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Nardini R, Pacchiarotti G, Svicher V, Salpini R, Bellocchi MC, Conti R, Sala MG, La Rocca D, Carioti L, Cersini A, Manna G, Scicluna MT. First National Prevalence in Italian Horse Population and Phylogenesis Highlight a Fourth Sub-Type Candidate of Equine Hepacivirus. Viruses 2024; 16:616. [PMID: 38675957 PMCID: PMC11054338 DOI: 10.3390/v16040616] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/18/2024] [Revised: 04/12/2024] [Accepted: 04/12/2024] [Indexed: 04/28/2024] Open
Abstract
Equine hepacivirus (EqHV, Flaviviridae, hepacivirus) is a small, enveloped RNA virus generally causing sub-clinical hepatitis with occasional fatalities. EqHV is reported in equids worldwide, but for Italy data are limited. To address this, a survey study was set up to estimate prevalence at a national level and among different production categories (equestrian; competition; work and meat; reproduction) and national macro-regions (North, Central, South, and Islands). Data obtained testing 1801 horse serum samples by Real-Time RT PCR were compared within the categories and regions. The NS3 fragment of the PCR-positive samples was sequenced by Sanger protocol for phylogenetic and mutational analysis. The tertiary structure of the NS3 protein was also assessed. The estimated national prevalence was 4.27% [1.97-6.59, 95% CI] and no statistical differences were detected among production categories and macro-regions. The phylogenesis confirmed the distribution in Italy of the three known EqHV subtypes, also suggesting a possible fourth sub-type that, however, requires further confirmation. Mutational profiles that could also affect the NS3 binding affinity to the viral RNA were detected. The present paper demonstrates that EqHV should be included in diagnostic protocols when investigating causes of hepatitis, and in quality control protocols for blood derived products due to its parental transmission.
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Affiliation(s)
- Roberto Nardini
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Giulia Pacchiarotti
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Valentina Svicher
- Department of Biology, University of Rome Tor Vergata, 00133 Rome, Italy;
| | - Romina Salpini
- Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy; (R.S.); (M.C.B.); (L.C.)
| | - Maria Concetta Bellocchi
- Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy; (R.S.); (M.C.B.); (L.C.)
| | - Raffaella Conti
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Marcello Giovanni Sala
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Davide La Rocca
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Luca Carioti
- Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy; (R.S.); (M.C.B.); (L.C.)
| | - Antonella Cersini
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | - Giuseppe Manna
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
| | | | - Maria Teresa Scicluna
- Istituto Zooprofilattico Sperimentale del Lazio e della Toscana “M. Aleandri”, 00178 Rome, Italy; (G.P.); (R.C.); (M.G.S.); (D.L.R.); (A.C.); (G.M.); (M.T.S.)
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Cavalleri JV, Korbacska‐Kutasi O, Leblond A, Paillot R, Pusterla N, Steinmann E, Tomlinson J. European College of Equine Internal Medicine consensus statement on equine flaviviridae infections in Europe. Vet Med (Auckl) 2022; 36:1858-1871. [DOI: 10.1111/jvim.16581] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2022] [Accepted: 10/19/2022] [Indexed: 11/13/2022]
Affiliation(s)
- Jessika‐M. V. Cavalleri
- Clinical Unit of Equine Internal Medicine, Department for Companion Animals and Horses University of Veterinary Medicine Vienna Vienna Austria
| | - Orsolya Korbacska‐Kutasi
- Clinical Unit of Equine Internal Medicine, Department for Companion Animals and Horses University of Veterinary Medicine Vienna Vienna Austria
- Department for Animal Breeding, Nutrition and Laboratory Animal Science University of Veterinary Medicine Budapest Hungary
- Hungarian Academy of Sciences—Szent Istvan University (MTA‐SZIE) Large Animal Clinical Research Group Üllő Dóra major Hungary
| | - Agnès Leblond
- EPIA, UMR 0346, Epidemiologie des maladies animales et zoonotiques, INRAE, VetAgro Sup University of Lyon Marcy l'Etoile France
| | - Romain Paillot
- School of Equine and Veterinary Physiotherapy Writtle University College Chelmsford UK
| | - Nicola Pusterla
- Department of Medicine and Epidemiology, School of Veterinary Medicine University of California Davis California USA
| | - Eike Steinmann
- Department of Molecular and Medical Virology, Faculty of Medicine Ruhr University Bochum Bochum Germany
| | - Joy Tomlinson
- Baker Institute for Animal Health Cornell University College of Veterinary Medicine Ithaca New York USA
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Gömer A, Delarocque J, Puff C, Nocke MK, Reinecke B, Baumgärtner W, Cavalleri JMV, Feige K, Steinmann E, Todt D. Dose-Dependent Hepacivirus Infection Reveals Linkage between Infectious Dose and Immune Response. Microbiol Spectr 2022; 10:e0168622. [PMID: 35993785 PMCID: PMC9602444 DOI: 10.1128/spectrum.01686-22] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2022] [Accepted: 08/03/2022] [Indexed: 12/31/2022] Open
Abstract
More than 70 million people worldwide are still infected with the hepatitis C virus 30 years after its discovery, underscoring the need for a vaccine. To develop an effective prophylactic vaccine, detailed knowledge of the correlates of protection and an immunocompetent surrogate model are needed. In this study, we describe the minimum dose required for robust equine hepacivirus (EqHV) infection in equids and examined how this relates to duration of infection, seroconversion, and transcriptomic responses. To investigate mechanisms of hepaciviral persistence, immune response, and immune-mediated pathology, we inoculated eight EqHV naive horses with doses ranging from 1-2 copies to 1.3 × 106 RNA copies per inoculation. We characterized infection kinetics, pathology, and transcriptomic responses via next generation sequencing. The minimal infectious dose of EqHV in horses was estimated at 13 RNA copies, whereas 6 to 7 copies were insufficient to cause infection. Peak viremia did not correlate with infectious dose, while seroconversion and duration of infection appeared to be affected. Notably, seroconversion was undetectable in the low-dose infections within the surveillance period (40 to 50 days). In addition, transcriptomic analysis revealed a nearly dose-dependent effect, with greater immune activation and inflammatory response observed in high-dose infections than in low-dose infections. Interestingly, inoculation with 6-7 copies of RNA that did not result in productive infection, but was associated with a strong immune response, similar to that observed in the high-dose infections. IMPORTANCE We demonstrate that the EqHV dose of infection plays an important role for inducing immune responses, possibly linked to early clearance in high-dose and prolonged viremia in low-dose infections. In particular, pathways associated with innate and adaptive immune responses, as well as inflammatory responses, were more strongly upregulated in high-dose infections than in lower doses. Hence, inoculation with low doses may enable EqHV to evade strong immune responses in the early phase and therefore promote robust, long-lasting infection.
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Affiliation(s)
- André Gömer
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
- Institute of Virology, University of Veterinary Medicine Hannover, Hanover, Germany
| | - Julien Delarocque
- Clinic for Horses, University of Veterinary Medicine Hannover, Hanover, Germany
| | - Christina Puff
- Department of Pathology, University of Veterinary Medicine Hannover, Hanover, Germany
| | - Maximilian K. Nocke
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
| | - Birthe Reinecke
- Institute of Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Hanover, Germany
| | - Wolfgang Baumgärtner
- Department of Pathology, University of Veterinary Medicine Hannover, Hanover, Germany
| | - Jessika M. V. Cavalleri
- Clinical Section of Equine Internal Medicine, Department of Companion Animals and Horses, University of Veterinary Medicine Vienna, Vienna, Austria
| | - Karsten Feige
- Clinic for Horses, University of Veterinary Medicine Hannover, Hanover, Germany
| | - Eike Steinmann
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
| | - Daniel Todt
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
- European Virus Bioinformatics Center (EVBC), Jena, Germany
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Pacchiarotti G, Nardini R, Scicluna MT. Equine Hepacivirus: A Systematic Review and a Meta-Analysis of Serological and Biomolecular Prevalence and a Phylogenetic Update. Animals (Basel) 2022; 12:2486. [PMID: 36230228 PMCID: PMC9558973 DOI: 10.3390/ani12192486] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2022] [Revised: 09/13/2022] [Accepted: 09/16/2022] [Indexed: 11/16/2022] Open
Abstract
Viral hepatitis has recently assumed relevance for equine veterinary medicine since a variety of new viruses have been discovered. Equine Hepacivirus (EqHV) is an RNA virus belonging to the Flaviviridae family that can cause subclinical hepatitis in horses, occasionally evolving into a chronic disease. EqHV, to date, is considered the closest known relative of human HCV. EqHV has been reported worldwide therefore assessing its features is relevant, considering both the wide use of blood products and transfusions in veterinary therapies and its similitude to HCV. The present review resumes the actual knowledge on EqHV epidemiology, risk factors and immunology, together with potential diagnostics and good practices for prevention. Moreover, adhering to PRISMA guidelines for systematic reviews a meta-analysis of serological and biomolecular prevalence and an updated phylogenetic description is presented as a benchmark for further studies.
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Durham AE. Association between forage mycotoxins and liver disease in horses. J Vet Intern Med 2022; 36:1502-1507. [PMID: 35792718 PMCID: PMC9308415 DOI: 10.1111/jvim.16486] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2022] [Accepted: 06/15/2022] [Indexed: 11/30/2022] Open
Abstract
Background Outbreaks of liver disease in horses are common but the etiology of most remains unknown. Forage mycotoxins have been suspected to be a cause. Objectives To examine the association between outbreaks of liver disease and the presence of mycotoxins in forage stored on the same premises. Animals Premises were identified where ≥4 horses were contemporaneously affected by liver disease, and a control group was formed from premises where ≥4 horses had been examined and found to have no evidence of liver disease. Methods Forage was collected from 29 case and 12 control premises. The forage was analyzed for mycotoxin content using a liquid chromatography/mass spectrometry method, targeting 54 mycotoxins. The presence and distribution of mycotoxins between case and control samples was compared. Results Mycotoxins were found in 23/29 (79%) case samples and 10/12 (83%) control samples (P > .99; relative risk, 0.93; 95% confidence interval [CI], 0.64‐1.75). Median (interquartile range [IQR]) total mycotoxin concentration was similar in case and control samples (85.8 μg/kg [1.6‐268] vs. 315 μg/kg [6.3‐860]; P = .16). Ten mycotoxins were found exclusively in case premises comprising fumonisin B1, 15‐acetyldeoxynivalenol, deoxynivalenol, zearalenone, aflatoxins B1 and G1, methylergonovine, nivalenol, verruculogen, and wortmannin. The median (IQR) concentration of fumonisin B1 was significantly higher in case versus control samples (0 μg/kg [0‐81.7] vs. 0 μg/kg [0‐0]; P = .04). Conclusions and Clinical Importance Several mycotoxins with known hepatotoxic potential were found, alone or in combination, exclusively at case premises, consistent with the hypothesis that forage‐associated mycotoxicosis may be a cause of outbreaks of liver disease in horses in the United Kingdom.
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Expanded Diversity and Host Range of Bovine Hepacivirus—Genomic and Serological Evidence in Domestic and Wild Ruminant Species. Viruses 2022; 14:v14071457. [PMID: 35891438 PMCID: PMC9319978 DOI: 10.3390/v14071457] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/24/2022] [Revised: 06/17/2022] [Accepted: 06/20/2022] [Indexed: 12/10/2022] Open
Abstract
The hepatitis C virus (HCV)-related bovine hepacivirus (BovHepV) can cause acute as well as persistent infections in cattle. The true clinical relevance of the virus is not yet known. As reliable antibody detection methods are lacking and prevalence studies have only been conducted in cattle and few countries to date, the true distribution, genetic diversity, and host range is probably greatly underestimated. In this study, we applied several RT-PCR methods and a nano-luciferase-based immunoprecipitation system (LIPS) assay to analyze bovine serum samples from Bulgaria as well as wild ruminant sera from Germany and the Czech Republic. Using these methods, BovHepV infections were confirmed in Bulgarian cattle, with viral genomes detected in 6.9% and serological reactions against the BovHepV NS3 helicase domain in 10% of bovine serum samples. Genetic analysis demonstrated co-circulation of highly diverse BovHepV strains in Bulgarian cattle, and three novel BovHepV subtypes within the genotype 1 could be defined. Furthermore, application of a nested RT-PCR led to the first description of a BovHepV variant (genotype 2) in a wild ruminant species. The results of this study significantly enhance our knowledge of BovHepV distribution, genetic diversity, and host range.
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Gömer A, Puff C, Reinecke B, Bracht S, Conze M, Baumgärtner W, Steinmann J, Feige K, Cavalleri JMV, Steinmann E, Todt D. Experimental cross-species infection of donkeys with equine hepacivirus and analysis of host immune signatures. ONE HEALTH OUTLOOK 2022; 4:9. [PMID: 35527255 PMCID: PMC9082851 DOI: 10.1186/s42522-022-00065-y] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 11/10/2021] [Accepted: 04/06/2022] [Indexed: 06/14/2023]
Abstract
BACKGROUND The Equine Hepacivirus (EqHV) is an equine-specific and liver-tropic virus belonging to the diverse genus of Hepaciviruses. It was recently found in a large donkey (Equus asinus) cohort with a similar seroprevalence (30%), but lower rate of RNA-positive animals (0.3%) compared to horses. These rare infection events indicate either a lack of adaptation to the new host or a predominantly acute course of infection. METHODS In order to analyze the susceptibility and the course of EqHV infection in donkeys, we inoculated two adult female donkeys and one control horse intravenously with purified EqHV from a naturally infected horse. Liver biopsies were taken before and after inoculation to study changes in the transcriptome. RESULTS Infection kinetics were similar between the equids. All animals were EqHV PCR-positive from day three. EqHV RNA-levels declined when the animals seroconverted and both donkeys cleared the virus from the blood by week 12. Infection did not have an impact on the clinical findings and no significant histopathological differences were seen. Blood biochemistry revealed a mild increase in GLDH at the time of seroconversion in horses, which was less pronounced in donkeys. Transcriptomic analysis revealed a distinct set of differentially expressed genes, including viral host factors and immune genes. CONCLUSION To summarize, our findings indicate that donkeys are a natural host of EqHV, due to the almost identical infection kinetics. The different immune responses do however suggest different mechanisms in reacting to hepaciviral infections.
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Affiliation(s)
- André Gömer
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
- Institute of Virology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany
| | - Christina Puff
- Department of Pathology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany
| | - Birthe Reinecke
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research, Hannover, Germany
| | - Stephanie Bracht
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research, Hannover, Germany
| | - Maria Conze
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany
| | - Wolfgang Baumgärtner
- Department of Pathology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany
| | - Jörg Steinmann
- Institute of Medical Microbiology, University of Hospital Essen, University of Duisburg-Essen, Essen, Germany
- Institute of Clinical Hygiene, Medical Microbiology and Infectiology, General Hospital Nürnberg, Paracelsus Medical University, Nürnberg, Germany
| | - Karsten Feige
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany
| | - Jessika M V Cavalleri
- Clinical Unit of Equine Internal Medicine, Department for Companion Animals and Horses, University of Veterinary Medicine Vienna (Vetmeduni), Vienna, Austria
| | - Eike Steinmann
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
| | - Daniel Todt
- Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany.
- European Virus Bioinformatics Center (EVBC), Jena, Germany.
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Bezerra CDS, Limeira CH, Monteiro dos Anjos D, Nogueira DB, Morais DDA, Falcão BMR, Alves CJ, Santos CDSAB, Silva MLCR, de Azevedo SS. Global prevalence of RNA-positive horses for hepacivirus (EqHV): systematic review and meta-analysis. J Equine Vet Sci 2022; 114:104003. [DOI: 10.1016/j.jevs.2022.104003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2022] [Revised: 04/18/2022] [Accepted: 04/27/2022] [Indexed: 10/18/2022]
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Chang WS, Rose K, Holmes EC. Meta-transcriptomic analysis of the virome and microbiome of the invasive Indian myna ( Acridotheres tristis) in Australia. One Health 2021; 13:100360. [PMID: 34917744 PMCID: PMC8666354 DOI: 10.1016/j.onehlt.2021.100360] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/14/2021] [Revised: 12/03/2021] [Accepted: 12/05/2021] [Indexed: 11/03/2022] Open
Abstract
Invasive species exert a serious impact on native fauna and flora and have become the target of eradication and management efforts worldwide. Invasive avian species can also be important pathogen reservoirs, although their viromes and microbiomes have rarely been studied. As one of the top 100 invasive pest species globally, the expansion of Indian mynas (Acridotheres tristis) into peri-urban and rural environments, in conjunction with increasing free-ranging avian agricultural practices, may increase the risk of microbial pathogens jumping species boundaries. Herein, we used a meta-transcriptomic approach to explore the microbes present in brain, liver and large intestine of 16 invasive Indian myna birds in Sydney, Australia. From this, we discovered seven novel viruses from the families Adenoviridae, Caliciviridae, Flaviviridae, Parvoviridae and Picornaviridae. Interestingly, each of the novel viruses identified shared less than 80% genomic similarity with their closest relatives from other avian species, indicative of a lack of detectable virus transmission between invasive mynas to native or domestic species. Of note, we also identified two coccidian protozoa, Isospora superbusi and Isospora greineri, from the liver and gut tissues of mynas. Overall, these data demonstrate that invasive mynas can harbor a diversity of viruses and other microorganisms such that ongoing pathogen surveillance in this species is warranted.
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Affiliation(s)
- Wei-Shan Chang
- School of Life and Environmental Sciences and School of Medical Sciences, The University of Sydney, Sydney, NSW 2006, Australia.,Sydney Institute for Infectious Diseases, School of Life and Environmental Sciences and School of Medical Sciences, The University of Sydney, Sydney, NSW 2006, Australia
| | - Karrie Rose
- Sydney Institute for Infectious Diseases, School of Life and Environmental Sciences and School of Medical Sciences, The University of Sydney, Sydney, NSW 2006, Australia.,Australian Registry of Wildlife Health, Taronga Conservation Society Australia, Mosman, NSW 2088, Australia
| | - Edward C Holmes
- School of Life and Environmental Sciences and School of Medical Sciences, The University of Sydney, Sydney, NSW 2006, Australia.,Sydney Institute for Infectious Diseases, School of Life and Environmental Sciences and School of Medical Sciences, The University of Sydney, Sydney, NSW 2006, Australia
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Altan E, Hui A, Li Y, Pesavento P, Asín J, Crossley B, Deng X, Uzal FA, Delwart E. New Parvoviruses and Picornavirus in Tissues and Feces of Foals with Interstitial Pneumonia. Viruses 2021; 13:v13081612. [PMID: 34452477 PMCID: PMC8402702 DOI: 10.3390/v13081612] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2021] [Revised: 07/20/2021] [Accepted: 07/27/2021] [Indexed: 12/12/2022] Open
Abstract
Six foals with interstitial pneumonia of undetermined etiology from Southern California were analyzed by viral metagenomics. Spleen, lung, and colon content samples obtained during necropsy from each animal were pooled, and nucleic acids from virus-like particles enriched for deep sequencing. The recently described equine copiparvovirus named eqcopivirus, as well as three previously uncharacterized viruses, were identified. The complete ORFs genomes of two closely related protoparvoviruses, and of a bocaparvovirus, plus the partial genome of a picornavirus were assembled. The parvoviruses were classified as members of new ungulate protoparvovirus and bocaparvovirus species in the Parvoviridae family. The picornavirus was classified as a new species in the Salivirus genus of the Picornaviridae family. Spleen, lung, and colon content samples from each foal were then tested for these viral genomes by nested PCR and RT-PCR. When present, parvoviruses were detected in both feces and spleen. The picornavirus, protoparvovirus, and eqcopivirus genomes were detected in the lungs of one animal each. Three foals were co-infected with the picornavirus and either a protoparvovirus, bocaparvovirus, or eqcopivirus. Two other foals were infected with a protoparvovirus only. No viral infection was detected in one animal. The complete ORFs of the first equine protoparvoviruses and bocaparvovirus, the partial ORF of the third equine picornavirus, and their detection in tissues of foals with interstitial pneumonia are described here. Testing the involvement of these viruses in fatal interstitial pneumonia or other equine diseases will require larger epidemiological and/or inoculation studies.
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Affiliation(s)
- Eda Altan
- Vitalant Research Institute, San Francisco, CA 94118, USA; (E.A.); (A.H.); (Y.L.); (X.D.)
- Department of Laboratory Medicine, University of California at San Francisco, San Francisco, CA 94118, USA
| | - Alvin Hui
- Vitalant Research Institute, San Francisco, CA 94118, USA; (E.A.); (A.H.); (Y.L.); (X.D.)
| | - Yanpeng Li
- Vitalant Research Institute, San Francisco, CA 94118, USA; (E.A.); (A.H.); (Y.L.); (X.D.)
- Department of Laboratory Medicine, University of California at San Francisco, San Francisco, CA 94118, USA
| | - Patricia Pesavento
- Department of Pathology Microbiology and Immunology, UC Davis, Davis, CA 95616, USA; (P.P.); (J.A.); (F.A.U.)
| | - Javier Asín
- Department of Pathology Microbiology and Immunology, UC Davis, Davis, CA 95616, USA; (P.P.); (J.A.); (F.A.U.)
- California Animal Health and Food Safety Laboratory System, UC Davis, Davis, CA 95616, USA;
| | - Beate Crossley
- California Animal Health and Food Safety Laboratory System, UC Davis, Davis, CA 95616, USA;
- Department of Medicine and Epidemiology, UC Davis, Davis, CA 95616, USA
| | - Xutao Deng
- Vitalant Research Institute, San Francisco, CA 94118, USA; (E.A.); (A.H.); (Y.L.); (X.D.)
- Department of Laboratory Medicine, University of California at San Francisco, San Francisco, CA 94118, USA
| | - Francisco A. Uzal
- Department of Pathology Microbiology and Immunology, UC Davis, Davis, CA 95616, USA; (P.P.); (J.A.); (F.A.U.)
- California Animal Health and Food Safety Laboratory System, UC Davis, Davis, CA 95616, USA;
| | - Eric Delwart
- Vitalant Research Institute, San Francisco, CA 94118, USA; (E.A.); (A.H.); (Y.L.); (X.D.)
- Department of Laboratory Medicine, University of California at San Francisco, San Francisco, CA 94118, USA
- Correspondence:
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Abbadi I, Lkhider M, Kitab B, Jabboua K, Zaidane I, Haddaji A, Nacer S, Matsuu A, Pineau P, Tsukiyama-Kohara K, Benjelloun S, Ezzikouri S. Non-primate hepacivirus transmission and prevalence: Novel findings of virus circulation in horses and dogs in Morocco. INFECTION GENETICS AND EVOLUTION 2021; 93:104975. [PMID: 34175479 DOI: 10.1016/j.meegid.2021.104975] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/16/2021] [Revised: 06/15/2021] [Accepted: 06/19/2021] [Indexed: 10/21/2022]
Abstract
Non-primate hepacivirus (NPHV) is a homolog of hepatitis C virus and has been isolated from dogs and horses. Data on NPHV prevalence and distribution are not complete, and there is a particular lack of reports from the African continent. The present study represents the first investigation of NPHV prevalence in horses and dogs in North Africa. Blood was collected from 172 horses and 36 dogs at different locations in Morocco, and screened for NPHV RNA using nested PCR targeting 5'UTR and NS3 regions and analyzed for anti-NPHV NS3 antibody using a Gaussia luciferase immunoprecipitation system-to determine seroprevalence. Eight sequences of the NS3 region isolated from positive serum samples were targeted for phylogenetic analysis. Horses and dogs showed respective NPHV RNA positivity rates of 10.5% and 5.5%, and seroprevalences of 65.7% and 8.33%. Juvenile horses appeared more susceptible to infection, with a 23.5% NHPV RNA positivity rate. Seropositivity was more extensive in mares than stallions (77.14% vs. 46.27%, p < 0.0001). Phylogenetically, that NPHV NS3 genes isolated from horses and dog are clustered together. The NPHV strains we detected showed no correlation with geographic location within Morocco. In conclusion, Moroccan horses showed much evidence of previous and/or current NPHV infection, with young age and female sex as noted potential risk factors. Interestingly, NPHV is circulating in dogs as well as horses, suggesting that it has crossed species barriers and that horses and dogs are potential vectors by which an ancestor to hepatitis C virus was transmitted into human populations.
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Affiliation(s)
- Islam Abbadi
- Virology Unit, Viral Hepatitis Laboratory, Institut Pasteur du Maroc, Casablanca, Morocco; Laboratory of Biosciences, School of Sciences and Technology, Mohammedia, Hassan II University of Casablanca, Morocco
| | - Mustapha Lkhider
- Laboratory of Biosciences, School of Sciences and Technology, Mohammedia, Hassan II University of Casablanca, Morocco
| | - Bouchra Kitab
- Transboundary Animal Diseases Centre, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan
| | | | - Imane Zaidane
- Virology Unit, Viral Hepatitis Laboratory, Institut Pasteur du Maroc, Casablanca, Morocco
| | - Asmaa Haddaji
- Virology Unit, Viral Hepatitis Laboratory, Institut Pasteur du Maroc, Casablanca, Morocco
| | - Sabrine Nacer
- Laboratory of Biosciences, School of Sciences and Technology, Mohammedia, Hassan II University of Casablanca, Morocco
| | - Aya Matsuu
- Transboundary Animal Diseases Centre, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan
| | - Pascal Pineau
- Unité "Organisation Nucléaire et Oncogenèse", INSERM U993, Institut Pasteur, Paris, France
| | - Kyoko Tsukiyama-Kohara
- Transboundary Animal Diseases Centre, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan
| | - Soumaya Benjelloun
- Virology Unit, Viral Hepatitis Laboratory, Institut Pasteur du Maroc, Casablanca, Morocco
| | - Sayeh Ezzikouri
- Virology Unit, Viral Hepatitis Laboratory, Institut Pasteur du Maroc, Casablanca, Morocco.
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12
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Nishizawa T, Usui R, Narabu Y, Takahashi M, Murata K, Okamoto H. Identification of a novel pegivirus in pet cats (Felis silvestris catus) in Japan. Virus Res 2021; 301:198452. [PMID: 33971193 DOI: 10.1016/j.virusres.2021.198452] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/24/2021] [Revised: 05/03/2021] [Accepted: 05/04/2021] [Indexed: 12/01/2022]
Abstract
We report a novel pegivirus in pet cats (Felis silvestris catus) in Japan. This virus was only 44.0-49.6 % identical to the reported viruses in the 11 current Pegivirus species and an unclassified pegivirus in dolphins within the entire protein-coding nucleotide sequence and was detected in 1.6 % of pet cats.
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Affiliation(s)
- Tsutomu Nishizawa
- Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, Shimotsuke, Tochigi 329-0498, Japan
| | - Reiko Usui
- Usui Dog and Cat Hospital, Utsunomiya, Tochigi 321-0136, Japan
| | - Yoko Narabu
- Narabu Animal Hospital, Mibu-machi, Shimotsuga-gun, Tochigi 321-0227, Japan
| | - Masaharu Takahashi
- Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, Shimotsuke, Tochigi 329-0498, Japan
| | - Kazumoto Murata
- Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, Shimotsuke, Tochigi 329-0498, Japan
| | - Hiroaki Okamoto
- Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, Shimotsuke, Tochigi 329-0498, Japan.
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13
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Mann S, Ramsay JD, Wakshlag JJ, Stokol T, Reed S, Divers TJ. Investigating the pathogenesis of high-serum gamma-glutamyl transferase activity in Thoroughbred racehorses: A series of case-control studies. Equine Vet J 2021; 54:39-51. [PMID: 33555643 DOI: 10.1111/evj.13435] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/19/2020] [Revised: 01/13/2021] [Accepted: 02/03/2021] [Indexed: 12/28/2022]
Abstract
BACKGROUND High-serum γ-Glutamyl Transferase (GGT) activity has been associated with and thought to be a marker of maladaptation to training and possibly poor performance in racehorses, but the cause is unknown. OBJECTIVES To investigate possible metabolic and infectious causes for the high GGT syndrome. STUDY DESIGN Pilot case-control study and nested case-control study. METHODS The case-control study in 2017 included 16 horses (8 cases and 8 controls with median [range] serum GGT 82 [74-148] and 22 [19-28] IU/L, respectively) from the same stable. In 2018, similar testing was performed in a nested case-control study that identified 27 case (serum GGT 50 ≥ IU/L)-control pairs from three stables for further testing. Serum liver chemistries, selenium measurements, viral PCR and metabolomics were performed. RESULTS No differences were found in frequency of detection of viral RNA/DNA or copy numbers for equine hepacivirus (EqHV) and parvovirus-hepatitis (EqPV-H) between cases and controls. Mild increases in hepatocellular injury and cholestatic markers in case vs control horses suggested a degree of liver disease in a subset of cases. Metabolomic and individual bile acid testing showed differences in cases compared with controls, including increased abundance of pyroglutamic acid and taurine-conjugated bile acids, and reduced abundance of Vitamin B6. Selenium concentrations, although within or above the reference intervals, were also lower in case horses in both studies. MAIN LIMITATIONS Observational study design did not allow us to make causal inferences. CONCLUSIONS We conclude that high GGT syndrome is likely a complex metabolic disorder and that viral hepatitis was not identified as a cause for this syndrome in this cohort of racehorses. Our results support a contribution of oxidative stress and cholestasis in its pathophysiology.
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Affiliation(s)
- Sabine Mann
- Department of Population Medicine and Diagnostic Sciences, Cornell College of Veterinary Medicine, Cornell University, Ithaca, NY, USA
| | - Joshua D Ramsay
- Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA
| | - Joseph J Wakshlag
- Department of Clinical Sciences, Cornell College of Veterinary Medicine, Cornell University, Ithaca, NY, USA
| | - Tracy Stokol
- Department of Population Medicine and Diagnostic Sciences, Cornell College of Veterinary Medicine, Cornell University, Ithaca, NY, USA
| | - Steven Reed
- Rood & Riddle Equine Hospital, Lexington, KY, USA
| | - Thomas J Divers
- Department of Clinical Sciences, Cornell College of Veterinary Medicine, Cornell University, Ithaca, NY, USA
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14
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Porter AF, Pettersson JHO, Chang WS, Harvey E, Rose K, Shi M, Eden JS, Buchmann J, Moritz C, Holmes EC. Novel hepaci- and pegi-like viruses in native Australian wildlife and non-human primates. Virus Evol 2020; 6:veaa064. [PMID: 33240526 PMCID: PMC7673076 DOI: 10.1093/ve/veaa064] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
The Flaviviridae family of positive-sense RNA viruses contains important pathogens of humans and other animals, including Zika virus, dengue virus, and hepatitis C virus. The Flaviviridae are currently divided into four genera-Hepacivirus, Pegivirus, Pestivirus, and Flavivirus-each with a diverse host range. Members of the genus Hepacivirus are associated with an array of animal species, including humans, non-human primates, other mammalian species, as well as birds and fish, while the closely related pegiviruses have been identified in a variety of mammalian taxa, also including humans. Using a combination of total RNA and whole-genome sequencing we identified four novel hepaci-like viruses and one novel variant of a known hepacivirus in five species of Australian wildlife. The hosts infected comprised native Australian marsupials and birds, as well as a native gecko (Gehyra lauta). From these data we identified a distinct marsupial clade of hepaci-like viruses that also included an engorged Ixodes holocyclus tick collected while feeding on Australian long-nosed bandicoots (Perameles nasuta). Distinct lineages of hepaci-like viruses associated with geckos and birds were also identified. By mining the SRA database we similarly identified three new hepaci-like viruses from avian and primate hosts, as well as two novel pegi-like viruses associated with primates. The phylogenetic history of the hepaci- and pegi-like viruses as a whole, combined with co-phylogenetic analysis, provided support for virus-host co-divergence over the course of vertebrate evolution, although with frequent cross-species virus transmission. Overall, our work highlights the diversity of the Hepacivirus and Pegivirus genera as well as the uncertain phylogenetic distinction between.
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Affiliation(s)
- Ashleigh F Porter
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - John H-O Pettersson
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - Wei-Shan Chang
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - Erin Harvey
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - Karrie Rose
- Australian Registry of Wildlife Health, Taronga Conservation Society Australia, Mosman 2088, Australia
| | - Mang Shi
- School of Medicine, Sun Yat-sen University, Guangzhou, China
| | - John-Sebastian Eden
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - Jan Buchmann
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
| | - Craig Moritz
- Research School of Biology, Centre for Biodiversity Analysis, Australian National University, Acton, ACT, Australia
| | - Edward C Holmes
- School of Life and Environmental Sciences and School of Medical Sciences, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney 2006, Australia
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15
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Tomlinson JE, Wolfisberg R, Fahnøe U, Sharma H, Renshaw RW, Nielsen L, Nishiuchi E, Holm C, Dubovi E, Rosenberg BR, Tennant BC, Bukh J, Kapoor A, Divers TJ, Rice CM, Van de Walle GR, Scheel TKH. Equine pegiviruses cause persistent infection of bone marrow and are not associated with hepatitis. PLoS Pathog 2020; 16:e1008677. [PMID: 32649726 PMCID: PMC7375656 DOI: 10.1371/journal.ppat.1008677] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/10/2020] [Revised: 07/22/2020] [Accepted: 06/02/2020] [Indexed: 12/20/2022] Open
Abstract
Pegiviruses frequently cause persistent infection (as defined by >6 months), but unlike most other Flaviviridae members, no apparent clinical disease. Human pegivirus (HPgV, previously GBV-C) is detectable in 1–4% of healthy individuals and another 5–13% are seropositive. Some evidence for infection of bone marrow and spleen exists. Equine pegivirus 1 (EPgV-1) is not linked to disease, whereas another pegivirus, Theiler’s disease-associated virus (TDAV), was identified in an outbreak of acute serum hepatitis (Theiler’s disease) in horses. Although no subsequent reports link TDAV to disease, any association with hepatitis has not been formally examined. Here, we characterized EPgV-1 and TDAV tropism, sequence diversity, persistence and association with liver disease in horses. Among more than 20 tissue types, we consistently detected high viral loads only in serum, bone marrow and spleen, and viral RNA replication was consistently identified in bone marrow. PBMCs and lymph nodes, but not liver, were sporadically positive. To exclude potential effects of co-infecting agents in experimental infections, we constructed full-length consensus cDNA clones; this was enabled by determination of the complete viral genomes, including a novel TDAV 3’ terminus. Clone derived RNA transcripts were used for direct intrasplenic inoculation of healthy horses. This led to productive infection detectable from week 2–3 and persisting beyond the 28 weeks of study. We did not observe any clinical signs of illness or elevation of circulating liver enzymes. The polyprotein consensus sequences did not change, suggesting that both clones were fully functional. To our knowledge, this is the first successful extrahepatic viral RNA launch and the first robust reverse genetics system for a pegivirus. In conclusion, equine pegiviruses are bone marrow tropic, cause persistent infection in horses, and are not associated with hepatitis. Based on these findings, it may be appropriate to rename the group of TDAV and related viruses as EPgV-2. Transmissible hepatitis in horses (Theiler’s disease) has been known for 100 years without knowledge of causative infectious agents. Recently, two novel equine pegiviruses (EPgV) were discovered. Whereas EPgV-1 was not associated to disease, the other was identified in an outbreak of acute serum hepatitis and therefore named Theiler’s disease-associated virus (TDAV). This finding was surprising since human and monkey pegiviruses typically cause long-term infection without associated clinical disease. Whereas no subsequent reports link TDAV to disease, the original association to hepatitis has not been formally examined. Here, we studied EPgV-1 and TDAV and found that their natural history of infection in horses were remarkably similar. Examination of various tissues identified the bone marrow as the primary site of replication for both viruses with no evidence of replication in the liver. To exclude potential effects of other infectious agents, we developed molecular full-length clones for EPgV-1 and TDAV and were able to initiate infection in horses using derived synthetic viral genetic material. This demonstrated long-term infection, but no association with hepatitis. These findings call into question the connection between TDAV, liver infection, and hepatitis in horses.
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Affiliation(s)
- Joy E. Tomlinson
- Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Raphael Wolfisberg
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
| | - Ulrik Fahnøe
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
| | - Himanshu Sharma
- Center for Vaccines and Immunity, Research Institute at Nationwide Children’s Hospital, Columbus, Ohio, United States of America
| | - Randall W. Renshaw
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Louise Nielsen
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
| | - Eiko Nishiuchi
- Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, United States of America
| | - Christina Holm
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
| | - Edward Dubovi
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Brad R. Rosenberg
- Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America
| | - Bud C. Tennant
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Jens Bukh
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
| | - Amit Kapoor
- Center for Vaccines and Immunity, Research Institute at Nationwide Children’s Hospital, Columbus, Ohio, United States of America
| | - Thomas J. Divers
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Charles M. Rice
- Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, United States of America
| | - Gerlinde R. Van de Walle
- Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
| | - Troels K. H. Scheel
- Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Hvidovre Hospital and Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
- Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, United States of America
- * E-mail:
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16
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Elia G, Caringella F, Lanave G, Martella V, Losurdo M, Tittarelli M, Colitti B, Decaro N, Buonavoglia C. Genetic heterogeneity of bovine hepacivirus in Italy. Transbound Emerg Dis 2020; 67:2731-2740. [PMID: 32426936 DOI: 10.1111/tbed.13628] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/27/2020] [Revised: 05/04/2020] [Accepted: 05/12/2020] [Indexed: 12/17/2022]
Abstract
Viruses similar to human hepatitis C virus (HCV) in the Hepacivirus genus have been identified in several animal hosts, including cattle. Since its first discovery in Germany, bovine hepacivirus (BovHepV) has been described in several countries globally. However, limited data are available on BovHepV epidemiology and genetic variability. The aim of this study was to investigate the prevalence and genetic diversity of BovHepV in Italy. Viral RNA was identified in 37 (0.15%) of 24,820 bovine sera, with titres ranging from 1.09 × 103 to 8.27 × 106 RNA copies/ml. Upon sequencing and phylogenetic analysis of the 5'UTR and NS3 genomic portions, the Italian BovHepV strains segregated into at least four distinct subtypes (A, B, C and F) that are also co-circulating globally.
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Affiliation(s)
- Gabriella Elia
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
| | | | - Gianvito Lanave
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
| | - Vito Martella
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
| | - Michele Losurdo
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
| | - Manuela Tittarelli
- Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale', Teramo, Italy
| | - Barbara Colitti
- Department of Veterinary Science, University of Torino, Grugliasco (Torino), Italy
| | - Nicola Decaro
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
| | - Canio Buonavoglia
- Department of Veterinary Medicine, University of Bari, Valenzano (Bari), Italy
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17
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Date T, Sugiyama M, Lkhagvasuren D, Wakita T, Oyunsuren T, Mizokami M. Prevalence of equine hepacivirus infection in Mongolia. Virus Res 2020; 282:197940. [PMID: 32259615 DOI: 10.1016/j.virusres.2020.197940] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/09/2019] [Revised: 01/05/2020] [Accepted: 03/15/2020] [Indexed: 01/17/2023]
Abstract
Equine hepacivirus (EHV) belongs to the hepacivirus A and is related to hepatitis C virus (HCV). This virus shows hepatic tropism and is known to chronically infect horses. EHV has been reported from various countries, but the prevalence in Mongolia, where large horse populations are pastured, remains unknown. This study collected serum samples from horses in six areas across Mongolia, in order to investigate the status of infection. The possibility of human infection was also examined. The results showed an infection rate among horses of about 40 % in all regions. However, no evidence of EHV viremia was found in human serum. A mutation characteristic of Mongolian EHV was found in the 5'-untranslated region of the viral sequence. Molecular phylogenetic trees for core, NS3, and NS5B sequences showed the formation of two clusters depending on the area from which samples were taken. The same results were obtained from molecular phylogenetic analyses using the full genome. From detailed calculations of genetic diversity calculated using the full genome, EHV appears divisible into two subgenotypes. Blood samples were collected again after a 7-month interval to examine infection persistence. Seventeen of 19 horses retested showed positive results for EHV after 7 months, suggesting a high rate of persistent infection. These results indicate a relatively higher frequency of EHV infection in Mongolia than in Europe or North America, with virus strains divided into at least two subgenotypes.
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Affiliation(s)
- Tomoko Date
- Genome Medical Sciences Project, National Center for Global Health and Medicine, 1-7-1 Kohnodai, Ichikawa, Chiba, 272-8516, Japan
| | - Masaya Sugiyama
- Genome Medical Sciences Project, National Center for Global Health and Medicine, 1-7-1 Kohnodai, Ichikawa, Chiba, 272-8516, Japan.
| | - Damdindorj Lkhagvasuren
- Laboratory of Molecular Biology, Institute of Biology, Mongolian Academy of Sciences, Peace av.54b, Bayanzurkh 3, Ulaanbaatar, 13330, Mongolia
| | - Takaji Wakita
- Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan
| | - Tsendsuren Oyunsuren
- Laboratory of Molecular Biology, Institute of Biology, Mongolian Academy of Sciences, Peace av.54b, Bayanzurkh 3, Ulaanbaatar, 13330, Mongolia
| | - Masashi Mizokami
- Genome Medical Sciences Project, National Center for Global Health and Medicine, 1-7-1 Kohnodai, Ichikawa, Chiba, 272-8516, Japan
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18
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de Albuquerque PPLF, Santos LHS, Antunes D, Caffarena ER, Figueiredo AS. Structural insights into NS5B protein of novel equine hepaciviruses and pegiviruses complexed with polymerase inhibitors. Virus Res 2020; 278:197867. [PMID: 31972246 DOI: 10.1016/j.virusres.2020.197867] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2019] [Revised: 01/15/2020] [Accepted: 01/17/2020] [Indexed: 01/09/2023]
Abstract
Infections produced by hepaciviruses have been associated with liver disease in horses. Currently, at least three viruses belonging to the Flaviviridae family are capable of producing a chronic infection in equines: non-primate hepacivirus (NPHV), Theiler's disease-associated virus (TDAV), and equine pegivirus (EPgV). The RNA-dependent RNA polymerases of viruses (RdRp) (NS5 protein), from the flavivirus family, use de novo RNA synthesis to initiate synthesis. The two antiviral drugs currently used to treat hepatitis C (HCV), sofosbuvir and dasabuvir, act on the viral NS5B polymerase as nucleoside and non-nucleoside inhibitors, respectively. Both drugs have shown significant clinical inhibition of viral response. In this work, we aimed to model the NS5B polymerase of the equine hepacivirus (EHCV) subtypes 1 and 2, TDAV and EPgV, to assess whether current direct-acting antiviral drugs against HCV interact with these proteins. Crystal structures of HCV-NS5B were used as templates for modeling target sequences in both conformations (open and closed). Also, molecular docking of sofosbuvir and dasabuvir were performed to predict their possible binding modes at the modeled NS5B polymerase binding sites. We observed that the NS5B models of the EHCV and EPgV shared well-conserved 3D structures to HCV-NS5B and other RdRps, suggesting functional conservation. Interactions of EHCV subtypes 1, 2 and TDAV polymerases with sofosbuvir showed a similar molecular interaction pattern compared to HCV-NS5B, while interactions with dasabuvir were less conserved. In silico studies of molecular interactions between these modeled structures and sofosbuvir suggest that this compound could be efficient in combating equine pathogens, thus contributing to animal welfare.
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Affiliation(s)
| | - Lucianna H S Santos
- Laboratório de Modelagem Molecular e Planejamento de Fármacos, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
| | - Deborah Antunes
- Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
| | - Ernesto Raul Caffarena
- Grupo de Biofísica Computacional e Modelagem Molecular, Programa de Computação Científica, Fiocruz, Rio de Janeiro, Brazil
| | - Andreza Soriano Figueiredo
- Laboratório de Desenvolvimento Tecnológico em Virologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil
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19
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Mrzljak A, Tabain I, Premac H, Bogdanic M, Barbic L, Savic V, Stevanovic V, Jelic A, Mikulic D, Vilibic-Cavlek T. The Role of Emerging and Neglected Viruses in the Etiology of Hepatitis. Curr Infect Dis Rep 2019; 21:51. [PMID: 31754812 DOI: 10.1007/s11908-019-0709-2] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
PURPOSE OF REVIEW In this review, we present the overview of emerging and neglected viruses associated with liver involvement. RECENT FINDINGS Hepatitis E virus (HEV) emerged in the last two decades, causing hepatitis in many parts of the world. Moreover, liver involvement was also described in some emerging arboviral infections. Many reports showed dengue-associated liver injury; however, chikungunya, West Nile, tick-borne encephalitis, and Zika virus are rarely associated with clinically manifest liver disease. In addition, some neglected highly prevalent viruses such as adenoviruses and parvovirus B19 are capable of causing hepatitis in specific population groups. Anelloviruses (torque teno virus/torque teno mini virus/torque teno midi virus, SEN virus), human bocavirus, pegiviruses, and lymphocytic choriomeningitis virus have shown a little potential for causing hepatitis, but their role in the etiology of liver disease remains to be determined. In addition to the well-known hepatotropic viruses, many emerging and neglected viruses have been associated with liver diseases. The number of emerging zoonotic viruses has been increasingly recognized. While zoonotic potential of HEV is well documented, the recent identification of new hepatitis-related animal viruses such as HEV strains from rabbits and camels, non-primate hepaciviruses in domestic dogs and horses, as well as equine and porcine pegivirus highlights the possible zoonotic transmission in the context of "One Health." However, zoonotic potential and hepatotropism of animal hepatitis viruses remain to be determined.
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Affiliation(s)
- Anna Mrzljak
- Department of Medicine, Merkur University Hospital, Salata 3b, 10000, Zagreb, Croatia.
- School of Medicine, University of Zagreb, Zagreb, Croatia.
| | - Irena Tabain
- Department of Virology, Croatian Institute of Public Health, Zagreb, Croatia
| | - Hrvoje Premac
- Department of Medicine, Varazdin General Hospital, Varazdin, Croatia
| | - Maja Bogdanic
- Department of Virology, Croatian Institute of Public Health, Zagreb, Croatia
| | - Ljubo Barbic
- Department of Microbiology and Infectious Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia
| | - Vladimir Savic
- Poultry Center, Laboratory for Virology and Serology, Croatian Veterinary Institute, Zagreb, Croatia
| | - Vladimir Stevanovic
- Department of Microbiology and Infectious Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia
| | - Ana Jelic
- Department of Medicine, Merkur University Hospital, Salata 3b, 10000, Zagreb, Croatia
| | - Danko Mikulic
- Department of Surgery, Merkur University Hospital, Zagreb, Croatia
| | - Tatjana Vilibic-Cavlek
- School of Medicine, University of Zagreb, Zagreb, Croatia
- Department of Virology, Croatian Institute of Public Health, Zagreb, Croatia
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20
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Badenhorst M, de Heus P, Auer A, Rümenapf T, Tegtmeyer B, Kolodziejek J, Nowotny N, Steinmann E, Cavalleri JMV. No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses. Viruses 2019; 11:v11111014. [PMID: 31683893 PMCID: PMC6893842 DOI: 10.3390/v11111014] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2019] [Revised: 10/28/2019] [Accepted: 10/29/2019] [Indexed: 12/21/2022] Open
Abstract
Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Concurrently, 386 serum samples from horses in eastern Austria were analyzed for EqHV-specific antibodies by luciferase immunoprecipitation system (LIPS) and for EqHV RNA by RT-qPCR. Additionally, liver-specific biochemistry parameters were compared between EqHV RNA-positive horses and EqHV RNA-negative horses. Phylogenetic analysis was conducted in comparison to previously published sequences from various origins. No EqHV RNA was detected in mosquito pools. Serum samples yielded an EqHV antibody prevalence of 45.9% (177/386) and RNA prevalence of 4.15% (16/386). EqHV RNA-positive horses had significantly higher glutamate dehydrogenase (GLDH) levels (p = 0.013) than control horses. Phylogenetic analysis showed high similarity between nucleotide sequences of EqHV in Austrian horses and EqHV circulating in other regions. Despite frequently detected evidence of EqHV infection in Austrian horses, no viral RNA was found in mosquitoes. It is therefore unlikely that mosquitoes are vectors of this flavivirus.
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Affiliation(s)
- Marcha Badenhorst
- University Equine Clinic - Internal Medicine, Department for Companion Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
| | - Phebe de Heus
- University Equine Clinic - Internal Medicine, Department for Companion Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
| | - Angelika Auer
- Institute of Virology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
| | - Till Rümenapf
- Institute of Virology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
| | - Birthe Tegtmeyer
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH) - Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Strasse 7, 30625 Hannover, Germany.
| | - Jolanta Kolodziejek
- Institute of Virology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
| | - Norbert Nowotny
- Institute of Virology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
- Department of Basic Medical Sciences, College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Building 14, Dubai Healthcare City, Dubai, UAE.
| | - Eike Steinmann
- Department of Molecular and Medical Virology, Ruhr-University Bochum, 44801 Bochum, Germany.
| | - Jessika-M V Cavalleri
- University Equine Clinic - Internal Medicine, Department for Companion Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
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21
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Human pegivirus 2 exhibits minimal geographic and temporal genetic diversity. Virology 2019; 539:69-79. [PMID: 31689572 DOI: 10.1016/j.virol.2019.10.012] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2019] [Revised: 10/17/2019] [Accepted: 10/21/2019] [Indexed: 01/18/2023]
Abstract
We applied an NGS based target capture approach to amplify HPgV-2 sequences from metagenomic libraries and enable full genome characterization. Despite expanded geographical sampling, sequence variability remains low, with diversity concentrated in approximately 3.3% of all amino acids. Serial samples from one HPgV-2 positive individual co-infected with comparable titers of HIV, HCV, and GBV-C showed that HPgV-2 remains highly stable over several weeks compared to other RNA viruses, despite a similarly error-prone polymerase. The consistent epidemiological association with and structural similarities to HCV, and the weak positive correlation of HCV and HPgV-2 titers shown here, suggests it may benefit from co-infection. While minimal selective pressure on HPgV-2 to evolve could suggest fitness, the rarity of HPgV-2 and the tight phylogenetic clustering of global strains likely indicates origination from a common source and a virus that is ill-suited to its host. Sporadic infections may explain the limited genetic diversity observed worldwide.
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22
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Viruses in Horses with Neurologic and Respiratory Diseases. Viruses 2019; 11:v11100942. [PMID: 31614994 PMCID: PMC6832430 DOI: 10.3390/v11100942] [Citation(s) in RCA: 22] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/29/2019] [Revised: 10/02/2019] [Accepted: 10/07/2019] [Indexed: 02/07/2023] Open
Abstract
Metagenomics was used to identify viral sequences in the plasma and CSF (cerobrospinal fluid) of 13 horses with unexplained neurological signs and in the plasma and respiratory swabs of 14 horses with unexplained respiratory signs. Equine hepacivirus and two copiparvoviruses (horse parvovirus-CSF and a novel parvovirus) were detected in plasma from neurological cases. Plasma from horses with respiratory signs contained the same two copiparvoviruses plus equine pegivirus D and respiratory swabs contained equine herpes virus 2 and 5. Based on genetic distances the novel copiparvovirus qualified as a member of a new parvovirus species we named Eqcopivirus. These samples plus another 41 plasma samples from healthy horses were tested by real-time PCRs for multiple equine parvoviruses and hepacivirus. Over half the samples tested were positive for one to three viruses with eqcopivirus DNA detected in 20.5%, equine hepacivirus RNA and equine parvovirus-H DNA in 16% each, and horse parvovirus-CSF DNA in 12% of horses. Comparing viral prevalence in plasma none of the now three genetically characterized equine parvoviruses (all in the copiparvovirus genus) was significantly associated with neurological and respiratory signs in this limited sampling.
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23
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Paim WP, Weber MN, Cibulski SP, da Silva MS, Puhl DE, Budaszewski RF, Varela APM, Mayer FQ, Canal CW. Characterization of the viral genomes present in commercial batches of horse serum obtained by high-throughput sequencing. Biologicals 2019; 61:1-7. [PMID: 31447377 DOI: 10.1016/j.biologicals.2019.08.005] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/17/2019] [Revised: 08/19/2019] [Accepted: 08/20/2019] [Indexed: 12/15/2022] Open
Abstract
Horses are often used as blood donors for commercial horse serum (HS) production and to manufacture biologicals. HS is an alternative for fetal bovine serum (FBS) used as a supplement for cell culture and vaccine production. Furthermore, HS is also frequently obtained in order to produce antisera toxins and pathogens. The advent of high-throughput sequencing (HTS) has promoted changes in virus detection, since previous knowledge of targets is not required. Thus, the present study aimed to describe the virome of five different batches of commercial HS from New Zealand (three batches) and Brazil and the United States (one batch each) using HTS. Each HS pool were processed and sequenced using an Illumina MiSeq platform. Sequences-related to viruses belonging to the Flaviviridae, Herpesviridae, and Parvoviridae families were detected. Particularly, equine hepacivirus (EqHV), equine pegivirus (EPgV), and Theiler's disease-associated virus (TDAV) were more frequent found in the batches analyzed. The presence of viral genomes in cell culture sera illustrates that these commercial sera can contain a mixture of different viruses and, therefore, can be regarded as potentially infectious for susceptible hosts. Moreover, the innocuity of commercial HS is important for the efficiency and security of diagnostics and the production of biological products.
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Affiliation(s)
- W P Paim
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil
| | - M N Weber
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil
| | - S P Cibulski
- Laboratório de Biotecnologia Cellular e Molecular, Centro de Biotecnologia-CBiotec, Universidade Federal da Paraíba (UFPB), Cidade Universitária, João Pessoa, PB, Brazil
| | - M S da Silva
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil
| | - D E Puhl
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil
| | - R F Budaszewski
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil
| | - A P M Varela
- Equipe de Virologia, Instituto de Ciências Básicas da Saúde, UFRGS, Porto Alegre, RS, Brazil
| | - F Q Mayer
- Laboratório de Biologia Molecular, Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Fundação Estadual de Pesquisa Agropecuária (FEPAGRO), Eldorado Do Sul, RS, Brazil
| | - C W Canal
- Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS), Brazil.
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24
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Genetic variability of porcine pegivirus in pigs from Europe and China and insights into tissue tropism. Sci Rep 2019; 9:8174. [PMID: 31160748 PMCID: PMC6547670 DOI: 10.1038/s41598-019-44642-0] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/29/2018] [Accepted: 05/21/2019] [Indexed: 12/29/2022] Open
Abstract
Pegiviruses belong to the family Flaviviridae and have been found in humans and other mammalian species. To date eleven different pegivirus species (Pegivirus A-K) have been described. However, little is known about the tissue tropism and replication of pegiviruses. In 2016, a so far unknown porcine pegivirus (PPgV, Pegivirus K) was described and persistent infection in the host, similar to human pegivirus, was reported. In this study, qRT-PCR, phylogenetic analyses and fluorescence in situ hybridization (FISH) were implemented to detect and quantify PPgV genome content in serum samples from domestic pigs from Europe and Asia, in tissue and peripheral blood mononuclear cell (PBMC) samples and wild boar serum samples from Germany. PPgV was detectable in 2.7% of investigated domestic pigs from Europe and China (viral genome load 2.4 × 102 to 2.0 × 106 PPgV copies/ml), while all wild boar samples were tested negative. Phylogenetic analyses revealed pairwise nucleotide identities >90% among PPgVs. Finally, PPgV was detected in liver, thymus and PBMCs by qRT-PCR and FISH, suggesting liver- and lymphotropism. Taken together, this study provides first insights into the tissue tropism of PPgV and shows its distribution and genetic variability in Europe and China.
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Tomlinson JE, Van de Walle GR, Divers TJ. What Do We Know About Hepatitis Viruses in Horses? Vet Clin North Am Equine Pract 2019; 35:351-362. [PMID: 31084975 DOI: 10.1016/j.cveq.2019.03.001] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Theiler disease (serum hepatitis or idiopathic acute hepatic necrosis) has long been suspected to have a viral etiology. Four viruses have been described in association with hepatitis in horses. Further investigation suggests equine pegivirus and Theiler disease-associated virus (a second pegivirus) are neither hepatotropic nor pathogenic. Nonprimate hepacivirus (NPHV) causes subclinical disease in experimental models and has been associated with hepatitis in some clinical cases. Equine parvovirus-hepatitis (EqPV-H) experimentally causes subclinical-to-clinical liver disease and is found in the vast majority of Theiler disease cases. EqPV-H is likely of clinical significance, whereas the significance of NPHV is unknown.
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Affiliation(s)
- Joy E Tomlinson
- Baker Institute for Animal Health, Cornell University College of Veterinary Medicine, 235 Hungerford Hill Road, Ithaca, NY 14853, USA.
| | - Gerlinde R Van de Walle
- Baker Institute for Animal Health, Cornell University College of Veterinary Medicine, 235 Hungerford Hill Road, Ithaca, NY 14853, USA
| | - Thomas J Divers
- Department of Clinical Sciences, Cornell University College of Veterinary Medicine, 930 Campus Road, Box25, Ithaca, NY 14853, USA
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Figueiredo AS, de Moraes MVDS, Soares CC, Chalhoub FLL, de Filippis AMB, Dos Santos DRL, de Almeida FQ, Godoi TLOS, de Souza AM, Burdman TR, de Lemos ERS, Dos Reis JKP, Cruz OG, Pinto MA. First description of Theiler's disease-associated virus infection and epidemiological investigation of equine pegivirus and equine hepacivirus coinfection in Brazil. Transbound Emerg Dis 2019; 66:1737-1751. [PMID: 31017727 DOI: 10.1111/tbed.13210] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/21/2019] [Revised: 03/31/2019] [Accepted: 04/17/2019] [Indexed: 02/06/2023]
Abstract
Recent advances in the study of equine pegivirus (EPgV), Theiler's disease-associated virus (TDAV) and equine hepacivirus (EqHV) highlight their importance to veterinary and human health. To gain some insight into virus distribution, possible risk factors, presence of liver damage and genetic variability of these viruses in Brazil, we performed a cross-sectional study of EPgV and TDAV infections using a simultaneous detection assay, and assessed EqHV coinfection in different horse cohorts. Of the 500 serum samples screened, TDAV, EPgV and EPgV-EqHV were present in 1.6%, 14.2% and 18.3%, respectively. EPgV-positive horses were present in four Brazilian states: Espírito Santo, Mato Grosso do Sul, Minas Gerais and Rio de Janeiro. Serum biochemical alterations were present in 40.4% of EPgV-infected horses, two of them presenting current liver injury. Chance of infection was 2.7 times higher in horses ≤5 years old (p = 0.0008) and 4.9 times higher in horses raised under intensive production systems (p = 0.0009). EPgV-EqHV coinfection was 75% less likely in horses older than 5 years comparatively to those with ≤5 years old (p = 0.047). TDAV-positive animals were detected in different horse categories without biochemical alteration. Nucleotide sequences were highly conserved among isolates from this study and previous field and commercial product isolates (≥88% identity). Tree topology revealed the formation of two clades (pp = 1) for both EPgV and TDAV NS3 partial sequences. In conclusion, the widespread presence of EPgV-RNA suggests an enzootic infection with subclinical viremia in Brazil. Horse management can influence virus spread. This first report of TDAV-infected horses outside the USA reveals the existence of subclinical viremic horses in distant geographical regions. EPgV and TDAV have similar circulating isolates worldwide. These findings contribute to global efforts to understand the epidemiology and pathogenesis of these equine viruses.
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Affiliation(s)
- Andreza Soriano Figueiredo
- Laboratório de Desenvolvimento Tecnológico em Virologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil
| | | | | | | | | | | | | | - Tatianne Leme Oliveira Santos Godoi
- Coordenação de Produção Integrada ao Ensino, Pesquisa e Extensão, Reitoria, Universidade Federal Rural do Rio de Janeiro, Rio de Janeiro, Brazil
| | - Aline Moreira de Souza
- Laboratório de Pesquisa Clínica e Diagnóstico Molecular Professor Marcílio Dias do Nascimento, Departamento de Patologia e Clínica Veterinária, Faculdade de Veterinária, Universidade Federal Fluminense, Niterói, Brazil
| | - Tatiana Rozental Burdman
- Laboratório de Hantaviroses e Rickettsioses, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil
| | | | | | | | - Marcelo Alves Pinto
- Laboratório de Desenvolvimento Tecnológico em Virologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil
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27
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Ramsay JD, Evanoff R, Mealey RH, Simpson EL. The prevalence of elevated gamma-glutamyltransferase and sorbitol dehydrogenase activity in racing Thoroughbreds and their associations with viral infection. Equine Vet J 2019; 51:738-742. [PMID: 30849186 DOI: 10.1111/evj.13092] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2018] [Accepted: 03/02/2019] [Indexed: 12/18/2022]
Abstract
BACKGROUND In racehorses, serum gamma-glutamyltransferase (GGT) activity is positively correlated with cumulative days in training and, when ≥100 IU/L, has been associated with poor performance. The prevalence of increased GGT activity in North American Thoroughbreds and its aetiopathogenesis are unknown. Four emerging viruses, pegivirus E (PgV E; equine pegivirus), hepacivirus A (HcV A; equine hepacivirus), pegivirus D (PgV D; Theiler's disease virus), and equine parvovirus-hepatitis (EqPV-H) have been identified in horses with clinical and subclinical hepatopathy. Available prevalence data indicate these viruses may commonly infect racehorses and contribute to increased liver enzyme activity in this population. OBJECTIVES To investigate the association between viral infection and increased liver enzyme activity in racing Thoroughbreds. STUDY DESIGN Cross-sectional study. METHODS Prerace blood samples were collected from 802 Thoroughbreds and tested for GGT and sorbitol dehydrogenase (SDH) activity, and the presence of PgV E, HcV A, PgV D and EqPV-H nucleic acid. RESULTS Increased SDH and/or GGT were detected in 56.2% of the 802 serum samples. The infection prevalence and relative risk (RR) of having concurrently increased liver enzyme activity were: PgV E = 18.2% (RR = 0.820, 95% CI = 0.662-0.978, P = 0.03), HcV A = 2.5% (RR = 1.132, 95% CI = 0.719-1.466, P = 0.6), PgV D = 0.5% (RR = 0.875, 95% CI = 0.165-1.598, P>0.9), EqPV-H = 2.9% (RR = 0.916, 95% CI = 0.564-1.266, P = 0.7). MAIN LIMITATIONS Longitudinal samples were not tested. CONCLUSIONS While viral infection was common among Thoroughbreds in this study, infection did not explain the high prevalence of increased liver enzyme activity. In fact, PgV E infection was associated with a reduced risk of having increased liver enzyme activity, indicating PgV E is unlikely to be a cause of hepatitis in horses. Importantly, like GGT, increased SDH activity was highly prevalent in this study, and provides additional evidence that hepatocellular injury was occurring in these horses.
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Affiliation(s)
- J D Ramsay
- Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA.,Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, Washington, USA
| | - R Evanoff
- Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA
| | - R H Mealey
- Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA
| | - E L Simpson
- Equine Medical & Surgical Group, Arcadia, California, USA
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28
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Rasche A, Sander AL, Corman VM, Drexler JF. Evolutionary biology of human hepatitis viruses. J Hepatol 2019; 70:501-520. [PMID: 30472320 PMCID: PMC7114834 DOI: 10.1016/j.jhep.2018.11.010] [Citation(s) in RCA: 50] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/22/2018] [Revised: 11/09/2018] [Accepted: 11/10/2018] [Indexed: 02/06/2023]
Abstract
Hepatitis viruses are major threats to human health. During the last decade, highly diverse viruses related to human hepatitis viruses were found in animals other than primates. Herein, we describe both surprising conservation and striking differences of the unique biological properties and infection patterns of human hepatitis viruses and their animal homologues, including transmission routes, liver tropism, oncogenesis, chronicity, pathogenesis and envelopment. We discuss the potential for translation of newly discovered hepatitis viruses into preclinical animal models for drug testing, studies on pathogenesis and vaccine development. Finally, we re-evaluate the evolutionary origins of human hepatitis viruses and discuss the past and present zoonotic potential of their animal homologues.
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Affiliation(s)
- Andrea Rasche
- Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Virology, 10117 Berlin, Germany,German Center for Infection Research (DZIF), Germany
| | - Anna-Lena Sander
- Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Virology, 10117 Berlin, Germany
| | - Victor Max Corman
- Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Virology, 10117 Berlin, Germany,German Center for Infection Research (DZIF), Germany
| | - Jan Felix Drexler
- Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Virology, 10117 Berlin, Germany; German Center for Infection Research (DZIF), Germany.
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29
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Divers TJ, Tennant BC, Kumar A, McDonough S, Cullen J, Bhuva N, Jain K, Chauhan LS, Scheel TKH, Lipkin WI, Laverack M, Trivedi S, Srinivasa S, Beard L, Rice CM, Burbelo PD, Renshaw RW, Dubovi E, Kapoor A. New Parvovirus Associated with Serum Hepatitis in Horses after Inoculation of Common Biological Product. Emerg Infect Dis 2019; 24:303-310. [PMID: 29350162 PMCID: PMC5782890 DOI: 10.3201/eid2402.171031] [Citation(s) in RCA: 65] [Impact Index Per Article: 10.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023] Open
Abstract
Equine serum hepatitis (i.e., Theiler's disease) is a serious and often life-threatening disease of unknown etiology that affects horses. A horse in Nebraska, USA, with serum hepatitis died 65 days after treatment with equine-origin tetanus antitoxin. We identified an unknown parvovirus in serum and liver of the dead horse and in the administered antitoxin. The equine parvovirus-hepatitis (EqPV-H) shares <50% protein identity with its phylogenetic relatives of the genus Copiparvovirus. Next, we experimentally infected 2 horses using a tetanus antitoxin contaminated with EqPV-H. Viremia developed, the horses seroconverted, and acute hepatitis developed that was confirmed by clinical, biochemical, and histopathologic testing. We also determined that EqPV-H is an endemic infection because, in a cohort of 100 clinically normal adult horses, 13 were viremic and 15 were seropositive. We identified a new virus associated with equine serum hepatitis and confirmed its pathogenicity and transmissibility through contaminated biological products.
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30
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Tomlinson JE, Kapoor A, Kumar A, Tennant BC, Laverack MA, Beard L, Delph K, Davis E, Schott Ii H, Lascola K, Holbrook TC, Johnson P, Taylor SD, McKenzie E, Carter-Arnold J, Setlakwe E, Fultz L, Brakenhoff J, Ruby R, Trivedi S, Van de Walle GR, Renshaw RW, Dubovi EJ, Divers TJ. Viral testing of 18 consecutive cases of equine serum hepatitis: A prospective study (2014-2018). J Vet Intern Med 2018; 33:251-257. [PMID: 30520162 PMCID: PMC6335536 DOI: 10.1111/jvim.15368] [Citation(s) in RCA: 43] [Impact Index Per Article: 6.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2018] [Accepted: 10/24/2018] [Indexed: 02/06/2023] Open
Abstract
Background Three flaviviruses (equine pegivirus [EPgV]; Theiler's disease–associated virus [TDAV]; non‐primate hepacivirus [NPHV]) and equine parvovirus (EqPV‐H) are present in equine blood products; the TDAV, NPHV, and EqPV‐H have been suggested as potential causes of serum hepatitis. Objective To determine the prevalence of these viruses in horses with equine serum hepatitis. Animals Eighteen horses diagnosed with serum hepatitis, enrolled from US referral hospitals. Methods In the prospective case study, liver, serum, or both samples were tested for EPgV, TDAV, NPHV, and EqPV‐H by PCR. Results Both liver tissue and serum were tested for 6 cases, serum only for 8 cases, and liver only for 4 cases. Twelve horses received tetanus antitoxin (TAT) 4‐12.7 weeks (median = 8 weeks), 3 horses received commercial equine plasma 6‐8.6 weeks, and 3 horses received allogenic stem cells 6.4‐7.6 weeks before the onset of hepatic failure. All samples were TDAV negative. Two of 14 serum samples were NPHV‐positive. Six of 14 serum samples were EPgV‐positive. All liver samples were NPHV‐negative and EPgV‐negative. EqPV‐H was detected in the serum (N = 8), liver (N = 4), or both samples (N = 6) of all 18 cases. The TAT of the same lot number was available for virologic testing in 10 of 12 TAT‐associated cases, and all 10 samples were EqPV‐H positive. Conclusions and Clinical Importance We demonstrated EqPV‐H in 18 consecutive cases of serum hepatitis. EPgV, TDAV, and NPHV were not consistently present. This information should encourage blood product manufacturers to test for EqPV‐H and eliminate EqPV‐H–infected horses from their donor herds.
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Affiliation(s)
- Joy E Tomlinson
- Department of Microbiology and Immunology, Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York
| | - Amit Kapoor
- Center for Vaccines and Immunity, Research Institute at Nationwide Children's Hospital, Columbus, Ohio
| | - Arvind Kumar
- Center for Vaccines and Immunity, Research Institute at Nationwide Children's Hospital, Columbus, Ohio
| | - Bud C Tennant
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York
| | - Melissa A Laverack
- New York State Animal Health Diagnostic Center, Cornell University, Ithaca, New York
| | - Laurie Beard
- Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas
| | - Katie Delph
- Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas
| | - Elizabeth Davis
- Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas
| | - Harold Schott Ii
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan
| | - Kara Lascola
- Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, Illinois
| | - Todd C Holbrook
- Department of Veterinary Clinical Sciences, Oklahoma State University, Stillwater, Oklahoma
| | - Philip Johnson
- Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, Missouri
| | - Sandra D Taylor
- Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, Indiana
| | - Erica McKenzie
- Department of Clinical Sciences, Oregon State University, Corvallis, Oregon
| | | | | | - Lisa Fultz
- Equine Medicine Specialists of South Florida, Wellington, Florida
| | | | - Rebecca Ruby
- Lloyd Veterinary Medical Center, Iowa State University, Ames, Iowa
| | - Sheetal Trivedi
- Center for Vaccines and Immunity, Research Institute at Nationwide Children's Hospital, Columbus, Ohio
| | - Gerlinde R Van de Walle
- Department of Microbiology and Immunology, Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York
| | - Randall W Renshaw
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York
| | - Edward J Dubovi
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York
| | - Thomas J Divers
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York
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Schlottau K, Fereidouni S, Beer M, Hoffmann B. Molecular identification and characterization of nonprimate hepaciviruses in equines. Arch Virol 2018; 164:391-400. [PMID: 30361815 DOI: 10.1007/s00705-018-4077-2] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/03/2018] [Accepted: 10/01/2018] [Indexed: 02/06/2023]
Abstract
Hepatitis C virus (HCV) is a positive-sense RNA virus belonging to the genus Hepacivirus, family Flaviviridae. Its genome has a length of 9.6 kb and encodes a single polyprotein flanked by two untranslated regions. HCV can cause liver cirrhosis and hepatocellular carcinoma, and approximately 2% of the world's population is chronically infected. The investigation of pathogenesis is complicated due to the lack of an animal model. The origin of this virus remains unclear, but in the last few years, relatives of HCV were initially identified in dogs and later in horses, rodents, bats and Old World monkeys. Non-primate hepacivirus (NPHV), which infects dogs and horses, is the closest relative to HCV. We established a pan-reactive "panHepaci"-RT-qPCR assay, which is able to detect human HCV as well as equine NPHV, and additionally, an equine-specific "equHepaci"-RT-qPCR for confirmation of positive results. Serum samples from 1158 clinically inconspicuous horses from Germany and several samples from other mammalian species were screened. We found 2.4% of the horses positive for hepacivirus RNA, and furthermore, the "panHepaci"-RT-qPCR assay also detected a hepacivirus in a donkey from Egypt. This virus had only 78% sequence identity in the E2 gene when compared to other known NPHVs. The established method could be useful for screening purposes, since it is likely that related hepaciviruses also occur in other species.
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Affiliation(s)
- Kore Schlottau
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493, Greifswald-Insel Riems, Germany
| | | | - Martin Beer
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493, Greifswald-Insel Riems, Germany
| | - Bernd Hoffmann
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493, Greifswald-Insel Riems, Germany.
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Identification and genetic characterization of a novel parvovirus associated with serum hepatitis in horses in China. Emerg Microbes Infect 2018; 7:170. [PMID: 30348940 PMCID: PMC6198012 DOI: 10.1038/s41426-018-0174-2] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/21/2018] [Revised: 09/13/2018] [Accepted: 09/17/2018] [Indexed: 12/14/2022]
Abstract
A novel equine parvovirus, equine parvovirus-hepatitis (EqPV-H), was first discovered in a horse that died of equine serum hepatitis in the USA in 2018. EqPV-H was shown to be a novel etiological agent associated with equine serum hepatitis. Following this initial report, no additional studies on EqPV-H have been published. In this study, a total of 143 serum samples were collected from racehorses at 5 separate farms in China and were analyzed to detect EqPV-H DNA via nested PCR. The results indicated a high prevalence of EqPV-H (11.9%, 17/143) in the studied animals. In addition, a remarkably high coinfection rate (58.8%, 10/17) with 2 equine flaviviruses (equine hepacivirus and equine pegivirus) was observed in the EqPV-H positive equines. However, all equines tested negative for Theiler’s disease-associated virus, an etiological agent associated with equine serum hepatitis. The genomes of six field EqPV-H strains were sequenced and analyzed, with the results indicating that the Chinese EqPV-H strains have low genetic diversity and high genetic similarity with the USA EqPV-H strain BCT-01. A phylogenetic analysis demonstrated that the Chinese EqPV-H strains clustered with BCT-01 in the genus Copiparvovirus but were distantly related to another equine parvovirus identified in horse cerebrospinal fluid. In addition, liver enzyme levels were detected in the EqPV-H positive serum samples, and all the values were in the normal range, indicating that infection can occur without concurrent liver disease. This study will promote an understanding of the geographical distribution, genetic diversity, and pathogenicity of EqPV-H.
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Kopper JJ, Schott HC, Divers TJ, Mullaney T, Huang L, Noland E, Smedley R. Theiler's disease associated with administration of tetanus antitoxin contaminated with nonprimate (equine) hepacivirus and equine parvovirus‐hepatitis virus. EQUINE VET EDUC 2018. [DOI: 10.1111/eve.12999] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Affiliation(s)
- J. J. Kopper
- Large Animal Clinical Sciences Michigan State University East Lansing Michigan USA
| | - H. C. Schott
- Large Animal Clinical Sciences Michigan State University East Lansing Michigan USA
| | - T. J. Divers
- Clinical Sciences Cornell University Ithaca New York USA
| | - T. Mullaney
- Pathobiology and Diagnostic Investigation Michigan State University East Lansing Michigan USA
| | - L. Huang
- Pathobiology and Diagnostic Investigation Michigan State University East Lansing Michigan USA
| | - E. Noland
- Pathobiology and Diagnostic Investigation Michigan State University East Lansing Michigan USA
| | - R. Smedley
- Veterinary Diagnostic Laboratory Michigan State University East Lansing Michigan USA
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Hepacivirus A Infection in Horses Defines Distinct Envelope Hypervariable Regions and Elucidates Potential Roles of Viral Strain and Adaptive Immune Status in Determining Envelope Diversity and Infection Outcome. J Virol 2018; 92:JVI.00314-18. [PMID: 29976666 DOI: 10.1128/jvi.00314-18] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2018] [Accepted: 06/12/2018] [Indexed: 12/12/2022] Open
Abstract
Hepacivirus A (also known as nonprimate hepacivirus and equine hepacivirus) is a hepatotropic virus that can cause both transient and persistent infections in horses. The evolution of intrahost viral populations (quasispecies) has not been studied in detail for hepacivirus A, and its roles in immune evasion and persistence are unknown. To address these knowledge gaps, we first evaluated the envelope gene (E1 and E2) diversity of two different hepacivirus A strains (WSU and CU) in longitudinal blood samples from experimentally infected adult horses, juvenile horses (foals), and foals with severe combined immunodeficiency (SCID). Persistent infection with the WSU strain was associated with significantly greater quasispecies diversity than that observed in horses who spontaneously cleared infection (P = 0.0002) or in SCID foals (P < 0.0001). In contrast, the CU strain was able to persist despite significantly lower (P < 0.0001) and relatively static envelope diversity. These findings indicate that envelope diversity is a poor predictor of hepacivirus A infection outcomes and could be dependent on strain-specific factors. Next, entropy analysis was performed on all E1/E2 genes entered into GenBank. This analysis defined three novel hypervariable regions (HVRs) in E2, at residues 391 to 402 (HVR1), 450 to 461 (HVR2), and 550 to 562 (HVR3). For the experimentally infected horses, entropy analysis focusing on the HVRs demonstrated that these regions were under increased selective pressure during persistent infection. Increased diversity in the HVRs was also temporally associated with seroconversion in some horses, suggesting that these regions may be targets of neutralizing antibody and may play a role in immune evasion.IMPORTANCE Hepacivirus C (hepatitis C virus) is estimated to infect 150 million people worldwide and is a leading cause of cirrhosis and hepatocellular carcinoma. In contrast, its closest relative, hepacivirus A, causes relatively mild disease in horses and is frequently cleared. The relationship between quasispecies evolution and infection outcome has not been explored for hepacivirus A. To address this knowledge gap, we examined envelope gene diversity in horses with resolving and persistent infections. Interestingly, two strain-specific patterns of quasispecies diversity emerged. Persistence of the WSU strain was associated with increased quasispecies diversity and the accumulation of amino acid changes within three novel hypervariable regions following seroconversion. These findings provided evidence that envelope gene mutation is influenced by adaptive immune pressure and may contribute to hepacivirus persistence. However, the CU strain persisted despite relative evolutionary stasis, suggesting that some hepacivirus strains may use alternative mechanisms to persist in the host.
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Lu G, Huang J, Li S. Genomic sequencing and characterization of Theiler's disease-associated virus identified in commercial equine sera in China. J Gen Virol 2018; 99:1221-1226. [PMID: 30041711 DOI: 10.1099/jgv.0.001114] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
Theiler's disease-associated virus (TDAV) could be the aetiological agent of Theiler's disease. Horses experimentally inoculated with equine plasma containing TDAV develop acute and chronic infections with viraemia. Since its first identification in 2013, TDAV has not been detected in equines in the epidemiological studies that have been conducted. Until now, only one genome sequence of TDAV (HorseA1_serum) had been obtained. In this study, we sequenced the genome of four TDAV strains (A/China, F/China, H/USA and I/USA) in commercial equine sera used for cell culture propagation in China using three rounds of RT-PCR. The PCR primers were designed based on the HorseA1_serum genome sequence. All four TDAV strains had a polyprotein gene that was 9567 nt long, the same nucleotide length as the polyprotein gene of HorseA1_serum. Sequence analysis demonstrated the genetic diversity of TDAV. The nucleotide similarity of the polyprotein genes of the TDAV strains ranged between 90.3 and 93.6 %, with a high amino acid similarity that ranged from 98.2 to 98.8 %. Phylogenetic analysis using the polyprotein gene showed that A/China, F/China, H/USA and I/USA were clustered together with HorseA1_serum in the genus Pegivirus D. This study enriches our knowledge of the genetic diversity of TDAV.
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Affiliation(s)
- Gang Lu
- 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, PR China
- 2Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, PR China
- 3Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong Province, PR China
| | - Ji Huang
- 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, PR China
- 2Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, PR China
- 3Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong Province, PR China
| | - Shoujun Li
- 2Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, PR China
- 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, PR China
- 3Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong Province, PR China
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Badenhorst M, Tegtmeyer B, Todt D, Guthrie A, Feige K, Campe A, Steinmann E, Cavalleri JMV. First detection and frequent occurrence of Equine Hepacivirus in horses on the African continent. Vet Microbiol 2018; 223:51-58. [PMID: 30173752 DOI: 10.1016/j.vetmic.2018.07.015] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/13/2018] [Revised: 07/20/2018] [Accepted: 07/20/2018] [Indexed: 01/30/2023]
Abstract
Since the discovery of equine hepacivirus (EqHV) in 2011, the virus has been detected in horse populations from more than twelve countries across five continents. EqHV seroprevalence has been reported to be as high as 61.8% and EqHV ribonucleic acid (RNA) prevalence to range between 0.9% and 34.1%. Molecular and serological indications of EqHV infection have never been reported in equids on the African continent. Therefore, investigation of EqHV prevalence in South African horses and subsequent viral genetic characterization contribute to a better understanding of the global epidemiology of this virus. In a cross-sectional study, serum samples from 454 Thoroughbred foals (aged 58-183 days) were analysed for anti-EqHV non-structural protein 3 (NS3)-specific antibodies (abs) with a luciferase immunoprecipitation system (LIPS) and for EqHV RNA by quantitative real-time polymerase chain reaction (qRT-PCR). Farms of origin (n = 26) were situated in South Africa's Western Cape Province. The associations between EqHV infection state and farm of origin, foal gender and foal age were subsequently described. Furthermore, nested PCRs were performed on parts of the 5'UTR, NS3 and NS5B genes of 17 samples. Samples were sequenced and phylogenetic analyses were conducted. The population's seroprevalence was 83.70% and RNA was detected in 7.93% of samples. Increasing foal age was associated with decreasing ab prevalence and increasing prevalence of EqHV RNA. Sequences from South African EqHV strains did not show in-depth clustering with published sequences of EqHV isolates from particular continents. In conclusion, EqHV is present in the South African Thoroughbred population and appears more prevalent than reported in other horse populations worldwide.
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Affiliation(s)
- Marcha Badenhorst
- Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, Pretoria, South Africa; Department for Companion Animals and Horses, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210, Vienna, Austria
| | - Birthe Tegtmeyer
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH) - Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Strasse 7, 30625, Hannover, Germany
| | - Daniel Todt
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH) - Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Strasse 7, 30625, Hannover, Germany; Department of Molecular and Medical Virology, Ruhr-University Bochum, 44801, Bochum, Germany
| | - Alan Guthrie
- Equine Research Centre, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, Pretoria, South Africa
| | - Karsten Feige
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559, Hannover, Germany
| | - Amely Campe
- Department of Biometry, Epidemiology and Information Processing (IBEI), WHO-Collaborating Centre for Research and Training for Health at the Human-Animal-Environment Interface, University of Veterinary Medicine Hannover, Foundation, Bünteweg 2, 30559, Hannover, Germany
| | - Eike Steinmann
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH) - Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Strasse 7, 30625, Hannover, Germany; Department of Molecular and Medical Virology, Ruhr-University Bochum, 44801, Bochum, Germany.
| | - Jessika M V Cavalleri
- Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, Pretoria, South Africa; Department for Companion Animals and Horses, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210, Vienna, Austria; Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559, Hannover, Germany.
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Schlottau K, Wernike K, Forth L, Holsteg M, Höper D, Beer M, Hoffmann B. Presence of two different bovine hepacivirus clusters in Germany. Transbound Emerg Dis 2018; 65:1705-1711. [PMID: 29971937 DOI: 10.1111/tbed.12930] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/23/2018] [Revised: 05/25/2018] [Accepted: 05/28/2018] [Indexed: 12/24/2022]
Abstract
During the last years, genetic information of hepaciviruses (family Flaviviridae), whose type species is the human hepatitis C virus, was detected in a wide range of primates and non-primate vertebrates. Here, samples collected from 263 German cattle kept in 22 different holdings were analysed for the presence of hepacivirus N (syn. bovine hepacivirus; BovHepV). One hundred eighty-six cattle that suffered from unspecific clinical signs such as fever and a reduced milk yield as well as 77 apparently healthy animals were included. A total of 39 cattle (14.8%) tested positive for BovHepV by real-time RT-PCR, but a correlation between clinical signs and virus infection could not be found. From 31 of the virus-positive samples, sequences of the NS3 coding region were generated and from two samples, viral sequences of the complete coding region were produced and compared to further European and African BovHepV sequences. Based on the NS3 genomic region, two distinct German BovHepV clusters were identified which differed between each other up to 20% at the nucleotide level, the diversity within the individual clusters reached up to 10%. Based on the full-length sequences, the newly detected virus variants group together with further German and African viruses in a sister relationship to other hepaciviruses from primates and further mammalians, but form distinct clusters within the BovHepV branch. In conclusion, highly diverse hepaciviruses were detected in German cattle further expanding the known phylogenetic diversity of the genus Hepacivirus.
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Affiliation(s)
- Kore Schlottau
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
| | - Kerstin Wernike
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
| | - Leonie Forth
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
| | - Mark Holsteg
- Chamber of Agriculture for North Rhine-Westphalia, Bovine Health Service, Bad Sassendorf, Germany
| | - Dirk Höper
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
| | - Martin Beer
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
| | - Bernd Hoffmann
- Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Greifswald - Insel Riems, Germany
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Tang W, Zhu N, Wang H, Gao Y, Wan Z, Cai Q, Yu S, Tang S. Identification and genetic characterization of equine Pegivirus in China. J Gen Virol 2018; 99:768-776. [PMID: 29658859 DOI: 10.1099/jgv.0.001063] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
In 2013, two new viruses, equine pegivirus (EPgV) and Theiler's disease-associated virus (TDAV), both belonging to the genus Pegivirus within the family Flaviviridae, were identified. To investigate the geographical distribution and genetic diversity of these two viruses in China, we screened EPgV and TDAV infection in imported race horses and Chinese work horses by using reverse-transcription polymerase chain reaction (RT-PCR). EPgV was detected in 10.8 % (8/74) of the total horses tested, with a prevalence of 5.8 and 22.7 % in the race horses and work horses, respectively. No TDAV infection was found. A near full-length genome sequence of EPgV was obtained that showed an identity of 89.5-90.6 % at the nucleotide level and 98.1-98.3 % at the amino acid level with an American strain, C0035, and another Chinese strain, LW/216, respectively. Phylogenetic analysis showed two different clusters of the sequences from the race horses and work horses, indicating a difference in virus origin. Our results demonstrated a higher positive rate of EPgV in the Chinese work horses than in the imported race horses, a moderate genetic diversity of EPgV strains worldwide and possibly no liver pathogenesis for EPgV infection.
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Affiliation(s)
- Weiping Tang
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Naling Zhu
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Haiying Wang
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Youwen Gao
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Zhengwei Wan
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Qundi Cai
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Shouyi Yu
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Shixing Tang
- Guangdong Provincial Key Laboratory of Tropical Disease Research, Southern Medical University, Guangzhou, Guangdong, PR China
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Van Nguyen D, Van Nguyen C, Bonsall D, Ngo TT, Carrique-Mas J, Pham AH, Bryant JE, Thwaites G, Baker S, Woolhouse M, Simmonds P. Detection and Characterization of Homologues of Human Hepatitis Viruses and Pegiviruses in Rodents and Bats in Vietnam. Viruses 2018; 10:v10030102. [PMID: 29495551 PMCID: PMC5869495 DOI: 10.3390/v10030102] [Citation(s) in RCA: 30] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/20/2018] [Revised: 02/19/2018] [Accepted: 02/23/2018] [Indexed: 12/20/2022] Open
Abstract
Rodents and bats are now widely recognised as important sources of zoonotic virus infections in other mammals, including humans. Numerous surveys have expanded our knowledge of diverse viruses in a range of rodent and bat species, including their origins, evolution, and range of hosts. In this study of pegivirus and human hepatitis-related viruses, liver and serum samples from Vietnamese rodents and bats were examined by PCR and sequencing. Nucleic acids homologous to human hepatitis B, C, E viruses were detected in liver samples of 2 (1.3%) of 157 bats, 38 (8.1%), and 14 (3%) of 470 rodents, respectively. Hepacivirus-like viruses were frequently detected (42.7%) in the bamboo rat, Rhizomys pruinosus, while pegivirus RNA was only evident in 2 (0.3%) of 638 rodent serum samples. Complete or near-complete genome sequences of HBV, HEV and pegivirus homologues closely resembled those previously reported from rodents and bats. However, complete coding region sequences of the rodent hepacivirus-like viruses substantially diverged from all of the currently classified variants and potentially represent a new species in the Hepacivirus genus. Of the viruses identified, their routes of transmission and potential to establish zoonoses remain to be determined.
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MESH Headings
- Animals
- Chiroptera/virology
- Genome, Viral
- Hepatitis Viruses/classification
- Hepatitis Viruses/genetics
- Hepatitis, Viral, Animal/diagnosis
- Hepatitis, Viral, Animal/epidemiology
- Hepatitis, Viral, Animal/virology
- Hepatitis, Viral, Human/diagnosis
- Hepatitis, Viral, Human/epidemiology
- Hepatitis, Viral, Human/virology
- Humans
- Phylogeny
- Public Health Surveillance
- RNA, Viral
- Rodentia/virology
- Vietnam/epidemiology
- Zoonoses/epidemiology
- Zoonoses/virology
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Affiliation(s)
- Dung Van Nguyen
- Nuffield Department of Medicine, University of Oxford, Oxford OX1 3SY, UK.
| | - Cuong Van Nguyen
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
| | - David Bonsall
- Nuffield Department of Medicine, University of Oxford, Oxford OX1 3SY, UK.
| | - Tue Tri Ngo
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
| | - Juan Carrique-Mas
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
- Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, Oxford University, Oxford OX3 7FZ, UK.
| | - Anh Hong Pham
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
| | - Juliet E Bryant
- Fondation Mérieux, Centre International de Recherche en Infectiologie (CIRI), 69365 Lyon CEDEX 07, France.
| | - Guy Thwaites
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
- Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, Oxford University, Oxford OX3 7FZ, UK.
| | - Stephen Baker
- Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit, Ho Chi Minh City 700000, Vietnam.
- Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, Oxford University, Oxford OX3 7FZ, UK.
- The London School of Hygiene & Tropical Medicine, London WC1E 7HT, UK.
| | - Mark Woolhouse
- Centre for Immunity, Infection and Evolution, University of Edinburgh, Edinburgh EH9 3FL, UK.
| | - Peter Simmonds
- Nuffield Department of Medicine, University of Oxford, Oxford OX1 3SY, UK.
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41
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Lu G, Huang J, Yang Q, Xu H, Wu P, Fu C, Li S. Identification and genetic characterization of hepacivirus and pegivirus in commercial equine serum products in China. PLoS One 2017; 12:e0189208. [PMID: 29216266 PMCID: PMC5720783 DOI: 10.1371/journal.pone.0189208] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2017] [Accepted: 11/21/2017] [Indexed: 01/29/2023] Open
Abstract
Equine hepacivirus (EqHV), equine pegivirus (EPgV) and Theiler’s disease-associated virus (TDAV) are three novel equine viruses in the family Flaviviridae. EqHV and EPgV have been identified to circulate in the equine population worldwide, whereas TDAV has not been detected in equines since the first reported case. To date, no studies have focused on investigating EqHV, EPgV and TDAV in commercial equine sera or equine blood products in China. Considering the potential threat of EqHV, EPgV and TDAV to biosecurity and considering their possible influences on research results, equine sera for cell culture propagation and pregnant mare serum gonadotropin (PMSG) were purchased from different companies in China and investigated for EqHV, EPgV and TDAV in this study. By performing nested PCR or two rounds of PCR targeting the viral NS3 gene, four serum samples were confirmed to be EqHV-, EPgV-, or TDAV-RNA positive; all of the PMSG samples were negative for these three viruses. Subsequent sequencing results indicated that the serum samples contained multiple viral variants of EqHV, EPgV or TDAV, and a genetic analysis based on the partial NS3 gene of the three equine viruses was performed. Our study is the first to confirm the presence of EqHV, EPgV and TDAV in equine sera for cell culture propagation that is commercially available in China and provides the first demonstration of the presence of TDAV in China.
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Affiliation(s)
- Gang Lu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Ji Huang
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Qiliang Yang
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Haibin Xu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Peixin Wu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Cheng Fu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
| | - Shoujun Li
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, People’s Republic of China
- Guangdong Technological Engineering Research Center for Pet Guangzhou, Guangdong Province, People’s Republic of China
- * E-mail:
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42
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Abstract
Hepaciviruses and pegiviruses constitute two closely related sister genera of the family Flaviviridae. In the past five years, the known phylogenetic diversity of the hepacivirus genera has absolutely exploded. What was once an isolated infection in humans (and possibly other primates) has now expanded to include horses, rodents, bats, colobus monkeys, cows, and, most recently, catsharks, shedding new light on the genetic diversity and host range of hepaciviruses. Interestingly, despite the identification of these many animal and primate hepaciviruses, the equine hepaciviruses remain the closest genetic relatives of the human hepaciviruses, providing an intriguing clue to the zoonotic source of hepatitis C virus. This review summarizes the significance of these studies and discusses current thinking about the origin and evolution of the animal hepaciviruses as well as their potential usage as surrogate models for the study of hepatitis C virus.
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Affiliation(s)
- Alex S Hartlage
- Center for Vaccines and Immunity, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205;
| | - John M Cullen
- North Carolina State University College of Veterinary Medicine, Raleigh, North Carolina 27606
| | - Amit Kapoor
- Center for Vaccines and Immunity, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205; .,Department of Pediatrics, College of Medicine and Public Health, Ohio State University, Columbus, Ohio 43210
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43
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Lu G, Fu C, Huang J, Xu H, Wu P, Ping X, Li S. Molecular characterization of a genetically divergent equine pegivirus strain identified in China. Arch Virol 2017; 163:249-252. [PMID: 29094242 DOI: 10.1007/s00705-017-3602-z] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2017] [Accepted: 08/31/2017] [Indexed: 01/24/2023]
Abstract
Equine pegivirus (EPgV) is a newly discovered equine virus, which is taxonomically classified in the Pegivirus genus of the Flaviviridae family. Until now, only the complete genome sequence of the first reported EPgV strain, from the USA (strain name: C0035) is available on online databases. Considering this, horse serum samples were collected from horses in China and screened for EPgV RNA by RT-PCR. One EPgV strain, LW/2016, was obtained and its near-complete genome sequence was acquired by standard PCR. Further analysis of its nucleotide sequence indicates LW/2016 is genetically divergent from C0035, with a nucleotide identity of 89.02%. These two viruses clustered into two independent branches following phylogenetic analysis of on the NS3 and NS5B genes. To our knowledge, this is the first report of genetic divergence in the EPgV genome.
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Affiliation(s)
- Gang Lu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Cheng Fu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Ji Huang
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Haibin Xu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Peixin Wu
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Xiaokun Ping
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China.,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China.,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China
| | - Shoujun Li
- College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, People's Republic of China. .,Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong, People's Republic of China. .,Guangdong Technological Engineering Research Center for Pet, Guangzhou, Guangdong, People's Republic of China.
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44
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Elia G, Lanave G, Lorusso E, Parisi A, Trotta A, Buono R, Martella V, Decaro N, Buonavoglia C. Equine hepacivirus persistent infection in a horse with chronic wasting. Transbound Emerg Dis 2017; 64:1354-1358. [PMID: 28707785 DOI: 10.1111/tbed.12679] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2017] [Indexed: 12/29/2022]
Abstract
Equine hepacivirus is the closest homologue of hepatitis C virus. Limited data on the clinical features of this infection are available. We report the identification of a horse with high-titre viremia by equine hepacivirus. Over a 15-month follow-up, the clinical signs and the viremic status persisted, suggesting a chronic evolution.
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Affiliation(s)
- G Elia
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - G Lanave
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - E Lorusso
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - A Parisi
- Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, Sezione di Putignano, Putignano, Bari, Italy
| | - A Trotta
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - R Buono
- Azienda Sanitaria Locale Bari, Bari, Italy
| | - V Martella
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - N Decaro
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
| | - C Buonavoglia
- Department of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy
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45
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Elia G, Lanave G, Lorusso E, Parisi A, Cavaliere N, Patruno G, Terregino C, Decaro N, Martella V, Buonavoglia C. Identification and genetic characterization of equine hepaciviruses in Italy. Vet Microbiol 2017; 207:239-247. [PMID: 28757030 DOI: 10.1016/j.vetmic.2017.07.004] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/13/2017] [Revised: 07/04/2017] [Accepted: 07/04/2017] [Indexed: 01/26/2023]
Abstract
Viruses similar to human hepatitis C virus, hepaciviruses, have been identified in various animal species. Equine hepacivirus (EqHV) is the closest relative of human hepaciviruses. Although detected worldwide, information on EqHV epidemiology, genetic diversity and pathogenicity is still limited. In this study we investigated the prevalence and genetic diversity of EqHV in Italian equids. The RNA of EqHV was detected in 91/1932 sera (4.7%) whilst it was not detectable in 134 donkey sera screened by a TaqMan-based quantitative assay. Upon sequencing and phylogenetic analysis of genomic portions located in the NS5B, 5'UTR and NS3 genes, the Italian EqHV strains segregated into two distinct clades that are also co-circulating globally, without apparent geographic restrictions.
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Affiliation(s)
- Gabriella Elia
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy.
| | - Gianvito Lanave
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy
| | - Eleonora Lorusso
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy
| | - Antonio Parisi
- Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, Foggia, Italy
| | - Nicola Cavaliere
- Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, Foggia, Italy
| | | | - Calogero Terregino
- Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Padova, Italy
| | - Nicola Decaro
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy
| | - Vito Martella
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy
| | - Canio Buonavoglia
- Dipartimento di Medicina Veterinaria, Università Aldo Moro di Bari, Valenzano, Italy
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46
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Abstract
Ben Sturgeon discusses Theiler's disease, one of the most common causes of acute hepatitis in horses.
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Affiliation(s)
- Ben Sturgeon
- Greenside Veterinary Practice, Greenside Farm, St Boswells, Melrose TD6 0AJ
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47
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Reichert C, Campe A, Walter S, Pfaender S, Welsch K, Ruddat I, Sieme H, Feige K, Steinmann E, Cavalleri JMV. Frequent occurrence of nonprimate hepacivirus infections in Thoroughbred breeding horses - A cross-sectional study for the occurrence of infections and potential risk factors. Vet Microbiol 2017; 203:315-322. [PMID: 28619163 DOI: 10.1016/j.vetmic.2017.03.030] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/08/2017] [Revised: 03/19/2017] [Accepted: 03/20/2017] [Indexed: 02/07/2023]
Abstract
Recently, several new hepaciviruses have been identified of which the nonprimate hepacivirus (NPHV) - the closest relative to hepatitis C virus (HCV) discovered to date - is highly prevalent in horses. However, potential risk factors for the transmission of NPHV among horses remain still unknown. Therefore, the objective of this study was to investigate the occurrence of NPHV infections in Thoroughbreds in northern and western Germany and to identify potential risk factors associated with NPHV infections. Using a cross-sectional study design, a total of 733 serum samples from Thoroughbred broodmares and stallions from northern and western Germany were analyzed for the presence of anti-NPHV nonstructural protein 3 (NS3) antibodies and NPHV RNA using the luciferase immunoprecipitation system (LIPS) and a quantitative real-time PCR, respectively. Information regarding signalment, stud farm, breeding history and international transportation history of each horse were collected and evaluated. A frequent occurrence of NPHV was found in the study population with 453 seropositive horses (61.8%) and 134 horses (18.3%) carrying NPHV RNA. Furthermore, statistical analysis revealed that the probability of being infected decreased for horses with a transportation history with increasing age by 20% each year. For horses that stayed in Germany no association between age and infection could be observed. In conclusion, the high occurrence of NPHV infections in breeding Thoroughbreds suggests circulating NPHV infections, endemic herds or persistent shedding in these animals and revealed the association of age and international transportation as risk factor for NPHV infections.
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Affiliation(s)
- Claudia Reichert
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover, Germany.
| | - Amely Campe
- Department of Biometry, Epidemiology and Information Processing, WHO-Collaborating Center for Research and Training for Health at the Human-Animal-Environment Interface, University of Veterinary Medicine Hannover, Foundation, Bünteweg 2, 30559 Hannover, Germany.
| | - Stephanie Walter
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
| | - Stephanie Pfaender
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
| | - Kathrin Welsch
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
| | - Inga Ruddat
- Department of Biometry, Epidemiology and Information Processing, WHO-Collaborating Center for Research and Training for Health at the Human-Animal-Environment Interface, University of Veterinary Medicine Hannover, Foundation, Bünteweg 2, 30559 Hannover, Germany.
| | - Harald Sieme
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover, Germany.
| | - Karsten Feige
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover, Germany.
| | - Eike Steinmann
- Institute of Experimental Virology, TWINCORE Center for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Feodor-Lynen-Str. 7, 30625 Hannover, Germany.
| | - Jessika M V Cavalleri
- Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover, Germany.
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48
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Kim HS, Moon HW, Sung HW, Kwon HM. First identification and phylogenetic analysis of equine hepacivirus in Korea. INFECTION GENETICS AND EVOLUTION 2017; 49:268-272. [PMID: 28161473 DOI: 10.1016/j.meegid.2017.01.030] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/24/2016] [Revised: 01/25/2017] [Accepted: 01/28/2017] [Indexed: 12/11/2022]
Abstract
Non-primate hepacivirus (NPHV) corresponds a group of isolates recently characterized in horses and dogs that present similar genomic organization and are closely related to hepatitis C virus. Since canine hapacivirus, NPHV identified in dogs, was first discovered in dogs in the United States, equine hepacivirus (EqHV, NPHV identified in horses) has been identified in horses in several countries. However, no epidemiological studies have investigated EqHV in horses in Korea. In this study, a total of 74 (n=74) serum samples collected from horses in four regions of Korea were tested for EqHV RNA using nested RT-PCR. Overall, 14 samples were identified as positive (18.9%) and further analyzed according to gender, age, breed, and region. There were high positive rates in males, young horses, and Thoroughbreds; however, these rates differed regionally. Sequencing of the partial NS3 region of 12 samples and the polyprotein encoding regions of two samples positive for EqHV RNA revealed that the Korean EqHV isolates shared approximately 85.3-99.6% and 97.7-100% homology at the nucleotide and deduced amino acid level, respectively. Phylogenetic analysis revealed that the partial NS3 genes clustered with sequences previously reported as NPHV. Notably, sequences of EqHV detected in horses in the same region showed sequence divergence. The sequences of the polyprotein encoding region of two representative EqHVs shared 83.9% and 95.7% homology with each other at the nucleotide and deduced amino acid level, respectively. Comparison of the sequences of polyprotein encoding regions of Korean EqHV isolates and hepaciviruses from different hosts revealed that the NS3 and NS5B regions were most conserved among hepaciviruses. The results of the present study demonstrate that there is a high positive rate of EqHV in Korea and provide significant information regarding the geographical distribution and genetic variability of Korean EqHV isolates that will help improve global epidemiology of EqHV.
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Affiliation(s)
- Ho-Seong Kim
- Laboratory of Veterinary Microbiology, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, Republic of Korea
| | - Hyun-Woo Moon
- Laboratory of Veterinary Microbiology, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, Republic of Korea
| | - Haan Woo Sung
- Laboratory of Veterinary Microbiology, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, Republic of Korea
| | - Hyuk Moo Kwon
- Laboratory of Veterinary Microbiology, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, Republic of Korea.
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49
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Ramsay JD. Science-in-brief: Equine viral hepatitis. Equine Vet J 2016; 49:138-140. [PMID: 27995661 DOI: 10.1111/evj.12652] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2016] [Accepted: 11/04/2016] [Indexed: 12/22/2022]
Affiliation(s)
- J D Ramsay
- Department of Veterinary Microbiology and Pathology, and Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, Washington, USA
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50
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Differential Infection Patterns and Recent Evolutionary Origins of Equine Hepaciviruses in Donkeys. J Virol 2016; 91:JVI.01711-16. [PMID: 27795428 DOI: 10.1128/jvi.01711-16] [Citation(s) in RCA: 35] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2016] [Accepted: 10/13/2016] [Indexed: 12/13/2022] Open
Abstract
The hepatitis C virus (HCV) is a major human pathogen. Genetically related viruses in animals suggest a zoonotic origin of HCV. The closest relative of HCV is found in horses (termed equine hepacivirus [EqHV]). However, low EqHV genetic diversity implies relatively recent acquisition of EqHV by horses, making a derivation of HCV from EqHV unlikely. To unravel the EqHV evolutionary history within equid sister species, we analyzed 829 donkeys and 53 mules sampled in nine European, Asian, African, and American countries by molecular and serologic tools for EqHV infection. Antibodies were found in 278 animals (31.5%), and viral RNA was found in 3 animals (0.3%), all of which were simultaneously seropositive. A low RNA prevalence in spite of high seroprevalence suggests a predominance of acute infection, a possible difference from the mostly chronic hepacivirus infection pattern seen in horses and humans. Limitation of transmission due to short courses of infection may explain the existence of entirely seronegative groups of animals. Donkey and horse EqHV strains were paraphyletic and 97.5 to 98.2% identical in their translated polyprotein sequences, making virus/host cospeciation unlikely. Evolutionary reconstructions supported host switches of EqHV between horses and donkeys without the involvement of adaptive evolution. Global admixture of donkey and horse hepaciviruses was compatible with anthropogenic alterations of EqHV ecology. In summary, our findings do not support EqHV as the origin of the significantly more diversified HCV. Identification of a host system with predominantly acute hepacivirus infection may enable new insights into the chronic infection pattern associated with HCV. IMPORTANCE The evolutionary origins of the human hepatitis C virus (HCV) are unclear. The closest animal-associated relative of HCV occurs in horses (equine hepacivirus [EqHV]). The low EqHV genetic diversity implies a relatively recent acquisition of EqHV by horses, limiting the time span for potential horse-to-human infections in the past. Horses are genetically related to donkeys, and EqHV may have cospeciated with these host species. Here, we investigated a large panel of donkeys from various countries using serologic and molecular tools. We found EqHV to be globally widespread in donkeys and identify potential differences in EqHV infection patterns, with donkeys potentially showing enhanced EqHV clearance compared to horses. We provide strong evidence against EqHV cospeciation and for its capability to switch hosts among equines. Differential hepacivirus infection patterns in horses and donkeys may enable new insights into the chronic infection pattern associated with HCV.
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