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Background  Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with  Leishmania  possibly involved in pathogen transmission.    Methods  Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for  Leishmania  infection screening using kinetoplastid DNA-PCR (kDNA-PCR) and nested Real time PCR for species identification.    Results  A total of 10,897 phlebotomines belonging to the genus  Lutzomyia  (58 species) and  Brumptomyia  (2 species) were collected. Our study confirmed the widespread distribution and abundance of  Lutzomyia (Trichophoromyia) spp . (24%),  Lu .  whitmani  (19.4%) and  Lu .  yucumensis  (15.8%) in the region. Analysis of Shannon diversity index indicates variability in sand fly composition across sites with Villa Primavera presenting the highest sand fly diversity and abundance.  Leishmania  screening by kDNA-PCR resulted in 45 positive pools collected from Flor de Acre (34 pools), Mavila (10 pools) and Arca Pacahuara (1 pool) and included 14 species:  Lu .  yucumensis ,  Lu .  aragoi ,  Lu .  sallesi ,  Lu .  sherlocki ,  Lu .  shawi ,  Lu .  walkeri ,  Lu nevesi ,  Lu .  migonei ,  Lu .  davisi ,  Lu .  carrerai ,  Lu .  hirsuta ,  Lu .  (Trichophoromyia) spp .,  Lu .  llanosmartinsi  and  Lu .  whitmani .  Lutzomyia sherlocki ,  Lu .  walkeri  and  Lu .  llanosmartinsi  had the highest infection rates (8%, 7% and 6%, respectively). We identified  Leishmania guyanensis  in two  Lu .  whitmani  pools, and  L .  braziliensis  in two  Lu .  llanosmartinsi  pools and one  Lu .  davisi  pool.    Conclusions  Based on our collections there is high sand fly diversity in Madre de Dios, with differences in sand fly abundance and species composition across sites. We identified 14 sand fly species naturally infected with  Leishmania  spp., having detected natural infection with  L . ( V .)  guyanensis  and  L . ( V .)  braziliensis  in three sand fly species. These results suggest the presence of several potential vectors that vary in their spatial and geographical distribution, which could explain the high prevalence of CL cases in this region.

Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon