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Background    Phlebotomus orientalis  Parrot (Diptera: Psychodidae) is the main vector of visceral leishmaniasis (VL) caused by  Leishmania donovani  in East Africa. Here we report on life cycle parameters and susceptibility to  L. donovani  of two  P. orientalis  colonies originating from different sites in Ethiopia: a non-endemic site in the lowlands - Melka Werer (MW), and an endemic focus of human VL in the highlands - Addis Zemen (AZ).    Methodology/Principal Findings  Marked differences in life-cycle parameters between the two colonies included distinct requirements for larval food and humidity during pupation. However, analyses using Random Amplified Polymorphic DNA (RAPD) PCR and DNA sequencing of cytB and COI mitochondrial genes did not reveal any genetic differences. F1 hybrids developed successfully with higher fecundity than the parental colonies. Susceptibility of  P. orientalis  to  L. donovani  was studied by experimental infections. Even the lowest infective dose tested (2×10 3  per ml) was sufficient for successful establishment of  L. donovani  infections in about 50% of the  P. orientalis  females. Using higher infective doses, the infection rates were around 90% for both colonies.  Leishmania  development in  P. orientalis  was fast, the presence of metacyclic promastigotes in the thoracic midgut and the colonization of the stomodeal valve by haptomonads were recorded in most  P. orientalis  females by day five post-blood feeding.    Conclusions  Both MW and AZ colonies of  P. orientalis  were highly susceptible to Ethiopian  L. donovani  strains. As the average volume of blood-meals taken by  P. orientalis  females are about 0.7 µl, the infective dose at the lowest concentration was one or two  L. donovani  promastigotes per sand fly blood-meal. The development of  L. donovani  was similar in both  P. orientalis  colonies; hence, the absence of visceral leishmaniasis in non-endemic area Melka Werer cannot be attributed to different susceptibility of local  P. orientalis  populations to  L. donovani .

plos neglected tropical diseases

Phlebotomus orientalis Sand Flies from Two Geographically Distant Ethiopian Localities: Biology, Genetic Analyses and Susceptibility to Leishmania donovani