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Ostertagia Ostertagi and Cooperia Oncophora

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Ostertagia Ostertagi and Cooperia Oncophora "OUIFMNJOUJDBDUJWJUZPGGPSBHFDIJDPSZ $JDIPSJVNJOUZCVT BOEmFMEFGmDBDZ PGJWFSNFDUJOBHBJOTUHBTUSPJOUFTUJOBM OFNBUPEFTJO%BOJTIDBUUMF IFTJT U 1I% .JHVFM1F×B&TQJOP[B Anthelmintic activity of forage chicory (Cichorium intybus) and field efficacy of ivermectin against gastrointestinal nematodes in Danish cattle Miguel Angel Peña-Espinoza PhD thesis Section for Bacteriology, Pathology and Parasitology National Veterinary Institute Technical University of Denmark Copenhagen 2016 Supervisors Ulrik Boas, Associate Professor Section for Immunology and Vaccinology National Veterinary Institute Technical University of Denmark Heidi Larsen Enemark, Senior Researcher Section for Parasitology Norwegian Veterinary Institute Stig Milan Thamsborg, Professor Department of Veterinary Disease Biology University of Copenhagen Table of Contents Preface .................................................................................................................................................. 1 Acknowledgments .............................................................................................................................. 2 Abstract ................................................................................................................................................ 4 Sammendrag ....................................................................................................................................... 7 Resumen ............................................................................................................................................ 10 Abbreviations .................................................................................................................................... 14 Introduction ...................................................................................................................................... 15 Chapter 1: Background .................................................................................................................... 16 1.1 Gastrointestinal (GI) nematodes of cattle: Ostertagia ostertagi and Cooperia oncophora...................................................................................................................................... 16 1.1.1 Life cycle ......................................................................................................................... 16 1.1.2 Pathophysiology ............................................................................................................ 18 1.1.3 Impact of GI nematodes on animal health and productivity .................................. 19 1.1.4 Immune response .......................................................................................................... 20 1.1.5 Epidemiology ................................................................................................................. 20 1.2. Control of GI nematodes of cattle with anthelmintics .................................................... 22 1.2.1 Ivermectin: mechanisms of action and expected in vivo efficacy in cattle ............ 22 1.2.2 Anthelmintic resistance in GI nematodes of cattle................................................... 23 1.2.3 Field detection of anthelmintic resistance in GI nematodes of cattle .................... 24 1.3 Alternative parasite control strategies ................................................................................ 25 1.4 Bioactive plants ...................................................................................................................... 26 1.4.1 Methods for evaluation of anthelmintic activity ....................................................... 27 1.5 Chicory ................................................................................................................................... 29 1.5.1 Effects of forage chicory on GI nematodes of ruminants ........................................ 30 1.5.2 Bioactive compounds in forage chicory ..................................................................... 35 1.5.3 Sesquiterpene lactones .................................................................................................. 35 Chapter 2. The present investigations ........................................................................................... 39 2.1 Objectives: .............................................................................................................................. 39 2.2 Materials and methods ......................................................................................................... 40 2.2.1. Field efficacy of ivermectin and prescription of anthelmintics in Danish cattle . 40 2.2.2. In vivo studies of the anthelmintic effects of forage chicory against GI nematodes in experimentally infected cattle ...................................................................... 44 2.2.3. In vitro studies of the anthelmintic activity of plant secondary metabolites of forage chicory .......................................................................................................................... 46 2.2.4 Parasite material and animals ...................................................................................... 47 2.2.5 Experimental feeds for in vivo studies and plant material for in vitro studies ..... 48 2.2.6 Parasitological analyses ................................................................................................ 50 Chapter 3: Manuscripts and preliminary studies − results and discussion .............................. 51 3.1. Field efficacy of anthelmintics in Danish cattle ............................................................... 51 3.1.1 Prescription of anthelmintics in Danish cattle: preliminary results ...................... 51 3.1.2 Reduced efficacy of ivermectin against GI nematodes in Danish cattle and comparison of methods to analyse FECR data (Manuscript I) ........................................ 53 3.2 Forage chicory selectively reduces O. ostertagi worm burdens in two independent experiments, while C. oncophora is unaffected (Manuscript II) ........................................... 56 3.3 In vitro studies of the anthelmintic activity of plant secondary metabolites of forage chicory ........................................................................................................................................... 58 3.3.1 In vitro studies of the direct anthelmintic effects of purified forage chicory extracts against Cooperia oncophora: preliminary investigations .................................... 58 3.3.2 Sesquiterpene lactone-containing extracts from two chicory cultivars show distinct chemical profiles and direct anthelmintic effects against O. ostertagi L1 and adult worms (Manuscript III) ............................................................................................... 59 Chapter 4: Conclusions ................................................................................................................... 61 Chapter 5: Perspectives and future research................................................................................. 63 Chapter 6: References ...................................................................................................................... 66 Chapter 7: Manuscripts ................................................................................................................... 84 7.1 Manuscript I ........................................................................................................................... 85 7.2 Manuscript II ...................................................................................................................... 125 7.3 Manuscript III ..................................................................................................................... 141 Preface The research work presented in this PhD thesis was conducted between October 2012 and December 2015 in the Section for Bacteriology, Pathology and Parasitology and Section for Immunology and Vaccinology at the National Veterinary Institute, Technical University of Denmark, and in the Section for Parasitology and Aquatic Diseases, Department of Veterinary Disease Biology, University of Copenhagen. The PhD studies were funded by the EMIDA ERA-NET project “Coping with anthelmintic resistance in ruminants” (CARES; 3405-11-0430/32) and CONICYT Chile (Becas Chile, segunda convocatoria 2009). The thesis is organized as a succinct review of gastrointestinal nematodes in cattle, anthelmintic resistance and the use of bioactive plants as an alternative helminth control approach, with focus on studies with forage chicory. Next, a brief description of the performed investigations is presented, including the experimental designs and methods, a summary of the major findings and conclusions, and final remarks on future research. The thesis culminates with the following scientific articles describing in detail the research work conducted during the PhD studies: Manuscript I. Efficacy of ivermectin against gastrointestinal nematodes of cattle in Denmark evaluated by different methods for analysis of faecal egg count reduction. International Journal for Parasitology: Drugs and Drug Resistance (in press). Manuscript II. Anthelmintic effects of forage chicory (Cichorium intybus) against gastrointestinal nematode parasites in experimentally infected cattle. Parasitology 2016, 143, 1279−1293. Manuscript III. Sesquiterpene lactone containing extracts from two cultivars of forage chicory
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