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The Health of Hedgehogs in Relation to Tick Infestation and Transmission of Pathogens by the Hedgehog Tick, Ixodes Hexagonus in Vitro

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The Health of Hedgehogs in Relation to Tick Infestation and Transmission of Pathogens by the Hedgehog Tick, Ixodes Hexagonus in Vitro The health of hedgehogs in relation to tick infestation and transmission of pathogens by the hedgehog tick, Ixodes hexagonus in vitro. Khyara van Hummel April 2014 – July 2014 P a g e | 1 The health of hedgehogs in relation to tick infestation and transmission of pathogens by the hedgehog tick, Ixodes hexagonus in vitro. Author: K.M.L.B. van Hummel Location: Utrecht Centre for Tick-borne Diseases (UCTD) Faculty of Veterinary Medicine Utrecht University Timeslot: April 2014 – July 2014 Supervisor: Prof. Dr. F. Jongejan The image on the front was taken by the author. P a g e | 2 Index Abstract .............................................................................................................................................5 Introduction ......................................................................................................................................6 Ixodes hexagonus ............................................................................................................................................... 6 In vitro feeding ................................................................................................................................................... 6 Objective of this research ................................................................................................................................... 7 Materials and method ........................................................................................................................8 Questionnaire ..................................................................................................................................................... 8 Ticks .................................................................................................................................................................... 8 Blood ................................................................................................................................................................... 8 Membranes ........................................................................................................................................................ 9 Feeding units ...................................................................................................................................................... 9 Bonbons ............................................................................................................................................................ 10 Attachment stimuli and odour test .................................................................................................................. 10 Water bath........................................................................................................................................................ 10 In vitro feeding procedure ................................................................................................................................ 10 DNA extraction ................................................................................................................................................. 11 PCR .................................................................................................................................................................... 11 Reverse Line Blot hybridization ........................................................................................................................ 12 Results ............................................................................................................................................. 13 Week 1 .............................................................................................................................................................. 13 Week 2 .............................................................................................................................................................. 13 Week 3 .............................................................................................................................................................. 14 Week 4 .............................................................................................................................................................. 16 Week 5 .............................................................................................................................................................. 16 Week 6 .............................................................................................................................................................. 18 Week 7 .............................................................................................................................................................. 19 Week 8 .............................................................................................................................................................. 21 Week 9 .............................................................................................................................................................. 22 Week 10 ............................................................................................................................................................ 24 Questionnaire ................................................................................................................................................... 26 .......................................................................................................................................................................... 27 P a g e | 3 RLB .................................................................................................................................................................... 27 Discussion ........................................................................................................................................ 28 Test feeding ...................................................................................................................................................... 28 Feeding 1 .......................................................................................................................................................... 28 Feeding 2 .......................................................................................................................................................... 28 Feeding 3 .......................................................................................................................................................... 28 Feeding 4 .......................................................................................................................................................... 28 Feeding 5 .......................................................................................................................................................... 28 Feeding 6 .......................................................................................................................................................... 28 Feeding 7, 8 and 9 ............................................................................................................................................ 29 Odour test ......................................................................................................................................................... 29 Questionnaire ................................................................................................................................................... 30 RLB .................................................................................................................................................................... 30 Conclusion ....................................................................................................................................... 31 References ....................................................................................................................................... 32 Appendix ......................................................................................................................................... 34 Appendix A ........................................................................................................................................................... 34 Enquête opvangcentra ..................................................................................................................................... 34 Appendix B ............................................................................................................................................................ 35 DNA EXTRACTION FROM TICKS ....................................................................................................................... 35 DNA EXTRACTION FROM BLOOD PROCEDURE ............................................................................................... 37 PCR RLB PROCEDURE ....................................................................................................................................... 39 REVERSE LINE BLOT HYBRIDIZATION PROCEDURE ......................................................................................... 41 IN VITRO FEEDING UNIT PREPARATION PROCEDURE ....................................................................................... 43 IN VITRO FEEDING BLOOD PREPARATION PROCEDURE ..................................................................................
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