apparent, the edges of a scalp lesion should 
be scraped with a sterile scalpel. Skin lesions 
should first be cleansed with 70% alcohol 
to reduce bacterial and saprophytic fungi. 
Scrapings should be made from the outer 
edges of skin lesions. In infections of the 
nails, the friable material beneath the edge 
of the nails should be scraped out, or por- 
tions of abnormal appearing nail should be 
scraped or clipped off, and saved for exami- 
nation and culture. 
Enclose hair specimens, skin scrapings, or 
nail clippings or scrapings in clean paper 
envelopes and label them with the patient’s 
name or specimen number. Enclose these 
envelopes in larger heavy paper envelopes 
for mailing to the laboratory. Do not put 
specimens in cotton plugged tubes, as the 
specimen may become trapped among the 
cotton fibers and lost. Do not put specimens 
into closed containers such as rubber 
stoppered tubes as this keeps the specimen 
moist and allows overgrowth of bacteria and 
saprophytic fungi. 
2. Subcutaneous fungus infections. Procedures for 
isolating the etiologic agents of chromoblastomy- 
cosis, mycetomas, and sporotrichosis are 
obvious. Granules, pus, or serosanguinous 
fluid from lesions or draining sinuses should be 
inoculated on several tubes of Sabouraud dex- 
trose agar with and without chloramphenicol and 
cycloheximide. 
3. Systemic fungus infections. Systemic mycoses 
are not contagious and most are chronic and 
evolve slowly. Since time is usually not of vital 
importance, it may be profitable to consult with 
a mycology laboratory to determine what type of 
specimen to collect in a given case, and how it 
should be handled. Examining unstained prepa- 
rations of clinical material as well as stained 
smears is of practical value in determining the 
appropriate types of media to inoculate and the 
correct incubation temperatures. It is usually 
wise to inoculate both simple media, such as 
Sabouraud dextrose agar, and enriched media, 
such as brain heart infusion agar. Each medium 
should be made with and without antibiotics. 
Infections with Histoplasma capsulatum and 
Coccidioides immitis present special problems 
to the laboratory worker. In the case of histo- 
plasmosis it is often difficult to isolate the 
causative fungus because the organisms are often 
few in number and tend to die out in the clinical 
materials. In the case of coccidioidomycosis the 
fungus agent is relatively easy to isolate. It is 
highly infectious, however, and many laboratory 
workers hesitate to attempt its isolation. Sug- 
gestions on methods for the isolation and 
handling of H. capsulatum and C. immitis are 
given below, 
a. Actinomycosis 
If isolates of suspected anaerobic to micro- 
aerophilic Actinomyces species are to be sent 
to a reference laboratory for identification, 
the following methods are recommended. 
(1) Liquid media 
(a) Inoculate culture into freshly 
boiled TST enriched* thioglycol- 
late broth. Incubate at 37° C. for 
3 to 4 days. If growth is apparent 
at that time, seal by pouring over 
the surface of the medium about 
3 ml. of a sterile melted mixture of 
paraffin and vaseline (50% paraf- 
fin & 50% vaseline).** Seal the 
top of the tube with a sterile 
rubber stopper, or use a screw-cap 
tube sealed with masking tape. 
(b) If Actinomyces Maintenance 
Broth*** is used, inoculate the 
tube of broth and incubate under 
pyrogallol-carbonate seal for 3 to 
4 days. If growth is apparent, 
transfer the liquid with the culture 
to a clean sterile tube, and seal 
with vaseline-paraffin mixture as 
descibed above. 
THE IMPORTANT POINTS 
TO REMEMBER ARE: 
(1) Actinomyces cultures should 
not be more than 3 to 4 days 
old before shipment. Send 
them by airmail if possible. 
(2) Liquid or semisolid cultures, 
should be sealed with vase- 
line-paraffin mixture before 
shipment. 
* Thioglycollate with TST 
Thioglycollate broth 
(with dextrose and indicator) 29.5 gms. 
Trypticase Soy broth 1.5 gm. 
Try ptose broth 1.25 gm. 
Distilled water 1000.00 ml. 
** A stock vaseline-paraffin mixture may be prepared by 
melting together equal amounts of each. After mixing 
well, it may be distributed in 3 ml. amounts in plugged 
tubes for sterilization. These may be stored for use as 
needed. 
*** This medium is recommended for fastidious strains, 
which do not grow well in thioglycollate broth. It is 
available in desiccated form from BBL. Division of Bio 
Quest, P. O. Box 175. Cockeysville. Md.. 21030. 
17 
