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2.1.14. Definition of sensitivity of cultures of mushrooms to antifungalnym to preparations.

Definition of sensitivity of cultures of mushrooms to antifungalnym to preparations spent a disco-diffuzionnym a method. Used disks, impregnirovannye antifungalnymi preparations manufactures HiMedia (India): Amphotericinum-in (100 U/disk), nystatin (100 U/disk), klot - rimazol (10 mkg/disk), flukonazol (10 mkg/disk), ketokonazol (10 mkg/disk).

For crops on sensitivity definition prepared suspensions of cultures

(L

Mushrooms in a normal saline solution with the maintenance 500x10 cells in 1 sm on an optical turbidity standard (without recalculation for the size of a cell of a mushroom). Crops were carried out on nutrient mediums in cups Petri in volume by of 2-3 sm3, and rocking in regular intervals distributed a suspension on all surface of a nutrient medium. Excess of a suspension merged. Cups maintained at a room temperature about 40 minutes After that impregnirovannye disks placed in cups Petri by means of a dispenser equally spaced. Crops inkubirovali at 36С, the account of results spent for 7 days of cultivation, measuring diameter of a zone of suppression of growth (ZPR) cultures round a disk. Results of measurement interpreted according to the corresponding table applied by the manufacturer to a set of disks: amfote - a ricin In: diameter ZPR to 14 mm - the culture is steady; more than 14 mm - it is sensitive; nystatin: diameter ZPR to 18 mm - the culture is steady; more than 18 mm - it is sensitive; Clotrimazolum, flukonazol, ketokonazol: diameter ZPR to 12 mm - the culture is steady; more than 12 mm - it is sensitive.

2.1.15. A lyophilization of cultures of mushrooms of sort Malassezia.

Works on a lyophilization of cultures of mushrooms were spent together with the manager. Department of lyophilization FGU VGNKI of JU.G.Oparin.

Preparation of a suspension of yeast cells for the subsequent lyophilization included: cultivation of cultures of mushrooms on nutrient mediums in matrovyh flasks during 7 days at 36С; I will eat mushroom cultures from a surface of a nutrient medium a spatula or a scraper; preparation of a suspension of mushroom cultures in the distilled water; mixing of a suspension of mushroom cultures with protective medium.

The used protective mediums: protective medium №4ВГНКИ (structure: a peptone - 5 %, Sorbitum - 5 %, gelatin of 1,3 %); protective medium for yeast mushrooms (structure: milk, an inosite-5 %).

Suspensions of mushroom cultures admixed with protective medium in the ratio 1:1 then syringes packed up in ampoules (wide type ШПВ-6 (ILT) or narrow type ШПВ-3 (IBB) in volume 1 Ampoules see pawned in the aluminium cartridge and subjected a frost at-50S. A lyophilization spent on equipment GT-2 (firm Finn-Aqua, Germany). After lyophilic drying of an ampoule soldered under vacuum.

The control liofilizirovannyh cultures of mushrooms included: definition of presence of vacuum in an ampoule on apparatus "Spark"; definition of a residual share of a moisture in an ampoule in accordance with GOST 24061-89; definition of presence of extraneous microorganisms by crops method resuspenzirovannyh liofilizirovannyh cultures on nutrient mediums MPA, MPB, Kitt-Tarotstsi, medium of Capek, medium Saburo, a mash-agar; definition of the general concentration of mushroom cells in an ampoule a method of calculation of mushroom cells in chamber Gorjaeva; definition of concentration of teleorgnic mushroom cells in an ampoule at crops resuspenzirovannyh liofilizirovannyh cultures on nutrient mediums in cups Petri a method of serial delutions with the subsequent account of evolved colonies of mushrooms; variability definition kulturalnyh and morphological properties of mushrooms before and after a lyophilization.

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A source: Ershov Peter Petrovich Etiologicheskaja. the importance of yeast mushrooms of sort Malassezia at dermal diseases of animals. The dissertation on competition of a scientific degree of the candidate of veterinary sciences Moscow - 2008. 2008

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  2. Sensitivity of measurements
  3. measuring of a threshold of sensitivity
  4. Studying assimiljativnyh properties of mushrooms of sort Malassezia on M Crespo [58].
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