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1942) so that the lower half of each strip was submerged. The tubes were plugged and autoclaved for 20 min at 15 lb pressure. Inoculation was by spore suspension and the tubes were incubated at 85°F for 10-14 days. Following incubation the strips were removed and washed in 1/1000 mercuric chloride, rinsed in water, and tested for breaking pressure. Wool was found to be less subject to fungus attack than cotton. Species attacking wool are limited, I. INTRODUCTION TO GENERAL METHODS 41 most are apparently dermatophytes such as Microsporum gypseum, or species related to dermatophytes.
Savory et al. (1967) considered the rather poorly defined clearance zone in this method to be a serious disadvantage, particularly when testing fungi with dark-coloured mycelia. They solved the problem by growing the fungus under test on a cellulose/mineral agar plate for about 14 days at 25°C. A new plate of the same medium was then poured and a series of spaced holes were cut in the agar using the Leach (Fig. 9) stainless steel cutter. The same cutter was used to cut plugs from the culture, and these plugs, with the surface mycelium uppermost, were inserted into the holes in the new plate.
It should be noted that culture pigmentation is directly related to and most strongly effected by the pH of the medium. Most fungi also require a high humidity for growth. H. dropped to approximately 90% (see Section X). 1. Pure and mixed cultures Although stress has bccn laid on the ncccssity of working with purc cultures, when one is dealing with sporulation and in particular when considering the production of perfect states it may be an advantage to use mixed cultures. In 1909 Heald and Pool found that Melanospora zamiae would produce perithecia only when grown with Nigrospora oryzae, Fusarium moniliforme or less successfully with Fusarium culmorum.
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