Background: In Peru, the information regarding sand fly vectors of leishmaniasis and bartonellosis in the Amazon region is limited. In this study, we carried out sand fly collections in Peruvian lowland and highland jungle areas using different trap type configurations and screened them forLeishmaniaandBartonellaDNA.Methodology/Principal findings: Phlebotomine sand flies were collected in Peruvian Amazon jungle and inter Andean regions using CDC light trap, UV and color LED traps, Mosquito Magnet trap, BG Sentinel trap, and a Shannon trap placed outside the houses.Leishmaniaspp. screening was performed by kDNA PCR and confirmed by a nested cytochrome B gene (cytB) PCR.Bartonellaspp. screening was performed by ITS PCR and confirmed by citrate synthase gene (gltA). The PCR amplicons were sequenced to identifyLeishmaniaandBartonellaspecies.UV and Blue LED traps collected the highest average number of sand flies per hour in low jungle; UV, Mosquito Magnet and Shannon traps in high jungle; and Mosquito Magnet in inter Andean region.Leishmania guyanensisinLutzomyia carrerai carreraiandL.naiffiinLu.hirsuta hirsutawere identified based oncytBsequencing.Bartonellaspp. related toBartonella bacilliformisinLu.whitmani,Lu.nevesi,Lu.hirsuta hirsutaandLu.sherlocki, and aBartonellasp. related toCandidatusB. rondoniensis inLu.nevesiandLu.maranonensiswere identified based ongltAgene sequencing.Conclusions/Significance: UV, Blue LED, Mosquito Magnet and Shannon traps were more efficient than the BG-Sentinel, Green, and Red LED traps. This is the first report ofL.naiffiand of two genotypes ofBartonellaspp. related toB.bacilliformisandCandidatusB. rondoniensis infecting sand fly species from the Amazon region in Peru.

The Peruvian-Brazilian border is a highly endemic tegumentary leishmaniasis region in South America. The interoceanic highway is a commercial route that connects Peru and Brazil through Madre de Dios and has raised concerns about its impact on previously undisturbed areas. In order to assess leishmaniasis transmission risk along this highway, we conducted a surveillance study of the sand fly populations in this area. Sand flies were collected between 2009 and 2010 along transects at 200 m, 600 m and 1000 m from six study sites located along the highway (Iberia, La Novia, Alto Libertad, El Carmen, Florida Baja, Mazuko and Mavila) and an undisturbed area (Malinowski). Collected specimens were identified based on morphology and non-engorged females of each species were pooled and screened by kinetoplast PCR to detect naturalLeishmaniainfections. A total of 9,023 specimens were collected belonging to 54 differentLutzomyiaspecies including the first report ofLu.gantieriin Peru. Four species accounted for 50% of all specimens (Lutzomyia carrerai carrerai,Lu.davisi,Lu.shawiandLu.richardwardi). El Carmen, Alto Libertad, Florida Baja and Malinowski presented higher Shannon diversity indexes (H = 2.36, 2.30, 2.17 and 2.13, respectively) than the most human disturbed sites of Mazuko and La Novia (H = 1.53 and 1.06, respectively). PCR detected 10 positive pools belonging toLu.carrerai carrerai,Lu.yuilli yuilli,Lu.hirsuta hirsuta,Lu.(Trichophoromyia)spp., andLu.(Lutzomyia)spp. Positive pools from 1,000 m transects had higher infectivity rates than those from 600 m and 200 m transects (9/169 = 5.3% vs 0/79 = 0% and 1/127 = 0.8%, p = 0.018). El Carmen, accounted for eight out of ten positives whereas one positive was collected in Florida Baja and Mazuko each. Our study has shown differences in sand fly diversity, abundance and species composition across and within sites. Multiple clusteredLutzomyiapools with naturalLeishmaniainfection suggest a complex, diverse and spotty role in leishmaniasis transmission in Madre de Dios, with increased risk farther from the highway.

Leishmaniasis and Carrion's disease are endemic neglected diseases in Peru. Both diseases are transmitted by phlebotomine sand flies competent for infection with Leishmania species and Bartonella bacilliformis. In Peru, 149 sand fly species have been identified. Knowledge regarding sand fly species diversity and distribution in leishmaniasis and Carrion's disease transmission areas has grown in the past 2 decades through multiple surveillance efforts in endemic regions, including unexplored low and high jungle areas. Furthermore, molecular testing approaches have been implemented to enhance surveillance of Leishmania and Bartonella species in sand fly populations from these endemic regions. This review emphasizes the importance of entomological surveillance in Peru for the characterization of sand fly species and the discovery of a great diversity of Leishmania and Bartonella species in sand fly populations from Andean and Amazonian regions.

Phlebotomine sand flies are blood-sucking dipterans widely distributed in tropical and subtropical areas of the Americas and are important vectors of leishmaniasis caused by Leishmania spp. parasites and Carrion’s disease caused by the bacteria Bartonella bacilliformis . Both are a significant economic burden in rural areas and a major risk to military personnel deployed to endemic areas. To better understand transmission of these pathogens and epidemiological trends, sand flies were collected from nine sites across the Ecuador-Peru border region in 2015 and 2017 and screened for Leishmania using PCR targeting kinetoplast DNA, and Bartonella using PCR targeting the 16S-23S internal transcribed spacer (ITS) region, citrate synthase ( gltA ) gene, and NADH dehydrogenase subunit G ( nuoG ) gene. A total of 548 sand flies belonging to 15 species and 2,711 sand flies belonging to 11 species were collected in Ecuador and Peru, respectively. Pintomyia (Pifanomyia) robusta was generally the most abundant species found across sites sampled in Ecuador and Peru. In the Chinchipe River basin, Pi. (Pif.) robusta , Pintomyia (Pifanomyia) maranonensis , and Lutzomyia (Helcocyrtomyia) castanea were collected on both sides in Zamora-Chinchipe, Ecuador, and Namballe, Peru. Of the 637 phlebotomine sand fly pools screened, no Leishmania positives were found; however, nine pools of Pi. (Pif.) robusta collected in Ecuador were positive for B. bacilliformis based on phylogenetic analysis of the gltA gene. One Pi. (Pif.) maranonensis from Peru and one Pi. (Pif.) robusta from Ecuador were positive for Bartonella DNA sequences that were close to Candidatus Bartonella rondoniensis based on gltA . This is the first reported detection of B. bacilliformis DNA in Pi. (Pif.) robusta , providing evidence for the role of this sand fly species in transmission of this pathogen at the Ecuador-Peru border.

Phlebotomine sand flies are the biological vectors of Leishmania spp. and host complex microbial communities that may modulate insect physiology, ecological adaptation, and vector competence. In Colombia, where leishmaniasis remains a significant public health concern, characterization of sand fly-associated microbiota may provide insights into host–microbe–parasite interactions and identify candidates for microbiome-informed control strategies. Here, we characterized the bac-terial and fungal microbiomes of field-collected female sand flies from three municipalities in Antioquia, Colombia. Specimens were identified using morphological characters and confirmed by cytochrome C oxidase subunit I sequencing. Bacterial and fungal community composition was assessed by amplicon sequencing of the 16S rRNA gene and the internal transcribed spacer 2 (ITS2) region, respectively. To determine natural parasite infection status, all specimens were screened for Leishmania by nested PCR targeting the cyt b gene. Five sand fly species were de-tected, including the recognized vector species Nyssomyia olmeca bicolor, Lutzomyia gomezi, and Psychodopygus panamensis, and the non-vectors Pressatia dysponeta and Pr. camposi. No specimen tested positive for Leishmania. Across taxa, bacterial communities were dominated by members of the phyla Proteobacteria, Firmicutes, and Actinobacteria. In contrast, Bacteroidetes exhibited significantly higher relative abundance in vector species than in non-vectors, suggesting possible associations between bacterial community structure and vector status. Fungal communities were primarily composed of Basidiomycota and Ascomycota, and beta-diversity analyses indicated significant compositional differentiation among genera. Collectively, these data demonstrate that sand flies from endemic areas of Colombia harbor taxonomically distinct bacterial and fungal assemblages, with detectable differences between vector and non-vector species.