The effects of condensed tannins against cattle nematodes
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1 The effects of condensed tannins against cattle nematodes Adam Novobilský 1, 2 Irene Mueller-Harvey 3 Stig Milan Thamsborg 2 1 (SLU), Department of Biomedicine and Veterinary Public Health, Uppsala, Sweden 2 University of Copenhagen, Department of Veterinary Disease Biology, Denmark 3 University of Reading, Department of Agriculture, ASRG Chemistry & Biochemistry Laboratory, Reading, United Kingdom
2 Anthelmintic plants as a novel approach for controlling parasitic nematodes in ruminants Plant secondary metabolites (PSM): glycosides, saponins, lactones, polyphenols the polyphenolic compounds group called condensed tannins
3 Tannin-containing plants with anthelmintic activity against sheep and goat nematodes Sulla (Hedysarum coronarium) Birdsfoot trefoil (Lotus corniculatus) Big trefoil (Lotus pedunculatus) Sedicea lespedeza (Lespedeza cuneata) Sainfoin (Onobrychis viciifolia)
4 EU Marie Curie Project HealthyHay Research of sainfoin (Onobrychis viciifolia) improvement of breeding, investigation of chemical composition, nutritional, environmental and antiparasitic benefits of sainfoin Aim: in vitro effects of tannin-containing plant extracts against cattle nematodes Onobrychis viciifolia
5 Material and Methods 3 plant extracts: Onobrychis viciifolia Lotus pedunculatus Lotus corniculatus Chemical analysis of plant extracts: PC/PD ratio, CT content, mdp, cis/trans ratio 2 cattle nematodes: Ostertagia ostertagi Cooperia oncophora 2 in vitro assays: Larval feeding inhibition assay (LFIA) Larval exsheathment assay (LEA)
6 Scheme of larval feeding inhibition assay 100 L1 larvae E. coli labelled with FITC incubation 2 h incubation 20 h plant extract Unfed larva fed larvae unfed larvae Fed larva inverted fluorescence microscope larvae on slide
7 Scheme of larval exsheathment assay Incubation of ensheathed L3 in extract (3 h) Washing with PBS Transferring of larve into wells containing hypochlorite sodium Stop reaction by adding Lugol solution after 15, 30, 45, and 60 min Counting of exsheathed and ensheathed larvae under microscope
8 Results: LFIA Concentrations of extract: 2.5, 10, 40, 160 µg/ml Lev = Levamisole (40 µg/ml) as positive control PBS as negative control 160 µg/ml of extract+pvpp (50 mg/ml) O.v.= O. viciifolia L.p. = L. pedunculatus L.c. = L. corniculatus
9 EC 50 in LFIA and tannin content and composition of three plant extracts Plant extract C. oncophora O. ostertagi CT mdp PC/PD cis/trans ratio EC 50 (95% CI) EC 50 (95% CI) mean mean PC : PD cis : trans (µg/ml) (µg/ml) g CT/100g ext. mean (%) mean (%) Onobrychis viciifolia 8.6 ( ) 7.7 ( ) : : 18.4 Lotus pedunculatus 2.6 ( ) 2.6 ( ) : : 20.9 Lotus corniculatus 12.8 ( ) 16.1 ( ) : : 37.6
10 Results: LEA
11 Discussion all three plant CT extracts act aganist L1 and L3 of O. ostertagi and C. oncophora different sensitivity of LFIA and LEA ranking according to effect in LFIA: 1. L. pedunculatus 2. O. viciifolia 3. L. corniculatus anthelmintic effect associeted with CT content and PC/PD ratio need of in vivo experiment in cattle combination of pasture management and plant anthelmitics or chemical anthelmintics = succesfull control of GIT nematodes in future
12 Acknowledgements Ron H. Brown - University of Reading, UK Elisabetta Stringano - University of Reading, UK Heidi L. Enemark - DTU, Denmark Yvonne Bartley - Moredun Research Institute, UK Fotini Manolaraki - Ecole Nationale Vétérinaire de Toulouse, France Herve Hoste - Ecole Nationale Vétérinaire de Toulouse, France Janina Demeler - Freie Universität Berlin, Germany Study was supported by the EU Marie Curie project HealthyHay (contract: MRTN-CT ). For more information, please, visit:
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