The sandfly fauna (Diptera: Psychodidae: Phlebotominae) of the Parque Estadual da Serra da Tiririca,Rio de Janeiro, Brazil

Cutaneous leishmaniasis (CL) in the state of Rio de Janeiro is sporadic and can be characterised as a peridomestic transmission that occurs in modified natural environments. The aim of this work was to study the fauna and ecological characteristics of sandflies in an environmentally protected area (the State Park of Serra da Tiririca) within the remnants of the Atlantic Forest in the municipalities of Niterói and Maricá and their possible relationship with leishmaniasis. Captures were performed using light traps during the night once a month for one year in both sylvatic environments and areas surrounding homes near the park. A total of 1,037 sandflies were captured, belonging to nine genera and 12 species: Evandromyia tupynambai (34.1%), Migonemyia migonei (20.6%), Brumptomyia cunhai (13.8%), Micropygomyia schreiberi (9.7%), Psathyromyia lanei (6.5%), Brumptomyia nitzulescui (5.7%), Evandromyia edwardsi (5.4%), Nyssomyia intermedia (2.8%), Evandromyia cortelezzii (0.6%), Pintomyia bianchigalatiae (0.5%), Lutzomyia longipalpis (0.2%) and Sciopemyia microps (0.1%). Both Mg. migonei and Ny. intermedia may be acting as vectors of CL in this area.     

The phlebotomine fauna (Diptera: Psychodidae) of Guaraí, state of Tocantins,with an emphasis on the putative vectors of American cutaneous leishmaniasis in rural settlement and periurban areas

Phlebotomine sandflies were captured in rural settlement and periurban areas of the municipality of Guaraí in the state of Tocantins (TO), an endemic area of American cutaneous leishmaniasis (ACL). Forty-three phlebotomine species were identified, nine of which have already been recognised as ACL vectors. Eleven species were recorded for the first time in TO. Nyssomyia whitmani was the most abundant species, followed by Evandromyia bourrouli, Nyssomyia antunesi and Psychodopygus complexus. The Shannon-Wiener diversity index and the evenness index were higher in the rural settlement area than in the periurban area. The evaluation of different ecotopes within the rural area showed the highest frequencies of Ev. bourrouli and Ny. antunesi in chicken coops, whereas Ny. whitmani predominated in this ecotope in the periurban area. In the rural settlement area, Ev. bourrouli was the most frequently captured species in automatic light traps and Ps. complexus was the most prevalent in Shannon trap captures. The rural settlement environment exhibited greater phlebotomine biodiversity than the periurban area. Ps. complexus and Psychodopygus ayrozai naturally infected with Leishmania (Viannia) braziliensis were identified. The data identified Ny. whitmani as a potential ACL vector in the periurban area, whereas Ps. complexus was more prevalent in the rural environment associated with settlements.     

Phlebotominae in peri‐domestic and forest environments inhabited by Alouatta caraya in northeastern Argentina

Multiple species of Phlebotominae are vectors of Leishmania (Protozoa: Trypanosomatidae), which causes visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL). To describe the Phlebotominae (Diptera: Psychodidae) related to the environments of black and gold howler monkeys Alouatta caraya (Humbodlt, 1812) (Primates: Atelidae), potential vectors were sampled in different landscapes and vertical strata of sleeping trees. Phlebotomine captured between December 2011 and March 2012 (2365 individuals) belonged to eight species, of which Nyssomyia neivai (Pinto, 1926) (61.4%) and Migonemyia migonei (França, 1920) (18.73%) were the most abundant, and Ny. withmani was recorded for the first time in the Chaco province. In the ‘peri‐domestic’ landscape, the phlebotomine were mainly captured in henhouses (78.7%), whereas the tree canopy in ‘rural’ and ‘wild’ landscapes yielded 31.2% and 29.1% of the phlebotomine, respectively. A significant association between the type of landscape and the species of phlebotomine was observed by multivariate analysis. Lutzomyia longipalpis (Lutz & Neiva, 1912) and Mg. migonei were associated with ‘peri‐domestic’ landscape, and Ny. neivai was associated with the ‘wild’ landscape. The results of this prospective study suggest that the interaction between phlebotomine and A. caraya could be a key factor with respect to understanding the epidemiology of leishmaniasis.     

Polymerase chain reaction‐based assay for the detection and identification of sand fly gregarines in Lutzomyia longipalpis,a vector of visceral leishmaniasis

Gregarines that parasitise phlebotomine sand flies belong to the genus Psychodiella and, even though they are highly host‐specific, only five species have been described to date. Their most outstanding features include the unique localisation of the oocysts in the accessory glands of the female host, which ensures contamination of the egg surface during oviposition, and the fact that they naturally parasitise the vectors of Leishmania, causal agent of leishmaniasis. The type species, Ps. chagasi, was first described in Lutzomyia longipalpis, vector of visceral leishmaniasis (VL), from Brazil. We recently reported Ps. chagasi sequences in Lu. longipalpis from Posadas (Misiones, Argentina), an endemic VL location where this gregarine had not been previously recorded. In order to analyse the incidence of Ps. chagasi infections in Lu. longipalpis from this location, the aim of this study was to develop a diagnostic assay for sand fly gregarine parasites in Lu. longipalpis. For this, we designed primers using the Ps. chagasi sequences we previously identified and performed an in vitro validation by PCR amplification of the original sand fly samples. Their specificity and sensitivity as diagnostic primers were subsequently confirmed by PCR reactions using total DNA extracted from naturally infected Lu. longipalpis from the same location (Posadas, Argentina).     

Molecular Detection of Leishmania in Phlebotomine Sand Flies (Diptera: Psychodidae) from a Cutaneous Leishmaniasis Focus at Xakriabá Indigenous Reserve,Brazil

Autochthonous cases of American cutaneous leishmaniasis (ACL) have been reported since 2001 in the Xakriabá Indigenous Reserve located in the municipality of São João das Missões in northern Minas Gerais state, Brazil. In order to study the presence of Leishmania DNA in phlebotomine sand flies, six entomological collections were carried out from July 2008 through July 2009, using 40 light traps placed in peridomicile areas of 20 randomly selected houses. From October 2011 through August 2012, another six collections were carried out with 20 light traps distributed among four trails (five traps per trail) selected for a previous study of wild and synanthropic hosts of Leishmania. A total of 4,760 phlebotomine specimens were collected belonging to ten genera and twenty-three species. Single female specimens or pools with up to ten specimens of the same locality, species and date, for Leishmania detection by molecular methods. Species identification of parasites was performed with ITS1 PCR-RFLP using HaeIII enzyme and genetic sequencing for SSU rRNA target. The presence of Leishmania DNA was detected in eleven samples from peridomicile areas: Lu. longipalpis (two), Nyssomyia intermedia (four), Lu. renei (two), Lu. ischnacantha, Micropygomyia goiana and Evandromyia lenti (one pool of each specie). The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one). The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively. The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.     

Phlebotomine sandflies (Diptera: Psychodidae) from Lábrea,state of Amazonas,Brazil, with a description of Evandromyia (Aldamyia) apurinan Shimabukuro,Figueira & Silva,sp. nov.

An entomological survey was conducted from July-December 2009 and September-December 2010, as part of the epidemiological monitoring of American cutaneous leishmaniasis (ACL) in the municipality of Lábrea, state of Amazonas (AM), Brazil. Sandflies were collected using CDC light traps installed in intra and peridomiciliary locations, as well as the border of forested areas around houses where autochthonous cases of ACL were recorded. A total of 510 sandflies belonging to 26 species were collected. The most abundant species was Nyssomyia antunesi (44.5%) followed by Evandromyia walkeri (10.6%) and Micropygomyia rorotaensis (9.8%). Here we also describe Evandromyia (Aldamyia) apurinan sp. nov. and report new records for Trichophoromyia flochi and Evandromyia sipani in AM and Brazil, respectively. Our results describe the composition of the sandfly fauna in the south of AM and suggest Ny. antunesi as the putative vector in the transmission of Leishmania in this area of the Amazon Region.     

Detection of Leishmania DNA in phlebotomines captured in Campo Grande, Mato Grosso do Sul, Brazil

Over the past years, leishmaniases have become a public health issue in the Brazilian state of Mato Grosso do Sul, particularly in Campo Grande, the state capital. The purpose of this study was to detect the presence of Leishmania DNA in the population of phlebotomine sandflies using DNA amplification by PCR. Insect captures were carried out from 4 pm. to 7 am for 4 consecutive days each month from October 2005 to September 2006 in 16 neighborhoods located in 7 urban regions of Campo Grande. Traps were placed indoors and in the vicinity of households. As many as 971 males and 203 females were collected. One hundred and five naturally fed females were identified and grouped as 1- to 4-specimen pools. DNA extraction was carried out using whole insects. Lutzomyia longipalpis predominated, accounting for 99.15% of the phlebotomines captured. Also found was Nyssomyia whitmani, the vector of tegumentary leishmaniasis. Abundance was greatest in the vicinity of households (69.8% of the phlebotomines captured). As revealed by PCR, parasites were present in 1.9% of the Leishmania spp. specimens investigated and confirmed for visceral leishmaniasis.     

Geographic distribution of phlebotomine sandfly species (Diptera: Psychodidae) in Central-West Brazil

This study updates the geographic distributions of phlebotomine species in Central-West Brazil and analyses the climatic factors associated with their occurrence. The data were obtained from the entomology services of the state departments of health in Central-West Brazil, scientific collections and a literature review of articles from 1962-2014. Ecological niche models were produced for sandfly species with more than 20 occurrences using the Maxent algorithm and eight climate variables. In all, 2,803 phlebotomine records for 127 species were analysed. Nyssomyia whitmani, Evandromyia lenti and Lutzomyia longipalpis were the species with the greatest number of records and were present in all the biomes in Central-West Brazil. The models, which were produced for 34 species, indicated that the Cerrado areas in the central and western regions of Central-West Brazil were climatically more suitable to sandflies. The variables with the greatest influence on the models were the temperature in the coldest months and the temperature seasonality. The results show that phlebotomine species in Central-West Brazil have different geographical distribution patterns and that climate conditions in essentially the entire region favour the occurrence of at least one Leishmania vector species, highlighting the need to maintain or intensify vector control and surveillance strategies.     

Polymorphic dinucleotide microsatellite loci in the sandfly Lutzomyia longipalpis (Diptera: Phlebotominae)

The sandfly Lutzomyia longipalpis, an important vector of visceral leishmaniasis in the New World, is believed to be a species complex. In an effort to better understand population dynamics and speciation in this vector we developed a panel of dinucleotide — (CA)_n— microsatellite loci using an enrichment technique. Eleven polymorphic loci that produced consistent allelic banding patterns were characterized using a laboratory population of L. longipalpis. These dinucleotide microsatellite loci were more polymorphic than trinucleotide microsatellites characterized in wild‐caught samples of two other sandfly species; the variability of these loci was unexpected because the laboratory flies were believed to be inbred.     

Genetic Divergence in the <Emphasis Type="Italic">cacophony</Emphasis> IVS6 Intron Among Five BrazilianPopulations of <Emphasis Type="Italic">Lutzomyia longipalpis</Emphasis>

Genes involved in the reproductive isolation are particularly useful as molecular markers in speciation studies. Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae), a putative species complex, is a vector of visceral leishmaniasis in Latin America. We isolated from this species a fragment homologous to cacophony, a Drosophila gene that encodes features of the lovesong, an acoustic signal that is important in the sexual isolation of closely related species and known to vary considerably among L. longipalpis putative siblings species. Using an intron of the sandfly cacophony as a marker, we analyzed the molecular variation and sequence divergence among five populations of L. longipalpis from Brazil, three allopatric (Jacobina, Lapinha and Natal) and two putative sympatric sibling species from the locality of Sobral. A high level of polymorphism was found and analysis of the data indicates that very little gene flow is occurring among the populations of Jacobina, Lapinha, and Natal. A high level of differentiation was also observed between the two putative sympatric species of Sobral, one of which seems to be the same sibling species found in Natal, while the other is somewhat more related to Jacobina and Lapinha. However, the amount of estimated gene flow among the Sobral siblings is about seven times higher than the previously estimated for period, another lovesong gene, perhaps indicating that introgression might be affecting cacophony more than period. The results suggest that L. longipalpis is not a single species in Brazil, but it is yet not clear whether the different populations studied deserve species status rather than representing an incipient speciation process.     

Spatiotemporal dynamics of Lutzomyia longipalpis and macro‐habitat characterization using satellite images in a leishmaniasis‐endemic city in Argentina

The spatiotemporal population dynamics of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) were evaluated in a city in Argentina in which visceral leishmaniasis is endemic. Over 14 sampling sessions, 5244 specimens of five species of Phlebotominae (Diptera: Psychodidae) were captured, of which 2458 (46.87%) specimens were L. longipalpis. Generalized linear models were constructed to evaluate the associations between L. longipalpis abundance and explanatory variables derived from satellite images. The spatial variable ‘stratum’ and the temporal variable ‘season’ were also included in the models. Three variables were found to have significant associations: the normalized difference vegetation index; land surface temperature, and low urban coverage. The last two of these were associated with L. longipalpis abundance only during summer and winter, respectively. This variation between seasons supports the development of models that include temporal variables because models of distributions of the abundance of a species may show different critical variables according to the climatic period of the year. Abundance decreased gradually towards the downtown area, which suggests that L. longipalpis responds to a meta‐population structure, in which rural–periurban source populations that persist over time may colonize adjacent areas. This information allows for a spatiotemporal stratification of risk, which provides public health authorities with a valuable tool to help optimize prevention measures against visceral leishmaniasis.     

Preliminary study on feeding preference of Nyssomyia neivai (Pinto) and Nyssomyia whitmani (Antunes & Coutinho) (Diptera: Psychodidae) in a rural area of the State of Paraná, South Brazil

Due to the importance of sandflies in the American cutaneous leishmaniasis epidemiology, we investigated the feeding preference of sandflies related to domestic animals common in domiciliary environments in endemic rural areas, where this disease occurs. The sandflies were collected with Falc?o light traps, installed in cages, each one containing one domestic animal kind (pig, dog, rabbit or chicken), in Recanto Marista, in Doutor Camargo municipality. Forty five collections were made, totaling 135h of collections per trap. A total of 2,854 sandflies of the following species were collected: Brumptomyia brumpti (Larousse), Evandromyia correalimai (Martins, Coutinho & Luz), Evandromyia cortelezzii (Brèthes), Expapillata firmatoi (Barreto, Martins & Pellegrino), Micropygomyia ferreirana (Barreto, Martins & Pellegrino), Migonemyia migonei (Fran?a), Nyssomyia neivai (Pinto), Nyssomyia whitmani (Antunes & Coutinho), Pintomyia fischeri (Pinto), Pintomyia monticola (Costa Lima), Pintomyia pessoai (Coutinho & Barreto), Psathyromyia lanei (Barreto & Coutinho) and Psathyromyia shannoni (Dyar). The dominant species was N. neivai. N. neivai and N. whitmani did not show any feeding preference towards the animals used as bait. It was concluded that N. neivai and N. whitmani are opportunist and, probably, the females adjust their feeding habits to the availability of hosts, suggesting their feeding eclecticism in the anthropic environments.     

Entomological surveys of Lutzomyia flaviscutellata and other vectors of cutaneous leishmaniasis in municipalities with records of Leishmania amazonensis within the Bragança region of Pará State,Brazil

In southeast Amazon, Lutzomyia (Nyssomyia) flaviscutellata is the incriminated vector of Leishmania (Leishmania) amazonensis, a causative agent of zoonotic cutaneous leishmaniasis (CL). The optimal methods for surveying Lu. flaviscutellata were investigated in the Bragança region, northeast Pará State, Brazil, selected for the presence of Le. amazonensis. The performances of modified Disney traps and CDC light traps were compared in four ecotopes within and around four village transects during the wet and dry seasons. The physiological age of female sand flies was estimated and natural infection by flagellates was evaluated by dissection. Disney traps were better for detecting the presence of Lu. flaviscutellata, while CDC traps performed well for detecting Lutzomyia (Nyssomyia) antunesi, suspected vector of Leishmania lindenbergi. The former was more abundant during the wet season, when female flies were naturally infected with Le. amazonensis. These findings identified the environments of local transmission. In order to improve surveys of Lu. flaviscutellata as part of integrated epidemiological surveillance of CL, our recommendations include focusing vector surveys with Disney traps on forest fragments where people work, during the seasonal peak of the vector. Further field studies are required to make model‐based predictions of seasonal variations in the vectorial capacity of vector populations.     

Impact of control measures and dynamics of sand flies in southern Brazil

We report the results of control measures introduced to reduce the density of sand flies in domiciles and subsequent monitoring of the effects of these measures on the sand fly populations. The most common species of sand flies were Nyssomyia neivai and Nyssomyia whitmani, which are naturally infected by Leishmania. A total of 268,382 (93.4%) sand flies were collected in ecotypes constructed with the aim of attracting sand flies, and 19,091 (6.6%) sand flies were collected in the ecotypes consisting of residences and other buildings. Human actions determine the growth or reduction of the sand fly population in human‐occupied space. Understanding the dynamics of sand flies in this environment can substantially contribute to the prevention of cutaneous leishmaniasis.     

Performance of light‐emitting diode traps for collecting sand flies in entomological surveys in Argentina

The performance of two light‐emitting diode traps with white and black light for capturing phlebotomine sand flies, developed by the Argentinean Leishmaniasis Research Network (REDILA‐WL and REDILA‐BL traps), were compared with the traditional CDC incandescent light trap. Entomological data were obtained from six sand fly surveys conducted in Argentina in different environments. Data analyses were conducted for the presence and the abundance of Lutzomyia longipalpis, Migonemyia migonei, and Nyssomyia whitmani (106 sites). No differences were found in presence/absence among the three types of traps for all sand fly species (p>0.05). The collection mean of Lu. longipalpis from the REDILA‐BL didn´t differ from the CDC trap means, nor were differences seen between the REDILA‐WL and the CDC trap collection means (p>0.05), but collections were larger from the REDILA‐BL trap compared to the REDILA‐WL trap (p<0.05). For Mg. migonei and Ny. whitmani, no differences were found among the three types of traps in the number of individuals captured (p>0.05). These results suggest that both REDILA traps could be used as an alternative capture tool to the original CDC trap for surveillance of these species, and that the REDILA‐BL will also allow a comparable estimation of the abundance of these flies to the CDC light trap captures. In addition, the REDILA‐BL has better performance than the REDILA‐WL, at least for Lu. longipalpis.     

Sandflies (Diptera: Psychodidae) collected with malaise trap in the Center-West of Minas Gerais State, Brazil

The aim of this paper was to verify the sand fly species collected with Malaise trap in the cities of Divinópolis and Pedra do Indaiá, both located in Minas Gerais state. Collections were carried out between November 2003 and October 2004. Thirty two specimens were captured belonging to seven species: Pintomyia monticola (Costa Lima), Nyssomyia whitmani (Antunes & Coutinho), Pintomyia christenseni (Young & Duncan), Evandromyia cortelezzii (Brèthes), Sciopemyia sordellii (Shannon & Del Ponte), Pintomyia pessoai (Coutinho & Barretto) and Psathyromyia aragaoi (Costa Lima).     

Spatial relations among environmental factors and phlebotomine sand fly populations (Diptera: Psychodidae) in central and southern Morocco

Phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae) are of considerable public health importance because of their ability to transmit several human parasites, mainly as vectors of Leishmania spp. Over the past decade, the epidemiological situation of cutaneous leishmaniasis (CL) has significantly increased with its geographic expansion to previously free areas and the emergence of overlapping foci of cutaneous leishmaniasis and visceral leishmaniasis (VL) in several provinces of Morocco. A total of 15,313 specimens was collected during this entomological survey. The genera Phlebotomus (57.38%) and Sergentomyia (42.62%) were identified. Sergentomyia minuta (22.01%) was the most prevalent species, followed by S. fallax (18.21%), Phlebotomus perniciosus (14.35%), P. papatasi (14.06%), P. sergenti (12.85%), P. longicuspis (10.74%), P. ariasi (2.68%), S. dreyfussi (1.53%), P. alexandri (1.31%), P. bergeroti (1.14%), S. christophersi (0.62%), S. africana (0.25%), P. chabaudi (0.14%), P. chadlii (0.05%), and P. kazeruni (0.04%). We aimed to determine current distribution of leishmaniases vectors, their ecological characteristics, and the significance of the predominant species at any bioclimate stage, altitude range, and soil texture in terms of the risk of leishmaniasis transmission.     

Assessing the importance of four sandfly species (Diptera: Psychodidae) as vectors of Leishmania mexicana in Campeche,Mexico

Localized cutaneous leishmaniasis represents a public health problem in many areas of Mexico, especially in the Yucatan Peninsula. An understanding of vector ecology and bionomics is of great importance in evaluations of the transmission dynamics of Leishmania parasites. A field study was conducted in the county of Calakmul, state of Campeche, during the period from November 2006 to March 2007. Phlebotomine sandfly vectors were sampled using Centers for Disease Control light traps, baited Disney traps and Shannon traps. A total of 3374 specimens were captured in the two villages of Once de Mayo (93.8%) and Arroyo Negro (6.1%). In Once de Mayo, the most abundant species were Psathyromyia shannoni, Lutzomyia cruciata, Bichromomyia olmeca olmeca and Psychodopygus panamensis (all: Diptera: Psychodidae). The Shannon trap was by far the most efficient method of collection. The infection rate, as determined by Leishmania mexicana‐specific polymerase chain reaction, was 0.3% in Once de Mayo and infected sandflies included Psy. panamensis, B. o. olmeca and Psa. shannoni. There were significant differences in human biting rates across sandfly species and month of sampling. Ecological niche modelling analyses showed an overall overlap of 39.1% for the four species in the whole state of Campeche. In addition, the finding of nine vector–reservoir pairs indicates a potential interaction. The roles of the various sandfly vectors in Calakmul are discussed.     

Leishmania infantum xenodiagnosis from vertically infected dogs reveals significant skin tropism

BackgroundDogs are the primary reservoir for human visceral leishmaniasis due to Leishmania infantum. Phlebotomine sand flies maintain zoonotic transmission of parasites between dogs and humans. A subset of dogs is infected transplacentally during gestation, but at what stage of the clinical spectrum vertically infected dogs contribute to the infected sand fly pool is unknown.Methodology/Principal findingsWe examined infectiousness of dogs vertically infected with L. infantum from multiple clinical states to the vector Lutzomyia longipalpis using xenodiagnosis and found that vertically infected dogs were infectious to sand flies at differing rates. Dogs with mild to moderate disease showed significantly higher transmission to the vector than dogs with subclinical or severe disease. We documented a substantial parasite burden in the skin of vertically infected dogs by RT-qPCR, despite these dogs not having received intradermal parasites via sand flies. There was a highly significant correlation between skin parasite burden at the feeding site and sand fly parasite uptake. This suggests dogs with high skin parasite burden contribute the most to the infected sand fly pool. Although skin parasite load and parasitemia correlated with one another, the average parasite number detected in skin was significantly higher compared to blood in matched subjects. Thus, dermal resident parasites were infectious to sand flies from dogs without detectable parasitemia.Conclusions/SignificanceTogether, our data implicate skin parasite burden and earlier clinical status as stronger indicators of outward transmission potential than blood parasite burden. Our studies of a population of dogs without vector transmission highlights the need to consider canine vertical transmission in surveillance and prevention strategies.