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Entwicklung Einer Software Zur Identifizierung Neuartiger Und
Entwicklung einer Software zur Identifizierung neuartiger und bekannter Infektionserreger in klinischen Proben Dissertation zur Erlangung des Doktorgrades an der Fakult¨at fur¨ Mathematik, Informatik und Naturwissenschaften Fachbereich Biologie der Universit¨at Hamburg vorgelegt von Malik Alawi Hamburg, 2020 Vorsitzender der Prufungskommission¨ Dr. PD Andreas Pommerening-R¨oser Gutachter Professor Dr. Adam Grundhoff Professor Dr. Stefan Kurtz Datum der Disputation 30. April 2021 Abstract Sequencing of diagnostic samples is widely considered a key technology that may fun- damentally improve infectious disease diagnostics. The approach can not only identify pathogens already known to cause a specific disease, but may also detect pathogens that have not been previously attributed to this disease, as well as completely new, previously unknown pathogens. Therefore, it may significantly increase the level of preparedness for future outbreaks of emerging pathogens. This study describes the development and application of methods for the identification of pathogenic agents in diagnostic samples. The methods have been successfully applied multiple times under clinical conditions. The corresponding results have been published within the scope of this thesis. Finally, the methods were made available to the scientific community as an open source bioinformatics tool. The novel software was validated by conventional diagnostic methods and it was compared to established analysis pipelines using authentic clinical samples. It is able to identify pathogens from different diagnostic entities and often classifies viral agents down to strain level. Furthermore, the method is capable of assembling complete viral genomes, even from samples containing multiple closely related viral strains of the same viral family. In addition to an improved method for taxonomic classification, the software offers functionality which is not present in established analysis pipelines. -
Using Cell Lines to Study Factors Affecting Transmission of Fish Viruses
Using cell lines to study factors affecting transmission of fish viruses by Phuc Hoang Pham A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for the degree of Doctor of Philosophy in Biology Waterloo, Ontario, Canada, 2014 ©Phuc Hoang Pham 2014 AUTHOR'S DECLARATION I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, as accepted by my examiners. I understand that my thesis may be made electronically available to the public. ii ABSTRACT Factors that can influence the transmission of aquatic viruses in fish production facilities and natural environment are the immune defense of host species, the ability of viruses to infect host cells, and the environmental persistence of viruses. In this thesis, fish cell lines were used to study different aspects of these factors. Five viruses were used in this study: viral hemorrhagic septicemia virus (VHSV) from the Rhabdoviridae family; chum salmon reovirus (CSV) from the Reoviridae family; infectious pancreatic necrosis virus (IPNV) from the Birnaviridae family; and grouper iridovirus (GIV) and frog virus-3 (FV3) from the Iridoviridae family. The first factor affecting the transmission of fish viruses examined in this thesis is the immune defense of host species. In this work, infections of marine VHSV-IVa and freshwater VHSV-IVb were studied in two rainbow trout cell lines, RTgill-W1 from the gill epithelium, and RTS11 from spleen macrophages. RTgill-W1 produced infectious progeny of both VHSV-IVa and -IVb. However, VHSV-IVa was more infectious than IVb toward RTgill-W1: IVa caused cytopathic effects (CPE) at a lower viral titre, elicited CPE earlier, and yielded higher titres. -
An Open Source Bioinformatics Tool for Fast, Systematic and Cohort Based
www.nature.com/scientificreports OPEN DAMIAN: an open source bioinformatics tool for fast, systematic and cohort based analysis of microorganisms in diagnostic samples Malik Alawi 1,2, Lia Burkhardt1, Daniela Indenbirken1, Kerstin Reumann1, Maximilian Christopeit3, Nicolaus Kröger3, Marc Lütgehetmann4, Martin Aepfelbacher4, Nicole Fischer4,5* & Adam Grundhof 1,5* We describe DAMIAN, an open source bioinformatics tool designed for the identifcation of pathogenic microorganisms in diagnostic samples. By using authentic clinical samples and comparing our results to those from established analysis pipelines as well as conventional diagnostics, we demonstrate that DAMIAN rapidly identifes pathogens in diferent diagnostic entities, and accurately classifes viral agents down to the strain level. We furthermore show that DAMIAN is able to assemble full-length viral genomes even in samples co-infected with multiple virus strains, an ability which is of considerable advantage for the investigation of outbreak scenarios. While DAMIAN, similar to other pipelines, analyzes single samples to perform classifcation of sequences according to their likely taxonomic origin, it also includes a tool for cohort-based analysis. This tool uses cross-sample comparisons to identify sequence signatures that are frequently present in a sample group of interest (e.g., a disease- associated cohort), but occur less frequently in control cohorts. As this approach does not require homology searches in databases, it principally allows the identifcation of not only known, but also completely novel pathogens. Using samples from a meningitis outbreak, we demonstrate the feasibility of this approach in identifying enterovirus as the causative agent. Nucleic acid based detection of pathogens has widely replaced culture based laboratory methods for the identif- cation of putative pathogens in samples from patients with infectious diseases1,2. -
A Novel Rhabdovirus Infecting Newly Discovered Nycteribiid Bat Flies
www.nature.com/scientificreports OPEN Kanyawara Virus: A Novel Rhabdovirus Infecting Newly Discovered Nycteribiid Bat Flies Received: 19 April 2017 Accepted: 25 May 2017 Infesting Previously Unknown Published: xx xx xxxx Pteropodid Bats in Uganda Tony L. Goldberg 1,2,3, Andrew J. Bennett1, Robert Kityo3, Jens H. Kuhn4 & Colin A. Chapman3,5 Bats are natural reservoir hosts of highly virulent pathogens such as Marburg virus, Nipah virus, and SARS coronavirus. However, little is known about the role of bat ectoparasites in transmitting and maintaining such viruses. The intricate relationship between bats and their ectoparasites suggests that ectoparasites might serve as viral vectors, but evidence to date is scant. Bat flies, in particular, are highly specialized obligate hematophagous ectoparasites that incidentally bite humans. Using next- generation sequencing, we discovered a novel ledantevirus (mononegaviral family Rhabdoviridae, genus Ledantevirus) in nycteribiid bat flies infesting pteropodid bats in western Uganda. Mitochondrial DNA analyses revealed that both the bat flies and their bat hosts belong to putative new species. The coding-complete genome of the new virus, named Kanyawara virus (KYAV), is only distantly related to that of its closest known relative, Mount Elgon bat virus, and was found at high titers in bat flies but not in blood or on mucosal surfaces of host bats. Viral genome analysis indicates unusually low CpG dinucleotide depletion in KYAV compared to other ledanteviruses and rhabdovirus groups, with KYAV displaying values similar to rhabdoviruses of arthropods. Our findings highlight the possibility of a yet- to-be-discovered diversity of potentially pathogenic viruses in bat ectoparasites. Bats (order Chiroptera) represent the second largest order of mammals after rodents (order Rodentia). -
Characterization of Viral Communities of Biting Midges and Identification
viruses Article Characterization of Viral Communities of Biting Midges and Identification of Novel Thogotovirus Species and Rhabdovirus Genus Sarah Temmam 1, Sonia Monteil-Bouchard 1, Catherine Robert 1, Jean-Pierre Baudoin 1, Masse Sambou 1, Maxence Aubadie-Ladrix 1, Noémie Labas 1, Didier Raoult 1,2, Oleg Mediannikov 1 and Christelle Desnues 1,* 1 Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63 CNRS 7278 IRD 198 INSERM U1095, Aix-Marseille Université, Marseille 13005, France; [email protected] (S.T.); [email protected] (S.M.-B.); [email protected] (C.R.); [email protected] (J.-P.B.); [email protected] (M.S.); [email protected] (M.A.-L.); [email protected] (N.L.); [email protected] (D.R.); [email protected] (O.M.). 2 Fondation IHU Méditerranée Infection, Pôle des Maladies Infectieuses et Tropicales Clinique et Biologique, Fédération de Bactériologie-Hygiène-Virologie, Centre Hospitalo-Universitaire Timone, Méditerranée Infection, Assistance Publique–Hôpitaux de Marseille, Marseille 13005, France * Correspondence: [email protected]; Tel.: +33-0-491324630; Fax: +33-0-491387772 Academic Editors: Johnson Mak, Peter Walker and Marcus Thomas Gilbert Received: 21 October 2015; Accepted: 1 March 2016; Published: 11 March 2016 Abstract: More than two thirds of emerging viruses are of zoonotic origin, and among them RNA viruses represent the majority. Ceratopogonidae (genus Culicoides) are well-known vectors of several viruses responsible for epizooties (bluetongue, epizootic haemorrhagic disease, etc.). They are also vectors of the only known virus infecting humans: the Oropouche virus. Female midges usually feed on a variety of hosts, leading to possible transmission of emerging viruses from animals to humans. -
Characterization of Farmington Virus, a Novel Virus from Birds That Is Distantly Related to Members of the Family Rhabdoviridae
Palacios et al. Virology Journal 2013, 10:219 http://www.virologyj.com/content/10/1/219 RESEARCH Open Access Characterization of Farmington virus, a novel virus from birds that is distantly related to members of the family Rhabdoviridae Gustavo Palacios1†, Naomi L Forrester2,3,4†, Nazir Savji5,7†, Amelia P A Travassos da Rosa2, Hilda Guzman2, Kelly DeToy5, Vsevolod L Popov2,4, Peter J Walker6, W Ian Lipkin5, Nikos Vasilakis2,3,4 and Robert B Tesh2,4* Abstract Background: Farmington virus (FARV) is a rhabdovirus that was isolated from a wild bird during an outbreak of epizootic eastern equine encephalitis on a pheasant farm in Connecticut, USA. Findings: Analysis of the nearly complete genome sequence of the prototype CT AN 114 strain indicates that it encodes the five canonical rhabdovirus structural proteins (N, P, M, G and L) with alternative ORFs (> 180 nt) in the N and G genes. Phenotypic and genetic characterization of FARV has confirmed that it is a novel rhabdovirus and probably represents a new species within the family Rhabdoviridae. Conclusions: In sum, our analysis indicates that FARV represents a new species within the family Rhabdoviridae. Keywords: Farmington virus (FARV), Family Rhabdoviridae, Next generation sequencing, Phylogeny Background Results Therhabdovirusesarealargeanddiversegroupofsingle- Growth characteristics stranded, negative sense RNA viruses that infect a wide Three litters of newborn (1–2 day old) ICR mice with aver- range of vertebrates, invertebrates and plants [1]. The family agesizeof10pupswereinoculated intracerebrally (ic) with Rhabdoviridae is currently divided into nine approved ge- 15–20 μl, intraperitoneally (ip) with 100 μlorsubcutane- nera (Vesiculovirus, Perhavirus, Ephemerovirus, Lyssavirus, ously (sc) with 100 μlofastockofVero-grownFARV(CT Tibrovirus, Sigmavirus, Nucleorhabdovirus, Cytorhabdovirus AN 114) virus containing approximately 107 plaque and Novirhabdovirus);however,alargenumberofanimal forming units (PFU) per ml. -
EU Position the EU Thanks the OIE and in General Supports the Adoption of This Modified User's Guide
Ref. Ares(2018)2526762 - 15/05/2018 Annex 2 Original: English February 2018 REPORT OF THE MEETING OF THE OIE AQUATIC ANIMAL HEALTH STANDARDS COMMISSION EU comment The EU would like to commend the OIE Aquatic Animal Health Standards Commission for its work and for having taken into consideration EU comments on the Aquatic Code and Manual submitted previously. A number of general comments on this report of the February 2018 meeting of the Aquatic Animals Commission as well as the intended positions of the EU on the draft Aquatic Code and Manual chapters proposed for adoption at the 86th OIE General Session are inserted in the text below, while specific comments are inserted in the text of the respective annexes to the report. The EU would like to stress again its continued commitment to participate in the work of the OIE and to offer all technical support needed by the Aquatic Animals Commission and its ad hoc groups for future work on the Aquatic Code and Manual. The OIE Aquatic Animal Health Standards Commission (hereinafter referred to as the Aquatic Animals Commission) met at OIE Headquarters in Paris from 14 to 21 February 2018. The list of participants is attached as Annex 1. The Aquatic Animals Commission thanked the following Member Countries for providing written comments on draft texts for the OIE Aquatic Animal Health Code (hereinafter referred to as the Aquatic Code) and OIE Manual of Diagnostic Tests for Aquatic Animals (hereinafter referred to as the Aquatic Manual) circulated after the Commission’s September 2017 meeting: Argentina, Australia, Canada, Chile, Chinese Taipei, Costa Rica, Fiji, Guatemala, Japan, Mexico, New Caledonia, Norway, Singapore, Switzerland, Thailand, the United States of America (USA) and the Member States of the European Union (EU). -
Aquatic Animal Viruses Mediated Immune Evasion in Their Host T ∗ Fei Ke, Qi-Ya Zhang
Fish and Shellfish Immunology 86 (2019) 1096–1105 Contents lists available at ScienceDirect Fish and Shellfish Immunology journal homepage: www.elsevier.com/locate/fsi Aquatic animal viruses mediated immune evasion in their host T ∗ Fei Ke, Qi-Ya Zhang State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China ARTICLE INFO ABSTRACT Keywords: Viruses are important and lethal pathogens that hamper aquatic animals. The result of the battle between host Aquatic animal virus and virus would determine the occurrence of diseases. The host will fight against virus infection with various Immune evasion responses such as innate immunity, adaptive immunity, apoptosis, and so on. On the other hand, the virus also Virus-host interactions develops numerous strategies such as immune evasion to antagonize host antiviral responses. Here, We review Virus targeted molecular and pathway the research advances on virus mediated immune evasions to host responses containing interferon response, NF- Host responses κB signaling, apoptosis, and adaptive response, which are executed by viral genes, proteins, and miRNAs from different aquatic animal viruses including Alloherpesviridae, Iridoviridae, Nimaviridae, Birnaviridae, Reoviridae, and Rhabdoviridae. Thus, it will facilitate the understanding of aquatic animal virus mediated immune evasion and potentially benefit the development of novel antiviral applications. 1. Introduction Various antiviral responses have been revealed [19–22]. How they are overcome by different viruses? Here, we select twenty three strains Aquatic viruses have been an essential part of the biosphere, and of aquatic animal viruses which represent great harms to aquatic ani- also a part of human and aquatic animal lives. -
Partitiviruses Infecting Drosophila Melanogaster and Aedes Aegypti Exhibit Efficient 2 Biparental Vertical Transmission 3 4 Shaun T
bioRxiv preprint doi: https://doi.org/10.1101/2020.06.01.128819; this version posted June 2, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Partitiviruses infecting Drosophila melanogaster and Aedes aegypti exhibit efficient 2 biparental vertical transmission 3 4 Shaun T. Cross1, Bernadette L. Maertens1, Tillie J. Dunham1, Case P. Rodgers1, Ali L. Brehm1, 5 Megan R. Miller1, Alissa M. Williams2, Brian D. Foy, Mark D. Stenglein1,* 6 7 1. Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine 8 and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA 9 2. Department of Biology, Colorado State University, Fort Collins, CO, USA 10 * Correspondence to: [email protected] 11 12 Abstract 13 14 Partitiviruses are segmented, multipartite dsRNA viruses that until recently were only known to 15 infect fungi, plants, and protozoans. Metagenomic surveys have revealed that partitivirus-like 16 sequences are also commonly associated with arthropods. One arthropod-associated partitivirus, 17 galbut virus, is extraordinarily common in wild populations of Drosophila melanogaster fruit 18 flies. To begin to understand the processes that underlie this virus’s high global prevalence, we 19 established colonies of wild-caught infected flies. Infection remained at stably high levels over 20 three years, with between 63-100% of individual flies infected. Galbut virus infects fly cells and 21 replicates in tissues throughout infected adults, including reproductive tissues and the gut 22 epithelium. -
Viruses 2015, 7, 456-479; Doi:10.3390/V7020456 OPEN ACCESS
Viruses 2015, 7, 456-479; doi:10.3390/v7020456 OPEN ACCESS viruses ISSN 1999-4915 www.mdpi.com/journal/viruses Article In Search of Pathogens: Transcriptome-Based Identification of Viral Sequences from the Pine Processionary Moth (Thaumetopoea pityocampa) Agata K. Jakubowska 1, Remziye Nalcacioglu 2, Anabel Millán-Leiva 3, Alejandro Sanz-Carbonell 1, Hacer Muratoglu 4, Salvador Herrero 1,* and Zihni Demirbag 2,* 1 Department of Genetics, Universitat de València, Dr Moliner 50, 46100 Burjassot, Spain; E-Mails: [email protected] (A.K.J.); [email protected] (A.S.-C.) 2 Department of Biology, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon, Turkey; E-Mail: [email protected] 3 Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” (IHSM-UMA-CSIC), Consejo Superior de Investigaciones Científicas, Estación Experimental “La Mayora”, Algarrobo-Costa, 29750 Málaga, Spain; E-Mail: [email protected] 4 Department of Molecular Biology and Genetics, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon, Turkey; E-Mail: [email protected] * Authors to whom correspondence should be addressed; E-Mails: [email protected] (S.H.); [email protected] (Z.D.); Tel.: +34-96-354-3006 (S.H.); +90-462-377-3320 (Z.D.); Fax: +34-96-354-3029 (S.H.); +90-462-325-3195 (Z.D.). Academic Editors: John Burand and Madoka Nakai Received: 29 November 2014 / Accepted: 13 January 2015 / Published: 23 January 2015 Abstract: Thaumetopoea pityocampa (pine processionary moth) is one of the most important pine pests in the forests of Mediterranean countries, Central Europe, the Middle East and North Africa. Apart from causing significant damage to pinewoods, T. -
The Discovery, Distribution and Diversity of DNA Viruses
bioRxiv preprint doi: https://doi.org/10.1101/2020.10.16.342956; this version posted March 17, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Title: The discovery, distribution and diversity of DNA viruses associated with Drosophila melanogaster in Europe Running title: DNA viruses of European Drosophila Key Words: DNA virus, Endogenous viral element, Drosophila, Nudivirus, Galbut virus, Filamentous virus, Adintovirus, Densovirus, Bidnavirus Authors: Megan A. Wallace 1,2 [email protected] 0000-0001-5367-420X Kelsey A. Coffman 3 [email protected] 0000-0002-7609-6286 Clément Gilbert 1,4 [email protected] 0000-0002-2131-7467 Sanjana Ravindran 2 [email protected] 0000-0003-0996-0262 Gregory F. Albery 5 [email protected] 0000-0001-6260-2662 Jessica Abbott 1,6 [email protected] 0000-0002-8743-2089 Eliza Argyridou 1,7 [email protected] 0000-0002-6890-4642 Paola Bellosta 1,8,9 [email protected] 0000-0003-1913-5661 Andrea J. Betancourt 1,10 [email protected] 0000-0001-9351-1413 Hervé Colinet 1,11 [email protected] 0000-0002-8806-3107 Katarina Eric 1,12 [email protected] 0000-0002-3456-2576 Amanda Glaser-Schmitt 1,7 [email protected] 0000-0002-1322-1000 Sonja Grath 1,7 [email protected] 0000-0003-3621-736X Mihailo Jelic 1,13 [email protected] 0000-0002-1637-0933 Maaria Kankare 1,14 [email protected] 0000-0003-1541-9050 Iryna Kozeretska 1,15 [email protected] 0000-0002-6485-1408 Volker Loeschcke 1,16 [email protected] 0000-0003-1450-0754 Catherine Montchamp-Moreau 1,4 [email protected] 0000-0002-5044-9709 Lino Ometto 1,17 [email protected] 0000-0002-2679-625X Banu Sebnem Onder 1,18 [email protected] 0000-0002-3003-248X Dorcas J. -
KHV) by Serum Neutralization Test
Downloaded from orbit.dtu.dk on: Nov 08, 2017 Detection of antibodies specific to koi herpesvirus (KHV) by serum neutralization test Cabon, J.; Louboutin, L.; Castric, J.; Bergmann, S. M.; Bovo, G.; Matras, M.; Haenen, O.; Olesen, Niels Jørgen; Morin, T. Published in: 17th International Conference on Diseases of Fish And Shellfish Publication date: 2015 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Cabon, J., Louboutin, L., Castric, J., Bergmann, S. M., Bovo, G., Matras, M., ... Morin, T. (2015). Detection of antibodies specific to koi herpesvirus (KHV) by serum neutralization test. In 17th International Conference on Diseases of Fish And Shellfish: Abstract book (pp. 115-115). [O-107] Las Palmas: European Association of Fish Pathologists. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. DISCLAIMER: The organizer takes no responsibility for any of the content stated in the abstracts.