Parasitological and molecular search for Leishmania natural infection in phlebotomine sand flies in peri-urban and rural sites of an Argentinean area endemic for tegumentary leishmaniasis

•This is the latest study on natural infection in the most endemic area of Tegumentary Leishmaniasis in Argentina.•Leishmania spp. natural infection was sought by gut dissection and duplex PCR.•A total of 1921 females were analyzed and all were negative for Leishmania infection.•The most abundant species was Nyssomyia neivai, followed by Migonemyia migonei.•The risk of transmission in the area is highlighted, since all the sand fly species captured are potential vectors. Leishmaniases are neglected tropical diseases caused by Leishmania spp. parasites transmitted by the bite of phlebotomine sand flies. In Argentina, the most endemic area of American tegumentary leishmaniasis (ATL) has been Orán department, Province of Salta, where Leishmania (Viannia) braziliensis prevails and Nyssomyia neivai is considered as its vector, although there is no accurate and sufficient information in this regard. The aim of this work was to search for natural infection by Leishmania spp. in sand flies from peri-urban and rural sites with ATL background in Orán department. For this, sand flies were caught at five sites; female sand flies captured with Shannon trap were dissected to microscopically examine their gut contents, while females captured with CDC traps were molecularly analyzed by duplex PCR with two primer pairs to simultaneously amplify kinetoplast DNA (kDNA) and mammalian actin. A total of 1921 females were captured, with Ny. neivai being the most abundant species (89%), followed by Migonemyia migonei (6%) and cortelezzii complex (3%). No natural infection was found in any of them neither by dissection nor by PCR, although the detection limit of kDNA PCR was up to 25 promastigotes. The absence of infected females in peri-urban sites suggest that the transmission did not take place in those environments during the study period. Future searches for natural infection should focus on rural settings to deepen knowledge and elucidate the role of the circulating sand fly species as all have been linked to ATL transmission at other sites. [Display omitted]