DOCSLIB.ORG

Epigenetic Regulation of Frog Innate Immunity

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Epigenetic Regulation of Frog Innate Immunity bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Epigenetic regulation of frog innate immunity: Discovery of 2 frog microRNAs associated with antiviral responses and 3 ranavirus infection using a Xenopus laevis skin epithelial-like 4 cell line 5 6 Lauren A. Todd1, Maxwell P. Bui-Marinos1, and Barbara A. Katzenback1,* 7 8 1Department of Biology, Faculty of Science, University of Waterloo, 200 University Avenue 9 West, Waterloo, Ontario, Canada, N2L 3G1. 10 11 Email addresses: 12 [email protected] (L.A.T) 13 [email protected] (M.P.B-M) 14 [email protected] (B.A.K) 15 16 *Corresponding author 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 17 Abstract 18 19 Epigenetic regulators such as microRNAs are emerging as conserved regulators of innate 20 antiviral immunity in vertebrates, yet their roles in amphibian antiviral responses remain 21 uncharacterized. We profiled changes in microRNA expressions in the Xenopus laevis skin 22 epithelial-like cell line Xela DS2 in response to poly(I:C) – an analogue of double-stranded viral 23 RNA and inducer of type I interferons – or frog virus 3 (FV3), an immunoevasive virus 24 associated with amphibian mortality events. We sequenced small RNA libraries generated from 25 untreated, poly(I:C)-treated, and FV3-infected cells. We detected 136 known X. laevis 26 microRNAs and discovered 133 novel X. laevis microRNAs. Sixty-five microRNAs were 27 differentially expressed in response to poly(I:C), many of which were predicted to target 28 regulators of antiviral pathways such as cGAS-STING, RIG-I/MDA-5, TLR signaling, and type I 29 interferon signaling, as well as products of these pathways (NF-ĸB-induced and interferon- 30 stimulated genes). In contrast, only 49 microRNAs were altered by FV3 infection, fewer of 31 which were predicted to interact with antiviral pathways. Interestingly, poly(I:C) treatment or 32 FV3 infection downregulated transcripts encoding factors of the host microRNA biogenesis 33 pathway. Our study is the first to suggest that host microRNAs regulate innate antiviral 34 immunity in frogs, and sheds light on microRNA-mediated mechanisms of immunoevasion by 35 FV3. 36 37 Keywords: Frog virus 3, microRNAs, antiviral immunity, poly(I:C), skin epithelial cells, 38 Xenopus laevis 2 bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 39 Introduction 40 Epigenetic mechanisms are emerging as key regulators of immune signaling in 41 vertebrates, and microRNAs (miRNAs) are a prime example of such a mechanism (Boosani and 42 Agrawal 2016). miRNAs are a class of small non-coding RNAs that function in post- 43 transcriptional regulation of gene expression. miRNA genes are transcribed in the nucleus by 44 RNA polymerase II and are processed into hairpin precursors (pre-miRNAs) by the RNase III 45 enzyme Drosha (Lee et al. 2002). Pre-miRNAs are exported to the cytoplasm where they are 46 further processed into mature miRNA duplexes by the RNase III enzyme Dicer (Lee et al. 2002). 47 One strand of the miRNA duplex is degraded, while the other guides the Argonaute-containing 48 RNA-induced silencing complex (RISC) to mRNAs which are complementary to the miRNA 49 guide (Lee et al. 2002). Direct interactions between the miRNA and mRNA [typically the 3’ 50 untranslated region (UTR)] induce degradation or inhibit translation of the mRNA (Lee et al. 51 2002). 52 Vertebrate miRNAs have been found to regulate the expression of genes involved in 53 antiviral signaling pathways that produce interferons (IFNs) and inflammatory cytokines in 54 response to pathogen infection (Boosani and Agrawal 2016). Host miRNAs have been shown to 55 regulate components of key pathogen sensing pathways such as the cytosolic RIG-I/MDA5 56 sensing pathway (Li et al. 2020), endosomal TLR3 pathway (Tili et al. 2007; Hou et al. 2009; 57 Imaizumi et al. 2010), endosomal/cell surface TLR4 pathway (Wendlandt et al. 2012), and 58 endosomal TLR7/8/9 pathways (Hou et al. 2009; Tang et al. 2009), as well as the JAK-STAT 59 signaling pathway that regulates the production of IFN-stimulated genes (ISGs) (Tang et al. 60 2009; Jarret et al. 2016). In some cases, these interactions promote efficient host immune 61 responses. However, viral infection can induce changes in host miRNA expression that detriment 3 bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 62 the host. For example, Coxsackievirus B3-induced miR-30a represses TRIM25 and RIG-I 63 function, resulting in enhanced viral replication (Li et al. 2020), vesicular stomatitis virus- 64 induced miR-146a represses type I IFN production and promotes viral replication by repressing 65 TRAF6, IRAK2, and IRAK1 (Hou et al. 2009), and hepatitis C virus upregulates miR-208b and 66 miR-499a-5p which enhances viral replication by dampening type I IFN signaling through 67 repression of the type I IFN receptor IFNAR1 (Jarret et al. 2016). miRNA-mediated regulation of 68 host immune responses is therefore complex, and while a growing number of functional studies 69 have been conducted, our understanding of the broad functions host miRNAs play in antiviral 70 responses remains limited. 71 Ranaviruses, type species frog virus 3 (FV3), are large double-stranded DNA viruses 72 causing emerging infectious diseases that threaten amphibian populations. Despite the 73 devastating amphibian morbidities and mortalities associated with ranaviral infection, our 74 understanding of frog antiviral defenses remains in its infancy, and the roles of frog miRNAs in 75 regulating these antiviral responses are largely unexplored. As frog skin is an important barrier to 76 pathogen entry (Varga et al. 2019), we have recently generated a Xenopus laevis dorsal skin 77 epithelial-like cell line (Xela DS2) (Bui-Marinos et al. 2020) that is permissive to FV3 (Bui- 78 Marinos et al. submitted) to facilitate our understanding of antiviral responses in frog skin 79 epithelial cells. All viruses, including FV3 (Doherty et al. 2016), produce viral double-stranded 80 RNA (dsRNA) at some point in their replication. Along with others, we have studied frog cell 81 antiviral responses through treatment with poly(I:C) (Sang et al. 2016; Wendel et al. 2018; Bui- 82 Marinos et al. 2020), a synthetic viral dsRNA analog and known inducer of type I IFNs. While 83 poly(I:C) is not a perfect mimic of viral dsRNA, as the sequence and length of viral dsRNA is 84 known to impact the induction of host antiviral responses (Poynter and DeWitte-Orr 2015), it 4 bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 85 permits comparisons of type I IFN responses across cell types and species. By modeling 86 “typical” antiviral responses in frogs using poly(I:C), we can further our understanding of how 87 miRNAs function to regulate effective antiviral responses. By comparing “typical” antiviral 88 miRNA responses in frog skin epithelial cells [modeled by poly(I:C)] to antiviral miRNA 89 responses to immunoevasive viruses such as FV3, we aim to elucidate how viral pathogens may 90 subvert epigenetic regulatory responses as direct or indirect mechanisms of immunoevasion. 91 In this study, we sought to perform initial investigations into the role of host miRNAs in 92 innate antiviral immune responses in frog skin epithelial cells. The goals of this study were to (1) 93 profile changes in miRNA expression in Xela DS2 skin epithelial-like cells during antiviral 94 responses to poly(I:C) and FV3, (2) determine the antiviral targets of differentially expressed 95 miRNAs, (3) compare normal antiviral miRNA responses modeled by poly(I:C) – a known 96 inducer of robust antiviral responses – to miRNA responses induced by FV3, an immunoevasive 97 virus, and (4) expand the number of currently annotated frog miRNAs through the discovery of 98 novel X. laevis miRNAs. 99 100 Materials and methods 101 Cell line maintenance 102 Xela DS2, a skin epithelial-like cell line previously generated from X. laevis dorsal skin 103 (Bui-Marinos et al. 2020), was maintained in seven parts Leibovitz’s L-15 (AL-15) medium 104 (Wisent, Mont-Saint-Hilaire, Canada) diluted with 3 parts sterile cell culture water to adjust for 105 amphibian osmolarity, and is herein referred to as amphibian L-15 (AL-15) medium. AL-15 106 medium was supplemented with 15% fetal bovine serum (FBS; lot #234K18; VWR, Radnor, 107 United States) and used to culture Xela DS2 cells at 26 °C in 75 cm2 plug-seal tissue-culture 5 bioRxiv preprint doi: https://doi.org/10.1101/2021.06.29.450442; this version posted July 1, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
Load more

Recommended publications
  • Changes to Virus Taxonomy 2004
    Arch Virol (2005) 150: 189–198 DOI 10.1007/s00705-004-0429-1 Changes to virus taxonomy 2004 M. A. Mayo (ICTV Secretary) Scottish Crop Research Institute, Invergowrie, Dundee, U.K. Received July 30, 2004; accepted September 25, 2004 Published online November 10, 2004 c Springer-Verlag 2004 This note presents a compilation of recent changes to virus taxonomy decided by voting by the ICTV membership following recommendations from the ICTV Executive Committee. The changes are presented in the Table as decisions promoted by the Subcommittees of the EC and are grouped according to the major hosts of the viruses involved. These new taxa will be presented in more detail in the 8th ICTV Report scheduled to be published near the end of 2004 (Fauquet et al., 2004). Fauquet, C.M., Mayo, M.A., Maniloff, J., Desselberger, U., and Ball, L.A. (eds) (2004). Virus Taxonomy, VIIIth Report of the ICTV. Elsevier/Academic Press, London, pp. 1258. Recent changes to virus taxonomy Viruses of vertebrates Family Arenaviridae • Designate Cupixi virus as a species in the genus Arenavirus • Designate Bear Canyon virus as a species in the genus Arenavirus • Designate Allpahuayo virus as a species in the genus Arenavirus Family Birnaviridae • Assign Blotched snakehead virus as an unassigned species in family Birnaviridae Family Circoviridae • Create a new genus (Anellovirus) with Torque teno virus as type species Family Coronaviridae • Recognize a new species Severe acute respiratory syndrome coronavirus in the genus Coro- navirus, family Coronaviridae, order Nidovirales
    [Show full text]
  • The DNA Virus Invertebrate Iridescent Virus 6 Is a Target of the Drosophila Rnai Machinery
    The DNA virus Invertebrate iridescent virus 6 is a target of the Drosophila RNAi machinery Alfred W. Bronkhorsta,1, Koen W. R. van Cleefa,1, Nicolas Vodovarb,2, Ikbal_ Agah Ince_ c,d,e, Hervé Blancb, Just M. Vlakc, Maria-Carla Salehb,3, and Ronald P. van Rija,3 aDepartment of Medical Microbiology, Nijmegen Centre for Molecular Life Sciences, Nijmegen Institute for Infection, Inflammation, and Immunity, Radboud University Nijmegen Medical Centre, 6500 HB Nijmegen, The Netherlands; bViruses and RNA Interference Group, Institut Pasteur, Centre National de la Recherche Scientifique, Unité de Recherche Associée 3015, 75015 Paris, France; cLaboratory of Virology, Wageningen University, 6708 PB Wageningen, The Netherlands; dDepartment of Genetics and Bioengineering, Yeditepe University, Istanbul 34755, Turkey; and eDepartment of Biosystems Engineering, Faculty of Engineering, Giresun University, Giresun 28100, Turkey Edited by Peter Palese, Mount Sinai School of Medicine, New York, NY, and approved October 19, 2012 (received for review April 28, 2012) RNA viruses in insects are targets of an RNA interference (RNAi)- sequently, are hypersensitive to virus infection and succumb more based antiviral immune response, in which viral replication inter- rapidly than their wild-type (WT) controls (11–14). mediates or viral dsRNA genomes are processed by Dicer-2 (Dcr-2) Small RNA cloning and next-generation sequencing provide into viral small interfering RNAs (vsiRNAs). Whether dsDNA virus detailed insights into vsiRNA biogenesis. In several studies in infections are controlled by the RNAi pathway remains to be insects, the polarity of the vsiRNA population deviates strongly determined. Here, we analyzed the role of RNAi in DNA virus from the highly skewed distribution of positive strand (+) over infection using Drosophila melanogaster infected with Invertebrate negative (−) viral RNAs that is generally observed in (+) RNA iridescent virus 6 (IIV-6) as a model.
    [Show full text]
  • Ecology and Pathology of Amphibian Ranaviruses
    Vol. 87: 243–266, 2009 DISEASES OF AQUATIC ORGANISMS Published December 3 doi: 10.3354/dao02138 Dis Aquat Org OPENPEN ACCESSCCESS REVIEW Ecology and pathology of amphibian ranaviruses Matthew J. Gray1,*, Debra L. Miller1, 2, Jason T. Hoverman1 1274 Ellington Plant Sciences Building, Center for Wildlife Health, Department of Forestry Wildlife and Fisheries, Institute of Agriculture, University of Tennessee, Knoxville, Tennessee 37996-4563, USA 2Veterinary Diagnostic and Investigational Laboratory, College of Veterinary Medicine, University of Georgia, 43 Brighton Road, Tifton, Georgia 31793, USA ABSTRACT: Mass mortality of amphibians has occurred globally since at least the early 1990s from viral pathogens that are members of the genus Ranavirus, family Iridoviridae. The pathogen infects multiple amphibian hosts, larval and adult cohorts, and may persist in herpetofaunal and oste- ichthyan reservoirs. Environmental persistence of ranavirus virions outside a host may be several weeks or longer in aquatic systems. Transmission occurs by indirect and direct routes, and includes exposure to contaminated water or soil, casual or direct contact with infected individuals, and inges- tion of infected tissue during predation, cannibalism, or necrophagy. Some gross lesions include swelling of the limbs or body, erythema, swollen friable livers, and hemorrhage. Susceptible amphi- bians usually die from chronic cell death in multiple organs, which can occur within a few days fol- lowing infection or may take several weeks. Amphibian species differ in their susceptibility to rana- viruses, which may be related to their co-evolutionary history with the pathogen. The occurrence of recent widespread amphibian population die-offs from ranaviruses may be an interaction of sup- pressed and naïve host immunity, anthropogenic stressors, and novel strain introduction.
    [Show full text]
  • Emerging Viral Diseases of Fish and Shrimp Peter J
    Emerging viral diseases of fish and shrimp Peter J. Walker, James R. Winton To cite this version: Peter J. Walker, James R. Winton. Emerging viral diseases of fish and shrimp. Veterinary Research, BioMed Central, 2010, 41 (6), 10.1051/vetres/2010022. hal-00903183 HAL Id: hal-00903183 https://hal.archives-ouvertes.fr/hal-00903183 Submitted on 1 Jan 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Vet. Res. (2010) 41:51 www.vetres.org DOI: 10.1051/vetres/2010022 Ó INRA, EDP Sciences, 2010 Review article Emerging viral diseases of fish and shrimp 1 2 Peter J. WALKER *, James R. WINTON 1 CSIRO Livestock Industries, Australian Animal Health Laboratory (AAHL), 5 Portarlington Road, Geelong, Victoria, Australia 2 USGS Western Fisheries Research Center, 6505 NE 65th Street, Seattle, Washington, USA (Received 7 December 2009; accepted 19 April 2010) Abstract – The rise of aquaculture has been one of the most profound changes in global food production of the past 100 years. Driven by population growth, rising demand for seafood and a levelling of production from capture fisheries, the practice of farming aquatic animals has expanded rapidly to become a major global industry.
    [Show full text]
  • Cefas PANDA Report
    Project no. SSPE-CT-2003-502329 PANDA Permanent network to strengthen expertise on infectious diseases of aquaculture species and scientific advice to EU policy Coordination Action, Scientific support to policies WP4: Report on the current best methods for rapid and accurate detection of the main disease hazards in aquaculture, requirements for improvement, their eventual standardisation and validation, and how to achieve harmonised implementation throughout Europe of the best diagnostic methods Olga Haenen*, Inger Dalsgaard, Jean-Robert Bonami, Jean-Pierre Joly, Niels Olesen, Britt Bang Jensen, Ellen Ariel, Laurence Miossec and Isabelle Arzul Work package leader & corresponding author: Dr Olga Haenen, CIDC-Lelystad, NL ([email protected]) PANDA co-ordinator: Dr Barry Hill, CEFAS, UK; www.europanda.net © PANDA, 2007 Cover image: Koi with Koi Herpes Virus Disease: enophthalmia and gill necrosis (M.Engelsma acknowl.) Contents Executive summary 5 Section 1 Introduction 7 1.1 Description of work 7 1.2 Deliverables 8 1.3 Milestones and expected results 9 1.4 Structure of the report and how to use it 9 1.5 General remarks and links with other WPs of PANDA 9 Section 2 Materials and methods 10 2.1 Task force 10 2.2 Network 10 2.3 Workshops and dissemination 10 2.4 Analysis of data 10 2.5 Why harmonization throughout Europe background and aim 11 2.6. CRL functions 11 Section 3 Results 12 3.1 Task force 12 3.2 Network 12 3.3 Workshops and dissemination 12 3.4 Analysis of data 14 Diseases/pathogens of fish 14 3.4.1 Epizootic haematopoietic necrosis
    [Show full text]
  • Evaluating Environmental DNA-Based Quantification of Ranavirus Infection
    Molecular Ecology Resources (2015) doi: 10.1111/1755-0998.12461 Evaluating environmental DNA-based quantification of ranavirus infection in wood frog populations EMILY M. HALL,* ERICA J. CRESPI,* CAREN S. GOLDBERG† andJESSE L. BRUNNER* *School of Biological Sciences, Washington State University, PO Box 644236, Pullman, WA 99164-4236, USA, †School of the Environment, Washington State University, PO Box 646410, Pullman, WA 99164-2812, USA Abstract A variety of challenges arise when monitoring wildlife populations for disease. Sampling tissues can be invasive to hosts, and obtaining sufficient sample sizes can be expensive and time-consuming, particularly for rare species and when pathogen prevalence is low. Environmental DNA (eDNA)-based detection of pathogens is an alternative approach to surveillance for aquatic communities that circumvents many of these issues. Ranaviruses are emerging pathogens of ectothermic vertebrates linked to die-offs of amphibian populations. Detecting ranavirus infections is critical, but nonlethal methods have the above issues and are prone to false negatives. We report on the feasibility and effectiveness of eDNA-based ranavirus detection in the field. We compared ranavirus titres in eDNA samples collected from pond water to titres in wood frog (Lithobates sylvaticus; n = 5) tadpoles in sites dominated by this one species (n = 20 pond visits). We examined whether ranavirus DNA can be detected in eDNA from pond water when infections are present in the pond and if viral titres detected in eDNA samples correlate with the prevalence or intensity of ranavirus infections in tadpoles. With three 250 mL water samples, we were able to detect the virus in all visits with infected larvae (0.92 diagnostic sensitivity).
    [Show full text]
  • Genome Analysis of Ranavirus Frog Virus 3Isolated from American Bullfrog
    www.nature.com/scientificreports OPEN Genome analysis of Ranavirus frog virus 3 isolated from American Bullfrog (Lithobates catesbeianus) in South America Marcelo Candido 1*, Loiane Sampaio Tavares1, Anna Luiza Farias Alencar2, Cláudia Maris Ferreira3, Sabrina Ribeiro de Almeida Queiroz1, Andrezza Maria Fernandes1 & Ricardo Luiz Moro de Sousa1 Ranaviruses (family Iridoviridae) cause important diseases in cold-blooded vertebrates. In addition, some occurrences indicate that, in this genus, the same virus can infect animals from diferent taxonomic groups. A strain isolated from a Ranavirus outbreak (2012) in the state of Sao Paulo, Brazil, had its genome sequenced and presented 99.26% and 36.85% identity with samples of Frog virus 3 (FV3) and Singapore grouper iridovirus (SGIV) ranaviruses, respectively. Eight potential recombination events among the analyzed sample and reference FV3 samples were identifed, including a recombination with Bohle iridovirus (BIV) sample from Oceania. The analyzed sample presented several rearrangements compared to FV3 reference samples from North America and European continent. We report for the frst time the complete genome of Ranavirus FV3 isolated from South America, these results contribute to a greater knowledge related to evolutionary events of potentially lethal infectious agent for cold-blooded animals. Among the major viral pathogens, worldwide distributed and recent history, Ranavirus (Rv) is highlighted, on which, studies in South America remain limited. Rv are part of the family Iridoviridae that is divided into fve genera, of which three are considered more relevant by infectious severity in aquatic and semi-aquatic animals: Lymphocystivirus, Megalocytivirus and Rv. Tey are enveloped and unenveloped viruses, showing double-stranded DNA whose genome ranges from 103 to 220 kbp.
    [Show full text]
  • Comparative Genomic Analyses and Expression of a Set of Overlapping Open Reading Frames of Frog Virus 3, Type Species of Ranavirus (Family Iridoviridae)
    Western Michigan University ScholarWorks at WMU Dissertations Graduate College 4-2006 Comparative Genomic Analyses and Expression of a Set of Overlapping Open Reading Frames of Frog Virus 3, Type Species of Ranavirus (Family Iridoviridae) Wendy Guat Hoon Tan Western Michigan University Follow this and additional works at: https://scholarworks.wmich.edu/dissertations Part of the Animal Sciences Commons, Biology Commons, and the Cell and Developmental Biology Commons Recommended Citation Tan, Wendy Guat Hoon, "Comparative Genomic Analyses and Expression of a Set of Overlapping Open Reading Frames of Frog Virus 3, Type Species of Ranavirus (Family Iridoviridae)" (2006). Dissertations. 995. https://scholarworks.wmich.edu/dissertations/995 This Dissertation-Open Access is brought to you for free and open access by the Graduate College at ScholarWorks at WMU. It has been accepted for inclusion in Dissertations by an authorized administrator of ScholarWorks at WMU. For more information, please contact [email protected]. COMPARATIVE GENOMIC ANALYSES AND EXPRESSION OF A SET OF OVERLAPPING OPEN READING FRAMES OF FROG VIRUS 3, TYPE SPECIES OF R A N A V IR U S (FAM ILY IRIDOVIRIDAE) by Wendy Guat Hoon Tan A Dissertation Submitted to the Faculty of The Graduate College in partial fulfillment of the requirements for the Degree of Doctor of Philosophy Department of Biological Sciences Dr. Karim Essani, Advisor Western Michigan University Kalamazoo, Michigan April 2006 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. COMPARATIVE GENOMIC ANALYSES AND EXPRESSION OF A SET OF OVERLAPPING OPEN READING FRAMES OF FROG VIRUS 3, TYPE SPECIES OF RANAVIRUS (FAMILY IRIDOVIRIDAE) Wendy Guat Hoon Tan, Ph.D.
    [Show full text]
  • A Field Guide to Common Wildlife Diseases and Parasites in the Northwest Territories
    A Field Guide to Common Wildlife Diseases and Parasites in the Northwest Territories 6TH EDITION (MARCH 2017) Introduction Although most wild animals in the NWT are healthy, diseases and parasites can occur in any wildlife population. Some of these diseases can infect people or domestic animals. It is important to regularly monitor and assess diseases in wildlife populations so we can take steps to reduce their impact on healthy animals and people. • recognize sickness in an animal before they shoot; •The identify information a disease in this or field parasite guide in should an animal help theyhunters have to: killed; • know how to protect themselves from infection; and • help wildlife agencies monitor wildlife disease and parasites. The diseases in this booklet are grouped according to where they are most often seen in the body of the Generalanimal: skin, precautions: head, liver, lungs, muscle, and general. Hunters should look for signs of sickness in animals • poor condition (weak, sluggish, thin or lame); •before swellings they shoot, or lumps, such hair as: loss, blood or discharges from the nose or mouth; or • abnormal behaviour (loss of fear of people, aggressiveness). If you shoot a sick animal: • Do not cut into diseased parts. • Wash your hands, knives and clothes in hot, soapy animal, and disinfect with a weak bleach solution. water after you finish cutting up and skinning the 2 • If meat from an infected animal can be eaten, cook meat thoroughly until it is no longer pink and juice from the meat is clear. • Do not feed parts of infected animals to dogs.
    [Show full text]
  • Downloaded from Transcriptome Shotgun Assembly (TSA) Database on 29 November 2020 (Ftp://Ftp.Ddbj.Nig.Ac.Jp/Ddbj Database/Tsa/, Table S3)
    viruses Article Discovery and Characterization of Actively Replicating DNA and Retro-Transcribing Viruses in Lower Vertebrate Hosts Based on RNA Sequencing Xin-Xin Chen, Wei-Chen Wu and Mang Shi * School of Medicine, Sun Yat-sen University, Shenzhen 518107, China; [email protected] (X.-X.C.); [email protected] (W.-C.W.) * Correspondence: [email protected] Abstract: In a previous study, a metatranscriptomics survey of RNA viruses in several important lower vertebrate host groups revealed huge viral diversity, transforming the understanding of the evolution of vertebrate-associated RNA virus groups. However, the diversity of the DNA and retro-transcribing viruses in these host groups was left uncharacterized. Given that RNA sequencing is capable of revealing viruses undergoing active transcription and replication, we collected previously generated datasets associated with lower vertebrate hosts, and searched them for DNA and retro-transcribing viruses. Our results revealed the complete genome, or “core gene sets”, of 18 vertebrate-associated DNA and retro-transcribing viruses in cartilaginous fishes, ray- finned fishes, and amphibians, many of which had high abundance levels, and some of which showed systemic infections in multiple organs, suggesting active transcription or acute infection within the host. Furthermore, these new findings recharacterized the evolutionary history in the families Hepadnaviridae, Papillomaviridae, and Alloherpesviridae, confirming long-term virus–host codivergence relationships for these virus groups.
    [Show full text]
  • Edna Increases the Detectability of Ranavirus Infection in an Alpine Amphibian Population
    viruses Technical Note eDNA Increases the Detectability of Ranavirus Infection in an Alpine Amphibian Population Claude Miaud 1,* ,Véronique Arnal 1, Marie Poulain 1, Alice Valentini 2 and Tony Dejean 2 1 CEFE, EPHE-PSL, CNRS, Univ. Montpellier, Univ Paul Valéry Montpellier 3, IRD, Biogeography and Vertebrate Ecology, 1919 route de Mende, 34293 Montpellier, France; [email protected] (V.A.); [email protected] (M.P.) 2 SPYGEN, 17 Rue du Lac Saint-André, 73370 Le Bourget-du-Lac, France; [email protected] (A.V.); [email protected] (T.D.) * Correspondence: [email protected]; Tel.: +33-(0)4-67-61-33-43 Received: 15 March 2019; Accepted: 4 June 2019; Published: 6 June 2019 Abstract: The early detection and identification of pathogenic microorganisms is essential in order to deploy appropriate mitigation measures. Viruses in the Iridoviridae family, such as those in the Ranavirus genus, can infect amphibian species without resulting in mortality or clinical signs, and they can also infect other hosts than amphibian species. Diagnostic techniques allowing the detection of the pathogen outside the period of host die-off would thus be of particular use. In this study, we tested a method using environmental DNA (eDNA) on a population of common frogs (Rana temporaria) known to be affected by a Ranavirus in the southern Alps in France. In six sampling sessions between June and September (the species’ activity period), we collected tissue samples from dead and live frogs (adults and tadpoles), as well as insects (aquatic and terrestrial), sediment, and water. At the beginning of the breeding season in June, one adult was found dead; at the end of July, a mass mortality of tadpoles was observed.
    [Show full text]
  • Viral and Bacterial Diseases of Amphibians
    CHAPTER 1 Viral and Bacterial Diseases of Amphibians Valentine Helllin~vay, Jesse Brunner, Rick Speare, and Lee Berger I. Introduction B. Frog Erythrocytic Virus C. Lucke Tumour Herpesvirus II. Viral Diseases D. Herpes-like Virus of Skin A. Ranaviruses E. Calicivirus 1. Taxonomy and Molecular Epidemiology F. Leucocyte Viruses 2. Biology III. Bacterial Diseases 3. Clinical Signs and Pathology A. Bacterial Septicaemia ("Red Leg") 4. Epidemiology B. Streptococcosis 5. Resistance to Infection C. Chlamydiosis 6. Transmission and Spread D. Mycobacteriosis 7. Diagnosis 8. Management IV. References 9. Discussion 1. INTRODUCTION A T least six groups of viruses have been reported to affect amphibians, including ficaliciviruses, herpesviruses, and iridoviruses (Johnson and Wellehall 2005). Only ranaviruses are known to cause widespread mass mortality and have been studied in detail; hence a review of this group of viruses forms the m~ority of this chapter. Various strains of ranavirus are found worldwide and some appear to have spread recently (Hyatt et al. 2000). Indicative of the broad host range of ranaviruses and their potentially devastating effects, ranaviral disease was listed by the World Organization for r\.nimal Health (OIE) as an internationally notifiable disease in 2008. Although their impacts 011 populations of declining species are a concern, there is currently no evidence that they have caused permanent declines or extinctions (Daszak et al. 2003). Nevertheless, because of their potential impacts on nai've populations, as well as OIl species that are facing multiple threats, it is important that the risk of spreading these pathogens is minimized (Cunningham et ai. 2007 a). There have been few investigations into other amphibian viruses.
    [Show full text]