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Isolation and Spread of Enteropathogenic Yersinia Spp. Throughout the Pork Production Chain

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Isolation and Spread of Enteropathogenic Yersinia Spp. Throughout the Pork Production Chain Isolation and spread of enteropathogenic Yersinia spp. throughout the pork production chain Inge Van Damme Dissertation submitted in fulfilment of the requirements for the degree of Doctor in Veterinary Sciences (PhD) 2013 Promotors: Prof. Dr. Lieven De Zutter Department of Veterinary Public Health and Food Safety Faculty of Veterinary Medicine Ghent University Prof. Dr. Dirk Berkvens Department of Biomedical Sciences Prince Leopold Institute of Tropical Medicine Members of the reading and examination committee Chairman Prof. Dr. Frank Gasthuys, dean Members of the reading committee Prof. Dr. Fredriksson-Ahomaa Dr. Martine Denis Prof. Dr. Marc Heyndrickx Members of the examination committee Dr. Nadine Botteldoorn Prof. Dr. Dominiek Maes ISBN: 978-90-5864-352-0 To cite this thesis Van Damme I. (2013). Isolation and spread of enteropathogenic Yersinia spp. throughout the pork production chain. Thesis submitted in fulfilment of the requirements for the degree of Doctor in Veterinary Sciences (PhD), Faculty of Veterinary Medicine, Ghent University. The author and promoters give the permission to consult and to copy parts of this work for personal use only. Any other use is subject to the Laws of Copyright. Permission to reproduce any material contained in this work should be obtained from the author. Table of contents List of abbreviations ......................................................................................................................... 5 General introduction ................................................................................................................. 7 1. Taxonomy and characteristics of Yersinia spp. ......................................................................... 9 1.1. The genus Yersinia ........................................................................................................ 9 1.2. Yersinia enterocolitica .................................................................................................. 9 1.3. Yersinia pseudotuberculosis ....................................................................................... 10 2. Pathogenesis and virulence genes of enteropathogenic Yersinia spp. .................................. 10 2.1. Chromosomal encoded virulence factors .................................................................. 11 2.2. The virulence plasmid ................................................................................................. 12 3. Yersiniosis in humans .............................................................................................................. 13 3.1. Clinical manifestations ................................................................................................ 13 3.2. Incidence and species/bioserotype distribution ........................................................ 15 3.3. Transmission routes ................................................................................................... 17 4. Enteropathogenic Yersinia spp. in animals and foods ............................................................ 18 4.1. Enteropathogenic Yersinia spp. in animals ................................................................ 18 4.2. Enteropathogenic Yersinia spp. in food ..................................................................... 25 5. Detection of enteropathogenic Yersinia spp. ......................................................................... 26 5.1. Isolation of enteropathogenic Yersinia spp. ............................................................... 26 5.2. Identification and characterization methods ............................................................. 30 5.3. Alternative methods for detection ............................................................................. 31 Aims ......................................................................................................................................... 33 Chapter 1 Recovery of human pathogenic Yersinia enterocolitica from pig tonsils - part 1 - enumeration and detection by enrichment versus direct plating ............................................... 37 1. Introduction ............................................................................................................................ 39 2. Materials and methods ........................................................................................................... 40 2.1. Sampling ..................................................................................................................... 40 2.2. Enumeration and isolation ......................................................................................... 40 2.3. Statistical analysis ....................................................................................................... 41 3. Results ..................................................................................................................................... 41 4. Discussion ................................................................................................................................ 43 Chapter 2 Recovery of human pathogenic Yersinia enterocolitica from pig tonsils - part 2 - effect of sampling and short isolation methodologies ............................................................... 45 1. Introduction ............................................................................................................................ 47 2. Materials and methods ........................................................................................................... 48 2.1. Sample collection and preparation ............................................................................ 48 2.2. Isolation and enumeration ......................................................................................... 48 2.3. Biochemical identification and biotyping ................................................................... 49 2.4. Molecular identification ............................................................................................. 49 2.5. Statistical analysis ....................................................................................................... 50 3. Results ..................................................................................................................................... 50 4. Discussion ................................................................................................................................ 52 Chapter 3 Recovery of human pathogenic Yersinia enterocolitica from pig carcasses and minced meat ............................................................................................................................ 55 1. Introduction ............................................................................................................................ 57 2. Materials and methods ........................................................................................................... 58 3 Table of contents 2.1. Recovery of human pathogenic Y. enterocolitica from naturally contaminated pig carcasses and minced meat samples ................................................................... 58 2.2. Recovery of Y. enterocolitica from artificially contaminated pork skin samples ....... 60 2.3. Statistical analysis ....................................................................................................... 61 3. Results ..................................................................................................................................... 61 3.1. Naturally contaminated pig carcasses and minced meat samples ............................ 61 3.2. Artificially contaminated pork skin samples .............................................................. 63 4. Discussion ................................................................................................................................ 63 Chapter 4 Contamination of freshly slaughtered pig carcasses with enteropathogenic Yersinia spp. and investigation of associated risk factors during slaughter ................................ 67 1. Introduction ............................................................................................................................ 69 2. Material and methods ............................................................................................................ 69 2.1. Sampling ..................................................................................................................... 69 2.2. Isolation and enumeration of enteropathogenic Yersinia spp. ................................. 70 2.3. Identification and virulence properties of enteropathogenic Yersinia spp. .............. 70 2.4. Statistical analyses...................................................................................................... 71 3. Results ..................................................................................................................................... 71 3.1. Presence of enteropathogenic Yersinia spp. in pigs at slaughter .............................. 71 3.2. Risk factors for carcass contamination ...................................................................... 73 4. Discussion ................................................................................................................................ 75 4.1. Presence of enteropathogenic Yersinia spp. in pigs and on pig carcasses at slaughter ....................................................................................................................
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