Lutzomyia Species Research Articles

Efficacy of imidacloprid/flumethrin collar in preventing canine leishmaniosis in Brazil.

The Leishmania infantum (synonym, Leishmania chagasi) causes life-threatening infection, namely canine leishmaniosis (CanL), which is a chronic zoonosis prevalent in various countries and spread by the bite of the infected Lutzomyia femalesandflyin South America. The objective of the study was to assess the effectiveness of a polymer matrix collar containing made up of10% imidacloprid and 4.5% flumethrin for the prevention of canineleishmaniosis from the hyperendemic region falling underAraçatuba municipality(Brazil). The research included a total of 146 dogs chosenfrom 75 households. Test were initiated via physical examination; weighing and biological sample collection (blood, popliteal lymph node and conjunctival swab) of these dogs were done in March 2018 (Day 0; GA, control =69, GB, treated=77) to initiate laboratory tests. Post-inclusion, the animals were monitored on the 120th, 240th, 360th and 480th days, respectively. The usage ofcollars continuedbetween 0 and480 days before being substituted in second (D240) and fourth (D480) follow-up visits. On the whole,25 dogs in GA(36.2%)and three in GB(3.9%) were found positive for L. infantum infection in a minimum of one diagnostic test used in the research. Therefore, the average collar effectiveness forprotection from L. infantum infection was 89.2% (p<.01). In the last follow-up, the average incidence density rate for GA was 30.7%, whereas for GB, it was 2.9%. The imidacloprid/flumethrin collars evaluated in the research were found to be safe and extremely efficient for the prevention of L. infantum infection throughLutzomyia species among the largepopulation of dogs in highly prone endemic regions. This is a dependable and efficient technique aimed at reducing the occurrence and propagation of this illness among the population of canines, which would eventually reduce the human-health-related hazards. In Brazil, Lutzomyia spp. is a leading vector of the infection; thus, the collar can be used to limit infection in dogs and humans.

Diversity, distribution and natural Leishmania infection of sand flies from communities along the Interoceanic Highway in the Southeastern Peruvian Amazon.

The Peruvian-Brazilian border is a highly endemic tegumentary leishmaniasis region in South America. The interoceanic highway is a commercial route that connects Peru and Brazil through Madre de Dios and has raised concerns about its impact on previously undisturbed areas. In order to assess leishmaniasis transmission risk along this highway, we conducted a surveillance study of the sand fly populations in this area. Sand flies were collected between 2009 and 2010 along transects at 200 m, 600 m and 1000 m from six study sites located along the highway (Iberia, La Novia, Alto Libertad, El Carmen, Florida Baja, Mazuko and Mavila) and an undisturbed area (Malinowski). Collected specimens were identified based on morphology and non-engorged females of each species were pooled and screened by kinetoplast PCR to detect natural Leishmania infections. A total of 9,023 specimens were collected belonging to 54 different Lutzomyia species including the first report of Lu. gantieri in Peru. Four species accounted for 50% of all specimens (Lutzomyia carrerai carrerai, Lu. davisi, Lu. shawi and Lu. richardwardi). El Carmen, Alto Libertad, Florida Baja and Malinowski presented higher Shannon diversity indexes (H = 2.36, 2.30, 2.17 and 2.13, respectively) than the most human disturbed sites of Mazuko and La Novia (H = 1.53 and 1.06, respectively). PCR detected 10 positive pools belonging to Lu. carrerai carrerai, Lu. yuilli yuilli, Lu. hirsuta hirsuta, Lu. (Trichophoromyia) spp., and Lu. (Lutzomyia) spp. Positive pools from 1,000 m transects had higher infectivity rates than those from 600 m and 200 m transects (9/169 = 5.3% vs 0/79 = 0% and 1/127 = 0.8%, p = 0.018). El Carmen, accounted for eight out of ten positives whereas one positive was collected in Florida Baja and Mazuko each. Our study has shown differences in sand fly diversity, abundance and species composition across and within sites. Multiple clustered Lutzomyia pools with natural Leishmania infection suggest a complex, diverse and spotty role in leishmaniasis transmission in Madre de Dios, with increased risk farther from the highway.

Characterization of the sand fly fauna in Barbalha, one of the municipalities with the highest leishmaniasis rates in Brazil.

Leishmaniases are a complex of sand fly-borne diseases that are considered a public health issue in several countries. Brazil presents high leishmaniases rates. The South of Ceará State, known as Cariri region, shows worrying statistics mainly on American tegumentary leishmaniasis. In Barbalha, which is one of the municipalities in this region, there is still a lack of studies regarding the local phlebotomine (Diptera: Psychodidae) fauna in order to help clarify the high rates. This study aimed to characterize such fauna by capturing sand flies with light traps during a four-year period. A total of 3730 sand flies were captured, of which 37.8% were females. Fourteen species were found: 13 of the Lutzomyia genus and one of the Brumptomyia genus. Of the Lutzomyia species, four were proven and five had potential involvement in leishmaniasis transmission. Lutzomyia longipalpis was the most common species (66.97%). This predominance, especially in the urban area, indicates its epidemiological importance and adaptation to environmental conditions modified by human activity. In fact, further studies are still required to accurately determine the behavioral features of these vectors in order to guide public health measurements towards its control and prevention.

Effects of forest degradation on the sand fly communities of northeast Brazil.

The richness and abundance of sand fly species were studied in northeastern Brazil in areas of leishmaniasis transmission. The study was carried out in two forest areas with different deforestation times for agricultural and livestock activities: one modified by long-term settlement (more than 50 years of occupation) and another less impacted by short-term settlement (10 years). The sand flies were captured with CDC light traps from 18:00 to 06:00 for three consecutive nights, once a month, from May, 2012 to April, 2014. The study captured 21,708 specimens and also 33 species of Lutzomyia and two of Brumptomyia. Species richness and abundance were higher in the more conserved area of short-term occupation (31 species; 61.7%) than in the more degraded area with long-term occupation (17 species; 38.3%). In the most conserved area, the species richness was higher in the forest fragment than in the rural settlement, whereas in the degraded area the richness was higher in the peri-domicile than in the forest. The diversity was higher in the degraded area forest. There were significant statistical differences when comparing the means of total abundance with the intra-domicile, peri-domicile, and forest environments. The average abundance was statistically higher in the peri-domicile compared to the forest (p = 0.009), but there were no statistically significant differences between intra-domicile-peri-domicile (p = 0.11) and forest-intra-domicile (p = 0.87). In conclusion, a change in vegetation cover negatively affects the richness and abundance of sand flies in the natural environment.

Development of Lutzomyia evansi immature stages in peridomiciliary environment in a leishmaniasis urban focus in the Colombian Caribbean

In the Caribbean region of Colombia, Lutzomyia evansi is recognized as the vector for Leishmania infantum and Leishmania braziliensis. Identifying breeding sites and surveying abundance of immature phlebotomine sand flies in urban foci of leishmaniasis are useful tool to design new vector control strategies. The objective of this study was to describe the natural breeding sites of Lu. evansi in peridomiciliary vegetation in a peri-urban area of the Colombian Caribbean region. Between 2013 and 2015, 466 microhabitats were sampled, collecting 621 kg of soil samples. The explored microhabitats were bases and tree holes, fallen trees, animal caves, leaf litter, domestic animal shelters, and the inside of dwellings. The immature phlebotomines were recovered by direct search under the stereoscope and incubation of soil samples. In total, 103 microhabitats, associated with 17 arboreal species, were identified as natural breeding sites. Of 422 immature sandflies detected, 98.6% were found in soils at the base of the trees. Eight species of the genus Lutzomyia were identified, of which Lu. evansi (52.6%) was the most abundant, followed by Lu. rangeliana, Lu. cayennensis cayennensis, Lu. atroclavata, Lu. micropyga, Lu. trinidadensis, Lu. dubitans and Lu. gomezi. The arboreal species Cordia alba was the most used by phlebotomines for the development of their immature stages. From 63 natural breeding sites identified 268 immatures were recovered including 176 Lu. evansi. The accumulated precipitation showed correlation (R2 = 0.643, p = 0.013) with the abundance of developmental stages, which increased in September and October. The natural breeding sites of Lu. evansi exhibited a local pattern of occurrence dependent on rainfall. The physicochemical analysis of the soil samples showed that the natural sites for C. alba were categorized as fertile loam soils. This is the first systematic study that estimates the temporal variation of immature sand flies in peridomiciliary vegetation in a peri-urban focus of leishmaniasis in Colombia.

Primera aproximación a la fauna flebotomínea (Psychodidae: Phlebotominae) del departamento de Atlántico

Dentro del género Lutzomyiaalgunas especies son vectores de la enfermedad llamada leishmaniasis, la cual tiene implicaciones en la salud pública, desarrollando diferentes manifestaciones clínicas como leishmaniasis cutánea, visceral y mucocutánea que afectan altamente al ser humano. El objetivo de este estudio fuerealizar un inventario de las especies del género Lutzomyia presentes en seis zonas del departamento del Atlántico. Se implementaron 3 técnicas de capturas tradicionales: trampas CDC entre 18:00-06:00, trampa Shannon entre 19:00-22:00 y búsqueda activa de flebotomíneos con aspiradores bucales en troncos, bases de árboles y otros sitios entre 09:00-11:00 y 16:00-17:30, en seis municipios del departamento del Atlántico, Colombia. Se capturaron diez especies flebotomíneas y se reporta por vez primera para el departamento del Atlántico el hallazgo de Lutzomyia longipalpis y Lutzomyia atroclavata. De las diez especies capturadas, Lutzomyia longipalpistiene antecedentes como vector de Leishmania infantumen Colombia y en otras regiones de Latinoamérica, además es considerada el principal vector de leishmaniasis visceral en las Américas. La especie más abundante y con mayor distribución espacial fue Lu. evansi, principal vector de leishmaniasis visceral en la región Caribe. En conclusión,la riqueza de la fauna flebotomínea del departamento de Atlántico se actualiza a doce especies, con el primer registro de Lu. longipalpisy Lu. atroclavata. También se reporta la fauna flebotomínea por primera vez en localidades del departamento de Atlántico como Campeche, Puerto Colombia, Luruaco, Ponedera y Campo de la Cruz.

Behavioral Aspects of Phlebotomine Sand Flies Associated with a Case of Cutaneous Leishmaniasis in Atlántico, Northern Colombia.

.After the first autochthonous case of cutaneous leishmaniasis was reported in the Atlántico department in the Caribbean region of Colombia, entomological sampling was conducted in the specific areas where the infection might have occurred. CDC traps were installed inside and outside dwellings in the peri-urban and rural areas of a settlement in the municipality of Luruaco. Sampling was performed during the night with protected human bait, and phlebotomine sand flies were actively sampled from potential diurnal resting sites within dwellings. Ten species of the genus Lutzomyia were identified; Lutzomyia evansi was the dominant species (78%) in the rural and peri-urban areas as well as in the different sampled habitats, followed by Lutzomyia panamensis and Lutzomyia gomezi. There was a 100% household infestation by Lu. evansi, and its indoor mean abundance was 13.3 sand flies/CDC trap/night. The indoor mean abundance of Lu. panamensis and Lu. gomezi was only 0.9 and 0.8 sand flies/CDC trap/night, respectively. Female Lu. evansi were collected with protected human bait, mostly in the peridomestic area, with sustained activity during the night and a slight increase in the activity from 19:00 to 23:00 hours. Of the total sand flies captured in the diurnal resting sites, 73.1% were collected from the walls of bedrooms and corresponded to Lu. evansi, Lutzomyia cayennensis cayennensis, and Lutzomyia trinidadensis. Owing to their vectorial importance, the species on which entomological surveillance should be focused are Lu. evansi, Lu. panamensis, and Lu. gomezi. The biting and resting behavior reported in this study will help guide vector prevention and the control of leishmaniasis within the study area.

Anthropophilic phlebotomine sand fly Lutzomyia species and search for the natural Leishmania infections in an area endemic for cutaneous leishmaniasis in Ecuador

By employing protected human bait landing and modified Shannon light trap, a total of 1924 phlebotomine sand fly Lutzomyia spp. were captured in an area from which L. (V.) guyanensis was reported as the causative parasite of cutaneous leishmaniasis (CL). The sand flies captured alive were dissected and identified at species level, based mainly on their spermathecae. At the same time, the sand flies dissected were searched for the Leishmania parasites by microscopic-test, and later on by PCR-test. No positive sand flies were detected by both tests, while considerable numbers of anthropophilic sand fly species of the genus Lutzomyia were observed as probable vectors of the Leishmania parasite in the areas. Those were eight species, Lu. robusta, Lu. trapidoi, Lu. maranonensis, Lu. gomezi, Lu. shannoni, Lu. migonei, Lu. punctigeniculata and Lu. spathotrichia. Among them, the first two species Lu. robusta and Lu. trapidoi were most dominant, suggesting probable vectors of the Leishmania parasite prevailing in the area. Lu. punctigeniculata and Lu. spathotrichia were for the first time recorded for the Manabí province, Ecuador. These findings provide basic information useful for future planning of the control and management of the disease in the areas, though further study to incriminate the vector sand fly remains.

Surveillance of phlebotomine fauna and Didelphis marsupialis (Didelphimorphia: Didelphidae) infection in an area highly endemic for visceral leishmaniasis in Colombia.

The study of the interaction between the parasite, the vector and the mammalian hosts, including man, allows to understand the behavior of the leishmaniases. To determine the presence of Lutzomyia species and to detect the Leishmania infection in Didelphis marsupialis in an endemic area for visceral leishmaniasis. Phlebotomine fauna and individuals of D. marsupialis were collected with CDC and Tomahawk™ traps, respectively. The species of Lutzomyia were identified using the Young and Duncan key (1994). Ear and tail biopsies and blood samples from D. marsupialis were taken to identify the Leishmania species by amplifying a fragment of the gene associated with the 70 kD heat shock protein. Seven Lutzomyia species were identified: Lu. evansi, Lu. gomezi, Lu. panamensis, Lu. dubitans, Lu. cayennensis cayennensis, Lu. rangeliana and Lu. trinidadensis. The first three species have epidemiological importance in Colombia because of their implications in the transmission of the Leishmania parasite. Sixty-five tissue samples from 19 D. marsupialis individuals were negative for Leishmania spp. The presence of the Lutzomyia species that have been identified as vectors for Leishmania inside and around houses in the village of El Bledo, in El Carmen de Bolívar represents a risk of infection. Furthermore, the presence of Lu. panamensis is reported for first time in El Carmen de Bolívar in Colombia. Although the lack of detection of Leishmania spp. in D. marsupialis samples may suggest that D. marsupialis does not play an important role in the transmission cycle of Leishmania in this region, it is necessary to carry out further longitudinal studies to confirm this hypothesis.

The salivary hyaluronidase and apyrase of the sand fly Sergentomyia schwetzi (Diptera, Psychodidae)

Current knowledge of sand fly salivary components has been based solely on Lutzomyia and Phlebotomus species which feed mainly on mammals; their hyaluronidases and apyrases were demonstrated to significantly affect blood meal intake and transmission of vector-borne pathogens. Members of the third sand fly genus Sergentomyia preferentially feed on reptiles but some of them are considered as Leishmania and arboviruses vectors; however, nothing is known about their salivary components that might be relevant for pathogens transmission. Here, marked hyaluronidase and apyrase activities were demonstrated in the saliva of a Sergentomyia schwetzi colony maintained on geckos. Hyaluronidase of S. schwetzi cleaved hyaluronan as the prominent substrate, and was active over a broad pH range from 4.0 to 8.0, with a sharp peak at pH 5.0. SDS PAGE zymography demonstrated the monomeric character of the enzyme, which remained active in reducing conditions. The apparent molecular weight of 43 kDa was substantially lower than in any sand fly species tested so far and may indicate relatively low grade of the glycosylation of the enzyme. The apyrase of S. schwetzi was typical strictly Ca2+ dependent Cimex-family apyrase. It was active over a pH range from 6.5 to 9.0, with a peak of activity at pH 8.5, and had an ATPase/ADPase ratio of 0.9. The apyrase activity increased during the first 3 days post-emergence, then reached a plateau and remained relatively constant until day 8. In comparison with a majority of Phlebotomus and Lutzomyia species tested to date, both the hyaluronidase and apyrase activities of S. schwetzi were relatively low, which may reflect an adaptation of this sand fly to blood feeding on non-mammalian hosts.

Bionomic aspects of Lutzomyia evansi and Lutzomyia longipalpis, proven vectors of Leishmania infantum in an endemic area of non-ulcerative cutaneous leishmaniasis in Honduras

BackgroundSome Lutzomyia species are the vectors of human leishmaniasis in the Americas. Visceral and cutaneous leishmaniasis are both endemic in the Pacific region of Honduras, but the non-ulcerative form is the more frequent clinical manifestation in this region, where Lutzomyia longipalpis is the most abundant and the only incriminated vector. Taxonomic identification and distribution studies of sand flies are important to understand the epidemiology and to control these neglected tropical diseases.ResultsHere, we identified more than 13,000 Lutzomyia specimens captured in Isla del Tigre, Honduras, through a classical morphological approach. The two most common species were Lutzomyia evansi and Lu. longipalpis, and this is the first report of three Lutzomyia species on this island. The blood meal source was successfully identified for five sand fly species. A barcode analysis using the cox1 mitochondrial marker proved to be effective in discriminating between species and seems to be a valuable tool for future epidemiological studies including a wider geographical area.ConclusionThis study updates the diversity and blood meal sources of Lutzomyia species in an island endemic for non-ulcerative cutaneous leishmaniasis in the Pacific region of Honduras, and determines the effectiveness of the barcoding approach to discriminate species, as a complementary tool to classical morphology.

Update on tegumentary leishmaniasis and carrion's disease vectors in Peru

Among approximately 190 species of Lutzomyia in Peru, only a small number have been identified as vectors of tegumentary leishmaniasis in Western and inter-Andean valleys. These include L. peruensis, L. verrucarum, L. tejadai, L. ayacuchensis, and L. pescei. In the Amazon region, L. yuilli yuilli, L. chagasi, L. davisi, and L. auraensis are naturally infected, among the subgenera Leishmania and Viannia. L. auraensis is newly reported as a potential vector of leishmaniasis in neotropical regions. Among the primary and most widely distributed vectors of human bartonellosis or Carrión's disease, L. verrucarum and L. peruensis are predominant in the Andean regions of northern, central, and southern Peru. Other potential vectors of Carrion's disease are L. serrana in the Monzon Valley, Huamalies, and Huanuco; L. pescei in Apurímac and Cusco; and L. robusta and L. maranonensis in Jaén, San Ignacio, and Utcubamba provinces, and the high forests of Peru. Because of the high prevalence of leishmaniasis and bartonellosis outside of known endemic areas in Peru, it is necessary to update data and distribution maps of these disease vectors. This may improve both prevention and control measures. Existing information about sandfly vectors in Peru is also provided in this article.

Molecular detection of Wolbachia pipientis in natural populations of sandfly vectors of Leishmania infantum in endemic areas: first detection in Lutzomyia longipalpis.

A polymerase chain reaction-based method was used to screen sandflies for infection with Wolbachia (Rickettsiales: Rickettsiaceae), an intracellular bacterial endosymbiont found in many arthropods and filarial hosts. Positive results were obtained in five of 200 field-collected sandflies and were confirmed by sequencing. All sandflies were Lutzomyia longipalpis (Diptera: Psychodidae) captured in a region endemic for visceral leishmaniasis in Brazil. This is the first study to identify Wolbachia infection in this Lutzomyia species, which is the main vector of leishmaniasis in the study area. The low infection rate found in this study (2.5%), together with the lack of detection of Wolbachia in previous studies and the diversity found in the sequences analysed, suggests horizontal transmission to these sandflies.

The role of wing geometric morphometrics in the identification of sandflies within the subgenus Lutzomyia.

The Lutzomyia subgenus (Diptera: Psychodidae) includes sibling species with morphologically indistinguishable females. The aims of this study were to analyse variations in the size and shape of wings of species within the Lutzomyia subgenus and to assess whether these analyses might be useful in their identification. Wings (n = 733) of 18 species deposited in Brazilian collections were analysed by geometric morphometrics, using other genera and subgenera as outgroups. Shape variation was summarized in multivariate analyses and differences in wing size among species were tested by analysis of variance. The results showed significant variation in the sizes and shapes of wings of different Lutzomyia species. Two clusters within the Lutzomyia subgenus were distinguished in analyses of both males and females. In Cluster 1 (Lutzomyia ischnacantha, Lutzomyia cavernicola, Lutzomyia almerioi, Lutzomyia forattinii, Lutzomyia renei and Lutzomyia battistinii), scores for correct reclassification were high (females, kappa = 0.91; males, kappa = 0.90), whereas in Cluster 2 (Lutzomyia alencari, Lutzomyia ischyracantha, Lutzomyia cruzi, Lutzomyia longipalpis, Lutzomyia gaminarai and Lutzomyia lichyi), scores for correct reclassification were low (females, kappa = 0.42; males, kappa = 0.48). Wing geometry was useful in the identification of some species of the Lutzomyia subgenus, but did not allow the identification of sibling species such as L.longipalpis and L.cruzi.

Insights into the sand fly saliva: Blood-feeding and immune interactions between sand flies, hosts, and Leishmania.

BackgroundLeishmaniases are parasitic diseases present worldwide that are transmitted to the vertebrate host by the bite of an infected sand fly during a blood feeding. Phlebotomine sand flies inoculate into the mammalian host Leishmania parasites embedded in promastigote secretory gel (PSG) with saliva, which is composed of a diverse group of molecules with pharmacological and immunomodulatory properties.Methods and findingsIn this review, we focus on 3 main aspects of sand fly salivary molecules: (1) structure and composition of salivary glands, including the properties of salivary molecules related to hemostasis and blood feeding, (2) immunomodulatory properties of salivary molecules and the diverse impacts of these molecules on leishmaniasis, ranging from disease exacerbation to vaccine development, and (3) use of salivary molecules for field applications, including monitoring host exposure to sand flies and the risk of Leishmania transmission. Studies showed interesting differences between salivary proteins of Phlebotomus and Lutzomyia species, however, no data were ever published on salivary proteins of Sergentomyia species.ConclusionsIn the last 15 years, numerous studies have characterized sand fly salivary proteins and, in parallel, have addressed the impact of such molecules on the biology of the host–sand fly–parasite interaction. The results obtained shall pave the way for the development of field-application tools that could contribute to the management of leishmaniasis in endemic areas.

DNA barcode for identification of immature stages of sand flies (Diptera: Psychodidae) collected from natural breeding sites.

Although phlebotomine sand flies breeding sites have been identified and recorded by several studies, the microhabitats exploited by these insects remain little-known and hard to find. In this context, the difficulty of finding immature stages, and the limited number of taxonomic studies to identify immature stages of phlebotomine sand flies, are considered the major obstacles when attempting a complete inventory of Lutzomyia species. The objective of this study is to validate Cytochrome Oxidase I (Barcode region) as a marker for the identification of immature stages of Lutzomyia species recovered from natural breeding sites in Colombia. Among 142 collected sand flies, 18 immature individuals that did not complete their life cycle were identified to species level through sequencing of the COI gene. Values of K2P genetic distance between 0.002-0.031 allowed the identification of larvae at species level. The bootstrap support values (96%) in the Neighbor-Joining dendrogram were consistent for the majority of the established MOTUS of Lutzomyia atroclavata, Lutzomyia micropyga, Lutzomyia serrana, Lutzomyia cayennensis, Lutzomyia rangeliana, Lutzomyia shannoni and some species of the genus Brumptomyia. The COI gene is validated as a marker for the identification of immature stages of the genus Lutzomyia.

Especies de flebotomíneos (Diptera: Psychodidae) recolectados en reservas naturales de las regiones del Darién y del Pacífico en Colombia

Introduction: The departments of Choco and Antioquia in Colombia show climatic and vegetation conditions favoring the establishment of vector species of the genus Lutzomyia and the transmission of Leishmania spp. to human populations entering conserved forest environments. Objective: To report the species of Phlebotomine sandflies present in three natural reserves in the Darien and Pacific regions of Colombia. Materials and methods: Sand flies were collected specifically in the natural reserves El Aguacate (Acandi, Choco), Nabuga (Bahia Solano, Choco) and Tulenapa (Carepa, Antioquia). Sand flies were collected with CDC light traps, active search in resting places and Shannon traps. The taxonomic determination of species was based on taxonomic keys. For some species of taxonomic interest, we evaluated the partial sequences of the 5’ region of COI gene. Results: A total of 611 adult sand flies were collected: 531 in Acandi, 45 in Carepa and 35 in Bahia Solano. Seventeen species of the genus Lutzomyia , three of the genus Brumptomyia and one of the genus Warileya were identified. The genetic distances (K2P) and grouping supported (>99%) in the neighbor joining dendrogram were consistent for most established molecular operational taxonomic units (MOTU) of the Aragaoi group and clearly confirmed the identity of Lu. coutinhoi . Conclusion: Species that have importance in the transmission of leishmaniasis in Acandi, Bahia Solano and Carepa were identified. The presence of Lu. coutinhoi was confirmed and consolidated in Colombia.

Molecular detection and identification of Wolbachia in three species of the genus Lutzomyia on the Colombian Caribbean coast

BackgroundThe hematophagous habits of insects belonging to the genus Lutzomyia (Diptera: Psychodidae), as well as their role as biological vectors of Leishmania species, make their presence an indication of infection risk. In the present study, seven species of Lutzomyia were identified and screened for natural infections with Wolbachia.MethodsCollection of sand flies was done in an endemic focus of leishmaniasis on the Colombian Caribbean coast (Department of Sucre, Ovejas municipality). DNA collected from Lutzomyia species was evaluated with PCR for wsp gene amplification to screen for bacterial infection.ResultsEndosymbiotic Wolbachia was found in three species: Lutzomyia c. cayennensis, Lutzomyia dubitans and Lutzomyia evansi. Two Wolbachia strains (genotypes) were found in Lutzomyia spp. These genotypes were previously unknown in dipteran insects. The wLev strain was found in Lutzomyia dubitans, L. c. cayennensis and L. evansi and the wLcy strain was found only in L. c. cayennensis.ConclusionsGenetic analysis indicated that the Wolbachia strains wLcy and wLev belong to the B Supergroup. This study provides evidence of infections of more than one strain of Wolbachia in L. c. cayennensis.

Environmental Niche Modelling of Phlebotomine Sand Flies and Cutaneous Leishmaniasis Identifies Lutzomyia intermedia as the Main Vector Species in Southeastern Brazil.

Cutaneous leishmaniasis (CL) is caused by a protozoan of the genus Leishmania and is transmitted by sand flies. The state of Espírito Santo (ES), an endemic area in southeast Brazil, has shown a considerably high prevalence in recent decades. Environmental niche modelling (ENM) is a useful tool for predicting potential disease risk. In this study, ENM was applied to sand fly species and CL cases in ES to identify the principal vector and risk areas of the disease. Sand flies were collected in 466 rural localities between 1997 and 2013 using active and passive capture. Insects were identified to the species level, and the localities were georeferenced. Twenty-one bioclimatic variables were selected from WorldClim. Maxent was used to construct models projecting the potential distribution for five Lutzomyia species and CL cases. ENMTools was used to overlap the species and the CL case models. The Kruskal–Wallis test was performed, adopting a 5% significance level. Approximately 250,000 specimens were captured, belonging to 43 species. The area under the curve (AUC) was considered acceptable for all models. The slope was considered relevant to the construction of the models for all the species identified. The overlay test identified Lutzomyia intermedia as the main vector of CL in southeast Brazil. ENM tools enable an analysis of the association among environmental variables, vector distributions and CL cases, which can be used to support epidemiologic and entomological vigilance actions to control the expansion of CL in vulnerable areas.

DNA barcoding to identify species of phlebotomine sand fly (Diptera: Psychodidae) in the mixed leishmaniasis focus of the Colombian Caribbean

Identification of the species of phlebotomine sand flies present in each focus of leishmaniasis is necessary to incriminate vectors and implement vector control strategies. Although the cytochrome oxidase I (COI) gene has been proposed as a barcode for the identification of animal species, less than 20% of New World phlebotomines have been characterized to date. In this study DNA barcoding was used to identify phlebotomine species of the mixed leishmaniasis focus in the Colombian Caribbean by means of three evolutionary models: Kimura’s two parameter (K2P) nucleotide substitution model, that of (Tamura and Nei, 1993) (TN93) and proportional sequence divergence (p-distances). A 681bp sequence of the COI gene was obtained from 66 individuals belonging to 19 species of the genus Lutzomyia (Lu. abonnenci, Lu. atroclavata, Lu. bicolor, Lu. carpenteri, Lu. cayennensis cayennensis, Lu. dubitans, Lu. evansi, Lu. gomezi, Lu. gorbitzi, Lu. longipalpis, Lu. micropyga, Lu. migonei, Lu. panamensis, Lu. (Psathyromyia) sp., Lu. rangeliana, Lu. serrana, Lu. shannoni, Lu. trinidadensis and Lu. venezuelensis) and one of Brumptomyia (B. mesai). The genetic divergence values for TN93 among individuals of the same species fluctuated up to 3.2% (vs. 2.9% for K2P and 2.8% for p-distances), while the values between species ranged from 8.8–43.7% (vs. 6.8–19.6% for K2P and 6.6–17.4% for p-distances). A dendrogram constructed by means of the Neighbor-Joining method grouped phlebotomines into 20 clusters according to species, with bootstrap values of up to 100% in those with more than one individual. However, loss of the phylogenetic signal of the gene COI was observed at the supraspecific level as a consequence of substitutional saturation. In conclusion, irrespective of the evolutionary model selected, all phlebotomines were correctly assigned to species, showing 100% concordance with morphological identification.