156 FUNGI: NON-PATHOGENIC AND PATHOGENIC. 



recognized by their large size, doubly contoured membrane, 

 and refractile granular contents, and perhaps by evidences of 

 budding at some portion of the cell wall. Staining of the pus 

 or scrapings is not so satisfactory as an examination of the fresh 

 material, but this can be carried out by making film^ and stain- 

 ing in an old solution of Loffler's methylene-blue or by Gram's 

 method, as described in Chapter III. 



Isolation in pure culture is most readily effected by the use 

 of saccharine media, such as glucose or maltose broth (or agar), 

 beerwort or beerwort agar or gelatin (see Chapter II.), yet in 

 spite of all efforts some cases yield negative results. Inoculation 

 of the media is made from pus, scrapings of the growth, or even 

 with very small pieces of excised tissue from the growing edge 

 of the lesion, and incubation at 37 C. is to be prolonged for two 

 weeks before any culture is declared negative. The appear- 

 ance of round or oval, budding and non-budding cells, alone 

 or mixed with branching or budding mycelial threads, together 

 with general characters described above, may be regarded as 

 of a positive nature. Animal inoculations should likewise be 

 carried out. 



