Recovery of Leishmania Promastigotes from Infected Sandflies: A Study in Phlebotomine Artificial Blood Feeding

Location: Peru

Alumni: Andrew Taniguchi

Faculty: Miguel Morales

Leishmaniasis is a disease caused by the trypanosomatid parasite Leishmania and manifests itself in a variety of symptoms from cutaneous ulcers to degradation of mucosal areas, and is fatal in certain cases when left untreated. It is a neglected tropical disease that continues to be a burden for many countries because of the limited amount of diagnostic tools and complex treatments with severe side effects. The parasite causing the disease features a digenic lifecycle and requires the sand fly as an essential intermediary host and vector to continue propagating. This project explores the methods used for infecting Phlebotomus papatasi, Lutzomyia verrucarum, and Lutzomyia peruensis sandflies with Leishmania promastigotes. The sand fly infections with Leishmania major and Leishmania peruviana were accomplished via artificial membrane feeding procedures using a variety of animal skin membranes, and both human and mouse blood to mimic conditions in which natural infections are transmitted.  Detailed instructions on how to prepare membranes, blood meals, and dissections are outlined for successful recovery of motile promastigotes from infected sandflies and their subsequent re-culture for future molecular assays of Leishmania promastigotes. These protozoa contain a sophisticated signaling cascade system of phosphorylated proteins to transduce environmental signals. It is because of these signaling transductions that Leishmania are able to respond to stress and adapt to their environments in order to survive. Future experiments aimed at extracting these phosphorylated proteins, and identifying the associated kinases, phosphatases, and their downstream targets represent new avenues for the discovery of novel drug targets and therapies designed to mitigate the signaling pathways of Leishmania, effectively haling the lifecycle of Leishmania and/or preventing clinical disease from manifesting itself in treated hosts or vectors.

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