197 results on '"Viral haemorrhagic septicaemia virus"'
Search Results
2. Comprehensive surveillance and molecular profiling of viral infections in cultured olive flounder (Paralichthys olivaceus) across Jeju Island, South Korea.
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Oh, Yeong Eun, Kim, Ye Ji, Lee, Young Juhn, Lee, Eung Jun, Jun, Lyu Jin, and Jeong, Joon Bum
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PARALICHTHYS , *VIRUS diseases , *FLATFISHES , *GENETIC variation , *OLIVE - Abstract
To effectively control viral diseases, ongoing research into virus detection and their genetic characteristics is imperative. We conducted an analysis of the prevalence of five species of viruses in olive flounder (Paralichthys olivaceus) farms in Jeju, specifically viral haemorrhagic septicaemia virus (VHSV), lymphocystis disease virus (LCDV), marine birnavirus (MABV), nervous necrosis virus (NNV), and flounder iridovirus (FLIV). Between July and December 2022, a total of 3282 olive flounders were monitored. VHSV and LCDV were detected in 116 and 173 olive flounders, respectively, confirming detection rates of 3.5% and 5.4%. The highest detection rate for VHSV occurred in December (12.5%), while for LCDV, it was in June (13.8%). MABV, NNV, and FLIV were not detected. Phylogenetic analysis of 19 VHSV isolates collected from 2010 to 2022 revealed that all VHSVs belonged to the IVa genotype based on the nucleoprotein gene. Mutations were identified, indicating genetic variability extending to protein mutations. Additionally, continuous and specific nucleotide mutations of VHSV were confirmed on Jeju Island, a central hub for the aquaculture of olive flounder. These observed mutations in VHSV are considered valuable markers for tracking future mutations in subsequent studies. LCDV isolates were classified under genotype II. The results of the phylogenetic analyses of VHSV, a negative-sense single-stranded RNA virus, and LCDV, a double-stranded DNA virus, provide insights into the mutation patterns of marine viruses. • VHSV and LCDV were detected in Jeju Island, South Korea. • The VHSV detected in Jeju has exhibited nucleotide mutations for 13 years. • Protein mutation identified in VHSV detected in Jeju. [ABSTRACT FROM AUTHOR]
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- 2024
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3. Molecular characterization and expression analysis of septin gene family and phagocytic function of recombinant septin 2, 3 and 8 of starry flounder (Platichthys stellatus).
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Sohn, Min-Young, Choi, Kwang-Min, Joo, Min-Soo, Kang, Gyoungsik, Woo, Won-Sik, Kim, Kyung-Ho, Son, Ha-Jeong, Lee, Jeong-Ho, Kim, Do-Hyung, and Park, Chan-Il
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GENE families , *AMINO acid sequence , *FLATFISHES , *PHAGOCYTOSIS , *G proteins , *CELL motility , *CELL polarity - Abstract
Septin is an evolutionarily conserved family of GTP-binding proteins. Septins are known to be involved in a variety of cellular processes, including cell division, chromosome separation, cell polarity, motility, membrane dynamics, exocytosis, apoptosis, phagocytosis, DNA damage responses, and other immune responses. In this study, the sequences of the septin gene family of starry flounder were obtained using NGS sequencing, and the integrity of the sequences was verified through cloning and sequencing. At first, the amino acid sequence was annotated using the cDNA sequence, and then, the gene sequence was verified through multiple sequence alignment and phylogenetic analyses using the related conserved sequences. The septin gene family was classified into three subgroups based on the phylogenetic analysis. High conservation within the domain and homology between the genes reported in different species were confirmed. The expression level of septin gene family mRNA in each tissue of healthy starry flounder was evaluated to confirm the tissue- and gene-specific expression levels. Additionally, as a result of the analysis of mRNA expression after simulated pathogen infection, significant expression changes and characteristics were confirmed upon infection with bacteria (Streptococcus parauberis PH0710) and virus (VHSV). Based on the current results and that of previous studies, to confirm the immunological function, Septin 2, 3, and 8 were produced as recombinant proteins based on the amino acid sequences, and their role in phagocytosis was further investigated. The results of this study indicate that septin gene family plays a complex and crucial role in the host immune response to pathogens of starry flounder. • We identified the septin gene family from starry flounder (Platichthys stellatus). • The expression level of septin gene family mRNA for each tissue in a healthy starry flounder was detected in all tissues used in the experiment, and tissue- and gene-specific expression levels could be confirmed. • Septin gene family mRNA expression levels were significantly regulated by S. parauberis and VHSV. • Based on the amino acid sequence, septin 2, 3, and 8 were produced as recombinant proteins, and phagocytosis was confirmed in rSeptin 2. [ABSTRACT FROM AUTHOR]
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- 2022
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4. First report of eosinophil peroxidase in starry flounder (Platichthys stellatus): Gene identification and gene expression profiling.
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Choi, Kwang-Min, Joo, Min-Soo, Kang, Gyoungsik, Woo, Won-Sik, Kim, Kyung Ho, Jeong, Son Ha, Son, Min Young, Kim, Do-Hyung, and Park, Chan-Il
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EOSINOPHILS , *PEROXIDASE , *GENE expression , *FLATFISHES , *AMINO acid sequence , *VIRUS diseases , *GENE expression profiling - Abstract
Eosinophils are granular leukocytes that are evolutionarily preserved in the innate immune system of some invertebrates and vertebrates, and these cells can directly remove invading microorganisms and secrete various cytokines, and are also involved in homeostasis. These eosinophils are made up of specific granular proteins that can be differentiated from other cells, and eosinophil peroxidase (EPX) is a peroxidase released only from eosinophils that plays an important role in maintaining the main function and homeostasis of eosinophils. We obtained the sequence information of EPX for the first time from the starry flounder (Platichthys stellatus), and predicted it by amino acid sequencing to confirm sequence alignment and phylogenetic characteristics with other species. Based on analysis of the expression characteristics of PsEPX mRNA in healthy P. stellatus , it was expressed at the highest level in peripheral blood lymphocytes (PBLs) and was also expressed at a relatively high level in the head kidney and intestine, which are immune-related tissues. After artificial infection with Streptococcus parauberis and viral haemorrhagic septicaemia virus, which are the causes of major pathogenic diseases, the expression level of PsEPX was significantly regulated, which showed specific characteristics of pathogens or tissues. These results suggest that PsEPX is an important component of the immune system of P. stellatus and is considered a basic research case for the study of the immunological function of eosinophils in fish. • We identified the eosinophil peroxidase (EPX) gene from starry flounder (Platichthys stellatus). • PsEPX mRNA was highly expressed in the PBLs, head kidney and intestine of healthy starry flounder. • PsEPX mRNA expression levels were significantly regulated by S. parauberis and VHSV. [ABSTRACT FROM AUTHOR]
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- 2021
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5. Kinetics of CD4‐1+ lymphocytes in brown trout after exposure to viral haemorrhagic septicaemia virus.
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Ashfaq, Hassan, Soliman, Hatem, Fajmann, Sabine, Sexl, Veronika, El‐Matbouli, Mansour, and Saleh, Mona
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BROWN trout , *MAJOR histocompatibility complex , *IMMUNE response , *LYMPHOCYTES , *AEROMONAS diseases , *SEPSIS - Abstract
T‐helper cells express CD4 as a co‐receptor that binds to major histocompatibility complex class II to synchronize the immune response against upcoming threats via mediating several cytokines. We have previously reported the presence of CD4 homologues in brown trout. The study of cellular immune responses in brown trout is limited by the availability of specific antibodies. We here describe the generation of a polyclonal antibody against CD4‐1 that allows for the investigation of CD4+ cells. We used this novel tool to study CD4+ cells in different tissues during viral haemorrhagic septicaemia infection (VHSV) using flow cytometric technique. Flow cytometric analyses revealed an enhanced level of surface CD4‐1 expression in the infected group in major lymphoid organs and in the intestine. These results suggest an important role for the T‐helper cells within the immune response against viruses, comparable to the immune response in higher vertebrates. [ABSTRACT FROM AUTHOR]
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- 2021
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6. Modifications of the nucleoprotein of viral haemorrhagic septicaemia virus showed gain of virulence in intraperitoneally infected rainbow trout.
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Alencar, Anna Luiza Farias, Kwon, Se Ryun, Rasmussen, Thomas Bruun, Mérour, Emilie, Olesen, Niels Jørgen, and Cuenca, Argelia
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RAINBOW trout , *RECOMBINANT viruses , *REVERSE genetics , *MARINE fishes , *INTRAPERITONEAL injections - Abstract
Viral haemorrhagic septicaemia virus (VHSV) is the cause of an important listed disease in European rainbow trout (Oncorhynchus mykiss) aquaculture and can be present in a wide range of fish species, including marine fish, which can act as viral reservoir. Recent studies revealed putative genetic virulence markers of VHSV to rainbow trout highlighting the roles of the nucleoprotein, phosphoprotein and non‐virion protein. Using reverse genetics, we produced recombinant viruses by introducing parts of or the entire nucleoprotein from a high‐virulent isolate VHSV into a low‐virulent backbone. Furthermore, we also made recombinant viruses by introducing residue modifications in the nucleoprotein that seem to play a role in virulence. Rainbow trout challenged with these recombinant viruses (rVHSVs) by intraperitoneal injection (IP) developed clinical signs and showed lower survival when compared to the parental rVHSV whereas fish challenged by immersion did not show clinical signs except for the high‐virulent control. The mutations did not influence the viral growth in cell culture. The recombinant viruses and parental recombinant were unable to replicate and show cytopathic effect in EPC cells whereas the high‐virulent control was well adapted in all the fish cell lines tested. We showed evidence that corroborates with the hypothesis that the nucleoprotein has virulence motifs associated with VHSV virulence in rainbow trout. [ABSTRACT FROM AUTHOR]
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- 2021
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7. Glycans with Antiviral Activity from Marine Organisms
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Grice, I. D., Mariottini, G. L., Kubiak, Jacek Z., Series Editor, and Kloc, Malgorzata, Series Editor
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- 2018
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8. Comparison of immune response between Escherichia coli‐derived recombinant subunit vaccine and formol‐inactivated whole particle vaccine against viral haemorrhagic septicaemia virus (VHSV) in rainbow trout.
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Tamer, Cuneyt, Albayrak, Harun, and Gumusova, Semra
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RAINBOW trout , *VIRAL vaccines , *ANTISENSE DNA , *SEPSIS , *IMMUNE response , *VIRAL antibodies - Abstract
Viral haemorrhagic septicaemia (VHS) is a serious disease of finfish with a high mortality rate (70%) worldwide. It is caused by infectious viral haemorrhagic septicaemia virus (VHSV). No commercial vaccine is available against VHSV so far. In the present study, we aimed to develop a sustainable recombinant subunit vaccine against a local isolate of VHSV belonging to the genogroup Ie. In order to develop a recombinant vaccine against VHSV genogroup Ie, a cDNA construct of the VHSV gG gene was amplified by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and cloned into an Escherichia coli expression vector (pET‐28a) to express recombinant protein products. In addition, a formol‐inactivated whole particle vaccine was prepared. Subsequently, rainbow trout (17–60 g) were vaccinated by injecting purified recombinant VHSV gG proteins with an adjuvant (montanide) and the formol‐inactivated whole particle vaccine and then challenged with moderately virulent VHSV. At the end of the trial, the mortality rate was found to be 10% (RPS 33.3%) in the recombinant vaccine group, while no mortality was detected (RPS 100%) in the formol‐inactivated whole particle vaccine group. The recombinant subunit vaccine did not provide as much protection as the inactivated whole particle vaccine, even if the desired antibody production and viral load decreased. [ABSTRACT FROM AUTHOR]
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- 2021
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9. Zebrafish (Danio rerio) larvae as a model for real‐time studies of propagating VHS virus infection, tissue tropism and neutrophil activity.
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Marana, Moonika Haahr, Schmidt, Jacob Günther, Biacchesi, Stéphane, Lorenzen, Niels, and Jørgensen, Louise von Gersdorff
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VIRAL tropism , *VIRUS diseases , *ZEBRA danio , *BRACHYDANIO , *LARVAE , *TROPISMS - Abstract
Viral haemorrhagic septicaemia virus (VHSV) is a negative‐sense single‐stranded RNA virus that infects more than 140 different fish species. In this study, zebrafish larvae were employed as in vivo model organisms to investigate progression of disease, the correlation between propagation of the infection and irreversibility of disease, cell tropism and in situ neutrophil activity towards the VHSV‐infected cells. A recombinant VHSV strain, encoding "tomato" fluorescence (rVHSV‐Tomato), was used in zebrafish to be able to follow the progress of the infection in the live host in real‐time. Two‐day‐old zebrafish larvae were injected into the yolk sac with the recombinant virus. The virus titre peaked 96 hr post‐infection in zebrafish larvae kept at 18°C, and correlated with 33% mortality and high morbidity among the larvae. By utilizing the transgenic zebrafish line Tg(fli1:GFP)y1 with fluorescently tagged endothelial cells, we were able to demonstrate that the virus initially infected endothelial cells lining the blood vessels. By observing the rVHSV‐Tomato infection in the neutrophil reporter zebrafish line Tg(MPX:eGFP)i114, we inferred that only a subpopulation of the neutrophils responded to the virus infection. We conclude that the zebrafish larvae are suitable for real‐time studies of VHS virus infections, allowing in vivo dissection of host–virus interactions at the whole organism level. [ABSTRACT FROM AUTHOR]
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- 2021
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10. Development of an oral vaccine using recombinant viral haemorrhagic septicaemia virus glycoproteins produced in tobacco
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Tae Geum Kim, Bong Jo Kang, Sang Ik Park, and Tae Jung Kim
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flounder ,oral vaccine ,plant expression ,viral haemorrhagic septicaemia virus ,Veterinary medicine ,SF600-1100 - Abstract
The viral haemorrhagic septicaemia virus (VHSV) causes high mortality in many marine and freshwater fish species, resulting in heavy economic losses in fish farming. Previously, cholera toxin B subunit (CTB)-fused recombinant viral haemorrhagic septicaemia virus glycoproteins (rec-VHSV-GPs) have been successfully expressed in tobacco, Nicotiana benthamiana. Here, we evaluated the potential of rec-VHSV-GPs as an oral vaccine against a live viral challenge. After immunisation of mice and fish (olive flounder, Paralichthys olivaceous) with those antigenic proteins in a feed additive form, the antibody titres were increased statistically, especially in the primed groups (P < 0.0001) in both the mouse and fish. After the viral challenge under low water temperature culture conditions (below 18 °C), the immunised fish were protected successfully against the challenge, showing a significantly lower mortality rate (P < 0.05). This result suggests that this plant-based immunisation system could induce an effective immune response. It could be used as a candidate to develop an oral vaccine for fish.
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- 2019
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11. Functional characterization and gene expression profile of perforin-2 in starry flounder (Platichthys stellatus).
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Choi, Kwang-Min, Cho, Dong-Hee, Joo, Min-Soo, Choi, Hye-Sung, Kim, Myoung Sug, Han, Hyun-Ja, Cho, Mi Young, Hwang, Seong Don, Kim, Do-Hyung, and Park, Chan-Il
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GENE expression profiling , *FLATFISHES , *PARALICHTHYS , *AMINO acid sequence , *RECOMBINANT proteins , *MESSENGER RNA - Abstract
The membrane attack complex/perforin (MACPF) superfamily consists of multifunctional proteins that form pores on the membrane surface of microorganisms to induce their death and have various immune-related functions. PFN2 is a perforin-like protein with an MACPF domain, and humans with deficient PFN2 levels have increased susceptibility to bacterial infection, which can lead to fatal consequences for some patients. Therefore, in this study, we confirmed the antimicrobial function of PFN2 in starry flounder (Platichthys stellatus). The molecular properties were confirmed based on the verified amino acid sequence of PsPFN2. In addition, the expression characteristics of tissue-specific and pathogen-specific PsPFN2 mRNA were also confirmed. The recombinant protein was produced using Escherichia coli , and the antimicrobial activity was then confirmed. The coding sequence of PFN2 (PsPFN2) in P. stellatus consists of 710 residues. The MACPF domain was conserved throughout evolution, as shown by multiple sequence alignment and phylogenetic analysis. PsPFN2 mRNA is abundantly distributed in immune-related organs such as the spleen and gills of healthy starry flounder, and significant expression changes were confirmed after artificial infection by bacteria or viruses. We cloned the MACPF domain region of PFN2 to produce a recombinant protein (rPFN2) and confirmed its antibacterial effect against a wide range of bacterial species and the parasite (Miamiensis avidus). • Perforin-2 (PFN2) from starry flounder (Platichthys stellatus) was identified. • PsPFN2 was highly expressed in the spleen, gills and PBLs of healthy starry flounder. • PsPFN2 mRNA expression levels were significantly regulated by S. parauberis and VHSV. • We confirmed the antimicrobial activities of rPsPFN2 against bacteria and M. avidus. [ABSTRACT FROM AUTHOR]
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- 2020
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12. VHSV IVb infection and autophagy modulation in the rainbow trout gill epithelial cell line RTgill‐W1.
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Liu, Juan‐Ting, Pham, Phuc H., Wootton, Sarah K., Bols, Niels C., and Lumsden, John S.
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RAINBOW trout , *EPITHELIAL cells , *CELL lines , *TYPE I interferons , *INTERFERON receptors , *WHITE spot syndrome virus , *VIRAL genes , *AUTOPHAGY - Abstract
Autophagy modulation influences the success of intracellular pathogens, and an understanding of the mechanisms involved might offer practical options to reduce the impact of infectious disease. Viral haemorrhagic septicaemia virus (VHSV) can cause high mortality and economic loss in some commercial fish species. VHSV IVb was used to infect a rainbow trout gill cell line, RTgill‐W1, followed by the treatment of the cells with different autophagy‐modulating reagents. LC3II protein using Western blot was significantly (p <.05) decreased for two days following VHSV infection, and immunofluorescence confirmed that LC3II‐positive intracytoplasmic puncta were also decreased. Infection with VHSV resulted in significantly decreased expression of the autophagy‐related (Atg) genes atg4, at12, atg13 and becn1 after one day using quantitative PCR. Both viral gene copy number and VHSV N protein were significantly decreased by treating the cells with autophagy‐blocking (chloroquine) and autophagy‐inhibiting reagents (deoxynivalenol and 3‐methyladenine) after three days, while autophagy induction (restricted nutrition and rapamycin) had limited effect. Only treatment of RTgill‐W1 with deoxynivalenol resulted in a significant increase in expression of type I interferon. Therefore, the suppression of autophagy initially occurs after VHSV IVb infection, but the modulation of autophagy can also inhibit VHSV IVb infection in RTgill‐W1 after three days. [ABSTRACT FROM AUTHOR]
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- 2020
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13. Whole‐genome next‐generation sequencing and phylogenetic characterization of viral haemorrhagic septicaemia virus in Korea.
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Hwang, Jee Youn, Ahn, Sang Jung, Kwon, Mun‐Gyeong, Seo, Jung Soo, Hwang, Seong Don, and Jee, Bo Young
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NUCLEOTIDE sequencing , *RNA replicase , *EXTRACELLULAR matrix proteins , *RAINBOW trout , *SEQUENCE analysis - Abstract
Whole‐genome next‐generation sequencing was used to investigate the local evolution of viral haemorrhagic septicaemia virus, a serious pathogen affecting economically important fish such as rainbow trout and turbot in Europe and olive flounder in Asia. Sequence analysis showed that all isolates were genotype IVa, but could be classified further into four subgroups (K1–K4). In addition, genomic regions encompassing the nucleoprotein, phosphoprotein, matrix protein and non‐virion protein genes, as well as the seven non‐coding regions, were relatively conserved, whereas glycoprotein and RNA‐dependent RNA polymerase genes were variable in the coding region. Taken together, the data demonstrate that whole‐genome next‐generation sequencing may be useful for future surveillance, prevention and control strategies against viral haemorrhagic septicaemia. [ABSTRACT FROM AUTHOR]
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- 2020
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14. Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus.
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Amtmann, Anette, Ahmed, Ibrahim, Zahner‐Rimmel, Petra, Mletzko, Adam, Jordan, Lisa Katharina, Oberle, Martin, Wedekind, Helmut, Christian, Jürgen, Bergmann, Sven Michael, and Becker, Anna Maria
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FISH pathogens , *SAND , *PERACETIC acid , *VIRUSES , *DETECTION limit - Abstract
In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease—Neutrase®—was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection. [ABSTRACT FROM AUTHOR]
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- 2020
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15. Isolation and identification of a viral haemorrhagic septicaemia virus (VHSV) isolate from wild largemouth bass Micropterus salmoides in China.
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Zhang, Wanwan, Li, Zelin, Xiang, Yangxi, Jia, Peng, Liu, Wei, Yi, Meisheng, and Jia, Kuntong
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LARGEMOUTH bass , *HEPATOMEGALY , *FATHEAD minnow , *SYMPTOMS , *VIRUSES - Abstract
Fish rhabdoviruses are a family of viruses responsible for large‐scale fish die‐offs worldwide. Here, we reported the isolation and identification of a member of rhabdoviruses from wild largemouth bass (Micropterus salmoides) in the coastal area of the Pearl River Estuary, China. This virus isolate was identified as viral haemorrhagic septicaemia virus (VHSV) by specific RT‐PCR. Furthermore, the virus (VHSVLB2018) was isolated by cell culture using fathead minnow cells and confirmed by RT‐PCR. Electron microscopy showed the presence of bullet‐shaped viral particles in the cytoplasm of infected cells. The complete sequencing of VHSVLB2018 confirmed that it was genome configuration typical of rhabdoviruses. Phylogenetic analysis based on whole‐genome sequences and G gene nucleotides sequences revealed that VHSVLB2018 was assigned to VHSV genogroup Ⅳa. The pathogenicity of VHSVLB2018 was determined in infection experiments using specific pathogen‐free largemouth bass juveniles. VHSVLB2018‐infected fish showed typical clinical signs of VHSV disease, including darkened skin, petechial haemorrhages and pale enlarged livers, with the cumulative mortalities reached 63.3%–93.3% by 7 days post‐infection. VHSVLB2018 was re‐isolated from dead fish and confirmed by RT‐PCR. Together, this is the first report of isolation and identification of a VHSV isolate from wild largemouth bass in China. [ABSTRACT FROM AUTHOR]
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- 2019
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16. Extensive literature review on vectors and reservoirs of AHL‐listed pathogens of fish
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European Food Safety Authority (efsa), Gnocchi, Marzia, Aires, Mariana, Alvarez, Julio, Arzul, Isabelle, Aznar, Inma, Bicout, Dominique, Carmosino, Ilaria, Drewe, Julian Ashley, Dharmaveer, Shetty, Bastuji, Bruno Garin, Karagianni, Anna Eleonora, Chueca, Miguel Ángel Miranda, Olesen, Niels Jørgen, Palaiokostas, Christos, Roberts, Helen, Nielsen, Søren Saxmose, Schiøtt, Morten, Sindre, Helen, Stone, David, Rusina, Alessia, Vendramin, Niccolo, Dhollander, Sofie, European Food Safety Authority (efsa), Gnocchi, Marzia, Aires, Mariana, Alvarez, Julio, Arzul, Isabelle, Aznar, Inma, Bicout, Dominique, Carmosino, Ilaria, Drewe, Julian Ashley, Dharmaveer, Shetty, Bastuji, Bruno Garin, Karagianni, Anna Eleonora, Chueca, Miguel Ángel Miranda, Olesen, Niels Jørgen, Palaiokostas, Christos, Roberts, Helen, Nielsen, Søren Saxmose, Schiøtt, Morten, Sindre, Helen, Stone, David, Rusina, Alessia, Vendramin, Niccolo, and Dhollander, Sofie
- Abstract
On request of the EU Commission, EFSA carried out an Extensive Literature Review (ELR) to provide a list of vector species or reservoirs species of pathogens of crustaceans, listed in Annex II to the AHL, aiming to update the Annex of Implementing Regulation (EU) 2018/1882. In this Technical Report, the detailed review protocol of the ELR and assessment of potential vector and reservoir species is described of the crustacean pathogens listed in Annex II to the AHL: Taura syndrome virus (TSV), Yellow head virus (YHV) or White spot syndrome virus (WSSV). In total 2,530 research publications were collected for abstract screening and from these, 110 were selected for further full text analysis. In the final data collection and assessment 34 relevant research publications were used for extracting information on vector and reservoir species of the above crustacean pathogens. The results for crustacean species for which scientific evidence indicates that a role as vector species or reservoir species is likely are presented as tables in the supplementary material of this report.
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- 2023
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17. Design and Evaluation of a Macroarray for Detection, Identification, and Typing of Viral Hemorrhagic Septicemia Virus (VHSV)
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Carmen López-Vázquez, Isabel Bandín, and Carlos P. Dopazo
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diagnosis ,viral haemorrhagic septicaemia virus ,Veterinary medicine ,SF600-1100 ,Zoology ,QL1-991 - Abstract
The viral hemorrhagic septicemia virus (VHSV) is the causative agent of an important disease in freshwater and marine fishes. Its diagnosis officially relies on the isolation of the virus in cell culture and its identification by serological or polymerase chain reaction (PCR) methodologies. Nowadays, reverse transcription real-time quantitative PCR (RT-qPCR) is the most widely employed technique for the detection of this virus and some studies have reported the validation of RT-qPCR procedures for the detection, typing, and quantification of VHSV isolates. However, although the efficacy of this technique is not in doubt, it can be cumbersome and even impractical when it comes to processing large numbers of samples, a situation in which cross-contamination problems cannot be ruled out. In the present study, we have designed and validated a macroarray for the simultaneous detection, typing, and quantification of VHSV strains. Its analytical sensitivity (5–50 TCID50/mL), analytical specificity (intra and intergroup), efficiency (E = 100.0–101.1) and reliability (repeatability and reproducibility with CV < 5%, and standard curves with R2 < 0.95) with strains from any VHSV genotype have been widely demonstrated. The procedure is based on the ‘binary multiplex RT-qPCR system (bmRT-qPCR)’ previously reported by the same team, applied to arrays of 96-well PCR strip tubes plates, which can be stored at −25 °C for three months and up to one year before their use, without significant loss of efficiency.
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- 2021
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18. Establishment and characterization of a fin tissue cell line derived from silver pomfret, Pampus argenteus.
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Li, Jianhuan, Jia, Peng, Chen, Xueji, Lai, Mingyan, Jin, Fanming, Liu, Wei, Yi, Meisheng, and Jia, Kuntong
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CELL lines , *CYTOCHROME b , *VIRUS diseases , *SILVER , *SEQUENCE analysis - Abstract
A cell line (PaF) derived from the fin tissue of silver pomfret (Pampus argenteus) was established and characterized in this study. The cell line has been subcultured for more than 50 times in Dulbecco's modified Eagle's medium (DMEM) containing 15% foetal bovine serum (FBS) since the initial primary culture. PaF cells grew well at temperatures from 24°C to 28°C in DMEM supplemented with 15% FBS. Partial amplification and sequence analysis of the cytochrome B gene indicated that PaF originated from silver pomfret. Cytogenetic analysis demonstrated that the modal chromosome number was 48. A significant cytopathic effect was observed in PaF cells during viral haemorrhagic septicaemia virus (VHSV) infection, and the VHSV replication was confirmed by qRT‐PCR and viral titre assays. In contrast, PaF cells were resistant to red‐spotted grouper nervous necrosis virus infection. Moreover, PaF cells could respond to VHSV and lipopolysaccharide treatments, as indicated by the expression of immune‐related genes, TLR5 and TLR9. In conclusion, the establishment of PaF cell line will provide an appropriate in vitro tool for the study of mechanisms of pathogen–silver pomfret interaction. [ABSTRACT FROM AUTHOR]
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- 2019
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19. Investigation of round goby viral haemorrhagic septicaemia outbreak in New York.
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Getchell, Rodman G., First, Erika J., Bogdanowicz, Steven M., Andrés, Jose A., Schulman, Adam T., Kramer, Jordan, Eckerlin, Geofrey E., Farrell, John M., and Marquis, Hélène
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NEOGOBIUS , *FISH kills , *NUCLEOTIDE sequence , *GOBIIDAE , *PARKS - Abstract
Eleven viral haemorrhagic septicaemia virus (VHSV) genotype IVb isolates were sequenced, and their genetic variation explored to determine the source of a VHS outbreak on the eastern shore of Cayuga Lake. An active fish kill of round gobies (Neogobius melanostomus, Pallas) was intensively sampled at King Ferry, NY and nearby Long Point State Park in May 2017. Gross lesions observed on 67 moribund round gobies and two rock bass (Ambloplites rupestris, Rafinesque) included moderately haemorrhagic internal organs and erythematous areas on the head, flank, and fins. RT‐qPCR tests for VHSV were positive for all 69 fish. Viral isolation on epithelioma papulosum cyprinid cells showed cytopathic effect characteristic of VHSV for six round goby samples from King Ferry. The complete nucleotide sequence of the VHSV IVb genomes of five Cayuga Lake round goby isolates were derived on an Illumina platform along with 2017 VHSV IVb isolates from round gobies collected from the following: Lake Erie near Dunkirk, NY; the St. Lawrence River near Clayton and Cape Vincent, NY; and Lake St. Lawrence near Massena, NY. The phylogenetic tree created from these aligned sequences and four other complete VHSV IVb genomes shows Cayuga Lake isolates are closely related to the Lake Erie isolates. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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20. Improvement of a diagnostic procedure in surveillance of the listed fish diseases IHN and VHS.
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Hoferer, Marc, Akimkin, Valerij, Skrypski, Julia, Sting, Reinhard, and Schütze, Heike
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DIAGNOSIS of fish diseases , *INFECTIOUS hematopoietic necrosis virus , *VIRAL hemorrhagic septicemia , *FISH disease epidemiology , *POLYMERASE chain reaction - Abstract
Infectious haematopoietic necrosis (IHN) and viral haemorrhagic septicaemia (VHS) are OIE‐listed and notifiable viral fish diseases which are controlled by eradication and surveillance programmes globally. The present study provides improved RT‐qPCR procedures based on recently described OIE protocols. Improvements comprise the design of a new TaqMan® probe, replacing a TaqMan® MGB probe that turned out to show impaired binding. Reason for this is SNPs detected in the nucleoprotein N gene sequences of IHNV strains targeted by the RT‐qPCR. Furthermore, the IHNV and VHSV RT‐qPCR assays were realized as one‐step and one‐run procedures supplemented by an endogenous control system. The IHNV and VHSV RT‐qPCR assays are characterized by a technical sensitivity of 19 and 190 gene equivalents (cRNA) and an analytical sensitivity of 2–7 and 13 TCID50/ml, respectively. For verification purposes, 105 IHNV and 165 VHSV isolates and several non‐targeted viral and bacterial pathogens were included and returned adequate results. However, in field samples divergent results left 14 samples of 154 undetected for IHNV and one sample of 127 for VHSV using cell culture. The study shows that RT‐qPCR assays ensure facilitated and reliable testing on IHNV and VHSV in eradication and surveillance programmes. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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21. Entry of Rhabdoviruses Into Animal Cells
- Author
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Regan, Andrew D., Whittaker, Gary R., Pöhlmann, Stefan, editor, and Simmons, Graham, editor
- Published
- 2013
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22. Validation of a novel one-step reverse transcription polymerase chain reaction method for detecting viral haemorrhagic septicaemia virus.
- Author
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Kim, Hyoung Jun, Cuenca, Argelia, and Olesen, Niels Jørgen
- Subjects
- *
SEPSIS , *NUCLEOPROTEINS , *FISH farming , *NUCLEOCAPSIDS , *CELL lines - Abstract
Viral haemorrhagic septicaemia (VHS) is one of the most serious viral diseases in salmonid and olive flounder farms. Various diagnostic methods for detecting VHS virus (VHSV) are described in the VHS chapter of the World Organization for Animal Health (OIE) Aquatic Diagnostic Manual. A conventional reverse transcription-PCR (cRT-PCR) targeting the viral nucleocapsid gene is recommended for the detection of VHSV and, to some extent, for genotypic classification. However, the recommended assay exhibits low sensitivity for the detection of VHSV genotype IVa isolates and often shows non-specific amplicons when the RNA template is extracted from non-infected fish cell lines. For these reasons, it is necessary to develop a new RT-PCR method for the foolproof detection of all VHSV genotypes and elimination of non-specific results. In this study, we selected five candidate primer sets that target the VHSV nucleoprotein (N) gene, and selected the most sensitive among them (3F/2R). We then established the optimal reaction conditions for these primers, and ensured that no non-specific amplification had occurred in the fish tissues, fish cell lines, or heterologous viruses. The analytical sensitivity of the novel cRT-PCR was compared to that of cell culture assays, real-time RT-PCR, and other cRT-PCR methods and was found to be as sensitive as or superior to the other methods for detecting all VHSV genotypes. Our newly developed cRT-PCR assay was tested with 80 isolates, representing a collection of all known VHSV genotypes worldwide. Clear and unique amplicons were amplified from all 80 VHSV isolates. The reproducibility, and partly the robustness, of the assay were confirmed by an inter-laboratory proficiency tests including nine laboratories. A high diagnostic sensitivity and specificity was confirmed on tissue material from affected fish. In conclusion a highly robust, sensitive and specific cRT-PCR for detection of VHSV was developed and validated. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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23. Kinetics of CD4‐1+ lymphocytes in brown trout after exposure to viral haemorrhagic septicaemia virus
- Author
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Veronika Sexl, Hatem Soliman, Sabine Fajmann, Hassan Ashfaq, Mona Saleh, and Mansour El-Matbouli
- Subjects
CD4-Positive T-Lymphocytes ,0301 basic medicine ,Trout ,Veterinary (miscellaneous) ,Aquatic Science ,Biology ,Immunofluorescence ,Microbiology ,Novirhabdovirus ,Fish Diseases ,03 medical and health sciences ,Brown trout ,Immune system ,Hemorrhagic Septicemia, Viral ,medicine ,Animals ,Viral haemorrhagic septicaemia virus ,Major Histocompatibility Complex Class II ,medicine.diagnostic_test ,04 agricultural and veterinary sciences ,Biomechanical Phenomena ,Kinetics ,Specific antibody ,030104 developmental biology ,Lymphatic system ,Polyclonal antibodies ,040102 fisheries ,biology.protein ,0401 agriculture, forestry, and fisheries - Abstract
T-helper cells express CD4 as a co-receptor that binds to major histocompatibility complex class II to synchronize the immune response against upcoming threats via mediating several cytokines. We have previously reported the presence of CD4 homologues in brown trout. The study of cellular immune responses in brown trout is limited by the availability of specific antibodies. We here describe the generation of a polyclonal antibody against CD4-1 that allows for the investigation of CD4+ cells. We used this novel tool to study CD4+ cells in different tissues during viral haemorrhagic septicaemia infection (VHSV) using flow cytometric technique. Flow cytometric analyses revealed an enhanced level of surface CD4-1 expression in the infected group in major lymphoid organs and in the intestine. These results suggest an important role for the T-helper cells within the immune response against viruses, comparable to the immune response in higher vertebrates.
- Published
- 2021
24. Fish Rhabdoviruses: Molecular Epidemiology and Evolution
- Author
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Hoffmann, B., Beer, M., Schütze, H., Mettenleiter, T. C., Compans, R.W., editor, Cooper, M.D., editor, Honjo, T., editor, Koprowski, H., editor, Melchers, F., editor, Oldstone, M.B.A., editor, Olsnes, S., editor, Potter, M., editor, Vogt, P.K., editor, Wagner, H., editor, and Fu, Zhen F., editor
- Published
- 2005
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- View/download PDF
25. Modern Biotechnology and Aquaculture
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Hayes, Ben, Andersen, Øivind, and Gjedrem, Trygve, editor
- Published
- 2005
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26. Comparison of immune response between Escherichia coli ‐derived recombinant subunit vaccine and formol‐inactivated whole particle vaccine against viral haemorrhagic septicaemia virus (VHSV) in rainbow trout
- Author
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Harun Albayrak, Cuneyt Tamer, and Semra Gumusova
- Subjects
Viral haemorrhagic septicaemia virus ,Protein subunit ,Aquatic Science ,Biology ,medicine.disease_cause ,Virology ,law.invention ,Immune system ,Viral haemorrhagic septicaemia virus VHSV ,law ,medicine ,Recombinant DNA ,Rainbow trout ,Escherichia coli - Published
- 2021
27. Pathogenicity trials regarding Turkish isolates of infectious pancreatic necrosis virus and viral haemorrhagic septicaemia virus in rainbow trout
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Hamza Kadi, Abdullah Cavunt, Emre Ozan, Harun Albayrak, Cuneyt Tamer, Bahadir Muftuoglu, and Yüksel Durmaz
- Subjects
Viral haemorrhagic septicaemia virus ,viruses ,Rainbow trout ,Aquatic Science ,Biology ,Pathogenicity ,Virology ,Infectious pancreatic necrosis virus - Abstract
Infectious pancreatic necrosis virus (IPNV) and viral haemorrhagic septicaemia virus (VHSV) are two of the major viral threats faced by the aquaculture industry in Turkey. The aim of our study was to investigate the pathogenicity of two Turkish viral strains isolated locally from the Bolu VHSV strain (Accession number: KM972678.1) and the HAH-4 IPNV strain (Accession number: KM972675). The titres of infectious virus were determined by virus titration tests using monolayer cultures of EPC cells to determine the challenge dose. The challenge trial was conducted with 40 rainbow trout (Oncorhynchus mykiss (Walbaum)) for each virus and control group. The infective dose of each virus was applied intraperitoneally as 1 x 10(7) of the tissue culture infective dose per ml. At the end of the trial period (day 21), all fish were examined for clinical signs and post-mortem changes. The average mortality rates for VHSV and IPNV were 36.6% and 33.3%, respectively. Necropsies performed on the deceased fish revealed the presence of IPNV only in fish that had been infected with IPNV, as determined using a real-time PCR method targeting the VP3 gene region of the virus. Similarly, VHSV was detected only in the fish infected with VHSV using a real-time PCR method targeting the gG gene region of the virus. In conclusion, the Bolu strain of VHSV and the HAH-4 strain of IPNV each has moderate pathogenicity in rainbow trout.
- Published
- 2020
28. Impact of feed restriction, chloroquine and deoxynivalenol on viral haemorrhagic septicaemia virus IVb in fathead minnow Pimephales promelas Rafinesque
- Author
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John S. Lumsden and Juan-Ting Liu
- Subjects
0301 basic medicine ,Veterinary (miscellaneous) ,medicine.medical_treatment ,Intraperitoneal injection ,Cyprinidae ,Aquatic Science ,Microbiology ,Novirhabdovirus ,Fish Diseases ,03 medical and health sciences ,Chloroquine ,biology.animal ,Autophagy ,Hemorrhagic Septicemia, Viral ,medicine ,Animals ,Restricted diet ,Survival rate ,Caloric Restriction ,Viral haemorrhagic septicaemia virus ,Bacterial disease ,biology ,food and beverages ,04 agricultural and veterinary sciences ,Minnow ,030104 developmental biology ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,Pimephales promelas ,Trichothecenes ,medicine.drug - Abstract
Autophagy can markedly alter host response to infectious disease, and several studies have demonstrated that a restricted diet or deoxynivalenol modulates autophagy and reduces mortality of fish due to bacterial disease. The picture is less clear for viral diseases of fish. Duplicate tanks of fathead minnow, Pimephales promelas Rafinesque, were fed a replete diet (control), 100 µM chloroquine, 5 µM deoxynivalenol, 10% (fasted) or 40% of a replete diet (pair-fed) for 2 weeks and then experimentally infected by intraperitoneal injection with 2 × 105 viral haemorrhagic septicaemia virus IVb. Survival from highest to lowest for the different treatments was as follows: deoxynivalenol (average 43.3%); control (40.0%); pair-fed (35.0%); fasted (33.3%); and chloroquine (21.7%). No treatment significantly altered the survival rate of fathead minnow after VHSV IVb infection when compared to controls; however, the fish fed with chloroquine had significantly lower survival rate than the fish fed deoxynivalenol (p
- Published
- 2020
29. In vivo study of viral haemorrhagic septicaemia virus and infectious pancreatic necrosis virus coexistence in Senegalese sole ( Solea senegalensis).
- Author
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López‐Vázquez, C, Alonso, M C, Dopazo, C P, and Bandín, I
- Subjects
- *
SEPSIS , *NECROSIS microbiology , *MIXED infections , *SOLEA senegalensis , *VIRUS diseases in fishes - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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30. Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus
- Author
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Ibrahim Ahmed, Lisa Katharina Jordan, Anette Amtmann, Sven Bergmann, Anna Maria Becker, Jürgen Christian, Martin Oberle, Adam Mletzko, Petra Zahner-Rimmel, and Helmut Wedekind
- Subjects
Proteases ,Veterinary (miscellaneous) ,medicine.medical_treatment ,Aquatic Science ,Solid material ,Biology ,Antiviral Agents ,Virus ,Novirhabdovirus ,Fish Diseases ,chemistry.chemical_compound ,Rhabdoviridae Infections ,ddc:570 ,Peracetic acid ,medicine ,Animals ,Koi herpesvirus ,Herpesviridae ,Viral haemorrhagic septicaemia virus ,Protease ,Herpesviridae Infections ,Viral Load ,Virology ,Titer ,chemistry ,Peptide Hydrolases - Abstract
In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease-Neutrase® -was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection.
- Published
- 2019
31. Modifications of the nucleoprotein of viral haemorrhagic septicaemia virus showed gain of virulence in intraperitoneally infected rainbow trout
- Author
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Emilie Mérour, Thomas Bruun Rasmussen, Niels Jørgen Olesen, Argelia Cuenca, Anna Luiza Farias Alencar, Se Ryun Kwon, DTU Aqua, National Institute of Aquatic Resources, Technical University of Denmark [Lyngby] (DTU), Sun Moon University, Statens Serum Institut [Copenhagen], Virologie et Immunologie Moléculaires (VIM (UR 0892)), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), NOVIMARK ERA- net anihwa, National Research Foundation of Korea (NRF) - Korea government (MSIT) 2017R1D1A1A09000992, and European Reference Laboratory for Fish and Crustacean Diseases
- Subjects
0301 basic medicine ,endocrine system ,animal structures ,Veterinary (miscellaneous) ,animal diseases ,viruses ,Virulence ,Aquatic Science ,Biology ,law.invention ,Cell Line ,Novirhabdovirus ,03 medical and health sciences ,Fish Diseases ,reverse genetics ,law ,viral haemorrhagic septicaemia virus ,Hemorrhagic Septicemia, Viral ,Animals ,14. Life underwater ,Cytopathic effect ,[SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health ,urogenital system ,Fishes ,virulence markers ,04 agricultural and veterinary sciences ,Virology ,rainbow trout ,Reverse genetics ,Nucleoprotein ,030104 developmental biology ,Nucleoproteins ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Cell culture ,Phosphoprotein ,Oncorhynchus mykiss ,040102 fisheries ,Recombinant DNA ,0401 agriculture, forestry, and fisheries ,Rainbow trout ,Injections, Intraperitoneal - Abstract
International audience; Viral haemorrhagic septicaemia virus (VHSV) is the cause of an important listed disease in European rainbow trout (Oncorhynchus mykiss) aquaculture and can be present in a wide range of fish species, including marine fish, which can act as viral reservoir. Recent studies revealed putative genetic virulence markers of VHSV to rainbow trout highlighting the roles of the nucleoprotein, phosphoprotein and non-virion protein. Using reverse genetics, we produced recombinant viruses by introducing parts of or the entire nucleoprotein from a high-virulent isolate VHSV into a low-virulent backbone. Furthermore, we also made recombinant viruses by introducing residue modifications in the nucleoprotein that seem to play a role in virulence. Rainbow trout challenged with these recombinant viruses (rVHSVs) by intraperitoneal injection (IP) developed clinical signs and showed lower survival when compared to the parental rVHSV whereas fish challenged by immersion did not show clinical signs except for the high-virulent control. The mutations did not influence the viral growth in cell culture. The recombinant viruses and parental recombinant were unable to replicate and show cytopathic effect in EPC cells whereas the high-virulent control was well adapted in all the fish cell lines tested. We showed evidence that corroborates with the hypothesis that the nucleoprotein has virulence motifs associated with VHSV virulence in rainbow trout.
- Published
- 2021
32. Differentially expressed genes after viral haemorrhagic septicaemia virus infection in olive flounder (Paralichthys olivaceus).
- Author
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Hwang, Jee Youn, Kwon, Mun-Gyeong, Seo, Jung Soo, Do, Jung Wan, Park, Myoung-Ae, Jung, Sung-Hee, and Ahn, Sang Jung
- Subjects
- *
VIRAL hemorrhagic septicemia , *GENE expression in viruses , *PARALICHTHYS , *COMMUNICABLE diseases in animals - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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33. Fish viruses make dsRNA in fish cells: characterization of dsRNA production in rainbow trout ( Oncorhynchus mykiss) cells infected with viral haemorrhagic septicaemia virus, chum salmon reovirus and frog virus 3.
- Author
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Doherty, L, Poynter, S J, Aloufi, A, and DeWitte‐Orr, S J
- Subjects
- *
REOVIRUS diseases , *RAINBOW trout , *DOUBLE-stranded RNA , *SEPSIS , *AQUACULTURE , *FISH viruses , *DISEASES - Abstract
The article provides an analysis of characterization of double-stranded RNA (dsRNA) production in rainbow trout cells infected with viral haemorrhagic septicaemia virus, chum salmon reovirus and frog virus 3. Topics include threat to aquaculture operations with fish viruses, use of immunocytochemistry to investigate rainbow trout cell lines infected with three fish viruses, and assessment of Viral dsRNA production in rainbow trout cell lines.
- Published
- 2016
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34. In vivo virulence of viral haemorrhagic septicaemia virus (VHSV) in rainbow trout Oncorhynchus mykiss correlates inversely with in vitro Mx gene expression.
- Author
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Cano, Irene, Collet, Bertrand, Pereira, Clarissa, Paley, Richard, Aerle, Ronny van, Stone, David, and Taylor, Nick G.H.
- Subjects
- *
RAINBOW trout , *FISH microbiology , *MICROBIAL virulence , *FISH genetics , *GENE expression , *HOSTS (Biology) , *IN vitro studies - Abstract
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 ( J 167), 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 J 167 (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, J 167 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 J 167. 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: J 167, 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 J 167-dose dependent inhibition of the luciferase activity. The data suggest that virulent VHSV isolates may interfere in the interferon pathways, potentially determining higher pathogenicity. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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35. Development of a multiplex RT-PCR assay for simultaneous detection of the major viruses that affect rainbow trout ( Oncorhynchus mykiss).
- Author
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Pinheiro, Ana, Volpe, Enrico, Principi, Donatella, Prosperi, Santino, and Ciulli, Sara
- Subjects
- *
STEELHEAD trout , *RAINBOW trout , *SALMONIDAE , *GENETIC vectors , *MICROORGANISMS - Abstract
The major viral diseases that affect rainbow trout ( Oncorhynchus mykiss) are viral haemorrhagic septicaemia, infectious haematopoietic necrosis, infectious pancreatic necrosis and sleeping disease. In the presented study, we developed a multiplex RT-PCR (mRT-PCR) assay for the simultaneous detection of these four rainbow trout viruses in a single assay. The choice of primers was carried out based on the expected size of the fragments, the temperature and time required for the amplification, and the specificity for the target sequence. Firstly, the method was optimised using reference strains of viral haemorrhagic septicaemia virus (VHSV), infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV) and sleeping disease virus (SDV) cultivated with permissive cell culture lines; subsequently, the method was used for the identification of these viral infections in rainbow trout samples. Twenty-two samples of rainbow trout, clinically suspected of having viruses, were analysed by the developed method to detect the presence of the four viruses, by directly analysing the animal tissues. The mRT-PCR method was able to efficiently detect the viral RNA in infected cell culture supernatants and in tissue samples, highlighting the presence of single infections as well as co-infections in rainbow trout samples. VHSV/SDV and IHNV/SDV co-infections were demonstrated for the first time in rainbow trout. The mRT-PCR method was revealed to be an accurate and fast method to support traditional diagnostic techniques in the diagnosis of major viral diseases of rainbow trout. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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- View/download PDF
36. Development of an oral vaccine using recombinant viral haemorrhagic septicaemia virus glycoproteins produced in tobacco
- Author
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TJ Kim, Si Park, BJ Kang, and TG Kim
- Subjects
chemistry.chemical_classification ,Viral haemorrhagic septicaemia virus ,General Veterinary ,chemistry ,law ,Recombinant DNA ,Biology ,Glycoprotein ,Virology ,law.invention - Published
- 2019
37. Designing an in‐house ELISA to detect antibody against viral haemorrhagic septicaemia virus using recombinant N protein in Iranian farmed rainbow trout ( Oncorhynchus mykiss )
- Author
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Masoud Reza Seyfi Abad Shapouri, Anahita Rezaie, Rahim Peyghan, Nastaran Shahbazian, and Roya Rahnama
- Subjects
Viral haemorrhagic septicaemia virus ,biology ,law ,biology.protein ,Recombinant DNA ,Rainbow trout ,Aquatic Science ,Antibody ,Virology ,law.invention - Published
- 2018
38. Design and Evaluation of a Macroarray for Detection, Identification, and Typing of Viral Hemorrhagic Septicemia Virus (VHSV)
- Author
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C López-Vázquez, Isabel Bandín, and Carlos P. Dopazo
- Subjects
040301 veterinary sciences ,diagnosis ,Virus ,Article ,law.invention ,0403 veterinary science ,03 medical and health sciences ,law ,Genotype ,viral haemorrhagic septicaemia virus ,lcsh:Zoology ,Multiplex ,Typing ,lcsh:QL1-991 ,Polymerase chain reaction ,030304 developmental biology ,0303 health sciences ,lcsh:Veterinary medicine ,General Veterinary ,biology ,04 agricultural and veterinary sciences ,biology.organism_classification ,Virology ,Reverse transcriptase ,Real-time polymerase chain reaction ,lcsh:SF600-1100 ,Animal Science and Zoology ,Viral hemorrhagic septicemia - Abstract
The viral hemorrhagic septicemia virus (VHSV) is the causative agent of an important disease in freshwater and marine fishes. Its diagnosis officially relies on the isolation of the virus in cell culture and its identification by serological or polymerase chain reaction (PCR) methodologies. Nowadays, reverse transcription real-time quantitative PCR (RT-qPCR) is the most widely employed technique for the detection of this virus and some studies have reported the validation of RT-qPCR procedures for the detection, typing, and quantification of VHSV isolates. However, although the efficacy of this technique is not in doubt, it can be cumbersome and even impractical when it comes to processing large numbers of samples, a situation in which cross-contamination problems cannot be ruled out. In the present study, we have designed and validated a macroarray for the simultaneous detection, typing, and quantification of VHSV strains. Its analytical sensitivity (5–50 TCID50/mL), analytical specificity (intra and intergroup), efficiency (E = 100.0–101.1) and reliability (repeatability and reproducibility with CV <, 5%, and standard curves with R2 <, 0.95) with strains from any VHSV genotype have been widely demonstrated. The procedure is based on the ‘binary multiplex RT-qPCR system (bmRT-qPCR)’ previously reported by the same team, applied to arrays of 96-well PCR strip tubes plates, which can be stored at −25 °C for three months and up to one year before their use, without significant loss of efficiency.
- Published
- 2021
39. Walleye Sander vitreus ( Mitchill) are relatively resistant to experimental infection with VHSV IVb and extant walleye strains vary in susceptibility.
- Author
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Grice, J, Reid, A, Peterson, A, Blackburn, K, Tubbs, L, Lord, S, Huber, P, Horricks, R, Dixon, B, Bols, N C, and Lumsden, J S
- Subjects
- *
FATHEAD minnow , *MEDICAL microbiology , *INFECTION , *PIMEPHALES , *FIBROBLASTS , *DNA polymerases - Abstract
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 (102-108 pfu/fish) or waterborne-exposed (w; 1.4 × 107 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 108 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). [ABSTRACT FROM AUTHOR]
- Published
- 2015
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40. 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.
- Author
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Wallace, I S, Donald, K, Munro, L A, Murray, W, Pert, C C, Stagg, H, Hall, M, and Bain, N
- Subjects
- *
MARINE fishes , *ATLANTIC salmon , *BRANCHIURA (Crustacea) , *NUCLEOCAPSIDS , *VIRUS diseases in fishes , *VIRAL replication , *CLEANER fishes - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
41. Do Imports of Rainbow Trout Carcasses Risk Introducing Viral Haemorrhagic Septicaemia Virus into England and Wales?
- Author
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Pearce, F. M., Oidtmann, B. C., Thrush, M. A., Dixon, P. F., and Peeler, E. J.
- Subjects
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RAINBOW trout , *ANIMAL carcasses , *HEMORRHAGIC septicemia , *RIVER channels , *DISEASE susceptibility , *LIQUID waste - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
42. Zebrafish (Danio rerio) larvae as a model for real-time studies of propagating VHS virus infection, tissue tropism and neutrophil activity
- Author
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Louise von Gersdorff Jørgensen, Stéphane Biacchesi, Jacob Günther Schmidt, Niels Lorenzen, Moonika Haahr Marana, Section of Parasitology and Aquatic Pathobiology, Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, IT University of Copenhagen-IT University of Copenhagen-Faculty of Health and Medical Sciences, IT University of Copenhagen-IT University of Copenhagen, Unit for Fish and Shellfish Diseases, National Institute of Aquatic Resources, Technical University of Denmark [Lyngby] (DTU)-Technical University of Denmark [Lyngby] (DTU), Virologie et Immunologie Moléculaires (VIM (UR 0892)), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Kirsten and Freddy Johansens Fond Det Frie Forskningsrad (DFF)9041-00227B
- Subjects
0301 basic medicine ,in vivo visualization ,neutrophil response ,animal structures ,Neutrophils ,Veterinary (miscellaneous) ,ved/biology.organism_classification_rank.species ,Aquatic Science ,Recombinant virus ,Tropism ,Virus ,Green fluorescent protein ,Novirhabdovirus ,03 medical and health sciences ,viral haemorrhagic septicaemia virus ,Hemorrhagic Septicemia, Viral ,Animals ,Model organism ,Zebrafish ,[SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health ,biology ,ved/biology ,fungi ,tissue tropism ,RNA virus ,04 agricultural and veterinary sciences ,biology.organism_classification ,Virology ,mortality ,Disease Models, Animal ,030104 developmental biology ,040102 fisheries ,Tissue tropism ,0401 agriculture, forestry, and fisheries ,infection kinetics - Abstract
International audience; Viral haemorrhagic septicaemia virus (VHSV) is a negative-sense single-stranded RNA virus that infects more than 140 different fish species. In this study, zebrafish larvae were employed as in vivo model organisms to investigate progression of disease, the correlation between propagation of the infection and irreversibility of disease, cell tropism and in situ neutrophil activity towards the VHSV-infected cells. A recombinant VHSV strain, encoding "tomato" fluorescence (rVHSV-Tomato), was used in zebrafish to be able to follow the progress of the infection in the live host in real-time. Two-day-old zebrafish larvae were injected into the yolk sac with the recombinant virus. The virus titre peaked 96 hr post-infection in zebrafish larvae kept at 18 degrees C, and correlated with 33% mortality and high morbidity among the larvae. By utilizing the transgenic zebrafish line Tg(fli1:GFP)(y1) with fluorescently tagged endothelial cells, we were able to demonstrate that the virus initially infected endothelial cells lining the blood vessels. By observing the rVHSV-Tomato infection in the neutrophil reporter zebrafish line Tg(MPX:eGFP)(i114) , we inferred that only a subpopulation of the neutrophils responded to the virus infection. We conclude that the zebrafish larvae are suitable for real-time studies of VHS virus infections, allowing in vivo dissection of host-virus interactions at the whole organism level.
- Published
- 2020
43. Presence of viral haemorrhagic septicaemia virus (VHSV) in the environment of virus-contaminated fish farms and processing plants
- Author
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Elina Välimäki, Anna-Maija Virtala, Pia Vennerström, Leena Maunula, Food Hygiene and Environmental Health, Helsinki One Health (HOH), Leena Maunula / Principal Investigator, Doctoral Programme in Food Chain and Health, Veterinary Biosciences, DAPHNE - Developing Assessment Practices in Higher Education, Teachers' Academy, Anna-Maija Kristiina Virtala / Principal Investigator, University Management, Veterinary Microbiology and Epidemiology, and Food and Environmental Virology Research Group
- Subjects
Veterinary medicine ,animal diseases ,UV-light ,413 Veterinary science ,law.invention ,Fish Diseases ,law ,SALMON-ANEMIA-VIRUS ,Hemorrhagic Septicemia, Viral ,WATER ,Polymerase chain reaction ,Finland ,RISK ,11832 Microbiology and virology ,0303 health sciences ,BLUE MUSSEL ,04 agricultural and veterinary sciences ,Mytilus ,3. Good health ,Processing plant ,PANCREATIC NECROSIS VIRUS ,DEPURATION ,Oncorhynchus mykiss ,Blue mussel ,animal structures ,Fish farming ,Fisheries ,RAINBOW-TROUT ,Aquatic Science ,Biology ,Environment ,Viral haemorrhagic septicaemia virus ,Virus ,Blue mussels ,Novirhabdovirus ,03 medical and health sciences ,Animals ,Ecology, Evolution, Behavior and Systematics ,Brackish water ,STABILITY ,030306 microbiology ,fungi ,PERSISTENCE ,Outbreak ,biology.organism_classification ,EVOLUTION ,Sea water ,Liquid waste ,13. Climate action ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,Rainbow trout - Abstract
After the first outbreak of viral haemorrhagic septicaemia virus (VHSV) in Finnish brackish water rainbow trout Oncorhynchus mykiss farms, infection spread rapidly between the farms. The infrastructure of fish farming did not take into account spreading of infectious fish diseases. To show the presence of VHSV in the environment, we tested seawater, sediment and wild blue mussels Mytilus edulis from VHSV-infected fish farms, and liquid waste from a processing plant that handled infected rainbow trout. Additionally, blue mussels were bath-challenged with VHSV (exposed to cultivated virus or naturally infected rainbow trout). To detect VHSV, virus isolation in cell culture and real-time reverse transcriptase polymerase chain reaction (qRT-PCR) were used. The virus or viral RNA was detected in sea water and in liquid waste from processing plants during wintertime when water temperature is close to 0°C and sunlight is sparse. VHSV did not appear to replicate in blue mussels in our study. Therefore, blue mussels were not considered relevant carriers of VHSV. However, traces of viral RNA were detected up to 29 d post challenge in mussels. Contact with water from processing plants handling VHSV-infected fish populations increases the risk of the disease spreading to susceptible fish populations, especially during cold and dark times of the year.
- Published
- 2020
44. Rhabdovirosis (viral haemorrhagic septicaemia virus)
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Megan D. Niner, Carol A. Stepien, and Douglas W. Leaman
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Viral haemorrhagic septicaemia virus ,surgical procedures, operative ,Biology ,Virology - Published
- 2020
45. Viral haemorrhagic septicaemia virus (VHSV) remains viable for several days but at low levels in the water flea Moina macrocopa
- Author
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Niels Jørgen Olesen and Takafumi Ito
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Disease reservoir ,040301 veterinary sciences ,animal diseases ,Aquatic Science ,Biology ,Viral haemorrhagic septicaemia virus ,Virus ,Microbiology ,Novirhabdovirus ,0403 veterinary science ,Genotype ,Hemorrhagic Septicemia, Viral ,Animals ,Incubation ,Ecology, Evolution, Behavior and Systematics ,Disease Reservoirs ,04 agricultural and veterinary sciences ,Cladocera ,biology.organism_classification ,Virology ,Gastrointestinal Contents ,Titer ,Viability ,Oncorhynchus mykiss ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,Rainbow trout ,Vector ,Rainbow trout oncorhynchus mykiss - Abstract
Viral haemorrhagic septicaemia virus (VHSV) Genotype IVb has been isolated from amphipods belonging to the genus Diporeia, but it has yet to be established whether crustacean zooplankton act as vectors of this virus for fish species. Therefore, we evaluated the viability of infectious VHSV in the water flea Moina macrocopa. VHSV was re-isolated from replicate groups of M. macrocopa that had been immersed with 108.0, 107.0, and 105.0 TCID50 ml-1 of VHSV (DK-3592B, Genotype Ia). Furthermore, 40 M. macrocopa that had been immersed with 108.0 TCID50 ml-1 of VHSV for 72 h had VHSV titers of 102.7-104.3 TCID50. Thus, VHSV was clearly taken up by M. macrocopa and remained viable in this crustacean for several days. However, no mortality was observed over a 28 d period in rainbow trout Oncorhynchus mykiss that were fed VHSV-contaminated M. macrocopa for 14 d, and we found that the virus titer significantly decreased after a 4 h incubation with pyloric caecal extracts from rainbow trout, indicating that passage through the gut is likely to result in a significant decrease in viral titer. This may explain why consumption of prey containing low levels of VHSV did not result in clinical VHS.
- Published
- 2017
46. Comparative susceptibility among three stocks of yellow perch, Perca flavescens ( Mitchill), to viral haemorrhagic septicaemia virus strain IVb from the Great Lakes.
- Author
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Olson, W, Emmenegger, E, Glenn, J, Winton, J, and Goetz, F
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YELLOW perch , *SEPSIS , *WATERBORNE infection , *WILDLIFE conservation , *VIRUS diseases - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
47. Development and validation of a novel Taqman-based real-time RT- PCR assay suitable for demonstrating freedom from viral haemorrhagic septicaemia virus.
- Author
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Jonstrup, S P, Kahns, S, Skall, H F, Boutrup, T S, and Olesen, N J
- Subjects
- *
FISH diseases , *SEPSIS , *CELL culture , *PHENOTYPES , *RHABDOVIRUSES - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
48. Using a Markov-Chain Monte-Carlo modelling approach to identify the relative risk to farmed Scottish Rainbow trout (Oncorhynchus mykiss) in a multi-sector industry of Viral Haemorrhagic Septicaemia Viruses from introduction and emergent sources
- Author
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Kilburn, R., Gregory, A., and Murray, A.G.
- Subjects
- *
MARKOV chain Monte Carlo , *RELATIVE medical risk , *RAINBOW trout industry , *SEPSIS , *ROBUST control , *PARAMETER estimation , *SENSITIVITY analysis - Abstract
Abstract: The risks of introduction of Viral Haemorrhagic Septicaemia Virus (VHSV) into farmed Scottish Rainbow trout (Oncorhynchus mykiss) was simulated using a simple but robust Markov-Chain Monte-Carlo (MCMC) modelling approach. Outputs from the models were subjected to sensitivity analysis to investigate the contribution towards these risks to our parameter assumptions. The aim was to identify the factors whereby Viral Haemorrhagic Septicaemia (VHS) outbreaks are likely to be most sensitive to and thereby most likely susceptible to control against VHSV genotype 1a (G1a) from continental Europe versus the emergence of VHSV genotype 1b (G1b) or VHSV genotype 3 (G3) from within native marine fish populations. Seven scenarios were tested for the three VHSV genotypes that represent different assumptions as to the epidemiology of VHS, different environmental conditions and or possible future expansion of aquaculture. Results from the MCMC model outputs and sensitivity analysis confirm that the greatest risk to fresh water rainbow trout are from VHSV G1a and G3 for marine rainbow trout. Sensitivity analysis revealed that probabilities of introduction and persistence of VHSV G1a in the freshwater environment and freshwater trout were the parameters that had the most significant effect on the model outputs. Atlantic salmon (Salmo salar) was not significantly affected in any of the scenarios and so salmon-associated parameters play very little role in the risk to freshwater rainbow trout. The scenarios are not intended to be absolute but are the best we have for exploring a range of assumptions that may put Scottish freshwater rainbow trout at risk of VHS. [Copyright &y& Elsevier]
- Published
- 2012
- Full Text
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49. Persistence of viral RNA in fish infected with VHSV-IVb at 15 °C and then moved to warmer temperatures after the onset of disease.
- Author
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Goodwin, A E, Merry, G E, and Noyes, A D
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SMALLMOUTH bass , *VIRUS diseases in fishes , *CHANNEL catfish , *INTRAPERITONEAL injections , *VIRAL hemorrhagic septicemia , *FISH mortality , *DISEASE prevalence , *DISEASES - Abstract
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 28 days post-infection, prevalence of infection was 66-100% depending on species and temperature, and VHSV-IVb was detected at 103-105 copies μg−1 host RNA. Our results show that qrt-RTPCR is a useful tool to investigate fish kills even 28 days after temperatures are elevated above those known to be permissive for VHSV replication. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
50. Development of an aquatic pathogen database (AquaPathogen X) and its utilization in tracking emerging fish virus pathogens in North America.
- Author
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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., and Kurath, G.
- Subjects
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PATHOGENIC microorganisms , *RHABDOVIRUSES , *NUCLEOTIDE sequence , *FISH viruses , *SEPSIS - Abstract
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 (MEAPIHNV) 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. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
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