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The promastigote, literally the body form with an anterior flagellum is 15-30 um in body length and 5mm in width; it is extracellular, motile, and grows and divides by longitudinal binary fission at 27 C in the sandfly. Promastigotes can be grown in vitro at 25 C temperature on NNN medium, which has a solid phase of blood agar and a liquid phase containing a physiologic salt solution. Liquid media that support promastigote growth are also available. Amastigotes usually are grown inside tissue culture cells and can also be grown extracellularly at 37 C under special conditions.
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Life Cycle
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Leishmania invade the phagocytic cells of the reticulo-endothelial system and macrophages, but are able to evade destruction in these phagocytic cells. Transmission is by the bite of female sandflies in the genera, Phleobotomus and Lutzomyia. Sandflies are pool feeders and when infected macrophages are taken up by the blood-feeding sandfly, the amastigotes transform into promastigotes.
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The promastigotes in the gut of the female sandfly attach to the midgut-hindgut epithelium where they divide by binary fission. Development in the sandfly takes 8-20 days after initial ingestion of blood, and after multiplication in the midgut-hindgut they move forward to the pharynx where they produce a partial or complete blockage of the sucking apparatus. (These are called metacyclic promastigote forms and the trigger for metacyclogenesis is a decline in tetrahydrobiopterin). When such a sandfly attempts to feed (one could say the fly clears its throat) it regurgitates a bolus of metacyclic promastigotes and this is injected into the bite wound. There is no infection of the sandflys salivary glands as in the case of the african trypanosomes in the tsetse fly.
The promastigotes are phagocytized by macrophages or other professional phagocytes, and transform into
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intracellular amastigotes. Although promastigotes specifically interact with the surface of the host cell they cannot actively penetrate the host cell (unlike T. cruzi) and thus infection is dependent on host cell phagocytosis.
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Study questions: 1. Can the entire life cycle of Leishmania sp. be studied in culture? 2. Why do you think that the parasites become promastigotes in the sandfly and why amastigotes in the vertebrate host?
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