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Haemoflagellates: commercially available liquid media for rapid cultivation

Published online by Cambridge University Press:  06 April 2009

L. D. Hendricks
Affiliation:
U.S. Army Medical Research Unit/WRAIR/Panama, Box 2011, Balboa Heights, Canal Zone
D. E. Wood
Affiliation:
Gorgas Memorial Laboratory, Apartado 6991, Panama 5, Republic of Panama
M. E. Hajduk
Affiliation:
U.S. Army Medical Research Unit/WRAIR/Panama, Box 2011, Balboa Heights, Canal Zone

Summary

The successful cultivation of a variety of haemoflagellates in three different liquid media is reported. These media include medium 199, Grace's insect tissue-culture medium and Schneider's drosophila medium, each in combination with 30% (v/v) foetal calf serum. These media were used to cultivate Old and New World species of visceral and cutaneous human Leishmania, as well as Leishmania species isolated from sandflies, rodents, and reptiles. Four strains of Trypanosoma cruzi, an isolate of T. rangeli and an isolate of T. lewisi have also been cultivated in these media. One or more of these media have been used to cultivate 121 strains of haemoflagellates, including at least 14 different species (11 Leishmania and 3 Trypanosoma) and many geographic isolates or strains. The Leishmania include L. braziliensis, L. peruviana, L. mexicana, L. tropica, L. donovani, L. chagasi, L. enriettii, L. hertigi, L. hoogstraali, L. adleri, and L. agamae. Using the Schneider's based medium, we have obtained primary isolates of both cutaneous and visceral Leishmania of man and of experimentally infected laboratory rodents and canines. Freeze-dried preparations of the Schneider's based medium that were reconstituted with distilled water after 24 months of storage at ambient temperature have proven to be suitable cultivation media. This feature makes the media valuable field tools.

The various species of human Leishmania cultivated in these media have in our experience demonstrated no differences in growth rate, viability after liquid nitrogen preservation, or infectivity for laboratory animals and tissue-culture cells compared with promastigotes derived from blood-agar cultivation.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1978

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References

REFERENCES

Berens, R. L., Brun, R. & Krassner, S. M. (1976). A simple monophasic medium for axenic culture of hemoflagellates. Journal of Parasitology 62, 360–5.CrossRefGoogle ScholarPubMed
Cross, G. A. M. & Manning, J. C. (1973). Cultivation of Trypanosoma brucei spp. in semi-defined and defined media. Parasitology 67, 315–33.CrossRefGoogle Scholar
Dwyer, D. M. (1972). A monophasic medium for cultivating Leishmania donovani in large numbers. Journal of Parasitology 58, 847–8.CrossRefGoogle ScholarPubMed
Hendricks, L. D. (1975). Liquid media for the rapid and quantitative cultivation of Leishmania. 50th Annual Meeting, the American Society of Parasitologists, p. 68 (Abstr.).Google Scholar
Lemma, A. & Schiller, E. L. (1964). Extracellular cultivation of the leishmanial bodies of species belonging to the protozoan genus Leishmania. Experimental Parasitology 15, 503–13.CrossRefGoogle Scholar
Mansour, N. S., Hady, J. & McConnell, E. (1973). A modified liquid medium for Leishmania. Journal of Parasitology 59, 1088–90.CrossRefGoogle ScholarPubMed
Nuernberger, S., Ramos, P. C. V. & Custodio, R. (1975). Visceral leishmaniasis in Honduras. Report of three proven cases and a suspected case. American Journal of Tropical Medicine and Hygiene 24, 917–20.CrossRefGoogle Scholar
O'daly, J. A. (1975). A new liquid medium for Trypanosoma (Schizotrypanum) cruzi. Journal of Protozoology 22, 265–70.CrossRefGoogle ScholarPubMed
Pan, C. T. (1968). Cultivation of the leishmaniform stage of Trypanosoma cruzi in cell-free media at different temperatures. American Journal of Tropical Medicine and Hygiene 17, 823–32.CrossRefGoogle ScholarPubMed
Pan, C. T. (1970). Cultivation and morphogenesis of Trypanosoma cruzi in cell-free media. Proceedings of the 2nd International Congress of Parasitology, Washington, D.C., 1970. Journal of Parasitology 56 (Suppl.), 258.Google Scholar
Steiger, R. F. & Steiger, E. (1976). A defined medium for cultivating Leishmania donovani and L. braziliensis. Journal of Parasitology 62, 1010–11.CrossRefGoogle ScholarPubMed
Trager, W. (1953). The development of Leishmania donovani in vitro at 37 °C. Effects of the kind of serum. Journal of Experimental Medicine 87, 177–88.CrossRefGoogle Scholar
Wonde, T. & Honigberg, B. M. (1971). Morphology and infectivity of Leishmania donovani cultivated in nonliving media at elevated temperatures. American Journal of Tropical Medicine and Hygiene 20, 828–38.CrossRefGoogle ScholarPubMed