Preparation and application of Beauveria bassiana BB-7 metabolites
A technology of Beauveria bassiana and BB-7, applied in the field of microorganisms, can solve the problems of short-acting, strong toxicity, frequent use, etc.
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Embodiment 1
[0029] Example 1: Screening of antifungal activity of metabolites of Beauveria bassiana
[0030] 1. Beauveria bassiana ( Beauveria bassiana ) BB-7 is an endophytic fungus with antifungal activity isolated from the root of Hosta japonica. Collect fresh and healthy roots of Hosta japonica, after surface disinfection of the root tissue of Hosta japonica, check the cleanliness of the ultra-clean workbench, check the rinse solution, and screen the aseptic tissue blocks by plant tissue imprinting method; cut the above-mentioned aseptic tissues on the aseptic workbench Form small pieces with a diameter of about 0.5 cm, place them in PDA medium, and culture them upside down in a constant temperature fungal incubator at 25 °C. When endophytic fungal mycelium grows outward along the tissue incision, use the tip mycelia picking method. Colonies with different shapes were picked, and after repeated purification, they were transferred to PDA slant medium for purification, numbering, and a...
Embodiment 2
[0043] Example 2: Screening of active antifungal polar parts of Beauveria bassiana fermentation broth
[0044] 1. Preparation of different solvent extraction parts of Beauveria bassiana fermentation broth: after freeze-drying Beauveria bassiana fermentation broth, disperse with appropriate amount of distilled water, put the dispersed sample in a separatory funnel, and use petroleum ether, chloroform, Ethyl acetate was extracted 1:1 (v / v), repeated 3 times, the extracts were combined, and the extracts were concentrated and dried under vacuum at 40°C to obtain petroleum ether, chloroform fractions, ethyl acetate fractions, and water fractions (lyophilized) .
[0045] 2. Determination of antifungal activity of different solvent extraction parts of Beauveria bassiana fermentation broth: dissolve the above four polar parts with 500 μL DMSO, dilute to 10 mg / mL with distilled water and add the melted PDA medium at 1:9 ( 40-50°C), so that the final concentration was 1 mg / mL, and the ...
Embodiment 3
[0052] Example 3: Preparation of Beauveria bassiana fermentation broth spray film agent and its antifungal animal pharmacodynamics evaluation
[0053] 1. Preparation of film-spraying agent: Select polyvinyl alcohol (PVA17-88) and polyvinylpyrrolidone (PVP-K30), which are commonly used in pharmacy, as film-forming materials, use water as solvent, and glycerin as humectant. Antioxidants and preservatives are added, and a certain amount of Beauveria bassiana fermented liquid freeze-dried powder is added, and after being screened by the preparation process, the film spraying agent is prepared.
[0054] 2. Animal drug efficacy test of spray film
[0055] 2.1 Preparation of test animals
[0056] Three days before the inoculation of the bacterial solution, the white rabbits were subcutaneously injected with dexamethasone sodium phosphate immunosuppressant at a dose of 2.5 mg / kg, once a day, and observed.
[0057] 2.2 Establishment of animal model of fungal infection
[0058] Rabbi...
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