Koch's Postulates

             5. Flame sterilize a teasing needle, cool, then pierce skin of disinfected fruit.
             6. Unscrew cap on Penicilium italicum culture tube with one hand and flame the mouth of the tube.
             7. Using the first three fingers of your writing hand, obtain a sterile applicator stick and remove a small sample of the fungus and smear over the puncture wound of the fruit.
             8. Flame tube and recap. Discard swabs appropriately.
             9. Obtain a second sample with a sterile applicator stick and smear over an unpunctured section of the fruit.
             10. Flame tube and recap. Discard swabs appropriately.
             11. Maintain a control for the experiment.
             12. Make observations periodically (make note of the fungus" odor, shrinkage, discoloration or growth characteristics).
             13.Make a slide of your inoculating organism.
             14. Disinfect all work surfaces.
             15. Partially unseal bag to allow quick but aseptic entrance for sample collection. .
             16. With sterile applicator stick collect a sample of spores (blue-green, moldy decay) from the infected fruit.
             17. Streak a Potato Dextrose Agar plate and incubate at room temperature for approximately 5 days.
             18. Observe growth of culture periodically with lid intact.
             19. Growth may appear as circular, a white fuzzy mass of mycelium or white specks that gradually change color, from white to blue-green. Observe with a steromicroscope.
             20. Repeat cleaning and washing procedure as in steps 1-4 with appropriate number and type of fruits.
             21. Reinoculate your fruits (hosts) as in steps 5-11.
             22. Stain a sample of the organism that you have reisolated and compare with starting culture.
             23. Observe and record periodically.
             24. Repeat procedures as in steps 14-19.
             25. Observe periodically.
             26. At the end of the experiment, again stain your organism.
             Results.
             Macro-.
             Several weeks after initial inoculation, a substantial growth was evident. The fungal growth had a distinct odor of mold. The same amount of growth was evident around the puncture-swabbed area as the unpunctured-swabbed area.