148 



STUDY AND IDENTIFICATION OF MOULDS 



Glycerine agar, bread paste, or potato media are all suitable, but the best medium 

 is that of Sabouraud: 



Maltose, 4 . o grams. 



Peptone, i . o gram. 



Agar, i . 5 grams. 



Water, 100.0 c.c. 



Make the reaction about +2. 



Before inoculating media with moulds, some recommend placing the material in 

 60% alcohol for one or two hours to kill the bacteria. The moulds withstand such 

 treatment. 



In cultivating moulds it is best to use small Erlenmeyer flasks, containing about 

 Y in. of media on the bottom, for the development of the colonies. In order to 

 separate the mould we may take the hair or scales on a sterile slide and cut them 

 into small fragments with a sterile knife. Then moisten a platinum loop from 

 the surface of an agar slant, touch a fragment with the loop, and when it adheres 

 transfer it to the agar slant. Make four or five inoculations on the surface and from 

 suitable growth, after four to seven days, inoculate the medium in the Erlenmeyer 

 flask. Esmarch roll cultures are better than flask ones. 



Plauth recommends receiving the mould material between two sterile glass slides. 

 Seal the edge of the slide with wax and place the preparation in a moist chamber for 

 four to seven days. From developing fungus growth inoculate the medium in the 

 Erlenmeyer flask. A Petri dish containing several layers of thoroughly moistened 

 filter-paper in top and bottom makes a satisfactory moist chamber. 



For the study of the morphology of Monilia in cultures, Boggs used 

 stab cultures of 15% gelatin. The growth in the tube was hardened 

 in 10% formalin, the glass cracked off and sections of the gelatin 

 column cut across at any desired level. These blocks were sectioned 

 with the freezing microtome; stained in dilute aqueous fuchsin (i to 

 30) for several hours; then differentiated in saturated solution of citric 

 acid until nearly decolorized. The sections were floated on slides, air 

 dried without blotting, cleared in xylol and mounted in balsam. 



For staining fungi in sections of tissue Busse recommends the folio wing method: 

 i. Haematoxylin, 10 to 15 minutes, then wash in tap water. 2. Carbol fuchsin 

 (i to 20) 30 minutes or over night. Decolorize in alcohol for a few minutes 

 then through absolute alcohol and xylol to mount in balsam. The moulds are red. 



