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Background  Almost 1000 cases of American cutaneous leishmaniasis have been registered yearly in Rondônia State, Brazil. Little is known about the  Leishmania  transmission cycle (vectors and reservoirs) in the state. This study aimed to evaluate sand fly fauna from two vertical stratification layers in order to identify potential vectors and their blood-meal sources.    Methods  The study was conducted in Jamari National Forest. Sand flies were collected in the canopy (15 m) and at ground level (1 m) using HP light traps during four months, February, April, August and October, 2018. Insects were identified to the species level, and females were subjected to DNA extraction and PCR targeting minicircle  k DNA and  hsp 70 (for  Leishmania  detection and species identification), and  cytb  (to identify blood-meal sources). Exploratory data analysis was used to determine mean of abundance and species richness between stratifications. The  hsp 70 and  cytb  sequences were analyzed and compared with sequences from GenBank.    Results  Overall, 68 species were identified from 15,457 individuals. On the Potosi trail, 7531 individuals of 49 species were collected; canopy captures totaled 6463 individuals of 46 species, while ground captures totaled 1068 individuals of 38 species. On the Santa Maria trail, 7926 individuals of 61 species were collected; canopy captures totaled 6136 individuals of 51 species, while ground captures totaled 1790 individuals of 53 species. A total of 23 pools were positive for  k DNA (canopy  n  = 21, ground  n  = 2). Only two samples were sequenced for  hsp 70 (both in canopy); one sequence exhibited similarity with  Leishmania braziliensis  ( Lutzomyia davisi  pool) and another with  L. naiffi  ( Lu. antunesi  pool). The  cytb  fragment was amplified in 11 of 86 samples. Sample sequencing identified  cytb  DNA from 5 blood-meal sources:  Micrastur gilvicollis ,  Psophia viridis ,  Tamandua tetradactyla ,  Homo sapiens  and  Choloepus didactylus .    Conclusions  Sand fly fauna is more diverse in the canopy than at ground level. Factors such as blood-meal sources, resting sites, and abiotic components probably contribute to high abundance in the canopy. Our results reinforce the possibility that  Lu. antunesi  and  Lu .  davisi  participate in  Leishmania  transmission in forest environments and may play an important role in transmission from sylvatic to human hosts.

parasites and vectors

Vertical stratification of sand fly diversity in relation to natural infections of Leishmania sp. and blood-meal sources in Jamari National Forest, Rondônia State, Brazil