Gut Microbiota Dynamics in Natural Populations of Pintomyia evansi under Experimental Infection with Leishmania infantum.

Pintomyia evansi is recognized by its vectorial competence in the transmission of parasites that cause fatal visceral leishmaniasis in rural and urban environments of the Caribbean coast of Colombia. The effect on and the variation of the gut microbiota in female P. evansi infected with Leishmania infantum were evaluated under experimental conditions using 16S rRNA Illumina MiSeq sequencing. In the coinfection assay with L. infantum, 96.8% of the midgut microbial population was composed mainly of Proteobacteria (71.0%), followed by Cyanobacteria (20.4%), Actinobacteria (2.7%), and Firmicutes (2.7%). In insect controls (uninfected with L. infantum) that were treated or not with antibiotics, Ralstonia was reported to have high relative abundance (55.1-64.8%), in contrast to guts with a high load of infection from L. infantum (23.4-35.9%). ASVs that moderately increased in guts infected with Leishmania were Bacillus and Aeromonas. Kruskal-Wallis nonparametric variance statistical inference showed statistically significant intergroup differences in the guts of P. evansi infected and uninfected with L. infantum (p < 0.05), suggesting that some individuals of the microbiota could induce or restrict Leishmania infection. This assay also showed a negative effect of the antibiotic treatment and L. infantum infection on the gut microbiota diversity. Endosymbionts, such as Microsporidia infections (<2%), were more often associated with guts without Leishmania infection, whereas Arsenophonus was only found in guts with a high load of Leishmania infection and treated with antibiotics. Finally, this is the first report that showed the potential role of intestinal microbiota in natural populations of P. evansi in susceptibility to L. infantum infection.

Description of the ovarian microbiota of Aedes aegypti (L) Rockefeller strain.

Aedes aegypti is one of the vectors responsible for transmitting the viruses that cause dengue, Zika and chikungunya in the human population. Mosquitoes have bacterial communities in different organs, mainly in the midgut, but to a lesser extent in their reproductive organs, such as the ovaries, where replication and vertical transmission is decisive for dengue virus. These bacteria also influence metabolic and physiological processes such as ingestion and digestion of blood. In this study, aerobic bacterial communities associated with ovaries of A. aegypti Rockefeller strain were determined, describing their potential function during ovocitary development. The groups of mosquitoes were separated into three treatments: diet with 10% sugar solution, diet with blood supply, and blood feeding combined with tetracycline. The ovaries were extracted from the mosquitoes, and then put in enriched culture media (blood and nutritive agar) by direct inoculation, for subsequent isolation and macroscopic and microscopic characterization of the colonies. The taxonomic determination of bacterial isolates was achieved by sequence analysis of the 16S rRNA gene. A higher bacterial load was observed in the sugar feeding group (6 × 10³ CFU/ml) in contrast to the group fed only with blood, with and without an antibiotic (4.03-4.04 × 10³CFU/ml; 4.85-5.04 × 10³CFU/ml). As a result, a total of 35 colonies were isolated, of which 80% were gram-negative and 20% gram-positive; 72% were lactose negative and 8% lactose positive. Of the total bacteria, 83% had gamma hemolysis, 17% alpha hemolysis, and none presented beta hemolysis. After phenotypic and biochemical characterization, 17 isolates were selected for molecular identification. Only phyla Actinobacteria and Proteobacteria were found. Bacteria associated with ovaries of A. aegypti were mainly identified as belonging to the Serratia and Klebsiella genera. Some bacteria (Serratia marcescens, Pantoea dispersa and Klebsiella oxytoca) have wide biotechnological potential due to their entomopathogenic power and their bioactivity against different pathogens.

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.

First record of the sand fly Warileya (Hertigia) hertigi from Antioquia, Colombia

Introduction: The genus Warileya is one of the least-known taxa of sandflies, comprising only nine species, i.e., W. (Warileya) phlebotomanica, W. (Hertigia) hertigi, W. (W.) rotundipennis, W. (W.) nigrosacculus, W. (W.) yungasi, W. (W.) fourgassiensis, W. (W.) lumbrerasi, W. (W.) euniceae and W. (W.) leponti. Objective: To document the presence of a species of the genus Warileya in Antioquia, Colombia. Materials and methods: Sandflies were collected in a cavern of the Cañón del Río Claro Natural Reserve, of the municipality of San Francisco, Antioquia department, Colombia. Phlebotomine sampling was carried out using a CDC light trap during three consecutive nights in May of 2008. Taxonomical determination was based on a revision of the type material of the species and through the use of standard keys for American sandflies. Results: Five male and two female sandflies were taxonomically identified as W. (H.) hertigi. In both sexes, the absence of setal scars in the anepisternum, proepimeron and clypeus; the presence of two transverse rows of setal scars in the tergites; and the short length of the vena gamma were notable. Conclusion: The finding of W. (H.) hertigi increases the number of sandfly species found in Antioquia department to 64. In total, 164 sandfly species have been recorded in Colombia.

Primer registro del flebotomíneo Warileya (Hertigia) hertigi en Antioquia, Colombia / First record of the sand fly Warileya (Hertigia) hertigi from Antioquia, Colombia

Resumen Introducción. El género Warileyaes uno de los taxones menos conocidos de los flebotomíneos, el cual está integrado por solo nueve especies: Warileya (Warileya) phlebotomanica, W. (Hertigia) hertigi, W. (W.) rotundipennis, W. (W.) nigrosacculus, W. (W.) yungasi, W. (W.) fourgassiensis, W. (W.) lumbrerasi, W. (W.) euniceae y W. (W.) leponti. Objetivo. Registrar el hallazgo de una especie del género Warileya en Antioquia, Colombia. Materiales y métodos. Los flebotomíneos se recolectaron en una caverna de la Reserva Natural Cañón del Río Claro del municipio de San Francisco, departamento de Antioquia, Colombia. El muestreo entomológico sehizo con una trampa de luz de tipo CDC, durante tres noches consecutivas, en mayo de 2008. La determinación taxonómica se basó en la revisión del material tipo de la especie y de las claves estándar para flebotomíneos americanos. Resultados. Se identificaron taxonómicamente cinco machos y dos hembras de flebotomíneos comoW. (H.) hertigi. En ambos sexos fue notoria la ausencia de cicatrices de setas en el anepisternón, el proepimerón y el clípeo, la presencia de dos hileras transversales de cicatrices de setas en los tergitos, y la longitud corta de la vena gamma. Conclusión. El hallazgo de W. (H.) hertigi incrementa a 64 el número de especies de flebotomíneos encontradas en el departamento de Antioquia. En total, 164 especies de flebotomíneos han sido registradas en Colombia.

Abstract Introduction: The genus Warileyais one of the least-known taxa of sandflies, comprising only nine species, i.e., W. (Warileya) phlebotomanica, W. (Hertigia) hertigi, W. (W.) rotundipennis, W. (W.) nigrosacculus, W. (W.) yungasi, W. (W.) fourgassiensis, W. (W.) lumbrerasi, W. (W.) euniceae and W. (W.) leponti. Objective: To document the presence of a species of the genus Warileya in Antioquia, Colombia. Materials and methods: Sandflies were collected in a cavern of the Cañóndel Río Claro Natural Reserve, of the municipality of San Francisco, Antioquia department, Colombia. Phlebotomine sampling was carried out using a CDC light trap during three consecutive nights in May of 2008. Taxonomical determination was based on a revision of the type material of the species and through the use of standard keys for American sandflies. Results: Five male and two female sandflies were taxonomically identified as W. (H.) hertigi. In both sexes, the absence of setal scars in the anepisternum, proepimeron and clypeus; the presence of two transverse rows of setal scars in the tergites; and the short length of the vena gamma were notable. Conclusion: The finding of W. (H.) hertigi increases the number of sandfly species found in Antioquia department to 64. In total, 164 sandfly species have been recorded in Colombia.

Composition and distribution of medically important phlebotomines (Diptera: Psychodidae) in the municipalities of Tierralta and Valencia (Córdoba, Colombia).

BACKGROUND & OBJECTIVES: Ecoepidemiological studies of cutaneous leishmaniasis and regular monitoring of Lutzomyia species have generated a knowledge base that can be used for control and prevention strategies targeted at the disease transmission dynamics in focal areas of Colombia. In this study, the presence and spatial distribution of phlebotomines of medical importance in the municipalities of Tierralta (El Loro and Tuis Tuis villages) and Valencia (Guadua and Mieles villages) were determined. METHODS: Entomological surveys were performed in 2015 (months of June, September and November) and samples were collected via CDC-traps located in intradomicilary and peridomiciliary areas in the municipalities of Tierralta and Valencia (Department of Córdoba, Colombia). Active searches were also carried out with a mouth aspirator to collect adult phlebotomines from resting sites. ANOVA and Kruskal-Wallis tests were performed to assess if the differences between the communities of phlebotomines. Spatial distribution maps of the Lutzomyia species were generated. RESULTS: A high species diversity of Lutzomyia was observed with a total of 1677 Lutzomyia individuals belonging to 12 species. Among these species, Lu. panamensis was the most abundant (80.18%). The composition of the intradomicilary and peridomiciliary phlebotomines varied significantly (F = 0.9962; df = 1; p = 0.02895). Species like Lu. carpenteri, Lu. camposi, Lu. dysponeta, Lu. atroclavata and Lu. yuilli yuilli were recorded for the first time in the Department of Córdoba, Colombia. INTERPRETATION & CONCLUSION: The spatial distribution shows that Lu. panamensis and Lu. gomezi are predominant and present in areas with high concentration of houses. This study provides basic information on new records of phlebotomines in the Department of Córdoba. The results suggest that greater vector-human contact occurs in the peridomiciliary environment and that a high number of Lutzomyia species associated with the transmission of leishmaniasis are present in Colombia.

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

BACKGROUND: The 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. METHODS: Collection 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. RESULTS: Endosymbiotic 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. CONCLUSIONS: Genetic 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.

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.

Structural differences in gut bacteria communities in developmental stages of natural populations of Lutzomyia evansi from Colombia's Caribbean coast.

BACKGROUND: Lutzomyia evansi, a phlebotomine insect endemic to Colombia's Caribbean coast, is considered to be the main vector of visceral and cutaneous leishmaniasis in the region. Although insects of this species can harbor pathogenic and non-pathogenic microorganisms in their intestinal microbiota, there is little information available about the diversity of gut bacteria present in Lutzomyia evansi. In this study, conventional microbiological methods and molecular tools were used to assess the composition of bacterial communities associated with Lutzomyia evansi guts in immature and adult stages of natural populations from the department of Sucre (Caribbean coast of Colombia). METHODS: Sand flies were collected from two locations (peri-urban and jungle biotype) in the Department of Sucre (Caribbean coast of Colombia). A total of 752 Lutzomyia evansi intestines were dissected. In this study, 125 bacterial strains were isolated from different culture media (LB Agar, MacConkey Agar). Different methods were used for bacterial identification, including ribosomal intergenic spacer analysis (RISA) and analysis of the 16S rRNA and gyrB gene sequences. The genetic profiles of the bacterial populations were generated and temporal temperature gradient gel electrophoresis (TTGE) was used to compare them with total gut DNA. We also used PCR and DNA sequence analysis to determine the presence of Wolbachia endosymbiont bacteria and Leishmania parasites. RESULTS: The culture-dependent technique showed that the dominant intestinal bacteria isolated belong to Acinetobacter, Enterobacter, Pseudomonas, Ochrobactrum, Shinella and Paenibacillus in the larval stage; Lysobacter, Microbacterium, Streptomyces, Bacillus and Rummeliibacillus in the pupal stage; and Staphylococcus, Streptomyces, Brevibacterium, Acinetobacter, Enterobacter and Pantoea in the adult stage. Statistical analysis revealed significant differences between the fingerprint patterns of the PCR-TTGE bands in bacterial communities from immature and adult stages. Additionally, differences were found in bacterial community structure in fed females, unfed females, males and larvae. The intestinal bacteria detected by PCR-TTGE were Enterobacter cloacae and Bacillus thuringiensis, which were present in different life stages of Lu. evansi, and Burkholderia cenocepacia and Bacillus gibsonii, which were detected only in the larval stage. Wolbachia and Leishmania were not detected in gut samples of Lutzomyia evansi. CONCLUSIONS: The analyses conducted using microbiological and molecular approaches indicated significant variations in the bacterial communities associated with the gut of Lu. evansi, depending on the developmental stage and food source. We propose that these elements affect microbial diversity in L. evansi guts and may in turn influence pathogen transmission to humans bitten by this insect.

Study on natural breeding sites of sand flies (Diptera: Phlebotominae) in areas of Leishmania transmission in Colombia.

BACKGROUND: The location of the microhabitats where immature phlebotomine sand flies of the genus Lutzomyia develop is one of the least-known aspects of this group of medically important insects. For this reason strategies of source reduction approach for their control have not been possible in contrast to other insect vectors (such as mosquitoes), because their juvenile stages in terrestrial microhabitats is difficult to detect. METHODS: Direct examination of soil samples, incubation of substrates and the use of emergence traps were the methods used to identify juvenile stages in 160 soil samples from urban and forest habitats within the foci of Leishmania transmission in Colombia. Immatures collected were identified subsequent from the rearing and emergence of adults using taxonomic keys or the analysis of the mitochondrial marker cytochrome oxidase I. Plant species associated with the natural breeding sites were identified and physicochemical properties of the soils were analyzed. RESULTS: A total of 38 (23.7%) sampling sites were identified as breeding sites, 142 phlebotomine sand flies were identified, belonging to 13 species of the genus Lutzomyia and two of Brumptomyia. The greatest numbers of immature were found within the tabular roots (51 immature sand flies from eight positive sites) and bases of trees (35 immature sand flies from 11 sites). The characterization and presence of the tree species (mainly Ceiba pentadra, Anacardium excelsum, Pseudosamanea guachapale) and the physicochemical properties (relative humidity and carbon/nitrogen ratio) of the soils associated with these breeding sites are significant factors in explaining the diversity and abundance of phlebotomine sand flies. CONCLUSIONS: Immature phlebotomine sand flies of the genus Lutzomyia in Colombia can be found in a wide variety of breeding sites rich in organic matter, high relative humidity and are associated with a typical vegetation of each locality. These results provide new perspectives for the study of the ecology of the genus Lutzomyia in Colombia and the development of vector control strategies.

[New records of phlebotomine sand flies (Diptera: Psychodidae) near the Amoya River in Chaparral, Tolima]. / Nuevos registros de flebotomíneos (Diptera: Psychodidae) en el área de influencia del río Amoyá en Chaparral, Tolima.

INTRODUCTION: In Colombia, the diversity of phlebotomine sand flies is high, with 162 recorded species, and which include vectors of Leishmania spp. OBJECTIVE: To identify the sand fly species of medically importance in the area of influence from Amoyá River Hydroelectric Project, Colombia. MATERIALS AND METHODS: Sand flies were collected with CDC light traps, Shannon traps and sticky traps, from 15 villages in Chaparral County,Tolima. RESULTS: A total of 1,077 adult sand fly specimens were collected. Thirteen species were found in the genus Lutzomyiaand one species in the genus Warileya.Among the Lutzomyia species, three species--Lutzomyia longiflocosa, Lutzomyia columbiana and Lutzomyia nuneztovari--are important for their epidemiological history.Lutzomyia suapiensis was a new record for Colombia, and Warileya rotundipennis was recorded for the first time in Tolima. CONCLUSIONS: This study contributed to an increased knowledge of Colombian sand flies in terms of (1) expanding the geographical distribution of members of the subfamily Phlebotominae, (2) gaining estimates of species-richness and species associations in central Colombia, and (3) providing a better understanding of epidemiology of leishmaniasis in the Chaparral area.

Description of Lutzomyia velezi, a new species of phlebotomine sand fly (Diptera: Psychodidae) from the Department of Antioquia, Colombia.

The phlebotomine sand fly Lutzomyia velezi sp.nov. was described and illustrated from male specimens collected by light trap in the Reserva Natural Cañon del Río Claro in the Central Cordillera of the Colombian Andes. The new species belongs to the series sanguinaria of the subgenus Helcocyrtomyia, which is represented in Colombia by Lutzomyia cirrita, Lutzomyia hartmanni, Lutzomyia sanguinaria, Lutzomyia scorzai, Lutzomyia sp. of Pichindé and Lutzomyia tortura. The new species can be differentiated from others of the subgenus by the combination of the following characteristics: long antennal ascoids, reaching level of the papilla, coxite with a single basal seta and fifth palpomere longer than or equal to the sum of the lengths of the third and fourth palpomeres.

Trece registros nuevos de Lutzomyia (Diptera: Psychodidae) para el departamento de Vichada, Orinoquia Colombiana / Thirteen new records of Lutzomyia (Diptera: Psychodidae) for the department of Vichada, Colombian Orinoquia

Se informa el primer hallazgo de Lutzomyia trinidadensis (Newstead, 1922), L. bettinii Feliciangeli, Ramírez-Pérez & Ramírez, 1988, L. flaviscutellata (Mangabeira, 1942), L. yuilli Young & Porter, 1972, L. saulensis (Floch & Abonnenc, 1944), L. runoides (Fairchild & Hertig, 1953), L. ayrozai (Barretto & Coutinho, 1940), L. claustrei Abonnenc, Leger & Fauran, 1979, L. walkeri (Newstead, 1914), L. preclara Young & Arias, 1984, L. (geniculata) sp., L. (squamiventris) sp., y L. (Evandromyia) sp., en el departamento de Vichada, Colombia. También se confirma la presencia de L. antunesi (Coutinho, 1939) y L. aragaoi (Costa Lima, 1932) en este departamento. Los flebotomíneos fueron recolectados en los municipios de Puerto Carreño, Cumaribo, y La Primavera, con trampas de luz tipo CDC, trampa Shannon y atrayente humano, y mediante búsqueda activa en lugares de reposo. Se discute la importancia epidemiológica de estas especies de flebotomíneos en relación con la leishmaniosis cutánea.

Lutzomyia trinidadensis (Newstead, 1922), L. bettinii Feliciangeli, Ramírez-Pérez & Ramírez, 1988, L. flaviscutellata (Mangabeira, 1942), L. yuilli Young & Porter, 1972, L. saulensis (Floch & Abonnenc, 1944), L. runoides (Fairchild & Hertig, 1953), L. ayrozai (Barretto & Coutinho, 1940), L. claustrei Abonnenc, Leger & Fauran, 1979, L. walkeri (Newstead, 1914), L. preclara Young & Arias, 1984, L. (geniculata) sp., L. (squamiventris) sp., and L. (Evandromyia) sp., are recorded for the first time in the department of Vichada, Colombia. In addition, the presence of L. antunesi (Coutinho, 1939) and L. aragaoi (Costa Lima, 1932) in this department is confirmed. Sand flies were collected in the municipalities of Puerto Carreño, Cumaribo, and La Primavera, using CDC light traps, Shannon trap, and human bait, and by active search in resting places. The epidemiological importance of these sand fly species is discussed in relation with the cutaneous leishmaniasis.

Description of Lutzomyia velezi, a new species of phlebotomine sand fly (Diptera: Psychodidae) from the Department of Antioquia, Colombia

The phlebotomine sand fly Lutzomyia velezi sp.nov. was described and illustrated from male specimens collected by light trap in the Reserva Natural Cañon del Río Claro in the Central Cordillera of the Colombian Andes. The new species belongs to the series sanguinaria of the subgenus Helcocyrtomyia, which is represented in Colombia by Lutzomyia cirrita, Lutzomyia hartmanni, Lutzomyia sanguinaria, Lutzomyia scorzai, Lutzomyia sp. of Pichindé and Lutzomyia tortura. The new species can be differentiated from others of the subgenus by the combination of the following characteristics: long antennal ascoids, reaching level of the papilla, coxite with a single basal seta and fifth palpomere longer than or equal to the sum of the lengths of the third and fourth palpomeres.

Nuevos registros de flebotomíneos (Diptera: Psychodidae), con el hallazgode Lutzomyia longipalpis (Lutz & Neiva, 1912), en los alrededoresde la Ciudad de Sincelejo, Colombia / New records of phlebotomine sandflies (Diptera: Psychodidae), with the report of Lutzomyia longipalpis (Lutz & Neiva, 1912), surrounding the City of Sincelejo, Colombia

Se registra por primera vez, para la Costa Caribe de Colombia, la presencia de Lutzomyia longipalpis (Lutz & Neiva, 1912), L. shannoni (Dyar, 1929), L. micropyga (Mangabeira, 1942) y L. atroclavata (Knab, 1913), en los alrededores de un núcleo urbano. Los flebotomíneos se recolectaron mediante búsqueda activa con trampas de succión y aspiradores bucales en árboles, en las proximidades de la ciudad de Sincelejo, capital del departamento de Sucre. Se enfatiza en las posibles implicaciones que se pueden derivar del hallazgo de L. longipalpis, debido a la capacidad vectorial de este flebotomíneo, vector natural de Leishmania infantum Nicolle, 1908, en América.

This is the first report of Lutzomyia longipalpis (Lutz & Neiva, 1912), L. shannoni (Dyar, 1929), L. micropyga (Mangabeira, 1942) and L. atroclavata (Knab, 1913) in the surroundings of an urban settlement in the Caribbean coast. Phlebotomine sandflies were collected by active search using suction devices and mouth aspirators in trees around the city of Sincelejo, the capital of the department of Sucre. Epidemiological importance derived from this finding is discussed due to the vectorial capacity of L. longipalpis, the natural vector of Leishmania infantum Nicolle, 1908 in America.

Análisis de la estructura primaria y secundaria del ARN de transferencia mitocondrial para serina en siete especies de Lutzomyia / Analysis of the primary and secondary structure of the mitochondrial serine transfer RNA in seven species of Lutzomyia

Introducción. Los insectos del género Lutzomyia son los responsables de la transmisión del parásito Leishmania spp. en América. La taxonomía de estos vectores se fundamenta en los caracteres morfológicos que exhiben los adultos, principalmente, en las estructuras pareadas de la cabeza y los genitales. Aunque estos caracteres permiten distinguir la mayoría de los taxones, la similitud en algunos subgéneros y grupos de especies pone límites a la identificación por criterios morfológicos. Objetivo. Evaluar la utilidad del ARN de transferencia mitocondrial para serina ARNtSer en la determinación taxonómica de Lutzomyia. Materiales y métodos. Se analizaron siete especies flebotomíneas, L. trinidadensis, L. panamensis, L. cayennensis cayennensis, L. dubitans, L. gomezi, L. rangeliana y L. evansi. A partir de cada individuo, se extrajo, amplificó y obtuvo la secuencia del gen mitocondrial que codifica para el ARNtSer, delimitado por los genes citocromo b y NAD deshidrogenasa uno. La estructura secundaria del ARNtSer se infirió teniendo como base las estructuras homólogas descritas en otros insectos del orden Diptera. Resultados. La longitud del gen ARNtSer osciló entre 66 pb en L. gomezi y 69 pb en L. trinidadensis. En el alineamiento nucleotídico de 70 posiciones, se detectaron 14 sitios polimórficos, incluyendo cuatro eventos indel. La mayoría de las sustituciones correspondieron a las lupas dihidrouridina, ribotimidina-pseudouridina-citosina y variable, así como al extremo basal del brazo anticodón. Conclusión. Los cambios en la secuencia primaria de nucleótidos y los rearreglos en la estructura secundaria del ARNtSer son potencialmente útiles para la discriminación taxonómica de las especies flebotomíneas estudiadas.

Introduction. Lutzomyia sand flies are involved in the transmission of the parasite Leishmania spp. in America. The taxonomy of these vectors is traditionally based on morphological features of the adult stage, particularly the paired structures of the head and genitalia. Although these characters are useful to distinguish most species of Lutzomyia, morphological identification may be complicated by the similarities within subgenera and species group. Objective. To evaluate the utility of mitochondrial serine transfer RNA tRNASer for taxonomic identification of Lutzomyia. Materials and methods. Seven sand fly species, each representing one of the 27 taxonomic subdivisions in genus Lutzomyia, were analyzed including L. trinidadensis (Oswaldoi group), L. (Psychodopygus) panamensis, L.(Micropygomyia) cayennensis cayennensis, L. dubitans (Migonei group), L. (Lutzomyia) gomezi, L. rangeliana (ungrouped) and L. evansi (Verrucarum group). The mitochondrial tRNASer gene, flanked by the cytochrome b and NAD dehydrogenase subunit one genes, was extracted, amplified and sequenced from each specimen. Secondary structure of the tRNASer was predicted by comparisons with previously described homologous structures from other dipteran species. Results. The tRNASer gene ranged in size from 66 base pairs in L. gomezi to 69 base pairs in L. trinidadensis. Fourteen polymorphic sites, including four insertion-deletion events, were observed in the aligned 70 nucleotide positions. The majority of the substitutions were located in the dihydrouridine, ribothymidine-pseudouridine-cytosine and variable loops, as well as in the basal extreme of the anticodon arm. Conclusion. Changes of primary sequence of the tRNASer provided useful molecular characters for taxonomic identification of the sand fly species under consideration.

[Analysis of the primary and secondary structure of the mitochondrial serine transfer RNA in seven species of Lutzomyia]. / Análisis de la estructura primaria y secundaria del ARN de transferencia mitocondrial para serina en siete especies de Lutzomyia.

INTRODUCTION: Lutzomyia sand flies are involved in the transmission of the parasite Leishmania spp. in America. The taxonomy of these vectors is traditionally based on morphological features of the adult stage, particularly the paired structures of the head and genitalia. Although these characters are useful to distinguish most species of Lutzomyia, morphological identification may be complicated by the similarities within subgenera and species group. OBJECTIVE: To evaluate the utility of mitochondrial serine transfer RNA tRNA Ser for taxonomic identification of Lutzomyia. MATERIALS AND METHODS: Seven sand fly species, each representing one of the 27 taxonomic subdivisions in genus Lutzomyia, were analyzed including L. trinidadensis (Oswaldoi group), L. (Psychodopygus) panamensis, L.(Micropygomyia) cayennensis cayennensis, L. dubitans (Migonei group), L. (Lutzomyia) gomezi, L. rangeliana (ungrouped) and L. evansi (Verrucarum group). The mitochondrial tRNA Ser gene, flanked by the cytochrome b and NAD dehydrogenase subunit one genes, was extracted, amplified and sequenced from each specimen. Secondary structure of the tRNA Ser was predicted by comparisons with previously described homologous structures from other dipteran species. RESULTS: The tRNA Ser gene ranged in size from 66 base pairs in L. gomezi to 69 base pairs in L. trinidadensis. Fourteen polymorphic sites, including four insertion-deletion events, were observed in the aligned 70 nucleotide positions. The majority of the substitutions were located in the dihydrouridine, ribothymidine-pseudouridine-cytosine and variable loops, as well as in the basal extreme of the anticodon arm. CONCLUSION: Changes of primary sequence of the tRNASer provided useful molecular characters for taxonomic identification of the sand fly species under consideration.