4.1. Phlebotomines collected
The large number of specimens collected in the AUT may be related to the ability of these sand flies to adapt to anthropogenic environments, and the availability of blood and ideal shelter for the maintenance and survival of sand flies in the AUT, corroborating the work described by Sanguinette et al. (2015)  in the state of Minas Gerais.
Among the most collected species Nyssomyia intermedia and Migonemyia migonei are already described species with a high degree of anthropophily and with a high capacity to adapt to the environment modified by man [15–17]. Nyssomyia whitmani, on the other hand, has been shown to be a species with greater tendency to the transitional environment, and adapting to the peridomicile [18, 19]. The large number of these species collected in Timóteo demonstrates that they are already adapted to the city's urban environment. And the fact that these species are of medical importance requires special attention from health authorities, such information can help in taking preventive and control measures.
Although it is not the first record of Lutzomyia longipalpis in the study area, it is important to emphasize that the capture of Lutzomyia longipalpis is not common, since in previous studies it was not identified in the urban area of the municipality [6, 9] which draws attention to the beginning of a possible adaptation of Lutzomyia longipalpis in the municipality, as demonstrated in Mato Grosso do Sul by Oliveira et al. (2006) .
4.2. Detection of Leishmania spp. DNA
Psychodopygus davisi and Pressatia choti were the only ones detected with Leishmania DNA in PERD despite having a low detection rate. Special attention should be given especially when dealing with wild areas, as there is already a record of the participation of Pressatia choti in the transmission of Leishmania in the Amazon region [21–23] and identification of Psychodopygus davisi with Leishmania braziliensis infection in the state of Acre . It is important to point out that there were records of TL cases of professionals and visitors after incursions on this trail of the PERD (unpublished data).
This is the first record of Leishmania braziliensis DNA detection in Pressatia choti. This species is not found among the vector species of Leishmania, however, it was the predominant species among those collected in the PERD (19.12%). These findings raise the hypothesis that this species may participate as a vector of Leishmania braziliensis in the PERD.
Among the species with the presence of Leishmania DNA in the AUT, detection rates were higher than those reported in other studies. Oliveira-Pereira et al. (2006)  in the state of Maranhão, found a rate of 0.80% in females of Nyssomyia whitmani. Neitzke-Abreu et al. (2014)  worked in endemic regions of TL in the state of Paraná and reported a detection rate of 1.12% in Nyssomyia whitmani. Carvalho et al. (2008) , in a study carried out in the state of Minas Gerais, found a rate of 0.83% in Nyssomyia whitmani.
Despite the large number of females collected from the Nyssomyia whitmani species, the highest rate of Leishmania DNA detection was from Migonemyia migonei, a species already recognized as a vector and which may be actively participating in the transmission cycle of Leishmania in the municipality, together with Nyssomyia whitmani and Nyssomyia intermedia. It is possible, therefore, to suggest that these species are responsible for the transmission of Leishmania in the municipality.
Regarding the species Evandromyia lenti (DNA detection rate of 14.29%), Evandromyia sallesi (DNA detection rate of 3.13%) and Micropygomyia quinquefer (DNA detection rate of 0.88%), all had only a positive sample, but as they presented a low number of captured specimens, the result of the DNA detection rate was high. This does not indicate that these species are responsible for the transmission of Leishmania in the area. However, we should not ignore these findings even though species are not reported as vectors. There are even other studies recording the natural infection of these species, which is not enough to define them as vectors.
Micropygomyia quinquefer has already been registered with Leishmania braziliensis DNA in a study carried out in Mato Grasso do Sul . However, this information should be viewed with caution, as this species belongs to a group of sand flies in which females feed on cold-blooded animals .
There has already been detection of Leishmania sp. DNA in Evandromyia lenti and Evandromyia sallesi in the state of Minas Gerais [30–32]. In Evandromyia sallesi, natural infection by Leishmania infantum .
The result obtained from Nyssomyia intermedia, found in urban areas with Leishmania guyanensis DNA, should be treated with caution, since the region found does not belong to the geographic distribution region of this species, although there is already a record of this parasite species in three sand fly species in the north of Minas Gerais Therefore, there is a need for studies that can better demonstrate the circulation of Leishmania guyanensis in this region.
This is the first report of de Migonemyia migonei, a specimen collected in urban areas, with Leishmania amazonensis DNA. Experimental infection data have shown that infection by Leishmania amazonensis in Migonemyia migonei is possible [34, 35]. It is important to mention that Leishmania amazonensis have been recorded in Minas Gerais in sand flies and dogs [31, 36–38]. Therefore, the present study suggests that Leishmania amazonensis may also be a possible agent of leishmaniasis in the municipality of Timóteo.
Our results show that in the municipality of Timóteo there are at least three species of Leishmania spp. with the possibility of maintenance of the life cycle, being able to have direct participation in cases of leishmaniasis both in the urban area and in the wild area (PERD) of the city, but with the possibility of different vectors between the two areas.
4.3. Blood source study
Among the samples collected in PERD, only the female sand fly with blood of Homo sapiens was not collected in the inner area of the PERD, this female sand fly was identified as Nyssomyia intermedia, collected near the forest, at a distance of 300 meters from the edge from the forest, the others were collected in more internal areas, at a distance of more than 1,000 meters from the edge of the forest. It is interesting to note that the edge of the forest is an area close to human habitation and the presence of human beings in this region is intense, which justifies the identification of a blood source in female Nyssomyia intermedia as Homo sapiens.
Also in PERD, the DNA of Dasyprocta leporina was identified in fed Pressatia choti females, while Dasypus novemcinctus and Sphiggurus villosus were identified in the blood of Psychodopygus davisi females. Due to the finding of Leishmania DNA in these sand fly species, the hypothesis that these mammals could act as Leishmania reservoirs in the PERD. Previous studies have already reported the presence of natural infection in these species of mammals, as in the case described by Lainson et al., (1981)  in a study developed in the state of Pará, reporting the presence of natural infection in Dasypus novemcinctus. Paiz et al. (2015)  performing a study in Botucatu state of São Paulo, with serological analysis detected anti-Leishmania spp. in Sphiggurus villosus.
The presence of a blood source by Homo sapiens in the urban area should be seen as normal, since the fed female sand flies were collected in the peri-domicile. These data are in agreement with studies carried out by Carvalho et al. (2017)  who observed that sand flies from urban areas feed to a greater extent on dogs, chickens and humans. In the present work, studies indicate a lower proportion of Homo sapiens compared to Gallus gallus (z = 4.276 p < 0.0001) and Sus scrofa (z = 1.715 p < 0.05).
The role of birds in the epidemiology of leishmaniasis has been widely discussed, even though they are considered refractory to Leishmania infection. Tanure et al., (2015)  in the municipality of Governador Valadares verified that 43.6% of Lutzomyia longipalpis fed in Gallus gallus. Sus scrofa, on the other hand, as a mammal, draws attention to a possible reservoir in urban areas having already been found infected by Leishmania sp. in Maranhão . Baum et al. (2015) , using molecular analysis in females of Nyssomyia intermedia collected in the state of Paraná, verified the presence of samples with DNA of Sus scrofa. The role of Gallus gallus and Sus scrofa, even though the latter is not proven as Leishmania reservoirs, are important, since these species help to maintain the life cycle of sand flies in urban areas. The results obtained from the source of infection are essential to help the adoption of more effective measures to prevent and control leishmaniasis.