investigated swine flu in 1920, had a suspicion that the cause of this disease might be a virus. Already in 1933, scientists from the National Institute for Medical Research in London - Wilson Smith, Sir Christopher Andrewes, and Sir Patrick Laidlaw - for the first time isolated the virus, which caused human flu. Then scientific community started the exhaustive research of influenza virus, and the great interest in this virus and its unique features is still active even today.

  3. Honeybee viruses in Uruguay.

    PubMed

    Antúnez, Karina; D'Alessandro, Bruno; Corbella, Eduardo; Ramallo, Gustavo; Zunino, Pablo

    2006-09-01

    Mortality of honeybees is a serious problem that beekeepers have to face periodically in Uruguay and worldwide. The presence of RNA viruses, in addition to other pathogens may be one of its possible causes. In this work, we detected Chronic bee paralysis virus, Acute bee paralysis virus, Black queen cell virus, Sacbrood virus and Deformed wing virus in samples of Uruguayan honeybees with or without Varroa destructor and Nosema apis. The detection of viruses in different provinces, simultaneous co-infection of colonies by several viruses and the fact that 96% of the samples were infected with one or more virus, indicates they are widely spread in the region.

  4. The Geometry of Viruses.

    ERIC Educational Resources Information Center

    Case, Christine L.

    1991-01-01

    Presented is an activity in which students make models of viruses, which allows them to visualize the shape of these microorganisms. Included are some background on viruses, the biology and geometry of viruses, directions for building viruses, a comparison of cells and viruses, and questions for students. (KR)

  5. The Geometry of Viruses.

    ERIC Educational Resources Information Center

    Case, Christine L.

    1991-01-01

    Presented is an activity in which students make models of viruses, which allows them to visualize the shape of these microorganisms. Included are some background on viruses, the biology and geometry of viruses, directions for building viruses, a comparison of cells and viruses, and questions for students. (KR)

  6. Measles virus.

    PubMed

    Naim, Hussein Y

    2015-01-01

    Measles was an inevitable infection during the human development with substantial degree of morbidity and mortality. The severity of measles virus (MV) infection was largely contained by the development of a live attenuated vaccine that was introduced into the vaccination programs. However, all efforts to eradicate the disease failed and continued to annually result in significant deaths. The development of molecular biology techniques allowed the rescue of MV from cDNA that enabled important insights into a variety of aspects of the biology of the virus and its pathogenesis. Subsequently these technologies facilitated the development of novel vaccine candidates that induce immunity against measles and other pathogens. Based on the promising prospective, the use of MV as a recombinant vaccine and a therapeutic vector is addressed.

  7. Plant Virus Metagenomics: Advances in Virus Discovery.

    PubMed

    Roossinck, Marilyn J; Martin, Darren P; Roumagnac, Philippe

    2015-06-01

    In recent years plant viruses have been detected from many environments, including domestic and wild plants and interfaces between these systems-aquatic sources, feces of various animals, and insects. A variety of methods have been employed to study plant virus biodiversity, including enrichment for virus-like particles or virus-specific RNA or DNA, or the extraction of total nucleic acids, followed by next-generation deep sequencing and bioinformatic analyses. All of the methods have some shortcomings, but taken together these studies reveal our surprising lack of knowledge about plant viruses and point to the need for more comprehensive studies. In addition, many new viruses have been discovered, with most virus infections in wild plants appearing asymptomatic, suggesting that virus disease may be a byproduct of domestication. For plant pathologists these studies are providing useful tools to detect viruses, and perhaps to predict future problems that could threaten cultivated plants.

  8. Zika Virus.

    PubMed

    Musso, Didier; Gubler, Duane J

    2016-07-01

    Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) in the genus Flavivirus and the family Flaviviridae. ZIKV was first isolated from a nonhuman primate in 1947 and from mosquitoes in 1948 in Africa, and ZIKV infections in humans were sporadic for half a century before emerging in the Pacific and the Americas. ZIKV is usually transmitted by the bite of infected mosquitoes. The clinical presentation of Zika fever is nonspecific and can be misdiagnosed as other infectious diseases, especially those due to arboviruses such as dengue and chikungunya. ZIKV infection was associated with only mild illness prior to the large French Polynesian outbreak in 2013 and 2014, when severe neurological complications were reported, and the emergence in Brazil of a dramatic increase in severe congenital malformations (microcephaly) suspected to be associated with ZIKV. Laboratory diagnosis of Zika fever relies on virus isolation or detection of ZIKV-specific RNA. Serological diagnosis is complicated by cross-reactivity among members of the Flavivirus genus. The adaptation of ZIKV to an urban cycle involving humans and domestic mosquito vectors in tropical areas where dengue is endemic suggests that the incidence of ZIKV infections may be underestimated. There is a high potential for ZIKV emergence in urban centers in the tropics that are infested with competent mosquito vectors such as Aedes aegypti and Aedes albopictus. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  9. Zika Virus

    PubMed Central

    2016-01-01

    SUMMARY Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) in the genus Flavivirus and the family Flaviviridae. ZIKV was first isolated from a nonhuman primate in 1947 and from mosquitoes in 1948 in Africa, and ZIKV infections in humans were sporadic for half a century before emerging in the Pacific and the Americas. ZIKV is usually transmitted by the bite of infected mosquitoes. The clinical presentation of Zika fever is nonspecific and can be misdiagnosed as other infectious diseases, especially those due to arboviruses such as dengue and chikungunya. ZIKV infection was associated with only mild illness prior to the large French Polynesian outbreak in 2013 and 2014, when severe neurological complications were reported, and the emergence in Brazil of a dramatic increase in severe congenital malformations (microcephaly) suspected to be associated with ZIKV. Laboratory diagnosis of Zika fever relies on virus isolation or detection of ZIKV-specific RNA. Serological diagnosis is complicated by cross-reactivity among members of the Flavivirus genus. The adaptation of ZIKV to an urban cycle involving humans and domestic mosquito vectors in tropical areas where dengue is endemic suggests that the incidence of ZIKV infections may be underestimated. There is a high potential for ZIKV emergence in urban centers in the tropics that are infested with competent mosquito vectors such as Aedes aegypti and Aedes albopictus. PMID:27029595

  10. Ebola Virus Disease

    MedlinePlus

    ... sheets Fact files Questions & answers Features Multimedia Contacts Ebola virus disease Fact sheet Updated January 2016 Key ... for survivors of Ebola virus disease Symptoms of Ebola virus disease The incubation period, that is, the ...

  11. Influenza (Flu) Viruses

    MedlinePlus

    ... Seasonal Avian Swine/Variant Pandemic Other Influenza (Flu) Viruses Language: English (US) Español Recommend on Facebook ... circulate and cause illness. More Information about Flu Viruses Types of Influenza Viruses Influenza A and B ...

  12. Respiratory Syncytial Virus (RSV)

    MedlinePlus

    ... and premature birth: Are you at risk? Zika virus and pregnancy Folic acid Medicine safety and pregnancy ... It's been added to your dashboard . Respiratory syncytial virus (RSV) is a common virus that infects the ...

  13. Computer Viruses: An Overview.

    ERIC Educational Resources Information Center

    Marmion, Dan

    1990-01-01

    Discusses the early history and current proliferation of computer viruses that occur on Macintosh and DOS personal computers, mentions virus detection programs, and offers suggestions for how libraries can protect themselves and their users from damage by computer viruses. (LRW)

  14. SAMPLING VIRUSES FROM SOIL

    EPA Science Inventory

    This chapter describes in detail methods for detecting viruses of bacteria and humans in soil. Methods also are presented for the assay of these viruses. Reference sources are provided for information on viruses of plants.

  15. Zika Virus and Pregnancy

    MedlinePlus

    ... Management Education & Events Advocacy For Patients About ACOG Zika Virus and Pregnancy Home For Patients Zika Virus ... Patient Education Pamphlets - Spanish Share: PEV002, September 2016 Zika Virus and Pregnancy There are risks to your ...

  16. Computer Viruses: An Overview.

    ERIC Educational Resources Information Center

    Marmion, Dan

    1990-01-01

    Discusses the early history and current proliferation of computer viruses that occur on Macintosh and DOS personal computers, mentions virus detection programs, and offers suggestions for how libraries can protect themselves and their users from damage by computer viruses. (LRW)

  17. Hanta virus (image)

    MedlinePlus

    Hanta virus is a distant cousin of Ebola virus, but is found worldwide. The virus is spread by human contact with rodent waste. Dangerous respiratory illness develops. Effective treatment is not yet ...

  18. Assessing the impact of climate change on disease emergence in freshwater fish in the United Kingdom.

    PubMed

    Marcos-López, M; Gale, P; Oidtmann, B C; Peeler, E J

    2010-10-01

    A risk framework has been developed to examine the influence of climate change on disease emergence in the United Kingdom. The fish immune response and the replication of pathogens are often correlated with water temperature, which manifest as temperature ranges for infection and clinical diseases. These data are reviewed for the major endemic and exotic disease threats to freshwater fish. Increasing water temperatures will shift the balance in favour of either the host or pathogen, changing the frequency and distribution of disease. A number of endemic diseases of salmonids (e.g. enteric red mouth, furunculosis, proliferative kidney disease and white spot) will become more prevalent and difficult to control as water temperatures increase. Outbreaks of koi herpesvirus in carp fisheries are likely to occur over a longer period each summer. Climate change also alters the threat level associated with exotic pathogens. The risk of viral haemorrhagic septicaemia (VHSV), infectious haematopoietic necrosis virus (IHNV) and spring viraemia of carp virus (SVCV) declines as infection generally only establishes when water temperatures are less than 14°C for VHSV and IHNV and 17°C for SCVC. The risk of establishment of other exotic pathogens (epizootic haematopoietic necrosis and epizootic ulcerative syndrome) increases. The spread of Lactococcus garvieae northwards in Europe is likely to continue, and thus is more likely to be both introduced and become established. Measures to reduce the threat of exotic pathogens need to be revised to account for the changing exotic diseases threat. Increasing water temperatures and the negative effects of extreme weather events (e.g. storms) are likely to alter the freshwater environment adversely for both wild and farmed salmonid populations, increasing their susceptibility to disease and the likelihood of disease emergence. For wild populations, surveillance and risk mitigation need to be focused on locations where disease emergence

  19. Virus Movement Maintains Local Virus Population Diversity

    SciTech Connect

    J. Snyder; B. Wiedenheft; M. Lavin; F. Roberto; J. Spuhler; A. Ortmann; T. Douglas; M. Young

    2007-11-01

    Viruses are the largest reservoir of genetic material on the planet, yet little is known about the population dynamics of any virus within its natural environment. Over a 2-year period, we monitored the diversity of two archaeal viruses found in hot springs within Yellowstone National Park (YNP). Both temporal phylogeny and neutral biodiversity models reveal that virus diversity in these local environments is not being maintained by mutation but rather by high rates of immigration from a globally distributed metacommunity. These results indicate that geographically isolated hot springs are readily able to exchange viruses. The importance of virus movement is supported by the detection of virus particles in air samples collected over YNP hot springs and by their detection in metacommunity sequencing projects conducted in the Sargasso Sea. Rapid rates of virus movement are not expected to be unique to these archaeal viruses but rather a common feature among virus metacommunities. The finding that virus immigration rather than mutation can dominate community structure has significant implications for understanding virus circulation and the role that viruses play in ecology and evolution by providing a reservoir of mobile genetic material.

  20. Virus movement maintains local virus population diversity.

    PubMed

    Snyder, Jamie C; Wiedenheft, Blake; Lavin, Matthew; Roberto, Francisco F; Spuhler, Josh; Ortmann, Alice C; Douglas, Trevor; Young, Mark

    2007-11-27

    Viruses are the largest reservoir of genetic material on the planet, yet little is known about the population dynamics of any virus within its natural environment. Over a 2-year period, we monitored the diversity of two archaeal viruses found in hot springs within Yellowstone National Park (YNP). Both temporal phylogeny and neutral biodiversity models reveal that virus diversity in these local environments is not being maintained by mutation but rather by high rates of immigration from a globally distributed metacommunity. These results indicate that geographically isolated hot springs are readily able to exchange viruses. The importance of virus movement is supported by the detection of virus particles in air samples collected over YNP hot springs and by their detection in metacommunity sequencing projects conducted in the Sargasso Sea. Rapid rates of virus movement are not expected to be unique to these archaeal viruses but rather a common feature among virus metacommunities. The finding that virus immigration rather than mutation can dominate community structure has significant implications for understanding virus circulation and the role that viruses play in ecology and evolution by providing a reservoir of mobile genetic material.

  1. Understanding viruses: Philosophical investigations.

    PubMed

    Pradeu, Thomas; Kostyrka, Gladys; Dupré, John

    2016-10-01

    Viruses have been virtually absent from philosophy of biology. In this editorial introduction, we explain why we think viruses are philosophically important. We focus on six issues (the definition of viruses, the individuality and diachronic identity of a virus, the possibility to classify viruses into species, the question of whether viruses are living, the question of whether viruses are organisms, and finally the biological roles of viruses in ecology and evolution), and we show how they relate to classic questions of philosophy of biology and even general philosophy. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Making a World of Difference: Recent USGS Contributions to the Nation.

    DTIC Science & Technology

    1998-01-01

    of a highly feared fish disease agent, viral hemorrhagic septicemia virus ( VHSV ). To safeguard the health of salmon and trout in North America...certain exotic fish pathogens. In 1988 and 1989, more than 5 million fish and eggs in western Washington were destroyed after scientists discovered VHSV ...that VHSV was confined to Europe, where it has caused high mortality and losses of over $40 million a year at rainbow trout farms. Scientists at the

  3. Disseminated intravascular coagulation with Fusobacterium necrophorum septicaemia.

    PubMed Central

    Potter, M. N.; Drysdale, H. C.; Price, P. A.; Buck, A. C.

    1988-01-01

    A 23 year old woman died within six hours of admission from acute disseminated intravascular coagulation. Fusobacterium necrophorum, a Gram negative anaerobic organism, was isolated as a single pathogen from the blood cultures. This association has not previously been reported. PMID:3174529

  4. Vaccination against enteric septicaemia of catfish

    USDA-ARS?s Scientific Manuscript database

    Edwardsiella ictaluri, the causative agent of enteric septicemia (ESC) of catfish, is one of the most economically important diseases of cultured channel catfish. In 2002, Wagner and coworkers reported that enteric septicemia of catfish (ESC) and columnaris (Flavobacterium columnaris) were the two m...

  5. Crystallization of viruses and virus proteins

    NASA Astrophysics Data System (ADS)

    Sehnke, Paul C.; Harrington, Melissa; Hosur, M. V.; Li, Yunge; Usha, R.; Craig Tucker, R.; Bomu, Wu; Stauffacher, Cynthia V.; Johnson, John E.

    1988-07-01

    Methods for crystallizing six isometric plant and insect viruses are presented. Procedures developed for modifying, purifying and crystallizing coat protein subunits isolated from a virus forming asymmetric, spheroidal particles, stabilized almost exclusively by protein-RNA interactions, are also discussed. The tertiary and quaternary structures of small RNA viruses are compared.

  6. Viruses and Virus Diseases of Rubus

    USDA-ARS?s Scientific Manuscript database

    Rubus species are propagated vegetatively and are subject to infection by viruses during development, propagation and fruit production stages. Reports of initial detection and symptoms of more than 30 viruses, virus-like diseases and phytoplasmas affecting Rubus spp. have been reviewed more than 20 ...

  7. The Tobacco Mosaic Virus.

    ERIC Educational Resources Information Center

    Sulzinski, Michael A.

    1992-01-01

    Explains how the tobacco mosaic virus can be used to study virology. Presents facts about the virus, procedures to handle the virus in the laboratory, and four laboratory exercises involving the viruses' survival under inactivating conditions, dilution end point, filterability, and microscopy. (MDH)

  8. The Tobacco Mosaic Virus.

    ERIC Educational Resources Information Center

    Sulzinski, Michael A.

    1992-01-01

    Explains how the tobacco mosaic virus can be used to study virology. Presents facts about the virus, procedures to handle the virus in the laboratory, and four laboratory exercises involving the viruses' survival under inactivating conditions, dilution end point, filterability, and microscopy. (MDH)

  9. Understanding Ebola Virus Transmission

    PubMed Central

    Judson, Seth; Prescott, Joseph; Munster, Vincent

    2015-01-01

    An unprecedented number of Ebola virus infections among healthcare workers and patients have raised questions about our understanding of Ebola virus transmission. Here, we explore different routes of Ebola virus transmission between people, summarizing the known epidemiological and experimental data. From this data, we expose important gaps in Ebola virus research pertinent to outbreak situations. We further propose experiments and methods of data collection that will enable scientists to fill these voids in our knowledge about the transmission of Ebola virus. PMID:25654239

  10. Understanding ebola virus transmission.

    PubMed

    Judson, Seth; Prescott, Joseph; Munster, Vincent

    2015-02-03

    An unprecedented number of Ebola virus infections among healthcare workers and patients have raised questions about our understanding of Ebola virus transmission. Here, we explore different routes of Ebola virus transmission between people, summarizing the known epidemiological and experimental data. From this data, we expose important gaps in Ebola virus research pertinent to outbreak situations. We further propose experiments and methods of data collection that will enable scientists to fill these voids in our knowledge about the transmission of Ebola virus.

  11. Virus-Vectored Influenza Virus Vaccines

    PubMed Central

    Tripp, Ralph A.; Tompkins, S. Mark

    2014-01-01

    Despite the availability of an inactivated vaccine that has been licensed for >50 years, the influenza virus continues to cause morbidity and mortality worldwide. Constant evolution of circulating influenza virus strains and the emergence of new strains diminishes the effectiveness of annual vaccines that rely on a match with circulating influenza strains. Thus, there is a continued need for new, efficacious vaccines conferring cross-clade protection to avoid the need for biannual reformulation of seasonal influenza vaccines. Recombinant virus-vectored vaccines are an appealing alternative to classical inactivated vaccines because virus vectors enable native expression of influenza antigens, even from virulent influenza viruses, while expressed in the context of the vector that can improve immunogenicity. In addition, a vectored vaccine often enables delivery of the vaccine to sites of inductive immunity such as the respiratory tract enabling protection from influenza virus infection. Moreover, the ability to readily manipulate virus vectors to produce novel influenza vaccines may provide the quickest path toward a universal vaccine protecting against all influenza viruses. This review will discuss experimental virus-vectored vaccines for use in humans, comparing them to licensed vaccines and the hurdles faced for licensure of these next-generation influenza virus vaccines. PMID:25105278

  12. The theoretical reliability of PCR-based fish viral diagnostic methods is critically affected when they are applied to fish populations with low prevalence and virus loads.

    PubMed

    Dopazo, C P; Moreno, P; Olveira, J G; Borrego, J J

    2017-09-15

    The reliability of PCR techniques is an important issue in viral diagnosis, and it is even crucial when they must be applied for detection of viruses in asymptomatic carriers. The problems will arise when the aim is to study fish wild populations, where the viral loads and prevalence values are extremely low. We have evaluated several PCR procedures employed by two laboratories for monitoring fish captured in several oceanographic campaigns performed in the Gulf of Cádiz. To evaluate the reliability of different diagnostics test used, we have re-analyzed fish samples that had been previously subjected to diagnosis for a surveillance of viruses performed in 2010-2011 in wild fish populations. The following parameters were employed: the clinical sensitivity (Ss), the clinical specificity (Sp), the predictive positive value (PPV), the predictive negative value (PNV), and the positive and negative likelihood ratio (LR(+) and LR(-) ). For viral nervious necrosis virus (VNNV), a RT-PCR procedure supplemented by nested PCR showed the highest values (100%) for all the parameters. For viral hemorrhagic septicemia virus (VHSV), the highest values were provided by RT-PCR supplemented by dot blot hybridization. In the case of Infectious pancreatic necrosis virus (IPNV), none of the procedures yielded 100% for any parameter. The results obtained for viral prevalence indicate: (i) that the conservation of the samples at -80 °C did not affect to the capacity of detection of the virus in the tissues, and (ii) that the reproducibility of the diagnosis can be affected by factors including the staff experience and/or the materials employed. Finally, the use of a combination of procedures in advised to ensure the maximum reliability of the diagnosis when it is applied to asymptomatic fish populations. This paper describes a strategy of combining diagnostic tests for the surveillance and monitoring of wild fish populations to reduce underestimation of the prevalence of viruses this

  13. Viruses Infecting Reptiles

    PubMed Central

    Marschang, Rachel E.

    2011-01-01

    A large number of viruses have been described in many different reptiles. These viruses include arboviruses that primarily infect mammals or birds as well as viruses that are specific for reptiles. Interest in arboviruses infecting reptiles has mainly focused on the role reptiles may play in the epidemiology of these viruses, especially over winter. Interest in reptile specific viruses has concentrated on both their importance for reptile medicine as well as virus taxonomy and evolution. The impact of many viral infections on reptile health is not known. Koch’s postulates have only been fulfilled for a limited number of reptilian viruses. As diagnostic testing becomes more sensitive, multiple infections with various viruses and other infectious agents are also being detected. In most cases the interactions between these different agents are not known. This review provides an update on viruses described in reptiles, the animal species in which they have been detected, and what is known about their taxonomic positions. PMID:22163336

  14. Viruses infecting reptiles.

    PubMed

    Marschang, Rachel E

    2011-11-01

    A large number of viruses have been described in many different reptiles. These viruses include arboviruses that primarily infect mammals or birds as well as viruses that are specific for reptiles. Interest in arboviruses infecting reptiles has mainly focused on the role reptiles may play in the epidemiology of these viruses, especially over winter. Interest in reptile specific viruses has concentrated on both their importance for reptile medicine as well as virus taxonomy and evolution. The impact of many viral infections on reptile health is not known. Koch's postulates have only been fulfilled for a limited number of reptilian viruses. As diagnostic testing becomes more sensitive, multiple infections with various viruses and other infectious agents are also being detected. In most cases the interactions between these different agents are not known. This review provides an update on viruses described in reptiles, the animal species in which they have been detected, and what is known about their taxonomic positions.

  15. Testing for Human Immunodeficiency Virus

    MedlinePlus

    ... education Fact Sheet PFS005: Testing for Human Immunodeficiency Virus AUGUST 2015 • Reasons for Getting Tested • Who Should ... For More Information • Glossary Testing for Human Immunodeficiency Virus Human immunodeficiency virus (HIV) is the virus that ...

  16. Viruses in the sea

    NASA Astrophysics Data System (ADS)

    Suttle, Curtis A.

    2005-09-01

    Viruses exist wherever life is found. They are a major cause of mortality, a driver of global geochemical cycles and a reservoir of the greatest genetic diversity on Earth. In the oceans, viruses probably infect all living things, from bacteria to whales. They affect the form of available nutrients and the termination of algal blooms. Viruses can move between marine and terrestrial reservoirs, raising the spectre of emerging pathogens. Our understanding of the effect of viruses on global systems and processes continues to unfold, overthrowing the idea that viruses and virus-mediated processes are sidebars to global processes.

  17. Viruses in the sea.

    PubMed

    Suttle, Curtis A

    2005-09-15

    Viruses exist wherever life is found. They are a major cause of mortality, a driver of global geochemical cycles and a reservoir of the greatest genetic diversity on Earth. In the oceans, viruses probably infect all living things, from bacteria to whales. They affect the form of available nutrients and the termination of algal blooms. Viruses can move between marine and terrestrial reservoirs, raising the spectre of emerging pathogens. Our understanding of the effect of viruses on global systems and processes continues to unfold, overthrowing the idea that viruses and virus-mediated processes are sidebars to global processes.

  18. Raspberry (Rubus spp.)-Viruses

    USDA-ARS?s Scientific Manuscript database

    There are several important virus diseases of raspberry and black raspberry in the Pacific Northwest. Pollen-borne viruses include Raspberry bushy dwarf virus and Strawberry necrotic shock virus (aka Tobacco streak virus –Rubus isolate or Black raspberry latent virus). Strawberry necrotic shock viru...

  19. Ebola (Ebola Virus Disease)

    MedlinePlus

    ... to Introduce a Vaccine against Ebola Ebola Virus Ecology and Transmission About Ebola Signs and Symptoms Symptoms ... Resources Videos Audio Infographics & Illustrations Factsheets Posters Virus Ecology Graphic File Formats Help: How do I view ...

  20. Quasispecies of dengue virus.

    PubMed

    Kurosu, Takeshi

    2011-12-01

    Pathogenic viruses have RNA genomes that cause acute and chronic infections. These viruses replicate with high mutation rates and exhibit significant genetic diversity, so-called viral quasispecies. Viral quasispecies play an important role in chronic infectious diseases, but little is known about their involvement in acute infectious diseases such as dengue virus (DENV) infection. DENV, the most important human arbovirus, is a causative agent of dengue fever (DF) and dengue hemorrhagic fever (DHF). Accumulating observations suggest that DENV exists as an extremely diverse virus population, but its biological significance is unclear. In other virus diseases, quasispecies affect the therapeutic strategies using drugs and vaccines. Here, I describe the quasispecies of DENV and discuss the possible role of quasispecies in the pathogenesis of and therapeutic strategy against DENV infection in comparison with other viruses such as Hepatitis C virus, human immunodeficiency virus type 1, and poliovirus.

  1. Respiratory syncytial virus (RSV)

    MedlinePlus

    Respiratory syncytial virus (RSV) is a very common virus that leads to mild, cold-like symptoms in adults and older healthy children. It can be more serious in young babies, especially those in certain high-risk groups.

  2. West Nile virus

    MedlinePlus

    ... believe West Nile virus is spread when a mosquito bites an infected bird and then bites a ... avoid getting West Nile virus infection after a mosquito bite. People in good health generally do not ...

  3. Hepatitis virus panel

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003558.htm Hepatitis virus panel To use the sharing features on this page, please enable JavaScript. The hepatitis virus panel is a series of blood tests used ...

  4. Human Parainfluenza Viruses

    MedlinePlus

    ... HPIVs Are Not the Same as Influenza (Flu) Viruses People usually get HPIV infections more often in ... hands, and touching objects or surfaces with the viruses on them then touching your mouth, nose, or ...

  5. Respiratory Syncytial Virus Infections

    MedlinePlus

    Respiratory syncytial virus (RSV) causes mild, cold-like symptoms in adults and older healthy children. It can cause serious problems in ... tests can tell if your child has the virus. There is no specific treatment. You should give ...

  6. Viruses and cancer

    SciTech Connect

    Rigby, P.W.J.; Wilkie, N.M.

    1985-01-01

    This book contains 14 selections. Some of the titles are: Immortalising gene(s) encoded by Epstein-Barr Virus; Adenovirus genes involved in transformation. What determines the oncogenic phenotype.; Oncogenesis by mouse mammary tumour virus; and Transforming ras genes.

  7. Live Virus Smallpox Vaccine

    MedlinePlus

    ... Submit What's this? Submit Button The Live Virus Smallpox Vaccine Language: English Español (Spanish) Recommend on Facebook ... the vaccinia virus. Who should NOT get the smallpox vaccine? People most likely to have side effects ...

  8. Advances in virus research

    SciTech Connect

    Maramorosch, K. ); Murphy, F.A. ); Shatkin, A.J. )

    1988-01-01

    This book contains eight chapters. Some of the titles are: Initiation of viral DNA replication; Vaccinia: virus, vector, vaccine; The pre-S region of hepadnavirus envelope proteins; and Archaebacterial viruses.

  9. Viruses and Breast Cancer

    PubMed Central

    Lawson, James S.; Heng, Benjamin

    2010-01-01

    Viruses are the accepted cause of many important cancers including cancers of the cervix and anogenital area, the liver, some lymphomas, head and neck cancers and indirectly human immunodeficiency virus associated cancers. For over 50 years, there have been serious attempts to identify viruses which may have a role in breast cancer. Despite these efforts, the establishment of conclusive evidence for such a role has been elusive. However, the development of extremely sophisticated new experimental techniques has allowed the recent development of evidence that human papilloma virus, Epstein-Barr virus, mouse mammary tumor virus and bovine leukemia virus may each have a role in the causation of human breast cancers. This is potentially good news as effective vaccines are already available to prevent infections from carcinogenic strains of human papilloma virus, which causes cancer of the uterine cervix. PMID:24281093

  10. Tumorigenic DNA viruses

    SciTech Connect

    Klein, G.

    1989-01-01

    The eighth volume of Advances in Viral Oncology focuses on the three major DNA virus groups with a postulated or proven tumorigenic potential: papillomaviruses, animal hepatitis viruses, and the Epstein-Bar virus. In the opening chapters, the contributors analyze the evidence that papillomaviruses and animal hepatitis viruses are involved in tumorigenesis and describe the mechanisms that trigger virus-host cell interactions. A detailed section on the Epstein-Barr virus (EBV) - comprising more than half the book - examines the transcription and mRNA processing patterns of the virus genome; the mechanisms by which EBV infects lymphoid and epithelial cells; the immunological aspects of the virus; the actions of EBV in hosts with Acquired Immune Deficiency Syndrome; and the involvement of EBV in the etiology of Burkitt's lymphoma.

  11. Virus Assembly and Maturation

    NASA Astrophysics Data System (ADS)

    Johnson, John E.

    2004-03-01

    We use two techniques to look at three-dimensional virus structure: electron cryomicroscopy (cryoEM) and X-ray crystallography. Figure 1 is a gallery of virus particles whose structures Timothy Baker, one of my former colleagues at Purdue University, used cryoEM to determine. It illustrates the variety of sizes of icosahedral virus particles. The largest virus particle on this slide is the Herpes simplex virus, around 1200Å in diameter; the smallest we examined was around 250Å in diameter. Viruses bear their genomic information either as positive-sense DNA and RNA, double-strand DNA, double-strand RNA, or negative-strand RNA. Viruses utilize the various structure and function "tactics" seen throughout cell biology to replicate at high levels. Many of the biological principles that we consider general were in fact discovered in the context of viruses ...

  12. Viruses and human cancer

    SciTech Connect

    Gallo, R.C.; Haseltine, W.; Klein, G.; Zur Hausen, H.

    1987-01-01

    This book contains papers on the following topics: Immunology and Epidemiology, Biology and Pathogenesis, Models of Pathogenesis and Treatment, Simian and Bovine Retroviruses, Human Papilloma Viruses, EBV and Herpesvirus, and Hepatitis B Virus.

  13. Avian influenza virus and Newcastle disease virus

    USDA-ARS?s Scientific Manuscript database

    Avian influenza virus (AIV) and Newcastle disease virus (NDV) severely impact poultry egg production. Decreased egg yield and hatchability, as well as misshapen eggs, are often observed during infection with AIV and NDV, even with low-virulence strains or in vaccinated flocks. Data suggest that in...

  14. Nairobi sheep disease virus/Ganjam virus.

    PubMed

    M D, Baron; B, Holzer

    2015-08-01

    Nairobi sheep disease virus (NSDV) is a tick-borne virus which causes a severe disease in sheep and goats, and has been responsible for several outbreaks of disease in East Africa. The virus is also found in the Indian subcontinent, where it is known as Ganjam virus. The virus only spreads through the feeding of competent infected ticks, and is therefore limited in its geographic distribution by the distribution of those ticks, Rhipicephalus appendiculata in Africa and Haemaphysalis intermedia in India. Animals bred in endemic areas do not normally develop disease, and the impact is therefore primarily on animals being moved for trade or breeding purposes. The disease caused by NSDV has similarities to several other ruminant diseases, and laboratory diagnosis is necessary for confirmation. There are published methods for diagnosis based on polymerase chain reaction, for virus growth in cell culture and for other simple diagnostic tests, though none has been commercialised. There is no established vaccine against NSDV, although cell-culture attenuated strains have been developed which show promise and could be put into field trials if it were deemed necessary. The virus is closely related to Crimean-Congo haemorrhagic fever virus, and studies on NSDV may therefore be useful in understanding this important human pathogen.

  15. Avian influenza virus

    USDA-ARS?s Scientific Manuscript database

    Avian influenza virus (AIV) is type A influenza, which is adapted to an avian host. Although avian influenza has been isolated from numerous avian species, the primary natural hosts for the virus are dabbling ducks, shorebirds, and gulls. The virus can be found world-wide in these species and in o...

  16. Blueberry latent spherical virus

    USDA-ARS?s Scientific Manuscript database

    ‘Blueray’ tissue was mechanically inoculated onto Chenopodium quinoa indicator plants as part of a study to determine virus presence in blueberries at Iwate University, Japan. Plants developed chlorosis indicative of virus presence and after virus purification and genome characterization it was dete...

  17. Computer Virus Protection

    ERIC Educational Resources Information Center

    Rajala, Judith B.

    2004-01-01

    A computer virus is a program--a piece of executable code--that has the unique ability to replicate. Like biological viruses, computer viruses can spread quickly and are often difficult to eradicate. They can attach themselves to just about any type of file, and are spread by replicating and being sent from one individual to another. Simply having…

  18. Respiratory Syncytial Virus

    MedlinePlus

    ... Your 1- to 2-Year-Old Respiratory Syncytial Virus KidsHealth > For Parents > Respiratory Syncytial Virus A A A What's in this article? About ... RSV When to Call the Doctor en español Virus respiratorio sincitial About RSV Respiratory syncytial (sin-SISH- ...

  19. Avian influenza virus

    USDA-ARS?s Scientific Manuscript database

    Avian influenza (AI) is caused by type A influenza virus, a member of the Orthomyxoviridae family. AI viruses are serologically categorized into 16 hemagglutinin (H1-H16) and 9 neuraminidase (N1-N9) subtypes. All subtypes have been identified in birds. Infections by AI viruses have been reported in ...

  20. Computer Virus Protection

    ERIC Educational Resources Information Center

    Rajala, Judith B.

    2004-01-01

    A computer virus is a program--a piece of executable code--that has the unique ability to replicate. Like biological viruses, computer viruses can spread quickly and are often difficult to eradicate. They can attach themselves to just about any type of file, and are spread by replicating and being sent from one individual to another. Simply having…

  1. Influenza viruses: an introduction.

    PubMed

    Kawaoka, Yoshihiro; Neumann, Gabriele

    2012-01-01

    We provide a brief introduction into the genome organization, life cycle, pathogenicity, and host range of influenza A viruses. We also briefly summarize influenza pandemics and currently available measures to control influenza virus outbreaks, including vaccines and antiviral compounds to influenza viruses.

  2. The taxonomy of viruses should include viruses.

    PubMed

    Calisher, Charles H

    2016-05-01

    Having lost sight of its goal, the International Committee on Taxonomy of Viruses has redoubled its efforts. That goal is to arrive at a consensus regarding virus classification, i.e., proper placement of viruses in a hierarchical taxonomic scheme; not an easy task given the wide variety of recognized viruses. Rather than suggesting a continuation of the bureaucratic machinations of the past, this opinion piece is a call for insertion of common sense in sorting out the avalanche of information already, and soon-to-be, accrued data. In this way information about viruses ideally would be taxonomically correct as well as useful to working virologists and journal editors, rather than being lost, minimized, or ignored.

  3. Blueberry (Vaccinium corymbosum)-Virus Diseases

    USDA-ARS?s Scientific Manuscript database

    At least six viruses have been found in highbush blueberry plantings in the Pacific Northwest: Blueberry mosaic virus, Blueberry red ringspot virus, Blueberry scorch virus, Blueberry shock virus, Tobacco ringspot virus, and Tomato ringspot virus. Six other virus and virus-like diseases of highbush b...

  4. [Viruses as biological weapons].

    PubMed

    Akçali, Alper

    2005-07-01

    The destruction made by nuclear, biological and chemical weapons used by governments and terrorist groups in the near history is posing anxiety and fear for human being. Rumour about the possible use of these agents leads to the development of serious negative effects on populations. Since there are no vaccine and therapy for most viral agents and cost of production as biological weapons is low, interest rate is rising for viruses. In this review, general characteristics, diagnosis, therapy and protective measures for viral agents such as variola virus, hemorrhagic fever viruses, encephalitis viruses, Hantaviruses and Nipah viruses, those can be used as biological weapon, have been summarized.

  5. Viruses and marine pollution.

    PubMed

    Danovaro, R; Armeni, M; Corinaldesi, C; Mei, M L

    2003-03-01

    This short review summarises the present knowledge on pollutant impacts on marine viruses, virus-host systems and their potential ecological implications. Excess nutrients from sewage and river effluents are a primary cause of marine eutrophication and mucilage formation, often related to the development of large viral assemblages. At the same time, hydrocarbons, polychlorinated biphenyl and pesticides alter ecosystem functioning and can determinate changes in the virus-host interactions, thus increasing the potential of viral infection. All these pollutants might have synergistic effects on the virus-host system and are able to induce prophage, thus increasing the impact of viruses on marine ecosystems.

  6. Viruses of asparagus.

    PubMed

    Tomassoli, Laura; Tiberini, Antonio; Vetten, Heinrich-Josef

    2012-01-01

    The current knowledge on viruses infecting asparagus (Asparagus officinalis) is reviewed. Over half a century, nine virus species belonging to the genera Ilarvirus, Cucumovirus, Nepovirus, Tobamovirus, Potexvirus, and Potyvirus have been found in this crop. The potyvirus Asparagus virus 1 (AV1) and the ilarvirus Asparagus virus 2 (AV2) are widespread and negatively affect the economic life of asparagus crops reducing yield and increasing the susceptibility to biotic and abiotic stress. The main properties and epidemiology of AV1 and AV2 as well as diagnostic techniques for their detection and identification are described. Minor viruses and control are briefly outlined. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Equine influenza virus.

    PubMed

    Landolt, Gabriele A

    2014-12-01

    For decades the horse has been viewed as an isolated or "dead-end" host for influenza A viruses, with equine influenza virus being considered as relatively stable genetically. Although equine influenza viruses are genetically more stable than those of human lineage, they are by no means in evolutionary stasis. Moreover, recent transmission of equine-lineage influenza viruses to dogs also challenges the horse's status as a dead-end host. This article reviews recent developments in the epidemiology and evolution of equine influenza virus. In addition, the clinical presentation of equine influenza infection, diagnostic techniques, and vaccine recommendations are briefly summarized.

  8. Avian influenza virus.

    PubMed

    Lee, Chang-Won; Saif, Yehia M

    2009-07-01

    Avian influenza viruses do not typically replicate efficiently in humans, indicating direct transmission of avian influenza virus to humans is unlikely. However, since 1997, several cases of human infections with different subtypes (H5N1, H7N7, and H9N2) of avian influenza viruses have been identified and raised the pandemic potential of avian influenza virus in humans. Although circumstantial evidence of human to human transmission exists, the novel avian-origin influenza viruses isolated from humans lack the ability to transmit efficiently from person-to-person. However, the on-going human infection with avian-origin H5N1 viruses increases the likelihood of the generation of human-adapted avian influenza virus with pandemic potential. Thus, a better understanding of the biological and genetic basis of host restriction of influenza viruses is a critical factor in determining whether the introduction of a novel influenza virus into the human population will result in a pandemic. In this article, we review current knowledge of type A influenza virus in which all avian influenza viruses are categorized.

  9. Virus Evolution Serial

    SciTech Connect

    Vassilevska, Tanya

    2015-01-05

    This is the first code, designed to run on a desktop, which models the intracellular replication and the cell-to-cell infection and demonstrates virus evolution at the molecular level. This code simulates the infection of a population of "idealized biological cells" (represented as objects that do not divide or have metabolism) with "virus" (represented by its genetic sequence), the replication and simultaneous mutation of the virus which leads to evolution of the population of genetically diverse viruses. The code is built to simulate single-stranded RNA viruses. The input for the code is 1. the number of biological cells in the culture, 2. the initial composition of the virus population, 3. the reference genome of the RNA virus, 4. the coordinates of the genome regions and their significance and, 5. parameters determining the dynamics of virus replication, such as the mutation rate. The simulation ends when all cells have been infected or when no more infections occurs after a given number of attempts. The code has the ability to simulate the evolution of the virus in serial passage of cell "cultures", i.e. after the end of a simulation, a new one is immediately scheduled with a new culture of infected cells. The code outputs characteristics of the resulting virus population dynamics and genetic composition of the virus population, such as the top dominant genomes, percentage of a genome with specific characteristics.

  10. Serodiagnosis for Tumor Viruses

    PubMed Central

    Morrison, Brian J.; Labo, Nazzarena; Miley, Wendell J.; Whitby, Denise

    2015-01-01

    The known human tumor viruses include the DNA viruses Epstein-Barr virus, Kaposi sarcoma herpesvirus, Merkel cell polyomavirus, human papillomavirus, and hepatitis B virus. RNA tumor viruses include Human T-cell lymphotrophic virus type-1 and hepatitis C virus. The serological identification of antigens/antibodies in plasma serum is a rapidly progressing field with utility for both scientists and clinicians. Serology is useful for conducting seroepidemiology studies and to inform on the pathogenesis and host immune response to a particular viral agent. Clinically, serology is useful for diagnosing current or past infection and for aiding in clinical management decisions. Serology is useful for screening blood donations for infectious agents and for monitoring the outcome of vaccination against these viruses. Serodiagnosis of human tumor viruses has improved in recent years with increased specificity and sensitivity of the assays, as well as reductions in cost and the ability to assess multiple antibody/antigens in single assays. Serodiagnosis of tumor viruses plays an important role in our understanding of the prevalence and transmission of these viruses and ultimately in the ability to develop treatments/preventions for these globally important diseases. PMID:25843726

  11. Virus transmission via food.

    PubMed

    Cliver, D O

    1997-01-01

    Viruses are transmitted to humans via foods as a result of direct or indirect contamination of the foods with human faeces. Viruses transmitted by a faecal-oral route are not strongly dependent on foods as vehicles of transmission, but viruses are important among agents of foodborne disease. Vehicles are most often molluscs from contaminated waters, but many other foods are contaminated directly by infected persons. The viruses most often foodborne are the hepatitis A virus and the Norwalk-like gastroenteritis viruses. Detection methods for these viruses in foods are very difficult and costly; the methods are not routine. Indicators that would rapidly and reliably suggest the presence of viral contamination of foods are still being sought. Contamination can be prevented by keeping faeces out of food or by treating vehicles such as water in order to inactivate virus that might be carried to food in this way. Virus cannot multiply in food, but can usually be inactivated by adequate heating. Other methods of inactivating viruses within a food are relatively unreliable, but viruses in water and on exposed surfaces can be inactivated with ultraviolet light or with strong oxidizing agents.

  12. Virus, Oncolytic virus and Human Prostate Cancer.

    PubMed

    Liu, Guang Bin; Zhao, Liang; Zhang, Lifang; Zhao, Kong-Nan

    2016-12-15

    Prostate cancer (PCa), a disease, is characterized by abnormal cell growth in the prostate - a gland in the male reproductive system. PCa is one of the leading causes of cancer death among men of all races. Although older age and a family history of the disease have been recognized as the risk factors of PCa, the cause of this cancer remains unclear. Mounting evidence suggests that infections with various viruses are causally linked to PCa pathogenesis. Published studies have provided strong evidence that at least two viruses (RXMV and HPV) contribute to prostate tumourigenicity and impact on the survival of patients with malignant PCa. Traditional therapies including chemotherapy and radiotherapy are unable to distinguish cancer cells from normal cells, which are a significant drawback and leads to toxicities for PCa patients undergoing treatment. So far, few other options are available for treating patients with advanced PCa. Virotherapy is being developed to be a novel therapy for cancers, which uses oncotropic and oncolytic viruses with their abilities to find and destroy malignant cells in the body. For PCa treatment, oncolytic virotherapy appears to be much more attractive, which uses live viruses to selectively kill cancer cells. Oncolytic viruses can be genetically engineered to induce cancer cell lysis through virus replication and expression of cytotoxic proteins. As oncolytic viruses are a relatively new class of anti-cancer immunotherapy agents, several important barriers still exist on the road to the use of oncolytic viruses for PCa therapy. In this review, we first discuss the controversy of the contribution of virus infection to PCa, and subsequently summarize the development of oncolytic virotherapy for PCa in the past several years.

  13. [The great virus comeback].

    PubMed

    Forterre, Patrick

    2013-01-01

    Viruses have been considered for a long time as by-products of biological evolution. This view is changing now as a result of several recent discoveries. Viral ecologists have shown that viral particles are the most abundant biological entities on our planet, whereas metagenomic analyses have revealed an unexpected abundance and diversity of viral genes in the biosphere. Comparative genomics have highlighted the uniqueness of viral sequences, in contradiction with the traditional view of viruses as pickpockets of cellular genes. On the contrary, cellular genomes, especially eukaryotic ones, turned out to be full of genes derived from viruses or related elements (plasmids, transposons, retroelements and so on). The discovery of unusual viruses infecting archaea has shown that the viral world is much more diverse than previously thought, ruining the traditional dichotomy between bacteriophages and viruses. Finally, the discovery of giant viruses has blurred the traditional image of viruses as small entities. Furthermore, essential clues on virus history have been obtained in the last ten years. In particular, structural analyses of capsid proteins have uncovered deeply rooted homologies between viruses infecting different cellular domains, suggesting that viruses originated before the last universal common ancestor (LUCA). These studies have shown that several lineages of viruses originated independently, i.e., viruses are polyphyletic. From the time of LUCA, viruses have coevolved with their hosts, and viral lineages can be viewed as lianas wrapping around the trunk, branches and leaves of the tree of life. Although viruses are very diverse, with genomes encoding from one to more than one thousand proteins, they can all be simply defined as organisms producing virions. Virions themselves can be defined as infectious particles made of at least one protein associated with the viral nucleic acid, endowed with the capability to protect the viral genome and ensure its

  14. Lifestyles of plant viruses

    PubMed Central

    Roossinck, Marilyn J.

    2010-01-01

    The vast majority of well-characterized eukaryotic viruses are those that cause acute or chronic infections in humans and domestic plants and animals. However, asymptomatic persistent viruses have been described in animals, and are thought to be sources for emerging acute viruses. Although not previously described in these terms, there are also many viruses of plants that maintain a persistent lifestyle. They have been largely ignored because they do not generally cause disease. The persistent viruses in plants belong to the family Partitiviridae or the genus Endornavirus. These groups also have members that infect fungi. Phylogenetic analysis of the partitivirus RNA-dependent RNA polymerase genes suggests that these viruses have been transmitted between plants and fungi. Additional families of viruses traditionally thought to be fungal viruses are also found frequently in plants, and may represent a similar scenario of persistent lifestyles, and some acute or chronic viruses of crop plants may maintain a persistent lifestyle in wild plants. Persistent, chronic and acute lifestyles of plant viruses are contrasted from both a functional and evolutionary perspective, and the potential role of these lifestyles in host evolution is discussed. PMID:20478885

  15. Postmortem stability of Ebola virus.

    PubMed

    Prescott, Joseph; Bushmaker, Trenton; Fischer, Robert; Miazgowicz, Kerri; Judson, Seth; Munster, Vincent J

    2015-05-01

    The ongoing Ebola virus outbreak in West Africa has highlighted questions regarding stability of the virus and detection of RNA from corpses. We used Ebola virus-infected macaques to model humans who died of Ebola virus disease. Viable virus was isolated <7 days posteuthanasia; viral RNA was detectable for 10 weeks.

  16. RNA Viruses Infecting Pest Insects

    USDA-ARS?s Scientific Manuscript database

    RNA viruses are viruses whose genetic material is ribonucleic acid (RNA). RNA viruses may be double or single-stranded based on the type of RNA they contain. Single-stranded RNA viruses can be further grouped into negative sense or positive-sense viruses according to the polarity of their RNA. Fur...

  17. Other Viruses and Viruslike Agents

    USDA-ARS?s Scientific Manuscript database

    The diseases reported under 'Virus and Virus-like Agents' in the first volume of this compendium, with the exception of Cherry rasp leaf virus and Rubus chinese seed-borne virus, should be considered oddities since there are no known type isolates available for these reported viruses. Without a po...

  18. Viruses within animal genomes.

    PubMed

    De Brognier, A; Willems, L

    2016-04-01

    Viruses and their hosts can co-evolve to reach a fragile equilibrium that allows the survival of both. An excess of pathogenicity in the absence of a reservoir would be detrimental to virus survival. A significant proportion of all animal genomes has been shaped by the insertion of viruses that subsequently became 'fossilised'. Most endogenous viruses have lost the capacity to replicate via an infectious cycle and now replicate passively. The insertion of endogenous viruses has contributed to the evolution of animal genomes, for example in the reproductive biology of mammals. However, spontaneous viral integration still occasionally occurs in a number of virus-host systems. This constitutes a potential risk to host survival but also provides an opportunity for diversification and evolution.

  19. Water system virus detection

    NASA Technical Reports Server (NTRS)

    Fraser, A. S.; Wells, A. F.; Tenoso, H. J. (Inventor)

    1978-01-01

    The performance of a waste water reclamation system is monitored by introducing a non-pathogenic marker virus, bacteriophage F2, into the waste-water prior to treatment and, thereafter, testing the reclaimed water for the presence of the marker virus. A test sample is first concentrated by absorbing any marker virus onto a cellulose acetate filter in the presence of a trivalent cation at low pH and then flushing the filter with a limited quantity of a glycine buffer solution to desorb any marker virus present on the filter. Photo-optical detection of indirect passive immune agglutination by polystyrene beads indicates the performance of the water reclamation system in removing the marker virus. A closed system provides for concentrating any marker virus, initiating and monitoring the passive immune agglutination reaction, and then flushing the system to prepare for another sample.

  20. DNA Virus Replication Compartments

    PubMed Central

    Schmid, Melanie; Speiseder, Thomas; Dobner, Thomas

    2014-01-01

    Viruses employ a variety of strategies to usurp and control cellular activities through the orchestrated recruitment of macromolecules to specific cytoplasmic or nuclear compartments. Formation of such specialized virus-induced cellular microenvironments, which have been termed viroplasms, virus factories, or virus replication centers, complexes, or compartments, depends on molecular interactions between viral and cellular factors that participate in viral genome expression and replication and are in some cases associated with sites of virion assembly. These virus-induced compartments function not only to recruit and concentrate factors required for essential steps of the viral replication cycle but also to control the cellular mechanisms of antiviral defense. In this review, we summarize characteristic features of viral replication compartments from different virus families and discuss similarities in the viral and cellular activities that are associated with their assembly and the functions they facilitate for viral replication. PMID:24257611

  1. Mechanical properties of viruses.

    PubMed

    de Pablo, Pedro J; Mateu, Mauricio G

    2013-01-01

    Structural biology techniques have greatly contributed to unveil the relationships between structure, properties and functions of viruses. In recent years, classic structural approaches are being complemented by single-molecule techniques such as atomic force microscopy and optical tweezers to study physical properties and functions of viral particles that are not accessible to classic structural techniques. Among these features are mechanical properties such as stiffness, intrinsic elasticity, tensile strength and material fatigue. The field of virus mechanics is contributing to materials science by investigating some physical parameters of "soft" biological matter and biological nano-objects. Virus mechanics studies are also starting to unveil the biological implications of physical properties of viruses. Growing evidence indicate that viruses are subjected to internal and external forces, and that they may have adapted to withstand and even use those forces. This chapter describes what is known on the mechanical properties of virus particles, their structural determinants, and possible biological implications, of which several examples are provided.

  2. Constructing computer virus phylogenies

    SciTech Connect

    Goldberg, L.A.; Goldberg, P.W.; Phillips, C.A.; Sorkin, G.B.

    1996-03-01

    There has been much recent algorithmic work on the problem of reconstructing the evolutionary history of biological species. Computer virus specialists are interested in finding the evolutionary history of computer viruses--a virus is often written using code fragments from one or more other viruses, which are its immediate ancestors. A phylogeny for a collection of computer viruses is a directed acyclic graph whose nodes are the viruses and whose edges map ancestors to descendants and satisfy the property that each code fragment is ``invented`` only once. To provide a simple explanation for the data, we consider the problem of constructing such a phylogeny with a minimal number of edges. In general, this optimization problem cannot be solved in quasi-polynomial time unless NQP=QP; we present positive and negative results for associated approximated problems. When tree solutions exist, they can be constructed and randomly sampled in polynomial time.

  3. Filamentous Influenza Viruses

    PubMed Central

    Badham, Matthew D.; Rossman, Jeremy S.

    2016-01-01

    Influenza A virus is a pathogen of global medical importance causing significant health and socio-economic costs every year. Influenza virus is an unusual pathogen in that it is pleomorphic, capable of forming virions ranging in shape from spherical to filamentous. Despite decades of research on the influenza virus, much remains unknown about the formation of filamentous influenza viruses and their role in the viral replication cycle. Here, we discuss what is known about influenza virus assembly and budding, focusing on the viral and host factors that are involved in the determination of viral morphology. Whilst the biological function of the filamentous morphology remains unknown, recent results suggest a role in facilitating viral spread in vivo. We discuss these results and speculate on the consequences of viral morphology during influenza virus infection of the human respiratory tract. PMID:28042529

  4. Viruses of lower vertebrates.

    PubMed

    Essbauer, S; Ahne, W

    2001-08-01

    Viruses of lower vertebrates recently became a field of interest to the public due to increasing epizootics and economic losses of poikilothermic animals. These were reported worldwide from both wildlife and collections of aquatic poikilothermic animals. Several RNA and DNA viruses infecting fish, amphibians and reptiles have been studied intensively during the last 20 years. Many of these viruses induce diseases resulting in important economic losses of lower vertebrates, especially in fish aquaculture. In addition, some of the DNA viruses seem to be emerging pathogens involved in the worldwide decline in wildlife. Irido-, herpes- and polyomavirus infections may be involved in the reduction in the numbers of endangered amphibian and reptile species. In this context the knowledge of several important RNA viruses such as orthomyxo-, paramyxo-, rhabdo-, retro-, corona-, calici-, toga-, picorna-, noda-, reo- and birnaviruses, and DNA viruses such as parvo-, irido-, herpes-, adeno-, polyoma- and poxviruses, is described in this review.

  5. Honey bee viruses.

    PubMed

    Chen, Yan Ping; Siede, Reinhold

    2007-01-01

    Viruses are significant threats to the health and well-being of the honey bee, Apis mellifera. To alleviate the threats posed by these invasive organisms, a better understanding of bee viral infections will be of crucial importance in developing effective and environmentally benign disease control strategies. Although knowledge of honey bee viruses has been accumulated considerably in the past three decades, a comprehensive review to compile the various aspects of bee viruses at the molecular level has not been reported. This chapter summarizes recent progress in the understanding of the morphology, genome organization, transmission, epidemiology, and pathogenesis of honey bee viruses as well as their interactions with their honey bee hosts. The future prospects of research of honey bee viruses are also discussed in detail. The chapter has been designed to provide researchers in the field with updated information about honey bee viruses and to serve as a starting point for future research.

  6. Viruses in Antarctic lakes

    NASA Technical Reports Server (NTRS)

    Kepner, R. L. Jr; Wharton, R. A. Jr; Suttle, C. A.; Wharton RA, J. r. (Principal Investigator)

    1998-01-01

    Water samples collected from four perennially ice-covered Antarctic lakes during the austral summer of 1996-1997 contained high densities of extracellular viruses. Many of these viruses were found to be morphologically similar to double-stranded DNA viruses that are known to infect algae and protozoa. These constitute the first observations of viruses in perennially ice-covered polar lakes. The abundance of planktonic viruses and data suggesting substantial production potential (relative to bacteria] secondary and photosynthetic primary production) indicate that viral lysis may be a major factor in the regulation of microbial populations in these extreme environments. Furthermore, we suggest that Antarctic lakes may be a reservoir of previously undescribed viruses that possess novel biological and biochemical characteristics.

  7. Viruses in Antarctic lakes

    NASA Technical Reports Server (NTRS)

    Kepner, R. L. Jr; Wharton, R. A. Jr; Suttle, C. A.; Wharton RA, J. r. (Principal Investigator)

    1998-01-01

    Water samples collected from four perennially ice-covered Antarctic lakes during the austral summer of 1996-1997 contained high densities of extracellular viruses. Many of these viruses were found to be morphologically similar to double-stranded DNA viruses that are known to infect algae and protozoa. These constitute the first observations of viruses in perennially ice-covered polar lakes. The abundance of planktonic viruses and data suggesting substantial production potential (relative to bacteria] secondary and photosynthetic primary production) indicate that viral lysis may be a major factor in the regulation of microbial populations in these extreme environments. Furthermore, we suggest that Antarctic lakes may be a reservoir of previously undescribed viruses that possess novel biological and biochemical characteristics.

  8. Zika virus - an overview.

    PubMed

    Zanluca, Camila; Dos Santos, Claudia Nunes Duarte

    2016-05-01

    Zika virus (ZIKV) is currently one of the most important emerging viruses in the world. Recently, it has caused outbreaks and epidemics, and has been associated with severe clinical manifestations and congenital malformations. However to date, little is known about the pathogenicity of the virus and the consequences of ZIKV infection. In this paper, we provide an overview of the current knowledge on ZIKV. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  9. Zika virus infection.

    PubMed

    Pougnet, Laurence; Thill, Chloé; Pougnet, Richard; Auvinet, Henri; Giacardi, Christophe; Drouillard, Isabelle

    2016-12-01

    A 21-year old woman from New-Caledonia had 40 ̊C fever with vomiting, arthralgia, myalgia, and measles-like rash. Etiological analyses showed primary infection with Zika virus. Because of severe clinical presentation, she was hospitalized in the intensive care unit of the Brest military Hospital. Zika virus is mainly transmitted by Aedes mosquitoes. If they settle in Metropolitan France, Zika virus might also spread there.

  10. Thermal Inactivation of Viruses

    DTIC Science & Technology

    1977-10-01

    Hammon. 1966. Studies on Japanese B encephalitis virus vaccines from tissue culture. VI. Development of a hamster kidney tissue culture inactivated... tissue culture passage, storage, temperature and drying on viability of SE polyoma virus. Exper. Biol. and Hed. Proc. of the Soc. for Exper. Biol...studies of heated tissue suspensions containing foot- and-mouth disease virus. Amer. J. Vet. Res. 20:510-521. Dupre’, M. V., and M. Frobisher. 1966

  11. The human oncogenic viruses

    SciTech Connect

    Luderer, A.A.; Weetall, H.H

    1986-01-01

    This book contains eight selections. The titles are: Cytogenetics of the Leukemias and Lymphomas; Cytogenetics of Solid Tumors: Renal Cell Carcinoma, Malignant Melanoma, Retinoblastoma, and Wilms' Tumor; Elucidation of a Normal Function for a Human Proto-Oncogene; Detection of HSV-2 Genes and Gene Products in Cervical Neoplasia; Papillomaviruses in Anogennital Neoplasms; Human Epstein-Barr Virus and Cancer; Hepatitis B Virus and Hepatocellular Carcinoma; and Kaposi's Sarcoma: Acquired Immunodeficiency Syndrome (AIDS) and Associated Viruses.

  12. Broadband Respiratory Virus Surveillance

    DTIC Science & Technology

    2011-10-01

    HSV – Herpes Simplex Virus LOD – Limit of Detection PCR – Polymerase Chain Reaction PIV – Parainfluenza viruses 37 PRMS – Pacific Regional Medical...the RVS assay was determined by testing 109 pre-characterized samples collected at TAMC. This included 20 adenovirus, 20 RSV, 20 PIV, 19 Herpes ... Simplex Virus (HSV) and 19 Enterovirus 7 positive as well as 11 HSV negative specimens as determined by the TAMC Department of Pathology’s current gold

  13. Water system virus detection

    NASA Technical Reports Server (NTRS)

    Fraser, A. S.; Wells, A. F.; Tenoso, H. J.

    1975-01-01

    A monitoring system developed to test the capability of a water recovery system to reject the passage of viruses into the recovered water is described. A nonpathogenic marker virus, bacteriophage F2, is fed into the process stream before the recovery unit and the reclaimed water is assayed for its presence. Detection of the marker virus consists of two major components, concentration and isolation of the marker virus, and detection of the marker virus. The concentration system involves adsorption of virus to cellulose acetate filters in the presence of trivalent cations and low pH with subsequent desorption of the virus using volumes of high pH buffer. The detection of the virus is performed by a passive immune agglutination test utilizing specially prepared polystyrene particles. An engineering preliminary design was performed as a parallel effort to the laboratory development of the marker virus test system. Engineering schematics and drawings of a fully functional laboratory prototype capable of zero-G operation are presented. The instrument consists of reagent pump/metering system, reagent storage containers, a filter concentrator, an incubation/detector system, and an electronic readout and control system.

  14. Viruses infecting marine molluscs.

    PubMed

    Arzul, Isabelle; Corbeil, Serge; Morga, Benjamin; Renault, Tristan

    2017-02-09

    Although a wide range of viruses have been reported in marine molluscs, most of these reports rely on ultrastructural examination and few of these viruses have been fully characterized. The lack of marine mollusc cell lines restricts virus isolation capacities and subsequent characterization works. Our current knowledge is mostly restricted to viruses affecting farmed species such as oysters Crassostrea gigas, abalone Haliotis diversicolor supertexta or the scallop Chlamys farreri. Molecular approaches which are needed to identify virus affiliation have been carried out for a small number of viruses, most of them belonging to the Herpesviridae and birnaviridae families. These last years, the use of New Generation Sequencing approach has allowed increasing the number of sequenced viral genomes and has improved our capacity to investigate the diversity of viruses infecting marine molluscs. This new information has in turn allowed designing more efficient diagnostic tools. Moreover, the development of experimental infection protocols has answered some questions regarding the pathogenesis of these viruses and their interactions with their hosts. Control and management of viral diseases in molluscs mostly involve active surveillance, implementation of effective bio security measures and development of breeding programs. However factors triggering pathogen development and the life cycle and status of the viruses outside their mollusc hosts still need further investigations.

  15. Rabies virus receptors.

    PubMed

    Lafon, Monique

    2005-02-01

    There is convincing in vitro evidence that the muscular form of the nicotinic acetylcholine receptor (nAChR), the neuronal cell adhesion molecule (NCAM), and the p75 neurotrophin receptor (p75NTR) bind rabies virus and/or facilitate rabies virus entry into cells. Other components of the cell membrane, such as gangliosides, may also participate in the entry of rabies virus. However, little is known of the role of these molecules in vivo. This review proposes a speculative model that accounts for the role of these different molecules in entry and trafficking of rabies virus into the nervous system.

  16. Viruses in artichoke.

    PubMed

    Gallitelli, Donato; Mascia, Tiziana; Martelli, Giovanni P

    2012-01-01

    Most of the 25 viruses found in globe artichoke (Cynara scolymus L.) and cardoon (Cynara cardunculus L.) were recorded from Europe and the Mediterranean basin, where they decrease both the productivity and the quality of the crop. Although, sometimes, these viruses are agents of diseases of different severity, most often their infections are symptomless. These conditions have contributed to spread virus-infected material since farmers multiply traditional artichoke types vegetatively with no effective selection of virus-free plants. This review reports the main properties of these viruses and the techniques used for their detection and identification. ELISA kits are commercially available for most of the viruses addressed in this review but have seldom been used for their detection in artichoke. Conversely, nucleic acid-based diagnostic reagents, some of which are commercially available, have successfully been employed to identify some viruses in artichoke sap. Control measures mainly use virus-free stocks for new plantations. A combined procedure of meristem-tip culture and thermotherapy proved useful for producing virus-free regenerants of the reflowering southern Italian cultivar Brindisino, which kept earliness and typical heads shape. Copyright © 2012 Elsevier Inc. All rights reserved.

  17. Chikungunya Virus, Southeastern France

    PubMed Central

    Caro, Valérie; Plumet, Sébastien; Thiberge, Jean-Michel; Souarès, Yvan; Failloux, Anna-Bella; Tolou, Hugues J.; Budelot, Michel; Cosserat, Didier; Leparc-Goffart, Isabelle; Desprès, Philippe

    2011-01-01

    In September 2010, autochthonous transmission of chikungunya virus was recorded in southeastern France, where the Aedes albopictus mosquito vector is present. Sequence analysis of the viral genomes of imported and autochthonous isolates indicated new features for the potential emergence and spread of the virus in Europe. PMID:21529410

  18. Viruses in reptiles.

    PubMed

    Ariel, Ellen

    2011-09-21

    The etiology of reptilian viral diseases can be attributed to a wide range of viruses occurring across different genera and families. Thirty to forty years ago, studies of viruses in reptiles focused mainly on the zoonotic potential of arboviruses in reptiles and much effort went into surveys and challenge trials of a range of reptiles with eastern and western equine encephalitis as well as Japanese encephalitis viruses. In the past decade, outbreaks of infection with West Nile virus in human populations and in farmed alligators in the USA has seen the research emphasis placed on the issue of reptiles, particularly crocodiles and alligators, being susceptible to, and reservoirs for, this serious zoonotic disease. Although there are many recognised reptilian viruses, the evidence for those being primary pathogens is relatively limited. Transmission studies establishing pathogenicity and cofactors are likewise scarce, possibly due to the relatively low commercial importance of reptiles, difficulties with the availability of animals and permits for statistically sound experiments, difficulties with housing of reptiles in an experimental setting or the inability to propagate some viruses in cell culture to sufficient titres for transmission studies. Viruses as causes of direct loss of threatened species, such as the chelonid fibropapilloma associated herpesvirus and ranaviruses in farmed and wild tortoises and turtles, have re-focused attention back to the characterisation of the viruses as well as diagnosis and pathogenesis in the host itself.

  19. West Nile Virus

    MedlinePlus

    ... spread by mosquitoes. Mosquitoes become infected by biting birds that carry the virus. People can get West Nile virus when an infected mosquito bites them. This happens most often in the warm-weather months of spring, summer and early fall. You ...

  20. Blueberry virus A

    USDA-ARS?s Scientific Manuscript database

    Leaf yellowing on highbush blueberry ‘Spartan’ prompted Isogai et al. to investigate whether a virus was the causal agent of the disorder. After double-stranded RNA extraction from symptomatic material they identified a single band of 17 Kb, indicative of virus infection. Shotgun cloning and sequenc...

  1. Diseases caused by viruses

    USDA-ARS?s Scientific Manuscript database

    Corn viruses are important disease agents worldwide that can be difficult to identify and diagnose. Previously undescribed viruses of corn also emerge periodically and their distributions and importance changes over time. The Compendium of Corn Diseases is a valuable tool for pre-diagnosis of diseas...

  2. Positive reinforcement for viruses.

    PubMed

    Vigant, Frederic; Jung, Michael; Lee, Benhur

    2010-10-29

    Virus-cell membrane fusion requires a critical transition from positive to negative membrane curvature. St. Vincent et al. (2010), in PNAS, designed a class of antivirals that targets this transition. These rigid amphipathic fusion inhibitors are active against an array of enveloped viruses.

  3. Rift Valley Fever Virus

    USDA-ARS?s Scientific Manuscript database

    Rift Valley fever virus (RVFV) is a mosquito-transmitted virus or arbovirus that is endemic in sub-Saharan Africa. In the last decade, Rift Valley fever (RVF) outbreaks have resulted in loss of human and animal life, as well as had significant economic impact. The disease in livestock is primarily a...

  4. Equine Arteritis Virus

    USDA-ARS?s Scientific Manuscript database

    03. Nidovirales : 03.004. Arteriviridae : 03.004.0. {03.004.0. unknown} : 03.004.0.01. Arterivirus : 03.004.0.01.001. Equine arteritis virus will be published online. The article details the phenotypic and genotypic makeup of equine arteritis virus (EAV), and summarizes its biological properties....

  5. Usutu Virus, Italy, 1996

    PubMed Central

    Bakonyi, Tamás; Rossi, Giacomo; Mani, Paolo; Nowotny, Norbert

    2013-01-01

    Retrospective analysis of archived tissue samples from bird deaths in the Tuscany region of Italy in 1996 identified Usutu virus. Partial sequencing confirmed identity with the 2001 Vienna strain and provided evidence for a much earlier introduction of this virus into Europe than previously assumed. PMID:23347844

  6. Avian influenza virus

    USDA-ARS?s Scientific Manuscript database

    Avian influenza virus (AIV) is type A influenza that is adapted to avian host species. Although the virus can be isolated from numerous avian species, the natural host reservoir species are dabbling ducks, shorebirds and gulls. Domestic poultry species (poultry being defined as birds that are rais...

  7. Zika Virus Disease.

    PubMed

    Slenczka, Werner

    2016-06-01

    The history of Zika virus disease serves as a paradigm of a typical emerging viral infection. Zika virus disease, a mosquito-borne flavivirus, was first isolated in 1947 in the Zika forest of Uganda. The same virus was also isolated from jungle-dwelling mosquitoes (Aedes [Stegomyia] africanus). In many areas of Africa and South Asia human infections with Zika virus were detected by both serology and virus isolation. About 80% of infections are asymptomatic, and in 20% a mostly mild disease with fever, rash, arthralgia, and conjunctivitis may occur. Fetal infections with malformations were not recorded in Africa or Asia. Zika virus was imported to northern Brazil possibly during the world soccer championship that was hosted by Brazil in June through July 2014. A cluster of severe fetal malformations with microcephaly and ocular defects was noted in 2015 in the northeast of Brazil, and intrauterine infections with Zika virus were confirmed. The dramatic change in Zika virus pathogenicity upon its introduction to Brazil has remained an enigma.

  8. Tobacco ringspot virus

    USDA-ARS?s Scientific Manuscript database

    Tobacco ringspot virus (TRSV), and its vector, the dagger nematodes (Xiphinema americanum and related species) are widely distributed throughout the world. Cucumber, melon, and watermelon are particularly affected by TRSV. Symptoms can vary with plant age, the strain of the virus, and environment...

  9. Papaya ringspot virus (Potyviridae)

    USDA-ARS?s Scientific Manuscript database

    Papaya ringspot virus, a member of the family Potyviridae, is single stranded RNA plant virus with a monocistronic genome of about 10,326 nucleotides that is expressed via a large polyprotein subsequently cleaved into functional proteins. It causes severe damage on cucurbit crops such as squash and...

  10. Virus separation using membranes.

    PubMed

    Grein, Tanja A; Michalsky, Ronald; Czermak, Peter

    2014-01-01

    Industrial manufacturing of cell culture-derived viruses or virus-like particles for gene therapy or vaccine production are complex multistep processes. In addition to the bioreactor, such processes require a multitude of downstream unit operations for product separation, concentration, or purification. Similarly, before a biopharmaceutical product can enter the market, removal or inactivation of potential viral contamination has to be demonstrated. Given the complexity of biological solutions and the high standards on composition and purity of biopharmaceuticals, downstream processing is the bottleneck in many biotechnological production trains. Membrane-based filtration can be an economically attractive and efficient technology for virus separation. Viral clearance, for instance, of up to seven orders of magnitude has been reported for state of the art polymeric membranes under best conditions.This chapter summarizes the fundamentals of virus ultrafiltration, diafiltration, or purification with adsorptive membranes. In lieu of an impractical universally applicable protocol for virus filtration, application of these principles is demonstrated with two examples. The chapter provides detailed methods for production, concentration, purification, and removal of a rod-shaped baculovirus (Autographa californica M nucleopolyhedrovirus, about 40 × 300 nm in size, a potential vector for gene therapy, and an industrially important protein expression system) or a spherical parvovirus (minute virus of mice, 22-26 nm in size, a model virus for virus clearance validation studies).

  11. Papaya Ringspot Virus

    USDA-ARS?s Scientific Manuscript database

    The term papaya ringspot virus (PRSV) was coined by Jensen in 1949, to describe a papaya disease in Hawaii. Later work showed that diseases such as papaya mosaic and watermelon mosaic virus-1 were caused by PRSV. The primary host range of PRSV is papaya and cucurbits, with Chenopium amaranticolor ...

  12. Zika Virus and Pregnancy.

    PubMed

    Stagg, Denise; Hurst, Helen M

    2016-01-01

    Recent outbreaks of Zika virus and reports linking infection in pregnant women with microcephaly in newborns have caused concern worldwide. Information has been evolving rapidly. Nurses and other clinicians, especially those who work with women of childbearing age, play a pivotal role in disseminating accurate information and identifying potential cases of Zika virus infection.

  13. Lethal mutagenesis of viruses.

    PubMed

    Perales, Celia; Martín, Verónica; Domingo, Esteban

    2011-11-01

    Lethal mutagenesis aims at extinguishing viruses by increased mutagenesis prompted by virus-specific mutagenic agents, mainly nucleoside analogues. It is derived from the error threshold relationship of quasispecies theory, and it is slowly finding its way towards a clinical application. We summarize the current situation of research in this field of antiviral therapy. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Virus excretion in smallpox

    PubMed Central

    Sarkar, J. K.; Mitra, A. C.; Mukherjee, M. K.; De, S. K.

    1973-01-01

    Throat swabs of 34 of 328 family contacts of 52 smallpox cases, examined 4-8 days after the onset of the disease in the family, were positive for variola virus. The log titre of virus per swab ranged from 2 to 3.95. A higher proportion of unvaccinated than of vaccinated contacts excreted the virus. Only 4 of the virus-positive contacts developed clinical smallpox; this occurred 5-7 days after their swabs were examined. Excretion of virus in the throats of these contacts, a few of whom were in the incubation period of the disease, suggests the possibility that they could have spread the infection. This possibility, if kept in mind, may help in tracing the source of infection or in determining the incubation period in a few instances when difficulty is experienced. PMID:4359679

  15. Hepatitis B Virus Biology

    PubMed Central

    Seeger, Christoph; Mason, William S.

    2000-01-01

    Hepadnaviruses (hepatitis B viruses) cause transient and chronic infections of the liver. Transient infections run a course of several months, and chronic infections are often lifelong. Chronic infections can lead to liver failure with cirrhosis and hepatocellular carcinoma. The replication strategy of these viruses has been described in great detail, but virus-host interactions leading to acute and chronic disease are still poorly understood. Studies on how the virus evades the immune response to cause prolonged transient infections with high-titer viremia and lifelong infections with an ongoing inflammation of the liver are still at an early stage, and the role of the virus in liver cancer is still elusive. The state of knowledge in this very active field is therefore reviewed with an emphasis on past accomplishments as well as goals for the future. PMID:10704474

  16. Electron tomography of viruses.

    PubMed

    Subramaniam, Sriram; Bartesaghi, Alberto; Liu, Jun; Bennett, Adam E; Sougrat, Rachid

    2007-10-01

    Understanding the molecular architectures of enveloped and complex viruses is a challenging frontier in structural biology. In these viruses, the structural and compositional variation from one viral particle to another generally precludes the use of either crystallization or image averaging procedures that have been successfully implemented in the past for highly symmetric viruses. While advances in cryo electron tomography of unstained specimens provide new opportunities for identification and molecular averaging of individual subcomponents such as the surface glycoprotein spikes on purified viruses, electron tomography of stained and plunge-frozen cells is being used to visualize the cellular context of viral entry and replication. Here, we review recent developments in both areas as they relate to our understanding of the biology of heterogeneous and pleiomorphic viruses.

  17. Mayaro virus proteins.

    PubMed

    Mezencio, J M; Rebello, M A

    1993-01-01

    Mayaro virus was grown in BHK-21 cells and purified by centrifugation in a potassium-tartrate gradient (5-50%). The electron microscopy analyses of the purified virus showed an homogeneous population of enveloped particles with 69 +/- 2.3 nm in diameter. Three structural virus proteins were identified and designated p1, p2 and p3. Their average molecular weight were p1, 54 KDa; p2, 50 KDa and p3, 34 KDa. In Mayaro virus infected Aedes albopictus cells and in BHK-21 infected cells we detected six viral proteins, in which three of them are the structural virus proteins and the other three were products from processing of precursors of viral proteins, whose molecular weights are 62 KDa, 64 KDa and 110 KDa. The 34 KDa protein was the first viral protein synthesized at 5 hours post-infection in both cell lines studied.

  18. Infectious Viral Quantification of Chikungunya Virus-Virus Plaque Assay.

    PubMed

    Kaur, Parveen; Lee, Regina Ching Hua; Chu, Justin Jang Hann

    2016-01-01

    The plaque assay is an essential method for quantification of infectious virus titer. Cells infected with virus particles are overlaid with a viscous substrate. A suitable incubation period results in the formation of plaques, which can be fixed and stained for visualization. Here, we describe a method for measuring Chikungunya virus (CHIKV) titers via virus plaque assays.

  19. The contribution of molecular epidemiology to the understanding and control of viral diseases of salmonid aquaculture.

    PubMed

    Snow, Michael

    2011-04-05

    Molecular epidemiology is a science which utilizes molecular biology to define the distribution of disease in a population (descriptive epidemiology) and relies heavily on integration of traditional (or analytical) epidemiological approaches to identify the etiological determinants of this distribution. The study of viral pathogens of aquaculture has provided many exciting opportunities to apply such tools. This review considers the extent to which molecular epidemiological studies have contributed to better understanding and control of disease in aquaculture, drawing on examples of viral diseases of salmonid fish of commercial significance including viral haemorrhagic septicaemia virus (VHSV), salmonid alphavirus (SAV) and infectious salmon anaemia virus (ISAV). Significant outcomes of molecular epidemiological studies include:Improved taxonomic classification of viruses. A better understanding of the natural distribution of viruses. An improved understanding of the origins of viral pathogens in aquaculture. An improved understanding of the risks of translocation of pathogens outwith their natural host range. An increased ability to trace the source of new disease outbreaks. Development of a basis for ensuring development of appropriate diagnostic tools. An ability to classify isolates and thus target future research aimed at better understanding biological function. While molecular epidemiological studies have no doubt already made a significant contribution in these areas, the advent of new technologies such as pyrosequencing heralds a quantum leap in the ability to generate descriptive molecular sequence data. The ability of molecular epidemiology to fulfil its potential to translate complex disease pathways into relevant fish health policy is thus unlikely to be limited by the generation of descriptive molecular markers. More likely, full realisation of the potential to better explain viral transmission pathways will be dependent on the ability to assimilate

  20. Virus discovery and recent insights into virus diversity in arthropods.

    PubMed

    Junglen, Sandra; Drosten, Christian

    2013-08-01

    Recent studies on virus discovery have focused mainly on mammalian and avian viruses. Arbovirology with its long tradition of ecologically oriented investigation is now catching up, with important novel insights into the diversity of arthropod-associated viruses. Recent discoveries include taxonomically outlying viruses within the families Flaviviridae, Togaviridae, and Bunyaviridae, and even novel virus families within the order Nidovirales. However, the current focusing of studies on blood-feeding arthropods has restricted the range of arthropod hosts analyzed for viruses so far. Future investigations should include species from other arthropod taxa than Ixodita, Culicidae and Phlebotominae in order to shed light on the true diversity of arthropod viruses.

  1. Virus-PEDOT Biocomposite Films

    PubMed Central

    Donavan, Keith C.; Arter, Jessica A.

    2012-01-01

    Virus-poly(3,4-ethylenedioxythiophene) (virus-PEDOT) biocomposite films are prepared by electropolymerizing 3,4-ethylenedioxythiophene (EDOT) in aqueous electrolytes containing 12 mM LiClO4 and the bacteriophage M13. The concentration of virus in these solutions, [virus]soln, is varied from 3 nM to 15 nM. A quartz crystal microbalance is used to directly measure the total mass of the biocomposite film during its electrodeposition. In combination with a measurement of the electrodeposition charge, the mass of the virus incorporated into the film is calculated. These data show that concentration of the M13 within the electropolymerized film, [virus]film, increases linearly with [virus]soln. The incorporation of virus particles into the PEDOT film from solution is efficient, resulting in a concentration ratio: [virus]film:[virus]soln ≈450. Virus incorporation into the PEDOT causes roughening of the film topography that is observed using scanning electron microscopy and atomic force microscopy (AFM). The electrical conductivity of the virus-PEDOT film, measured perpendicular to the plane of the film using conductive tip AFM, decreases linearly with virus loading, from 270 μS/cm for pure PE-DOT films to 50 μS/cm for films containing 100 μM virus. The presence on the virus surface of displayed affinity peptides did not significantly influence the efficiency of incorporation into virus-PEDOT biocomposite films. PMID:22856875

  2. Realms of the Viruses Online

    ERIC Educational Resources Information Center

    Liu, Dennis

    2007-01-01

    Viruses have evolved strategies for infecting all taxa, but most viruses are highly specific about their cellular host. In humans, viruses cause diverse diseases, from chronic but benign warts, to acute and deadly hemorrhagic fever. Viruses have entertaining names like Zucchini Yellow Mosaic, Semliki Forest, Coxsackie, and the original terminator,…

  3. Computer Viruses: Pathology and Detection.

    ERIC Educational Resources Information Center

    Maxwell, John R.; Lamon, William E.

    1992-01-01

    Explains how computer viruses were originally created, how a computer can become infected by a virus, how viruses operate, symptoms that indicate a computer is infected, how to detect and remove viruses, and how to prevent a reinfection. A sidebar lists eight antivirus resources. (four references) (LRW)

  4. A Virus in Turbo Pascal.

    ERIC Educational Resources Information Center

    Teleky, Heidi Ann; And Others

    1993-01-01

    Addresses why the authors feel it is not inappropriate to teach about viruses in the how-to, hands-on fashion. Identifies the special features of Turbo Pascal that have to be used for the creation of an effective virus. Defines virus, derives its structure, and from this structure is derived the implemented virus. (PR)

  5. A Virus in Turbo Pascal.

    ERIC Educational Resources Information Center

    Teleky, Heidi Ann; And Others

    1993-01-01

    Addresses why the authors feel it is not inappropriate to teach about viruses in the how-to, hands-on fashion. Identifies the special features of Turbo Pascal that have to be used for the creation of an effective virus. Defines virus, derives its structure, and from this structure is derived the implemented virus. (PR)

  6. Realms of the Viruses Online

    ERIC Educational Resources Information Center

    Liu, Dennis

    2007-01-01

    Viruses have evolved strategies for infecting all taxa, but most viruses are highly specific about their cellular host. In humans, viruses cause diverse diseases, from chronic but benign warts, to acute and deadly hemorrhagic fever. Viruses have entertaining names like Zucchini Yellow Mosaic, Semliki Forest, Coxsackie, and the original terminator,…

  7. Computer Viruses: Pathology and Detection.

    ERIC Educational Resources Information Center

    Maxwell, John R.; Lamon, William E.

    1992-01-01

    Explains how computer viruses were originally created, how a computer can become infected by a virus, how viruses operate, symptoms that indicate a computer is infected, how to detect and remove viruses, and how to prevent a reinfection. A sidebar lists eight antivirus resources. (four references) (LRW)

  8. ICTV virus taxonomy profile: dicistroviridae

    USDA-ARS?s Scientific Manuscript database

    Dicistroviridae is a family of small non-enveloped viruses with RNA genomes of approximately 8-10 kilobases in length. All members infect arthropod hosts with some having devastating economic consequences, such as Acute bee paralysis virus, Kashmir bee virus, and Israeli acute paralysis virus towar...

  9. Enteric hepatitis viruses

    PubMed Central

    Tahaei, Seyed Mohammad Ebrahim; Zali, Mohammad Reza

    2012-01-01

    Hepatitis viruses are infectious agents that can infect liver and cause inflammation. The infection triggers immune response against infected cells that leads to the destruction of hepatic cells. This destruction has two consequences: leaking ALT and AST liver enzymes which increases during the course of disease and accumulation of bilirubin- a red pigmented compound released from dead red cells- which causes the yellow coloration of eyes and skin. These viruses transmit through diverse routes i.e. blood transfusion, sexual contacts and consuming water or food contaminated by feces. Enteric hepatitis viruses use the latter route for transmission; hence their outbreaks are more common in underdeveloped countries. There are currently two distinguished enteric hepatitis viruses, hepatitis A and hepatitis E. These viruses belong to different family of viruses and their epidemiological characteristics are different. These infections can be diagnosed by an ELISA for IgM antibody. A vaccine has been developed in last decade of twentieth century for hepatitis A virus, which is administered mostly in the developed world i.e. U.S and Japan. Treatment for these infections is mostly supportive; however, in the case of fulminant hepatitis the liver transplantation might be necessary. PMID:24834192

  10. Respiratory RNA Viruses.

    PubMed

    Hodinka, Richard L

    2016-08-01

    Acute upper and lower respiratory infections are a major public health problem and a leading cause of morbidity and mortality worldwide. At greatest risk are young children, the elderly, the chronically ill, and those with suppressed or compromised immune systems. Viruses are the predominant cause of respiratory tract illnesses and include RNA viruses such as respiratory syncytial virus, influenza virus, parainfluenza virus, metapneumovirus, rhinovirus, and coronavirus. Laboratory testing is required for a reliable diagnosis of viral respiratory infections, as a clinical diagnosis can be difficult since signs and symptoms are often overlapping and not specific for any one virus. Recent advances in technology have resulted in the development of newer diagnostic assays that offer great promise for rapid and accurate detection of respiratory viral infections. This chapter emphasizes the fundamental characteristics and clinical importance of the various RNA viruses that cause upper and lower respiratory tract diseases in the immunocompromised host. It highlights the laboratory methods that can be used to make a rapid and definitive diagnosis for the greatest impact on the care and management of ill patients, and the prevention and control of hospital-acquired infections and community outbreaks.

  11. Enteric hepatitis viruses.

    PubMed

    Tahaei, Seyed Mohammad Ebrahim; Mohebbi, Seyed Reza; Zali, Mohammad Reza

    2012-01-01

    Hepatitis viruses are infectious agents that can infect liver and cause inflammation. The infection triggers immune response against infected cells that leads to the destruction of hepatic cells. This destruction has two consequences: leaking ALT and AST liver enzymes which increases during the course of disease and accumulation of bilirubin- a red pigmented compound released from dead red cells- which causes the yellow coloration of eyes and skin. These viruses transmit through diverse routes i.e. blood transfusion, sexual contacts and consuming water or food contaminated by feces. Enteric hepatitis viruses use the latter route for transmission; hence their outbreaks are more common in underdeveloped countries. There are currently two distinguished enteric hepatitis viruses, hepatitis A and hepatitis E. These viruses belong to different family of viruses and their epidemiological characteristics are different. These infections can be diagnosed by an ELISA for IgM antibody. A vaccine has been developed in last decade of twentieth century for hepatitis A virus, which is administered mostly in the developed world i.e. U.S and Japan. Treatment for these infections is mostly supportive; however, in the case of fulminant hepatitis the liver transplantation might be necessary.

  12. Giant viruses infecting algae.

    PubMed

    Van Etten, J L; Meints, R H

    1999-01-01

    Paramecium bursaria chlorella virus (PBCV-1) is the prototype of a family of large, icosahedral, plaque-forming, double-stranded-DNA-containing viruses that replicate in certain unicellular, eukaryotic chlorella-like green algae. DNA sequence analysis of its 330, 742-bp genome leads to the prediction that this phycodnavirus has 376 protein-encoding genes and 10 transfer RNA genes. The predicted gene products of approximately 40% of these genes resemble proteins of known function. The chlorella viruses have other features that distinguish them from most viruses, in addition to their large genome size. These features include the following: (a) The viruses encode multiple DNA methyltransferases and DNA site-specific endonucleases; (b) PBCV-1 encodes at least part, if not the entire machinery to glycosylate its proteins; (c) PBCV-1 has at least two types of introns--a self-splicing intron in a transcription factor-like gene and a splicesomal processed type of intron in its DNA polymerase gene. Unlike the chlorella viruses, large double-stranded-DNA-containing viruses that infect marine, filamentous brown algae have a circular genome and a lysogenic phase in their life cycle.

  13. Influenza virus isolation.

    PubMed

    Krauss, Scott; Walker, David; Webster, Robert G

    2012-01-01

    The isolation of influenza viruses is important for the diagnosis of respiratory diseases in lower animals and humans, for the detection of the infecting agent in surveillance programs, and is an essential element in the development and production of vaccine. Since influenza is caused by a zoonotic virus it is necessary to do surveillance in the reservoir species (aquatic waterfowls), intermediate hosts (quails, pigs), and in affected mammals including humans. Two of the hemagglutinin (HA) subtypes of influenza A viruses (H5 and H7) can evolve into highly pathogenic (HP) strains for gallinaceous poultry; some HP H5 and H7 strains cause lethal infection of humans. In waterfowls, low pathogenic avian influenza (LPAI) isolates are obtained primarily from the cloaca (or feces); in domestic poultry, the virus is more often recovered from the respiratory tract than from cloacal samples; in mammals, the virus is most often isolated from the respiratory tract, and in cases of high pathogenic avian influenza (HPAI) from the blood and internal organs of infected birds. Virus isolation procedures are performed by inoculation of clinical specimens into embryonated eggs (primarily chicken eggs) or onto a variety of primary or continuous tissue culture systems. Successful isolation of influenza virus depends on the quality of the sample and matching the appropriate culture method to the sample type.

  14. Ocular tropism of respiratory viruses.

    PubMed

    Belser, Jessica A; Rota, Paul A; Tumpey, Terrence M

    2013-03-01

    Respiratory viruses (including adenovirus, influenza virus, respiratory syncytial virus, coronavirus, and rhinovirus) cause a broad spectrum of disease in humans, ranging from mild influenza-like symptoms to acute respiratory failure. While species D adenoviruses and subtype H7 influenza viruses are known to possess an ocular tropism, documented human ocular disease has been reported following infection with all principal respiratory viruses. In this review, we describe the anatomical proximity and cellular receptor distribution between ocular and respiratory tissues. All major respiratory viruses and their association with human ocular disease are discussed. Research utilizing in vitro and in vivo models to study the ability of respiratory viruses to use the eye as a portal of entry as well as a primary site of virus replication is highlighted. Identification of shared receptor-binding preferences, host responses, and laboratory modeling protocols among these viruses provides a needed bridge between clinical and laboratory studies of virus tropism.

  15. Ocular Tropism of Respiratory Viruses

    PubMed Central

    Rota, Paul A.; Tumpey, Terrence M.

    2013-01-01

    SUMMARY Respiratory viruses (including adenovirus, influenza virus, respiratory syncytial virus, coronavirus, and rhinovirus) cause a broad spectrum of disease in humans, ranging from mild influenza-like symptoms to acute respiratory failure. While species D adenoviruses and subtype H7 influenza viruses are known to possess an ocular tropism, documented human ocular disease has been reported following infection with all principal respiratory viruses. In this review, we describe the anatomical proximity and cellular receptor distribution between ocular and respiratory tissues. All major respiratory viruses and their association with human ocular disease are discussed. Research utilizing in vitro and in vivo models to study the ability of respiratory viruses to use the eye as a portal of entry as well as a primary site of virus replication is highlighted. Identification of shared receptor-binding preferences, host responses, and laboratory modeling protocols among these viruses provides a needed bridge between clinical and laboratory studies of virus tropism. PMID:23471620

  16. Viruses in reptiles

    PubMed Central

    2011-01-01

    The etiology of reptilian viral diseases can be attributed to a wide range of viruses occurring across different genera and families. Thirty to forty years ago, studies of viruses in reptiles focused mainly on the zoonotic potential of arboviruses in reptiles and much effort went into surveys and challenge trials of a range of reptiles with eastern and western equine encephalitis as well as Japanese encephalitis viruses. In the past decade, outbreaks of infection with West Nile virus in human populations and in farmed alligators in the USA has seen the research emphasis placed on the issue of reptiles, particularly crocodiles and alligators, being susceptible to, and reservoirs for, this serious zoonotic disease. Although there are many recognised reptilian viruses, the evidence for those being primary pathogens is relatively limited. Transmission studies establishing pathogenicity and cofactors are likewise scarce, possibly due to the relatively low commercial importance of reptiles, difficulties with the availability of animals and permits for statistically sound experiments, difficulties with housing of reptiles in an experimental setting or the inability to propagate some viruses in cell culture to sufficient titres for transmission studies. Viruses as causes of direct loss of threatened species, such as the chelonid fibropapilloma associated herpesvirus and ranaviruses in farmed and wild tortoises and turtles, have re-focused attention back to the characterisation of the viruses as well as diagnosis and pathogenesis in the host itself. 1. Introduction 2. Methods for working with reptilian viruses 3. Reptilian viruses described by virus families 3.1. Herpesviridae 3.2. Iridoviridae 3.2.1 Ranavirus 3.2.2 Erythrocytic virus 3.2.3 Iridovirus 3.3. Poxviridae 3.4. Adenoviridae 3.5. Papillomaviridae 3.6. Parvoviridae 3.7. Reoviridae 3.8. Retroviridae and inclusion body disease of Boid snakes 3.9. Arboviruses 3.9.1. Flaviviridae 3.9.2. Togaviridae 3.10. Caliciviridae

  17. [Ebola virus disease].

    PubMed

    Nazimek, Katarzyna; Bociaga-Jasik, Monika; Bryniarski, Krzysztof; Gałas, Aleksander; Garlicki, Aleksander; Gawda, Anna; Gawlik, Grzegorz; Gil, Krzysztof; Kosz-Vnenchak, Magdalena; Mrozek-Budzyn, Dorota; Olszanecki, Rafał; Piatek, Anna; Zawilińska, Barbara; Marcinkiewicz, Janusz

    2014-01-01

    Ebola is one of the most virulent zoonotic RNA viruses causing in humans haemorrhagic fever with fatality ratio reaching 90%. During the outbreak of 2014 the number of deaths exceeded 8.000. The "imported" cases reported in Western Europe and USA highlighted the extreme risk of Ebola virus spreading outside the African countries. Thus, haemorrhagic fever outbreak is an international epidemiological problem, also due to the lack of approved prevention and therapeutic strategies. The editorial review article briefly summarizes current knowledge on Ebola virus disease epidemiology, etiology, pathogenesis, clinical presentation, diagnosis as well as possible prevention and treatment.

  18. Genome of Horsepox Virus

    PubMed Central

    Tulman, E. R.; Delhon, G.; Afonso, C. L.; Lu, Z.; Zsak, L.; Sandybaev, N. T.; Kerembekova, U. Z.; Zaitsev, V. L.; Kutish, G. F.; Rock, D. L.

    2006-01-01

    Here we present the genomic sequence of horsepox virus (HSPV) isolate MNR-76, an orthopoxvirus (OPV) isolated in 1976 from diseased Mongolian horses. The 212-kbp genome contained 7.5-kbp inverted terminal repeats and lacked extensive terminal tandem repetition. HSPV contained 236 open reading frames (ORFs) with similarity to those in other OPVs, with those in the central 100-kbp region most conserved relative to other OPVs. Phylogenetic analysis of the conserved region indicated that HSPV is closely related to sequenced isolates of vaccinia virus (VACV) and rabbitpox virus, clearly grouping together these VACV-like viruses. Fifty-four HSPV ORFs likely represented fragments of 25 orthologous OPV genes, including in the central region the only known fragmented form of an OPV ribonucleotide reductase large subunit gene. In terminal genomic regions, HSPV lacked full-length homologues of genes variably fragmented in other VACV-like viruses but was unique in fragmentation of the homologue of VACV strain Copenhagen B6R, a gene intact in other known VACV-like viruses. Notably, HSPV contained in terminal genomic regions 17 kbp of OPV-like sequence absent in known VACV-like viruses, including fragments of genes intact in other OPVs and approximately 1.4 kb of sequence present only in cowpox virus (CPXV). HSPV also contained seven full-length genes fragmented or missing in other VACV-like viruses, including intact homologues of the CPXV strain GRI-90 D2L/I4R CrmB and D13L CD30-like tumor necrosis factor receptors, D3L/I3R and C1L ankyrin repeat proteins, B19R kelch-like protein, D7L BTB/POZ domain protein, and B22R variola virus B22R-like protein. These results indicated that HSPV contains unique genomic features likely contributing to a unique virulence/host range phenotype. They also indicated that while closely related to known VACV-like viruses, HSPV contains additional, potentially ancestral sequences absent in other VACV-like viruses. PMID:16940536

  19. Virus templated metallic nanoparticles.

    PubMed

    Aljabali, Alaa A A; Barclay, J Elaine; Lomonossoff, George P; Evans, David J

    2010-12-01

    Plant viruses are considered as nanobuilding blocks that can be used as synthons or templates for novel materials. Cowpea mosaic virus (CPMV) particles have been shown to template the fabrication of metallic nanoparticles by an electroless deposition metallization process. Palladium ions were electrostatically bound to the virus capsid and, when reduced, acted as nucleation sites for the subsequent metal deposition from solution. The method, although simple, produced highly monodisperse metallic nanoparticles with a diameter of ca. ≤35 nm. CPMV-templated particles were prepared with cobalt, nickel, iron, platinum, cobalt-platinum and nickel-iron.

  20. Fighting cancer with viruses

    NASA Astrophysics Data System (ADS)

    Ferreira, S. C.; Martins, M. L.; Vilela, M. J.

    2005-01-01

    One of the most promising strategies to treat cancer is attacking it with viruses. Viruses can kill tumor cells specifically or act as carriers that deliver normal genes into cancer cells. A model for virotherapy of cancer is investigated and its predictions are in agreement with results obtained from experimental tumors. Furthermore, the model reveals an oscillatory (periodic or aperiodic) response of tumor cells and virus populations which may make clinical prognosis difficult. These results suggest the need for new in vivo and in vitro experiments aiming to detect this oscillatory response.

  1. Zika Virus Infection.

    PubMed

    Shirley, Debbie-Ann T; Nataro, James P

    2017-08-01

    In less than 2 years since entry into the Americas, we have witnessed the emergent spread of Zika virus into large subsets of immunologically naïve human populations and then encountered the devastating effects of microcephaly and brain anomalies that can arise from in utero infection with the virus. Diagnostic evaluation and management of affected infants continues to evolve as our understanding of Zika virus rapidly advances. The development of a safe and effective vaccine holds the potential to attenuate the spread of infection and limit the impact of congenital infection. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Zika virus in Asia.

    PubMed

    Duong, Veasna; Dussart, Philippe; Buchy, Philippe

    2017-01-01

    Zika virus (ZIKV) is an emerging mosquito-borne virus that was first isolated from a sentinel rhesus monkey in the Zika Forest in Uganda in 1947. In Asia, the virus was isolated in Malaysia from Aedes aegypti mosquitoes in 1966, and the first human infections were reported in 1977 in Central Java, Indonesia. In this review, all reported cases of ZIKV infection in Asia as of September 1, 2016 are summarized and some of the hypotheses that could currently explain the apparently low incidence of Zika cases in Asia are explored.

  3. Zika virus: An overview

    PubMed Central

    Rawal, Gautam; Yadav, Sankalp; Kumar, Raj

    2016-01-01

    The Zika virus has been in the news for quite some time due to the ongoing recent outbreak in the Southern America, which started in December 2015. It has been declared a public health emergency by the World Health Organization in February 2016 owing to its association with the congenital deformities, particularly microcephaly in infants borne to the infected mothers. The rapid spread of the virus throughout the United States of America and subsequently to Asia has raised serious international concerns. Its spread to countries neighboring India is a serious threat to the Indian population. This review article gives an overview about the virus, its diagnosis, clinical features, and the management. PMID:28217576

  4. Development of high-yield influenza B virus vaccine viruses.

    PubMed

    Ping, Jihui; Lopes, Tiago J S; Neumann, Gabriele; Kawaoka, Yoshihiro

    2016-12-20

    The burden of human infections with influenza A and B viruses is substantial, and the impact of influenza B virus infections can exceed that of influenza A virus infections in some seasons. Over the past few decades, viruses of two influenza B virus lineages (Victoria and Yamagata) have circulated in humans, and both lineages are now represented in influenza vaccines, as recommended by the World Health Organization. Influenza B virus vaccines for humans have been available for more than half a century, yet no systematic efforts have been undertaken to develop high-yield candidates. Therefore, we screened virus libraries possessing random mutations in the six "internal" influenza B viral RNA segments [i.e., those not encoding the major viral antigens, hemagglutinin (HA) and neuraminidase NA)] for mutants that confer efficient replication. Candidate viruses that supported high yield in cell culture were tested with the HA and NA genes of eight different viruses of the Victoria and Yamagata lineages. We identified combinations of mutations that increased the titers of candidate vaccine viruses in mammalian cells used for human influenza vaccine virus propagation and in embryonated chicken eggs, the most common propagation system for influenza viruses. These influenza B virus vaccine backbones can be used for improved vaccine virus production.

  5. Development of high-yield influenza B virus vaccine viruses

    PubMed Central

    Ping, Jihui; Lopes, Tiago J. S.; Neumann, Gabriele; Kawaoka, Yoshihiro

    2016-01-01

    The burden of human infections with influenza A and B viruses is substantial, and the impact of influenza B virus infections can exceed that of influenza A virus infections in some seasons. Over the past few decades, viruses of two influenza B virus lineages (Victoria and Yamagata) have circulated in humans, and both lineages are now represented in influenza vaccines, as recommended by the World Health Organization. Influenza B virus vaccines for humans have been available for more than half a century, yet no systematic efforts have been undertaken to develop high-yield candidates. Therefore, we screened virus libraries possessing random mutations in the six “internal” influenza B viral RNA segments [i.e., those not encoding the major viral antigens, hemagglutinin (HA) and neuraminidase NA)] for mutants that confer efficient replication. Candidate viruses that supported high yield in cell culture were tested with the HA and NA genes of eight different viruses of the Victoria and Yamagata lineages. We identified combinations of mutations that increased the titers of candidate vaccine viruses in mammalian cells used for human influenza vaccine virus propagation and in embryonated chicken eggs, the most common propagation system for influenza viruses. These influenza B virus vaccine backbones can be used for improved vaccine virus production. PMID:27930325

  6. Simian hemorrhagic fever virus

    USDA-ARS?s Scientific Manuscript database

    This book chapter describes the taxonomic classification of Simian hemorrhagic fever virus (SHFV). Included are: host, genome, classification, morphology, physicochemical and physical properties, nucleic acid, proteins, lipids, carbohydrates, geographic range, phylogenetic properties, biological pro...

  7. How rigid are viruses

    NASA Astrophysics Data System (ADS)

    Hartschuh, R. D.; Wargacki, S. P.; Xiong, H.; Neiswinger, J.; Kisliuk, A.; Sihn, S.; Ward, V.; Vaia, R. A.; Sokolov, A. P.

    2008-08-01

    Viruses have traditionally been studied as pathogens, but in recent years they have been adapted for applications ranging from drug delivery and gene therapy to nanotechnology, photonics, and electronics. Although the structures of many viruses are known, most of their biophysical properties remain largely unexplored. Using Brillouin light scattering, we analyzed the mechanical rigidity, intervirion coupling, and vibrational eigenmodes of Wiseana iridovirus (WIV). We identified phonon modes propagating through the viral assemblies as well as the localized vibrational eigenmode of individual viruses. The measurements indicate a Young’s modulus of ˜7GPa for single virus particles and their assemblies, surprisingly high for “soft” materials. Mechanical modeling confirms that the DNA core dominates the WIV rigidity. The results also indicate a peculiar mechanical coupling during self-assembly of WIV particles.

  8. Respiratory Syncytial Virus (RSV)

    MedlinePlus

    ... common virus that infects the lungs and breathing passages. Almost all babies get it before the age ... infection that causes swelling in the smallest air passages in the lungs Pneumonia, an infection in one ...

  9. VIRUS instrument enclosures

    NASA Astrophysics Data System (ADS)

    Prochaska, T.; Allen, R.; Mondrik, N.; Rheault, J. P.; Sauseda, M.; Boster, E.; James, M.; Rodriguez-Patino, M.; Torres, G.; Ham, J.; Cook, E.; Baker, D.; DePoy, Darren L.; Marshall, Jennifer L.; Hill, G. J.; Perry, D.; Savage, R. D.; Good, J. M.; Vattiat, Brian L.

    2014-08-01

    The Visible Integral-Field Replicable Unit Spectrograph (VIRUS) instrument will be installed at the Hobby-Eberly Telescope† in the near future. The instrument will be housed in two enclosures that are mounted adjacent to the telescope, via the VIRUS Support Structure (VSS). We have designed the enclosures to support and protect the instrument, to enable servicing of the instrument, and to cool the instrument appropriately while not adversely affecting the dome environment. The system uses simple HVAC air handling techniques in conjunction with thermoelectric and standard glycol heat exchangers to provide efficient heat removal. The enclosures also provide power and data transfer to and from each VIRUS unit, liquid nitrogen cooling to the detectors, and environmental monitoring of the instrument and dome environments. In this paper, we describe the design and fabrication of the VIRUS enclosures and their subsystems.

  10. Viruses causing gastroenteritis.

    PubMed

    Wilhelmi, I; Roman, E; Sánchez-Fauquier, A

    2003-04-01

    Acute gastro