Régulation des populations de Nématodes gastro-intestinaux ...

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Résultats for 1h at 37°C with 200 µL of 5% skimmed milk PBS-T (PBS-TSM). Subsequently the PBS- TSM was discarded and the plates dried, before adding 100 µL of serum diluted in PBS-TSM or undiluted mucus to each well for 1h at 37°C. After three washes with PBS-T (the third being of 5 minutes), the plates were subsequently incubated for 1,5h with 100 µL of Donkey anti-sheep IgG HRP (Sigma, reference A3415) (IgG determination) or for two periods of 1h with the first (Mouse IgG1 anti-IgA bovine/ovine, Serotec, reference MCA628) and the second (Goat anti-mouse IgG1 HRP, Serotec, reference STAR81P) conjugates separated by three washings in PBS-T (IgA determination). Finally, the plates were washed three times with PBS-T and 100 µL of 2-2’-azino-bis (3-ethylbenylthiazoline-6-sulphonic acid, ABTS) in 100 mM citrate buffer (Sigma, reference 104.4) containing 0.01% H2O2 was added to each well. The colour was allowed to develop for 1h at 37°C. To stop the reaction, the plates were placed 15 min at 4°C. The optical density was measured at 405 nm using a Microplate Reader (System Dias, Dynatech). 2.5. Histology At necropsy, two tissue samples from duodenum (between 50 and 100 cm from the pylorus) were taken ; the first one was fixed in a 10% formalin for conventional histology and immunohistochemistry (IHC), and the second was frozen at –70°C for frozen IHC. 2.5.1. Conventional histology and IHC with paraffin-embedded samples Duodenal tissue was embedded in paraffin wax and tissue sections of 3 µm were mounted on glass slides. Eosinophils and globule leukocytes were counted on haematoxylin/eosin stained slides, whereas mast cells were counted on Giemsa stained slides. Immunohistochemistry was performed using antibodies already described for cell phenotyping on paraffin-embedded tissues in sheep (Lacroux et al., 2006). This antibody set allows specific labelling of macrophages/monocytes (mouse anti-human CD68 antibody, 140
Résultats clone Ki-M6, Serotec, 1:150 dilution), B lymphocytes (mouse anti-human CD20-like antibody, clone BLA-36, Novocastra, 1:50 dilution), or T lymphocytes (rabbit anti-human CD3 antibody, A 0542, DAKO, 1:100 dilution). 2.5.2. IHC on frozen samples Five µm sections of duodenal tissue were obtained using a Leika cryomicrotome. Slides were fixed for 20 min in an acetone bath then directly processed for IHC with the same method as for paraffin-embedded samples, using monoclonal antibodies directed against CD4 (mouse anti-ovine CD4, SEROTEC, reference MCA2213, 1:600 dilution) or CD8 (mouse anti-bovine CD8, SEROTEC, reference MCA837G, 1:800 dilution) antigens. For each cell type, cells were counted on ten randomly selected optical fields at high magnification (x400), equally distributed between deep and superficial lamina propria. The results were expressed as a sum of the ten fields. 2.6. Statistical analysis Comparisons between the three groups (previously-infected, primary-infected and uninfected control) and between the necropsy dates within a group was tested using the non- parametric tests of Kruskall-Wallis and Mann-Withney (SYSTAT Software). P0.217 (P0.283 (P
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N° d’ordre : 2346 THESE présent
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A mon très cher et respecté « co
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4.3.3. Mise en place et rôle des e
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ANNEXE A L’ARTICLE « HAEMONCHUS
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Un problème économique … et sci
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1. GENERALITES SUR LES STRONGLES GA
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1.2. Le cycle biologique des Tricho
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Famille Molécule Noms déposés Be
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3.1. Tarir les sources de contamina
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Contexte bibliographique Le tableau
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*La vaccination contre Trichostrong
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3.2.3. La sélection de la résista
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LIEU DE L’ETUDE RACES ETUDIEES /
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*Sélection au sein d’une même r
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Contexte bibliographique sont pas r
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*Réduction de la taille des vers a
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4.3.5. Régulation de la réponse i
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Contexte bibliographique d’infest
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Contexte bibliographique Figure n°
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Objectifs des travaux de thèse CHA
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Objectifs des travaux de thèse Dan
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1. MODELES EXPERIMENTAUX 1.1. Les a
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Procédures expérimentales mais n
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Procédures expérimentales 2.3. Et
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Page 107 and 108: Résultats CHAPITRE V : RESULTATS 1
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Page 111 and 112: Vet. Res. 37 (2006) 1-16 © INRA, E
Page 113 and 114: ut no clinical signs of coccidiosis
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Page 125 and 126: populations in sheep exposed to con
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Page 129 and 130: Résultats Les corrélations entre
Page 131 and 132: Article II : Immune responses in la
Page 133 and 134: 1. INTRODUCTION Résultats Due to t
Page 135 and 136: 2. MATERIAL AND METHODS 2.1. Experi
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Page 151 and 152: REFERENCES Résultats Balic, A., Bo
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Page 155 and 156: Table 1 : Schematic description of
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Page 169 and 170: 1 - INTRODUCTION Résultats The con
Page 171 and 172: 2 - MATERIALS AND METHODS 2.1. Expe
Page 173 and 174: Résultats of water. Larvae were co
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Table 3 : Total worm burden, length
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Epg Epg 30000 25000 20000 15000 100
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Discussion générale CHAPITRE VI :
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Discussion générale effectrices,
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Discussion générale chez des ovin
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Discussion générale connaissance
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Références bibliographiques Adams
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