Several parameters should be addressed before incriminating a vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain parasite infection under laboratory conditions. This study evaluated the vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis–infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis–infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of vectorial capacity for L. infantum transmission. Moreover, it was also permissive to L. amazonensis.

Standardization of the methods for extraction of DNA from sand flies is essential for obtaining high efficiency during subsequent molecular analyses, such as the new sequencing methods. Information obtained using these methods may contribute substantially to taxonomic, evolutionary, and eco-epidemiological studies. The aim of the present study was to standardize and compare two methods for the extraction of genomic DNA from sand flies for obtaining DNA in sufficient quantities for next-generation sequencing. Sand flies were collected from the municipalities of Campo Grande, Camapuã, Corumbá and Miranda, state of Mato Grosso do Sul, Brazil. Three protocols using a silica column-based commercial kit (ReliaPrep™ Blood gDNA Miniprep System kit, Promega), and three protocols based on the classical phenol-chloroform extraction method (Uliana et al., 1991), were compared with respect to the yield and quality of the extracted DNA. DNA was quantified using a Qubit 2.0 fluorometer. The presence of sand fly DNA was confirmed by PCR amplification of the IVS6 region (constitutive gene), followed by electrophoresis on a 1.5% agarose gel. A total of 144 male specimens were analyzed, 72 per method. Significant differences were observed between the two methods tested. Protocols 2 and 3 of phenol-chloroform extraction presented significantly better performance than all commercial kit extraction protocols tested. For phenol-chloroform extraction, protocol 3 presented significantly better performance than protocols 1 and 2. The IVS6 region was detected in 70 of 72 (97.22%) samples extracted with phenol, including all samples for protocols 2 and 3. This is the first study on the standardization of methods for the extraction of DNA from sand flies for application to next-generation sequencing, which is a promising tool for entomological and molecular studies of sand flies.

Biological and ecological relations among vectors and their pathogens are important to understand the epidemiology of vector-borne diseases. Camapuã is an endemic area for visceral and tegumentary leishmaniasis. The aim of this study was to characterize the sandfly fauna present in Camapuã , MS, Brazil. Sand flies were collected every fortnight from May 2014 to April 2015 using automatic light traps in the domicile and peridomicile of twelve neighborhoods and forest. The collected specimens were identified based on morphology according to the valid identification keys. In total, 2005 sandflies of five genera and nine species were collected. Nyssomyia whitmani and Lutzomyia cruzi were the most abundant species. Males were more abundant, with a male-to-female ratio of 2.14. The highest diversity was observed in peripheral neighborhood, with abundant plant cover. The peridomicile presented greater abundance of sandflies, with the predominance of Ny. whitmani . No significant correlation between the absolute frequencies of the most abundant species and the precipitation variable was observed; however, there was a predominance of Lu. cruzi in the rainy season. We observed a high frequency of sandflies in urban area, especially vector species. The presence of Nyssomyia whitmani and Lutzomyia cruzi indicate the necessity for health surveillance in the municipality. Additional method of collection such as sticky trap is also recommended for appropriate faunestic study.

The presence, abundance, and distribution of sandflies are strongly influenced by climate and environmental changes. This study aimed to describe the sandfly fauna in an intense transmission area for visceral leishmaniasis and to evaluate the association between the abundance of Lutzomyia longipalpis sensu lato (Lutz & Neiva 1912) (Diptera: Psychodidae) and climatic variables. Captures were carried out 2 yr (July 2017 to June 2019) with automatic light traps in 16 sites of the urban area of Campo Grande, Mato Grosso do Sul state. The temperature (°C), relative humidity (%), precipitation (mm3), and wind speed (km/h) were obtained by a public domain database. The Wilcoxon test compared the absolute frequencies of the species by sex. The association between climatic variables and the absolute frequency of Lu. longipalpis s.l. was assessed using the Spearman’s correlation coefficient. A total of 1,572 sandflies into four species were captured. Lutzomyia longipalpis s.l. was the most abundant species and presented a significant correlation with the average temperature, humidity, and wind speed in different periods. Lutzomyia longipalpis s.l. was captured in all months, showing its plasticity in diverse weather conditions. We emphasize the importance of regular monitoring of vectors and human and canine cases, providing data for surveillance and control actions to continue to be carried out in the municipality.

Visceral leishmaniasis (VL) is a neglected vector-borne disease associated with socioeconomic and environmental issues. In Brazil, epidemics of VL have occurred in major cities since 1980. Applied models for medical and epidemiological research have been used to assess the distribution and characteristics of disease endpoints and identify and characterize potential risk factors. This study described the demographic features of VL and modeled the spatio-temporal distribution of human VL cases and their relationship with underlying predicitve factors using generalized additive models. We conducted an ecological study covering an 18-year period from the first report of an autochthonous case of VL in Campo Grande, state of Mato Grosso do Sul, in 2001 to 2018. The urban area of the city has 74 neighborhoods, and they were the units of analysis of our work. Socioeconomic and demographic data available from Brazilian public databases were considered as covariables. A total of 1,855 VL cases were reported during the study period, with an annual mean incidence rate of 13.23 cases per 100,000 population and a cumulative crude incidence of 235.77 per 100,000 population. The results showed the rapid transition from epidemic to endemic and the centrifugal dispersal pattern of the disease. Moreover, the model highlighted that the urban quality of life index, which is calculated based on income, education, housing conditions, and environmental sanitation data, plays a role in VL occurrence. Our findings highlighted the potential for improving spatio-temporal segmentation of control measures and the cost-effectiveness of integrated disease management programs as soon as VL is difficult to control and prevent and has rapid geographical dispersion and increased incidence rates.