, Influence du pH du liquide de mise en suspension des (1,3)-?-D-glucanes sur leur dosage par le LAL

, i et le tampon PBS en l'absence de traitement, après broyage cellulaire et après broyage cellulaire combiné au traitement au Tween 80. Pour toutes les répétitions effectuées dans l'eau p.p.i, il n'y a pas de (1,3)-?-D-glucanes dosés. La décontamination par sonication des flacons contaminés pendant 2h dans de l'éthanol a permis de diminuer la concentration en (1,3)-?-D-glucanes en dessous de la limite de détection

, En revanche, pour les répétions réalisées dans du tampon PBS, des concentrations de glucanes de 4, p.5

, ± 0,43 pg.mL -1 en absence de traitement ; 2,94 ± 1,22 pg.mL -1 après le broyage seul, pp.3-078

. Pg, mL -1 après broyage suivi d'un traitement au Tween sont mesurées. Le tampon PBS est donc contaminé en (1,3)-?-D-glucanes

. La-mesure-du-ph-dans-les, deux liquides de mise en suspension montre une différence de pH entre l'eau p.p.i et le tampon PBS (figure 28) En effet, l'eau p.p.i est acide (pH=5,0 ± 0,3) tandis que le tampon PBS est neutre, Le pH de chaque solution reste stable après traitement

, utilisation de ces deux liquides est judicieuse pour étudier l'influence éventuelle du pH sur le dosage des (1,3)-?-D-glucanes par la

, Dans l'eau p.p.i, la concentration de glucanes mesurée en absence de traitement est de 1, p.38

. Pg, mL -1 (figure 29) Cette concentration est augmentée d'un facteur 80 après broyage cellulaire et d'un facteur supérieur à 115 après broyage cellulaire suivi d'un, p.75

, La répétabilité du dosage est améliorée après traitement des suspensions. Le coefficient de variation diminue de 19,31% sans traitement à 4

P. Dans-le-tampon, 16 ± 0,38 pg.mL -1 lorsque la suspension n'est pas traitée Après broyage cellulaire, une augmentation de la concentration mesurée d'un facteur 11 est observée. Cette augmentation est maximum après le broyage cellulaire suivi du traitement au Tween (facteur 47) Les coefficients de variation sont plus importants que dans l'eau, ils sont d'environ 13% Il ne varie pas selon les traitements

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, Annexe 1 : composition des milieux et solutions utilisés dans le cadre du stage Milieu MEA

M. Le-milieu and . Est-un-milieu-utilisé-pour-la-recherche, la culture sur milieu gélosé, l'isolement ou le dénombrement des moisissures et des levures Dans le cadre du stage, le milieu MEA est utilisé pour la culture de P.brevicompactum. Au laboratoire, ce milieu est préparé à partir d'un mélange de poudres pré-dosées

, Peptone mycologique 5 g

, Agar 15 g

, Milieu YPD (Yeast Peptone Dextrose)

, Le milieu YPD est un milieu riche utilisé pour la croissance des levures contenant : ? Bacto yeast extract 10 g

, Bacto peptone 20 g