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Intramammary Infections with Coagulase-Negative Staphylococcus Species

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Intramammary Infections with Coagulase-Negative Staphylococcus Species Printing of this thesis was financially supported by Printed by University Press, Zelzate ISBN number: 9789058642738 INTRAMAMMARY INFECTIONS WITH COAGULASE-NEGATIVE STAPHYLOCOCCUS SPECIES IN BOVINES - MOLECULAR DIAGNOSTICS AND EPIDEMIOLOGY - KARLIEN SUPRÉ 2011 PROMOTORS/PROMOTOREN Prof. dr. Sarne De Vliegher Faculteit Diergeneeskunde, UGent Prof. dr. Ruth N. Zadoks Royal (Dick) School of Veterinary Studies, University of Edinburgh; Moredun Research Institute, Penicuik, Schotland Prof. dr. Freddy Haesebrouck Faculteit Diergeneeskunde, UGent MEMBERS OF THE EXAMINATION COMMITTEE/LEDEN VAN DE EXAMENCOMMISSIE Prof. dr. dr. h. c. Aart de Kruif Voorzitter van de examencommissie Prof. dr. Mario Vaneechoutte Faculteit Geneeskunde en Gezondheidswetenschappen, UGent Dr. Margo Baele Directie Onderzoeksaangelegenheden, UGent Dr. Lic. Luc De Meulemeester MCC-Vlaanderen, Lier Prof. dr. Geert Opsomer Faculteit Diergeneeskunde, UGent Prof. dr. Marc Heyndrickx Instituut voor Landbouw en Visserijonderzoek (ILVO), Melle Dr. Suvi Taponen University of Helsinki, Finland Prof. dr. Ynte H. Schukken Cornell University, Ithaca, USA INTRAMAMMARY INFECTIONS WITH COAGULASE-NEGATIVE STAPHYLOCOCCUS SPECIES IN BOVINES - MOLECULAR DIAGNOSTICS AND EPIDEMIOLOGY - KARLIEN SUPRÉ Department of Reproduction, Obstetrics, and Herd Health Faculty of Veterinary Medicine, Ghent University Dissertation submitted in the fulfillment of the requirements for the degree of Doctor in Veterinary Sciences, Faculty of Veterinary Medicine, Ghent University INTRAMAMMAIRE INFECTIES MET COAGULASE-NEGATIEVE STAPHYLOCOCCUS SPECIES BIJ MELKVEE - MOLECULAIRE DIAGNOSTIEK EN EPIDEMIOLOGIE - KARLIEN SUPRÉ Vakgroep Voortplanting, Verloskunde en Bedrijfsdiergeneeskunde Faculteit Diergeneeskunde, Universiteit Gent Proefschrift voorgedragen tot het behalen van de graad van Doctor in de Diergeneeskundige Wetenschappen aan de Faculteit Diergeneeskunde, Universiteit Gent Uit de oude doos Onzindelijk gewonnen melk of melk van zieke koeien kan oorzaak zijn van verschillende ziekten; frisse, blozende, gezonde kleintjes kunnen er hun gezondheid bij inschieten, voor gans hun leven een knak krijgen. Het is een sociale plicht te streven naar een gezonde veestapel en zindelijke melkwinning.” Ook uit welbegrepen eigenbelang moeten de landbouwers streven naar het winnen van gezonde kwaliteitsmelk. Wie in de algemene vooruitgang ten achter blijft is gedoemd te verdwijnen. Het werk dat we goed doen geeft ons een voldoening, waarop we recht hebben. Deze arbeidsvreugde moet het dagelijks brood zijn, dat onze honger naar genot op een edele en voortreffelijke wijze stilt. Als we ons werk goed doen, verheffen we ons zelf en verheffen we ook onze stand. Dat is de echte fierheid, de welbegrepen standsfierheid. Uit: “Melk en de melkverzorging op de hoeve”, Uitgave van de Boerinnenbond, Leuven, 1947, 2de uitgave. List of abbreviations AFLP Amplified fragment length polymorphism ATCC American Type Culture Collection BMSCC Bulk milk somatic cell count CCM Czech Collection of Microorganisms CCUG Culture Collection of the University of Göteborg, Sweden CE Capillary electrophoresis CFU Colony forming units CLSI Clinical and Laboratory Standards Institute CM Clinical mastitis CNRS Centre National de Référence des Staphylocoques CNS Coagulase-negative staphylococci CPS Coagulase-positive staphylococci DHI Dairy herd improvement DIM Days-in-milk DNA Desoxyribonucleic acid DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen hsp60 Heat shock protein 60 IMI Intramammary infection LMG Laboratory for Microbiology, Ghent University LnqSCC Natural logarithm of quarter milk somatic cell count LnSCC Natural logarithm of somatic cell count MLST Multilocus sequence typing MMUL Milking machine unit liner MSG Milkers’ skin or gloves NCBI National Center for Biotechnology Information NCTC National Collection of Type Cultures NMC National Mastitis Council, a global organization for mastitis control and milk quality PCR Polymerase chain reaction PFGE Pulsed-field gel electrophoresis PPV Positive predictive value qSCC Quarter milk somatic cell count RNA Ribonucleic acid rpoB β-subunit of RNA polymerase rRNA Ribosomal ribonucleic acid SCC Somatic cell count SCM Subclinical mastitis TA Teat apex tDNA-PCR Transfer RNA-intergenic spacer-PCR tuf Elongation factor tu Table of contents Chapter 1 Preface ..................................................................................................................................................................... 1 Chapter 2 General introduction .................................................................................................................................... 5 Chapter 3 Aims and outline of this thesis ..................................................................................... 45 Chapter 4 Molecular identification of coagulase-negative staphylococci .............................. 49 Use of transfer RNA-intergenic spacer polymerase chain reaction combined with capillary electrophoresis to identify coagulase-negative Staphylococcus species originating from bovine milk and teat apices Chapter 5 Description of new Staphylococcus species originating from dairy cows .... 65 Chapter 5.1 Staphylococcus devriesei sp. nov., isolated from teat apices and milk of dairy cows .......................................................................................................................................................... 67 Chapter 5.2 Staphylococcus agnetis sp. nov., a coagulase-variable species from bovine subclinical and mild clinical mastitis ........................................................................................... 87 Chapter 6 Epidemiology of coagulase-negative Staphylococcus species and species- specific impact on udder health .................................................................................................. 105 Chapter 6.1 Some coagulase-negative Staphylococcus species are affecting udder health more than others ....................................................................................................................................... 107 Chapter 6.2 Distribution of coagulase-negative Staphylococcus species isolated from parlour-associated niches is herd-dependent and differs from that in intramammary infections ............................................................................................ 133 Chapter 7 General discussion ................................................................................................................................... 151 Chapter 8 Summary .......................................................................................................................................................... 205 Chapter 9 Samenvatting ................................................................................................................................................ 211 Chapter 10 Curriculum Vitae and publications ............................................................................................ 217 Chapter 11 Dankwoord .................................................................................................................................................... 225 CHAPTER 1 PREFACE Karlien Supré Department of Reproduction, Obstetrics, and Herd Health Faculty of Veterinary Medicine Ghent University [email protected] Preface The evolving society is subject to a variety of new challenges, and in a way, this is also true for agriculture in all its aspects. The constant urge for food safety and food security, due to the global population growth, determines the way food is currently produced. This is also influencing the dairy industry since milk consumption increases. The discussion on global warming also affects agriculture, given that it accounts for about 14 % of the greenhouse gas emission. The dairy sector generates 4 % of the greenhouse gas emission, with methane as the most important contributor (FAO, 2010). During the last decades, a higher efficiency in dairy farming helped to lower emissions (FAO, 2010) although there still is a clear margin for improvement. Besides, the image of animal production is under pressure, because of animal welfare issues and inconsiderate use of antimicrobials. In order to live up to these high standards, dairy farmers are now urged to put more focus on prevention rather than cure of diseases affecting productivity, profitability, animal welfare, and drug use. Cost-effective production is key as consumers have high expectations but are not eager to pay more for dairy products with added value. A variety of metabolic disorders and infectious diseases hinder profitability of dairy farming, with mastitis being the most important (Bradley, 2002). Added to the financial losses, animal discomfort due to clinical mastitis has to be considered as well. The majority of mastitis cases is due to a bacterial infection such as by Staphylococcus aureus, Streptococcus species, or Escherichia coli. While we have been successful in combating some of these bacteria (e. g. incidence of Streptococcus agalactiae IMI has been brought to a minimum in many countries), another group of bacteria, the coagulase-negative Staphylococcus (CNS) species, has appeared on the forefront. The CNS have become the most prevalent group of mastitis bacteria in many areas of the world, but research has been hampered by the plethora of different species within the group and unsound identification tools. 3 CHAPTER 2 GENERAL INTRODUCTION Karlien Supré Department
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