Effect of environmental variables on the incidence of Visceral Leishmaniasis in Brazil and Colombia.

Visceral Leishmaniasis (VL) is the most severe of the three forms of Leishmaniasis. In the Americas, Brazil and Colombia present more than 90 % of the cases in the region. Our aim in this research was to estimate the association of the incidence rate of Visceral Leishmaniasis with the following environmental variables: the percentage of area suitable for the vector Lutzomyia longipalpis, the episodes of La Niña and El Niño, the Brazilian and Colombian biomes. Epidemiological data were obtained from the Brazilian Notifiable Diseases Information System and the Colombian National Public Health Surveillance System. Environmental data were downloaded from the NASA Giovanni web app, the Modis Sensor database, and the meteorological agencies of Australia, Japan, and the United States of America. Records of the presence of Lu. longipalpis were obtained from public databases and previous studies. As a result, the incidence per 10,000 inhabitants with LEBS for each El Niño-Southern Oscillation (ENSO) episode showed the largest values during El Niño 2015-2016, mainly in Brazil's Northeast and Central regions and the Northeast region of Colombia. Compared with the Neutral 2012-2014 episode, the episodes of El Niño 2015-2016 and La Niña 2010-2011 showed an average increase in the monthly incidence rate of VL, and the average increase was higher during El Niño 2015-2016 (aIRR = 2.304 vs.1.453) We found a positive association between the incidence rate of VL and the El Niño 2015-2016 episode and an impressive% of area suitable for the vector Lu. longipalpis in the Amazon region.An increase of 1 % in the area suitable for the vector Lu. longipalpis leads to an average rise of 0.8 % in the monthly incidence rate of VL. Our study shows a possible association between VL incidence and ENSO, with the most considerable incidence rates observed during El Niño 2015-2016 in Brazil's Northeast and Central regions and the Northeast region of Colombia. The present study is very important to better understand the Visceral Leishmaniasis transmission dynamics.

Molecular and morphometric study of Brazilian populations of Psychodopygus davisi.

In this study, we analysed the molecular and morphometric differences of several populations of the putative sand fly vector Psychodopygus davisi (Root, 1934) (Diptera, Psychodidae, Phlebotominae) in Brazil. We amplified the 658 base pair fragments of the DNA barcoding region-cytochrome c oxidase subunit 1 (COI) gene-for 57 specimens of P. davisi and three specimens of Psychodopygus claustrei (Abonnenc, Léger & Fauran, 1979). We merged our data with public sequences of the same species available from GenBank. Then, the combined dataset-87 sequences and 20 localities-was analysed using population structure analysis and different species delimitation approaches. Geometric morphometry of wings was performed for 155 specimens of P. davisi populations from the North, Midwest and Southeast Brazilian regions, analysing the differences in centroid sizes and canonical variates. Molecular analysis indicated high intraspecific genetic distance values for P. davisi (maximum p distance = 5.52%). All algorithms identified P. davisi and P. claustrei as distinct molecular taxonomic units, despite the low interspecific distance (p distance to the nearest neighbour = 4.79%). P. davisi sequences were split into four genetic clusters by population structure analysis and at least five genetic lineages using intermediate scenarios of the species delimitation algorithms. The species validation analysis of BPP strongly supported the five-species model in our dataset. We found high genetic diversity in this taxon, which is in agreement with its wide geographic distribution in Brazil. Furthermore, the wing analysis showed that specimens from the Southeast Region of Brazil are different from those in the North and the Midwest. The evolutionary patterns of P. davisi populations in Brazil suggest the presence of candidate species, which need to be validated in future studies using a more comprehensive approach with both genomic data and morphological characters.

New sand fly (Diptera, Psychodidae) records and COI DNA barcodes in the state of Maranhão, Eastern Amazon, Brazil.

The sand fly fauna and the usefulness of the DNA barcoding fragment of the cytochrome c oxidase subunit I (COI) gene were accessed in a forest fragment in the municipality of Governador Newton Bello, state of Maranhão, Brazil. We performed entomological collections in three independent campaigns in May and October 2021, and January 2023. Sand flies were morphologically-identified and then DNA barcoded. Sequences were deposited and analyzed in the BOLD System Database, and various species delimitation algorithms, to assess whether DNA sequences merge into taxonomic units in accordance with nominal species. In total, 1,524 sand flies were collected, comprising 32 nominal species. Nyssomyia antunesi was the most abundant species (31.5 %), followed by Psychodopygus davisi (27 %). We reported for the first time in the state of Maranhão, the presence of Lutzomyia evangelistai, Lutzomyia sherlocki, Pressatia equatorialis, and Psathyromyia barrettoi. We amplified and analyzed 67 COI barcodes of 23 species, which were merged with conspecific sequences extracted from GenBank. The maximum intraspecific p distances ranged from 0.0 % to 14.74 %, while the distances to the nearest neighbor varied from 1.67 % to 13.64 %. The phylogenetic gene tree and species delimitation tools clustered sequences into well-supported clades/clusters for each nominal species, except for Pressatia choti/Pr. equatorialis, which have the lowest interspecific genetic distance (1.67 %). We sequenced for the first time COI barcodes of Brumptomyia brumpti, Evandromyia monstruosa, Micropygomyia rorotaensis, Micropygomyia pilosa, Pintomyia christenseni, Pintomyia pacae, Pr. equatorialis, Pa. barrettoi, and Psathyromyia hermanlenti, which will be useful for further molecular identification and classification proposals of Neotropical species. This study updated the current list of the sand fly fauna for the state of Maranhão to 97, and demonstrated that COI barcodes are useful for specific identification.

First Detection and Molecular Analysis of Leishmania infantum DNA in Sand Flies of Kosovo.

Phlebotomine sand flies (Diptera: Psychodidae) are the principal vectors of phleboviruses and Leishmania spp., the causative agents of leishmaniases. The Mediterranean sand fly fauna is diverse, and leishmaniasis, mainly caused by Leishmania infantum, is endemic in the Balkan countries. Despite recent entomological surveys, only some districts of Kosovo have been sampled for sand flies, with no proof/confirmation of L. infantum. This study aimed to gain further insights into the species composition of natural sand fly populations in previously unsampled districts and areas in Kosovo without reports of leishmaniasis and to detect Leishmania DNA in sand flies. A sand fly survey was conducted in 2022 in all seven districts of Kosovo. Collected females were screened for Leishmania DNA by PCR. Positive samples were sequenced and subjected to maximum likelihood analysis with reference sequences for further molecular characterization. The trapping activities at 114 different localities resulted in 3272 caught specimens, comprising seven sand fly species of two genera, namely Phlebotomus neglectus, Ph. perfiliewi, Ph. tobbi, Ph. papatasi, Ph. simici, Ph. balcanicus and Sergentomyia minuta. Leishmania infantum DNA was detected in three individual sand flies of Ph. neglectus and Ph. perfiliewi. This study provides the most extensive sand fly survey in Kosovo and reports the first record of L. infantum DNA in sand flies, indicating autochthonous circulation of L. infantum.

Trichophoromyia auraensis: evidence for cryptic species and first record in the state of Maranhão, Brazil.

Trichophoromyia auraensis (Mangabeira, 1942) (Diptera, Psychodidae, Phlebotominae) has a wide geographic distribution in the western region of the Amazon biome, where it is a putative Leishmania vector. Here, we reported for the first time a population of this species in the Brazilian state of Maranhão, in the eastern Amazon, from which we DNA-barcoded and compared with previously processed specimens from Acre State, in the western Amazon. For this, we analyzed the DNA barcoding fragment (658 bp) of the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer 2 (ITS2) of Trichophoromyia species using phylogenetic gene trees, and species delimitation algorithms. The analyses of COI barcodes showed high values of genetic distance (mean K2P = 5.17) and well-supported clades/MOTUs for the eastern and western populations of T. auraensis, which may indicate a possible complex of cryptic species. The western population of this taxon merged with the close-related sand fly Trichophoromyia velezbernali Posada-López, Galvis and Galati, 2018 from Colombia, which may be associated with the recent speciation history and introgression between these populations. These evidences should be evaluated with a more comprehensive sampling in terms of analyzed populations and molecular markers.

Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.

Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The ß-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.

Morphological and DNA-based description of Trichophoromyia peixotoi n. sp. (Diptera: Psychodidae), a new sand fly species from the Brazilian Amazon.

BACKGROUND: Phlebotomine sand flies of the genus Trichophoromyia Barretto, 1962 are of great relevance to public health as vectors of Leishmania protozoans. A new phlebotomine species named Trichophoromyia peixotoi n. sp. is here described based on both male morphology and COI DNA barcodes. METHODS: The sand fly specimens were collected in the Parque Nacional da Amazônia (PNA), situated in the municipality of Itaituba, state of Pará, Brazil. Morphological description was done based on 10 male specimens. Five specimens were DNA barcoded for the COI gene. RESULTS: The morphological and molecular analyses allowed the delimitation of this new species from others of Trichophoromyia. Trichophoromyia peixotoi n. sp. is closely related to other species with aedeagal ducts > 4 times the length of the sperm pump, from which it may be distinguished by the gonocoxite bristles and paramere shape. CONCLUSIONS: The description of T. peixotoi n. sp. brings the number of species of Trichophoromyia to 45, including 24 for Brazil. The integrative taxonomy effort through the analysis of COI barcodes proved to be effective in the species delimitation of some Trichophoromyia spp.

Population dynamics of Nyssomyia whitmani (Diptera: Psychodidae) in domestic and peridomestic environments in Northeast Argentina, a tegumentary leishmaniasis outbreak area.

In the present study, the temporal dynamics of the main vector of Leishmania braziliensis, Nyssomyia whitmani, was measured by monthly captures of phlebotominae sandflies during 5 consecutive years (from 2011 to 2016) in the Paranaense region of South America. The captures were performed in environments where the human-vector contact risk is high: domiciliary and peridomiciliary environments in a rural area endemic of tegumentary leishmaniasis. Nyssomyia whitmani was recorded as the dominant species of the phlebotominae ensemble in all domiciliary and peridomiciliary environments (House, Chicken Shed, Pigsty, and Forest Edge). Using generalized additive models, intra- and interannual fluctuations were observed, modulated by meteorological variables such as the minimum temperature and the accumulated precipitation 1 wk prior to capture. The installation of a pigsty by the farmer during the study period allowed us to observe and describe the so-called "pigsty effect" where the Ny. whitmani population was spatially redistributed, turning the pigsty as the environment that obtained the highest phlebotominae record counts, thus maintaining the farm overall abundance, supporting the idea that the environmental management of the peridomicile could have an impact on the reduction of epidemiological risk by altering the spatial distribution of the phlebotominae ensemble in the environments.

Improving the COI DNA barcoding library for Neotropical phlebotomine sand flies (Diptera: Psychodidae).

Sand fly species are traditionally identified using morphological traits, though this method is hampered by the presence of cryptic species. DNA barcoding is a widely used tool in the case of insects of medical importance, where it is necessary to know quickly which species are present in a transmission area. Here, we assess the usefulness of mitochondrial cytochrome c oxidase subunit I (COI) DNA barcoding as a practical tool for species identification, correct assignment of isomorphic females, and to evaluate the detection of cryptic diversity that occurs in the same species. A fragment of the COI gene was used to generate 156 new barcode sequences for sand flies from different countries of the Neotropical region, mainly Colombia, which had been identified morphologically as 43 species. The sequencing of the COI gene allowed the detection of cryptic diversity within species and correctly associated isomorphic females with males identified by morphology. The maximum intraspecific genetic distances ranged from 0 to 8.32% and 0 to 8.92% using uncorrected p distances and the Kimura 2-parameter (K2P) model, respectively. The minimum interspecific distance (nearest neighbor) for each species ranged from 1.5 to 14.14% and 1.51 to 15.7% using p and K2P distances, respectively. Three species had more than 3% maximum intraspecific distance: Psychodopygus panamensis, Micropygomyia cayennensis cayennensis, and Pintomyia evansi. They also were split into at least two molecular operational taxonomic units (MOTUs) each, using different species delimitation algorithms. Regarding interspecific genetic distances, the species of the genera Nyssomyia and Trichophoromyia generated values lower than 3% (except Nyssomyia ylephiletor and Ny. trapidoi). However, the maximum intraspecific distances did not exceed these values, indicating the presence of a barcode gap despite their proximity. Also, nine sand fly species were DNA barcoded for the first time: Evandromyia georgii, Lutzomyia sherlocki, Ny. ylephiletor, Ny. yuilli pajoti, Psathyromyia punctigeniculata, Sciopemyia preclara, Trichopygomyia triramula, Trichophoromyia howardi, and Th. velezbernali. The COI DNA barcode analysis enabled the correct delimitation of several Neotropical sand fly species from South and Central America and raised questions about the presence of cryptic species for some taxa, which should be further assessed.

Diversity and temporal distribution of sand flies in an endemic area of cutaneous leishmaniasis in Centre-West Colombia.

The community structure of sand flies indicates the level of adaptation of vector species in a region, and in the context of vector management and control, this information allows for identifying the potential risks of pathogen transmission. This study aimed to analyze sand fly diversity and spatial-temporal distribution in an endemic area of cutaneous leishmaniasis. The study was carried out in the Carrizales hamlet (Caldas), between September 2019 and October 2021. The monthly distribution of sand fly species was evaluated through collections with CDC traps. Shannon and evenness indices were calculated and used to compare species frequencies at each house. The association between climatic variables and the frequency of sand flies was evaluated using Spearman's correlation. A total of 6,265 females and 1,958 males belonging to 23 species were found. Low diversity and evenness were observed, with the dominance of Nyssomyia yuilli yuilli (Young & Porter). Ecological and diversity indices did not reveal differences between the houses. The sand fly community was composed of 3 dominant species, Ny. yuilli yuilli, Psychodopygus ayrozai (Barretto & Coutinho), and Ps. panamensis (Shannon), representing 75.8% of the total catches. No statistical association was found between the absolute frequency of sand flies, rainfall, and temperature. The results show one dominant species, this fact has epidemiological relevance since density influences parasite-vector contact. The high densities of sand flies recorded in peri- and intradomiciliary areas highlight the necessity of periodic monitoring of vector populations and control activities to reduce the risk of Leishmania transmission in this endemic area.

Phlebotomine Fauna (Diptera: Psychodidae) and Infection by Leishmania spp. in Forest Fragments of a University Campus, Western Amazon.

Forest fragments consist of important ecosystems for the maintenance of sand fly populations and Leishmania hosts. This study sought to identify the phlebotomine fauna and its infection by Leishmania spp. in forest fragments on the campus of the Federal University of Acre (UFAC), Western Amazon. Monthly collections with CDC traps were carried out from March 2020 to June 2021, in four forest fragments of UFAC. Male and female insects were processed and identified at species level. A sample of females was subjected to polymerase chain reaction (PCR) analysis to verify the presence Leishmania DNA. In total, 465 specimens were collected, of which 238 were males and 227 were females. The most frequent species were Nyssomyia antunesi (Coutinho, 1939) (47.3%), Trichophoromyia sp. (Mangabeira, 1942) (18.70%), and Ny. whitmani (Antunes & Coutinho, 1939) (8.81%). Molecular analysis detected the presence of Leishmania (Ross, 1903) DNA in a specimen of Ny. antunesi, and another one of Evandromyia walker (Newstead, 1914). The forest fragments of the university campus harbor a diverse sand fly fauna with the presence of Leishmania DNA in these insects, in addition to the presence of other species considered incriminated vectors of Leishmania parasites.

Sciopemyia sordellii in the Neotropical region: distribution, biology, and ecology.

The species Sciopemyia sordellii (Diptera, Phlebotominae) is widely distributed throughout the American continent. This raises doubts about their identity as a single biological entity and points to the occurrence of species complex. Here, we review the geographic distribution of S. sordellii in the Neotropical region, as well as data on its biology and ecology. Records were obtained from literature and from specimens deposited in biological collections. In total, 149 records were analysed, from 12 countries in America. However, after taxonomic revision, some specimens were wrongly identified as S. sordellii, especially those from Peru, putting in doubt the occurrence of the species in that country. In Brazil, it has been recorded in all five geographic regions and collected in different biomes, besides to other ecotypes such as caves. Furthermore, its presence in an anthropized environment has also been reported. Some studies have also diagnosed Leishmania DNA in S. sordellii, indicating that parasite circulation is being maintained between the reservoir hosts. The data evaluated in our study allowed us to provide updated information on the distribution of S. sordellii in America, which may be useful in the taxonomy of this species.

Nyssomyia whitmani (Diptera: Psychodidae) Activity From Sunset to Sunrise: A Two-Year Study in a Rural Area in Northeast Argentina.

We model the nocturnal patterns of the main vector of Leishmania braziliensis (Vianna) in the Paranaense region of South America, Nyssomyia whitmani (Antunes & Coutinho). Occurrence and abundance variation were analyzed by hourly periods, and in relation to sunrise and sunset to avoid differences in luminosity at fixed times throughout the year. The possible role of climatic and environmental variables on nocturnal activity curves was explored. A rotating light trap was operated biweekly in the peridomestic henhouse of a farm in Puerto Iguazú, from May 2016 to May 2018. Ny. whitmani, is the predominant Phlebotominae species (6,720 specimens captured), and is present throughout the night and seasons. Generalized additive models of nocturnal abundance of Ny. whitmani show a bell shape with an acrophase in the early evening. Models of abundance distribution as a function of total abundance showed significant nonlinear variations, only for the tertile of highest vector abundance, in the curves by hourly periods, and related to dawn, while female abundance relative to total abundance showed an asymptotic increasing curve, suggesting density-dependent events with abundance thresholds. Finally, temperature was the variable that best explained the pattern of abundance distribution of Ny. whitmani during the night, although triggers for scotophase activity due to internal clocks or luminosity could not be ruled out. The relationship of activity and abundance with climatic variables would also explain the nights of extraordinary abundance, and therefore of greater probability of vector-human and vector-reservoir contacts.

First detection of Leishmania DNA in Lutzomyia longipalpis sensu lato (Diptera: Phlebotominae) in southem Mexico.

Background & objectives: Lutzomyia longipalpis sensu lato is an important vector of Leishmania infantum, the causative agent of visceral leishmaniasis (VL) in Latin America. In Mexico, this species has been recorded in endemic areas of leishmaniasis transmission, but it has never been detected as infected with Leishmania sp. This study aimed to explore the presence of Leishmania DNA in Lutzomyia longipalpis s.l. from samples collected with a human baited trap from an endemic region of leishmaniasis in southeastern Mexico. Methods: This is a prospective study where a total of 45 specimens of Lu. longipalpis s.l. collected in two sites of Yucatan state with records of leishmaniasis were tested. The nuclear ribosomal Internal Transcribed Spacer was amplified for the detection of Leishmania DNA. Results: Two females were positive for Leishmania DNA. None of the specimens positive for parasite DNA were found fed or gravid. Our finding represents the first record of infection by Leishmania in Lu. longipalpis s.l. for the country. Interpretation & conclusion: More studies are necessary to understand the potential role of this vector species in the transmission cycle of the causative agent of leishmaniasis in the southeastern and other regions of Mexico.

Cutaneous Leishmaniasis Emergence in Southeastern Mexico: The Case of the State of Yucatan.

Environmental changes triggered by deforestation, urban expansion and climate change are present-day drivers of the emergence and reemergence of leishmaniasis. This review describes the current epidemiological scenario and the feasible influence of environmental changes on disease occurrence in the state of Yucatan, Mexico. Relevant literature was accessed through different databases, including PubMed, Scopus, Google, and Mexican official morbidity databases. Recent LCL autochthonous cases, potential vector sandflies and mammal hosts/reservoirs also have been reported in several localities of Yucatan without previous historical records of the disease. The impact of deforestation, urban expansion and projections on climate change have been documented. The current evidence of the relationships between the components of the transmission cycle, the disease occurrence, and the environmental changes on the leishmaniasis emergence in the state shows the need for strength and an update to the intervention and control strategies through a One Health perspective.

Artificial blood-feeding of phlebotomines (Diptera: Psychodidae: Phlebotominae): is it time to repurpose biological membranes in light of ethical concerns?

BACKGROUND: The aims of the present study were to evaluate and compare the efficacy of blood-feeding in phlebotomines through industrially processed membranes from the small intestine of pigs (used for the production of commercial sausages) and the skin of euthanized chicks. METHODS: Laboratory-bred Lutzomyia longipalpis and different field-caught phlebotomine species were subjected to the artificial feeding systems under similar conditions. Paired tests were performed using the control (skin from euthanized chicks) and test membranes (pig small intestine). The feeding rates were compared by paired t-test, and Pearson correlation was used to examine the relationship between the thickness of the membranes and feeding rate. RESULTS: The feeding rate was greater with the test membrane than with the control membrane for L. longipalpis (t-test, t = -3.3860, P = 0.0054) but not for the most frequent field-caught species, Nyssomyia antunesi (t-test, t = 0.7746, P = 0.4535). The average thicknesses of the control and test membranes were 184 ± 83 µm and 34 ± 12 µm, respectively (Mann-Whitney U-test, U = 0.00, Z = 2.8823, P = 0.0039); however, there was no correlation between feeding rate and membrane thickness. A moderate positive correlation was observed between the number of phlebotomines that fed and the total number of phlebotomines in the cage for each type of membrane and for each species. CONCLUSIONS: The test membrane is a viable alternative for the artificial blood-feeding of phlebotomines, and is thus a potential substitute for the skin of animals that are euthanized for this purpose. Feeding rate was independent of membrane thickness.

Molecular Detection of Leishmania (Leishmania) mexicana in Sandflies from the State of Yucatan, Mexico.

Background: Leishmaniases are a group of vector-borne zoonotic diseases of public health relevance within the tropical and subtropical regions of the world. The state of Yucatan is a vulnerable and receptive area to localized cutaneous leishmaniasis (LCL) due to its proximity to the high-transmission endemic states of Campeche and Quintana Roo. Autochthonous cases of LCL caused by Leishmania (Leishmania) mexicana have been documented in the state, showing a geographical expansion of the disease. Materials and Methods: Using CO2-supplemented Centers for Disease Control and Prevention light traps and Shannon traps, we captured anthropophilic sandflies in the surroundings of a locality with recent records of autochthonous cases of LCL. Sandflies carrying Leishmania DNA were evidenced using PCR. Results: A total of 140 Phlebotominae (Diptera: Psychodidae) females of four species were captured: Lutzomyia (Tricholateralis) cruciata (Coquillett), Psathyromyia (Psathyromyia) shannoni (Dyar), Lutzomyia (Lutzomyia) longipalpis (Lutz and Neiva), and Dampfomyia (Coromyia) deleoni (Fairchild and Hertig). Molecular results showed that 6.1% (95% confidence interval [CI] = 2.3-12.9%) of Lu. cruciata and 43.8% (95% CI = 19.8-70.1%) of Pa. shannoni showed evidence of carrying L. (L.) mexicana DNA. Conclusion: We provide evidence of anthropophilic sandflies carrying L. mexicana DNA in a municipality with recorded autochthonous cases of LCL caused by this parasite species in the state of Yucatan, suggesting the emergence of new focus of LCL in Mexico.

Abundance of Lutzomyia longipalpis and Nyssomyia whitmani, the Leishmania spp. vectors in northeastern of Argentina: Are spatial and temporal changing patterns consistence?

Lutzomyia longipalpis and Nyssomyia whitmani were incriminated as vectors of Leishmania sp. Spatial heterogeneity together with seasonal changes in abundance constitute important elements for the understanding of the dynamics of vector populations, and there are fundamental for the development of adequate prevention and control strategies. The aim of this work was to compare the spatial and seasonal abundance of Lu. longipalpis and Ny. whitmani at a city spatial scale between two periods separated by three years. To study the spatial distribution, we compared the abundance distribution of these species at two warm times, 2011 with 2014. Respect to inter-annual seasonal abundance changes, we compared the four seasons of the year between two periods (2011-2012 vs 2014-2016). The spatial distribution for both species were found to be distributed mainly in the same areas of the city in both periods. We change for: Respect to the seasonal pattern of abundance, we observed that seasonal patterns showed changes between periods. Our study defines the 'where' and 'when' implement the actions to mitigate leishmaniasis cases.

First identification of Lutzomyia longipalpis in an area of visceral leishmaniasis transmission in central Rio Grande do Sul State, southern Brazil.

Sand flies are hematophagous dipterans and are considered vectors of the parasites belonging to the genus Leishmania, which can infect mammals, including humans, and subsequently cause leishmaniasis. Rio Grande do Sul State (RS), southern Brazil, was considered free of this disease for the last few decades. However, several autochthonous cases of visceral leishmaniasis (VL) have been recorded in different municipalities in recent years. Since 2017, cases of canine visceral leishmaniasis (CVL) have been reported in the municipality of Santa Maria, located in the central region of RS. In 2021, two cases of human VL were confirmed in the city, resulting in one death. To capture the possible vectors involved in the disease transmission cycle, Centers for Disease Control (CDC) modified light traps were installed in the peridomestic residential areas. These points were situated in two city districts where cases of CVL had been reported from January 2021 to January 2022. In the 41 sample collections of sand flies, 9 specimens of Lutzomyia longipalpis were identified, confirming the municipality as an area of VL transmission, according to Brazilian sanitary standards. The spread of Leishmania spp. has been occurring efficiently, resulting in a negative impact on the One Health. This study reports the first detection of L. longipalpis in the central region of RS.

Blood Feeding Sources of Nyssomyia antunesi (Diptera: Psychodidae): A Suspected Vector of Leishmania (Kinetoplastida: Trypanosomatidae) in the Brazilian Amazon.

Present work aimed to identify blood feeding sources and attempt to detect Leishmania DNA in Nyssomyia antunesi, suspected vector of Leishmania sp., from a park in the urban center of Belém, the capital of Pará State, in the Brazilian Amazon. Entire bodies and gut contents of Ny. antunesi engorged females, previously captured in the urban park with Centers for Disease Control (CDC) light traps and aspiration on tree bases, were subjected to Leishmania and vertebrate DNA detection through amplification of the Leishmania mini-exon and vertebrate cytochrome b (cyt b) gene regions, respectively. The quality of DNA extraction from entire bodies was ensured through amplification of the dipteran cyt b region. The vertebrate cyt b amplicons were sequenced and compared with those available on GenBank. A maximum likelihood phylogenetic tree was constructed to assess the clustering patterns of these sequences. Leishmania DNA was not detected. The sequences of 13 vertebrate cyt b amplicons were considered informative, exhibiting similarity and clustering with the following six vertebrate species: Dasyprocta leporina (1), Cuniculus paca (1), Tamandua tetradactyla (4), Choloepus didactylus (4), Pteroglossus aracari aracari (2), Homo sapiens (1). The samples of D. leporina and C. paca were obtained from the CDC canopy, whereas the others were by aspiration from tree bases. The present results revealed the eclectic and opportunist blood-feeding behavior of Ny. antunesi, with birds and mammals, these last ones acting as potential reservoirs for Leishmania species, distributed throughout the vertical forest strata.