PROGRESS IN PROTOZOOLOGY

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for the development of methods to control the transmission of these
amoebae, especially the human pathogen, E. histolytica. It is important
to develop methods to induce encystation under the unique conditions
in axenic cultures.
Leishmania
Dr. B. M. Honigberg presented the paper on the cultivation of
Leishmania submitted by Dr. Larry Hendricks who was unable
to attend the Congress. The cultivation methods of the two developmental
stages of this parasite were dealt with separately.
(1) Promastigotes
Methods to cultivate promastigote forms morphologically similar to
those found in the sandfly have existed for many years. This extracellular,
flagellated organism grows optimally at 30°C. The early workers
used monophasic or biphasic blood-containing media which have
been superceded by the semi-defined or defined liquid media more
suitable for assessing the condition and growth of the parasites and
for immunological or biochemical investigations.
Commercially available insect media of Grace; Mitsuhashi and Maramorosch,
and Schneider (Hendricks et al. 1978, Chi Ids et al.
1978) and the mammalian cell media, e.g., Eagle's MEM and TC 199
(B e r e n s et al. 1976) can generate satisfactory yields of promastigotes
of a wide variety of Leishmania sspp. These media available commercially,
and prepackaged lyophilized with 30% foetal bovine serum in
serum vials, have been useful for the diagnosis of both cutaneous and
visceral leishmaniasis from both the old and new world. The insect media
can support the large scale production of promastigotes for metabolic
studies. Several culture systems used for this purpose include the
chemostat (S c h a e f e r et al. 1970) and the fermenter (E n d e r s
et al. 1977). A typical bulk culture system consisted of promastigotes
grown in 9-liter quantities of Schneider's Drosophila medium supplemented
with 30% foetal bovine serum. Yields of up to 5 X 10 7 organisms/ml
produced 16 ml of packed cells of several strains of L. braziliensis
and L. mexicana. The tsetse fly-trypanosome medium SM (C u nn
i n g h a m 1977) mixed 1 : 1 with RPMI 1640 and enriched with 30%
foetal bovine serum has been used routinely for large scale production
of promastigotes (5—10 ml of packed cells) of strains of L. donovani from
East Africa, L. chagasi from Central America and cutaneous leishmania
from Central and South America and the Middle East.
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