SALINE
IODINE
MIF
IRON
TRICHROME* CBE
FLAGELLA &
colorless colorless
dark red
black
red
gray-
AXONEMES
black
EGGS AND
natural
tan to
pink to
gray to
red
gray-
LARVAE
color
dark brown
dark red
black
black
* Small cyst nuclei may stain blue with trichrome.
2-21. CULTIVATION OF PARASITES
The cultivation of parasites has proven to be a beneficial technique in the study of
human parasitology. The time, effort, and money spent in these techniques have
prevented the methodology from being utilized in the routine medical parasitology
laboratory. Teaching institutions have a constant need for study material, and therefore,
have more of a need for the procedures enumerated below.
a. Boeck and Drbohlav's Medium (modified). This is a semi-solid egg-based
medium that requires an overlay with Locke's solution. It is used for most intestinal
protozoans (excluded: Balantidium coli and Giardia lamblia).
(1)
Reagents.
(a) Locke's solution (modified).
Sodium chloride (NaCl) ..................................... 8.0 gm
Calcium chloride (CaCl2.2H20) .......................... 0.2 gm
Potassium chloride (KCl)................................... 0.2 gm
Magnesium chloride (MgCl2.6H20) .................... 0.01 gm
Disodium hydrogen phosphate (Na2HPO4) ....... 2.0 gm
Sodium bicarbonate (NaHCO3) ......................... 0.4 gm
Potassium dihydrogen phosphate (KH2PO4) ..... 0.3 gm
Distilled water .................................................... 1,000.0 ml
1 Add the salts in the order listed to 500 milliliters of distilled water
in a 2,000 milliliter Erlenmeyer flask and mix well. Slowly add the remaining 500 milliliters
of water and mix until all the salts are dissolved.
2 Boil the solution for 10 minutes (a white precipitate
should be formed).
3 Allow to cool at room temperature, filter, and autoclave for 15
minutes at 121 C (15 pounds).
MD0841
2-56