(e) When sputum cannot be obtained, 
gastric washings may be sub- 
mitted. They should be collected 
in a sterile bottle containing 
chloramphenicol. 
(f) Spinal fluid is taken only if there 
is cerebral meningeal involvement. 
It is collected in a tube or bottle 
containing antibiotic. 
(2) Animal inoculation 
Intraperitoneal inoculation of mice 
with clinical materials greatly increases 
the probability of demonstrating the 
presence of H. capsulatum. Sacrifice the 
animals after 2 weeks and culture the 
liver and spleen on cycloheximide agar 
at room temperature and on blood agar 
without antibiotics at 37° C. 
d. Fluorescent Antibody (FA) Staining in 
Mycology 
In certain reference laboratories, FA tech- 
nics are regularly used in conjunction with 
conventional methods of identification. 
Currently these procedures have been most 
successfully applied in medical mycology in 
the identification and detection of Sporo- 
trichum schenckii, Blastomyces dermatitidis, 
Cryptococcus neoj ormans, H. capsulatum, 
and C. immitis. 
Specimens for examination by the fluorescent 
antibody technique are obtained from such 
varied sources as blood, spinal fluid, sputum, 
lesion exudates, sternal marrow, scrapings 
from edges of ulcers and abscesses, and 
aspirated fluid from abscesses and tissue. If 
the laboratory is close by, such specimens 
can be submitted directly to the laboratory. 
If these materials have to be shipped to a 
distant laboratory, smears should be pre- 
pared for shipment. Sputum should be 
enzymatically digested (by parasentin or 
trypsin) and smears made from the centri- 
fuged sediment. Such smears are made 
directly within etched circular (1 cm. di.) 
areas of glass slides and are then allowed to 
air dry. These preparations are fixed by heat. 
Other materials such as exudates or tissues 
do not require digestion. These smears are 
made directly within the etched areas of 
glass slides, are allowed to air dry and are 
then fixed by heat. Spinal fluid or gastric 
washings should be centrifuged and smears 
made from the sediment. 
PARASITIC DISEASES 
The diseases caused by animal parasites may be 
conveniently considered as blood, intestinal, or tissue 
infections; and laboratory diagnosis may be made by 
direct examination of the specimen or indirectly by 
serologic tests. The specimens submitted for labora- 
tory examination may be smears of bone marrow, 
liver, spleen, or whole blood, aspirates from lesions, 
vaginal or urethral secretions, bile, sputum, duodenal 
drainage, anal swabs, feces, or portions of the 
parasite itself. 
1. Blood Parasites. Proper collection and handling 
of specimens to be examined for blood parasites 
are important since inadequate or poor samples 
may lead to erroneous conclusions. Not all the 
organisms usually grouped as blood parasites are 
diagnosed from blood. In certain instances, 
spinal or peritoneal fluid, aspirates and biopsies 
of organs and tissues are also used. The speci- 
men to be obtained depends on the location of 
the parasite or its diagnostic stage in the body. 
Table 1 shows the location of parasites or their 
diagnostic stage within the body. 
TABLE 1 
Location of Farasites or their 
Diagnostic Stage within the Body 
PERIPHERAL BLOOD OTHER 
Within 
RBC 
In 
Plasma 
Within 
Leuco- 
cytes 
Lx: 
00 
Bone 
Marrow 
Lymph 
Nodes 
Spinal 
Fluid 
Plasmodia spp. (4) 
+ 
+ 
Trypanosoma gambi- 
ense and T. 
rhodesiense 
+ 
+ 
+ 
Trypanosoma cruzi 
+ 
+ 
Leishmania donovani 
+ 
+ 
+ 
Leishmania tropica 
and L. brasiliensis 
+ 
Filaria-microfilariae 
(5 spp.) 
+ 
+* 
+ *' 
Onchocerca volvulus 
+ 
Toxoplasma gondii 
+ 
+ 
+ 
# Loa loa microfilariae may be found in calabar swellings 
*’ > Wuchereria bancrofti and Brugia malayi microfilariae 
a. Blood specimens 
Two types of specimens are collected for 
recognition of those parasites whose diag- 
nostic stages are found in peripheral blood, 
(1) dried blood films for staining and (2) 
19 
