A new approach to improve acoustic trapping effectiveness for Aedes aegypti (Diptera: Culicidae).

Monitoring mosquito populations is essential to designing and implementing control strategies. Recent strategies based on releasing biologically modified mosquitoes have increased the need to effectively monitor mosquito abundance. Unfortunately, existing surveillance traps are of limited value due to their high cost and low capture rates. Here, we report the results of experiments designed to evaluate the effectiveness of an acoustic trap prototype. Stimuli synthesized from recordings of Ae. aegypti wingbeat signals and pure tones were evaluated as attractants to males in indoor and semi-field conditions. Overall, the acoustic trap´s efficacy differed significantly between indoor and semi-field conditions. After two hours of indoor recapture, ∼69% of males were collected from acoustic traps broadcasting pure tones while ∼78% of males were collected using synthesized wingbeat signals. Under semi-field conditions, however, acoustic traps collected less than ∼1.7% of the males released. Increasing the intensity of the signals up to 90 dB (SPL re. 20 uPa at 1 m from the trap) did not improve the capture rate under semi-field conditions. Overall, our results indicate that acoustic signals synthesized from recordings of wingbeats can be used to enhance capture of male Ae. aegypti.

Brotes emergentes de leptospirosis del Amazonas colombiano / Emerging leptospirosis outbreaks in the Colombian Department of Amazonas

Se describe por primera vez una serie de nueve casos con clínica indicativa de leptospirosis en el municipio Puerto Nariño en el departamento Amazonas, Colombia. Se muestran evidencias serológicas de exposición con Rickettsia del grupo de las fiebres manchadas. Los casos fueron clínicamente considerados como síndrome febril de origen desconocido. Se descartó infección por dengue y malaria. El diagnóstico de Leptospira se realizó mediante el método de reacción en cadena de la polimerasa en tiempo real. Igualmente, se detectó la presencia de anticuerpos contra rickettsias del grupo de las fiebres manchadas por inmunofluorescencia Indirecta. Finalmente, se realiza revisión del tema(AU)

A description is provided for the first time of a series of nine cases with a clinical examination suggestive of leptospirosis in the municipality of Puerto Nariño, Department of Amazonas, Colombia. Serological evidence is presented of exposure to Rickettsia, spotted fever group. The cases were clinically considered as febrile syndrome of unknown origin. Infection with dengue or malaria was ruled out. Diagnosis of leptospirosis was achieved by real-time polymerase chain reaction. Additionally, indirect immunofluorescence detected the presence of antibodies against rickettsia, spotted fever group. Finally, a review was conducted about the topic(AU)

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

Among vector-borne diseases malaria is the leading cause of morbidity in the world, with more than 200 million cases per year and a large number of deaths. The techniques traditionally used for the detection of Plasmodium in humans and Anopheles mosquitoes include microscopy, IRMA, ELISA, antibody or molecular assays, and anopheline dissection. However, these techniques are limited by their requirement of skilled personnel, low sensitivity or long processing times. A PCR-based high-resolution melting (PCR-HRM) analysis was developed for the detection and identification of P. falciparum, P. vivax and P. malariae that infect humans and Anopheles. In 41 human samples PCR-HRM detected 14 samples positive for P. vivax, 17 for P. falciparum, three for P. malariae, three mixed infections for P. vivax/P. malariae and four negative samples. Whereas benchmarking assays of microscopy and nested PCR had false positive detections. Additionally, PCR-HRM was able to detect natural infection with Plasmodium spp. in An. darlingi and An. mattogrossensis. The PCR-HRM presented is the first single assay developed for the detection and identification of P. vivax, P. falciparum and/or P. malariae in human and Anopheles. This method improves on currently available assays as it is easy-to-use, rapid, sensitive and specific with a low risk of contamination.

Presence of Aedes (Stegomyia) aegypti (Linnaeus, 1762) and its natural infection with dengue virus at unrecorded heights in Colombia.

INTRODUCTION: Aedes aegypti is the main vector of urban yellow fever, dengue, chikungunya and Zika viruses. The biogeographical distribution of this species has expanded due to global warming, and socioeconomic and cultural factors. The changes in the altitudinal distribution patterns of this vector and its natural infection are priority fields of research to develop entomological, virological and public health surveillance strategies.  OBJECTIVE: To evaluate the presence of A. aegypti and its natural infection with dengue virus in altitudes above 1.800 meters above sea level in two peripheral municipalities of the Valle de Aburrá, Antioquia, Colombia.  MATERIALS AND METHODS: Twenty-one ovitraps were set in the municipalities of Bello and San Pedro de los Milagros, at altitudes ranging from 1.882 to 2.659 masl. Emerged adults caught in the ovitraps were tested by RT-PCR for dengue virus detection.  RESULTS: We collected 367 A. aegypti adults, seven of which were found as high as 2.302 masl in Tierradentro, Bello. We detected serotype 2 dengue infection in 12 A. aegypti specimens collected in the neighbourhood of París, in Bello, at 1.984 masl.  CONCLUSION: We recorded A. aegypti at 2.302 masl, so far the highest altitudinal record in Colombia for this vector. Furthermore, mosquitoes collected at 1.984 masl were positive for dengue virus. These findings are significant as they identify regions in Colombia at risk of potential autochthonous transmission of dengue and other arboviruses by A. aegypti.

Presencia de Aedes ( Stegomyia ) aegypti (Linnaeus, 1762) y su infección natural con el virus del dengue en alturas no registradas para Colombia / Presence of Aedes ( Stegomyia ) aegypti (Linnaeus, 1762) and its natural infection with dengue virus at unrecorded heights in Colombia

Introducción. Aedes aegypti es el principal vector de fiebre amarilla urbana, dengue, chikungunya y zika. Se ha demostrado que la distribución biogeográfica de esta especie se ha expandido debido al calentamiento global y a factores socioeconómicos y culturales. Los cambios en los patrones de la distribución altitudinal de este vector y su infección con el virus son prioridades de la investigación encaminada a desarrollar estrategias de vigilancia entomológica y virológica en salud pública. Objetivo. Evaluar la presencia de A. aegypti y su infección natural por el virus del dengue en alturas superiores a los 1.800 msnm en dos municipios periféricos del Valle de Aburrá, Antioquia, Colombia. Materiales y métodos. Se instalaron 21 ovitrampas en los municipios de Bello y San Pedro de los Milagros, en un rango altitudinal de 1.882 a 2.659 msnm. Los adultos que emergieron de las ovitrampas se evaluaron con reacción en cadena de la polimerasa en tiempo real (RT-PCR) para la detección del virus del dengue. Resultados. Se recolectaron 367 adultos de A. aegypti , siete de los cuales se encontraron a una altitud de 2.302 msnm en Tierradentro, Bello. Se detectaron 12 especímenes de A. aegypti positivos para dengue serotipo 2 en el barrio París de Bello, a 1.984 msnm. Conclusión. Por primera vez se registró A. aegypti a 2.302 msnm, la mayor altitud registrada para este vector en Colombia. De igual forma, se encontró infección con el virus del dengue a 1.984 msnm. Estos hallazgos son significativos, ya que determinan regiones de Colombia con riesgo potencial de transmisión autóctona de dengue y otros arbovirus por A. aegypti .

Introduction: Aedes aegypti is the main vector of urban yellow fever, dengue, chikungunya and Zika viruses. The biogeographical distribution of this species has expanded due to global warming, and socioeconomic and cultural factors. The changes in the altitudinal distribution patterns of this vector and its natural infection are priority fields of research to develop entomological, virological and public health surveillance strategies. Objective: To evaluate the presence of A. aegypti and its natural infection with dengue virus in altitudes above 1.800 meters above sea level in two peripheral municipalities of the Valle de Aburrá, Antioquia, Colombia. Materials and methods: Twenty-one ovitraps were set in the municipalities of Bello and San Pedro de los Milagros, at altitudes ranging from 1.882 to 2.659 masl. Emerged adults caught in the ovitraps were tested by RT-PCR for dengue virus detection. Results: We collected 367 A. aegypti adults, seven of which were found as high as 2.302 masl in Tierradentro, Bello. We detected serotype 2 dengue infection in 12 A. aegypti specimens collected in the neighbourhood of París, in Bello, at 1.984 masl. Conclusion: We recorded A. aegypti at 2.302 masl, so far the highest altitudinal record in Colombia for this vector. Furthermore, mosquitoes collected at 1.984 masl were positive for dengue virus. These findings are significant as they identify regions in Colombia at risk of potential autochthonous transmission of dengue and other arboviruses by A. aegypti .

Morphological and molecular study of Symphyla from Colombia.

The symphylans are a poorly studied group. In Colombia the number of symphylan species is unknown with only Scutigerellaimmaculata (Symphyla: Scutigerellidae) being reported previously. The aim of this research was to collect and identify the symphylan pests of flower crops in Colombia. Morphological descriptions showed that our specimens shared more than one of the characters that define different genera within Scutigerellidae. The COI barcode haplotype showed interspecific level genetic divergence with Scutigerellacauseyae (at least 23%) and Hanseniella sp. (22%). Furthermore, our Colombian symphylans shared the same COI haplotype as some Symphyla found in Cameroon indicating a wide geographical distribution of this taxon. Our results suggest the presence of a new genus or subgenus in the class Symphyla.

Primer registro de Anopheles (Nyssorhynchus) oswaldoi B y Anopheles (Nys) albitarsis F en la cuenca del río Caura, Estado Bolívar, Venezuela / First record of anopheles (Nyssorhynchus) oswaldoi B y anopheles (Nys) albi-tarsis F from the Caura river basin, Bolívar State, Venezuela

Se registran por primera vez anopheles albitarsis F y anopheles oswaldoi B recolectadas en localidades de los municipios Sucre y Cedeño del estado Bolívar. Los análisis morfométricos de hembras adultas mostraron que los caracteres analizados en las manchas claras y oscuras de la vena Costa del ala y los tarsómeros de las patas posteriores para An. albitarsis F caen dentro de los rangos reportados para An. marajora sensu lato, mientras que para An. oswaldoi B los rangos de variación de los caracteres medidos están dentro del rango reportado para An. oswaldoi s.l., An. oswaldoi sensu stricto de Brasil y An. konderi. Sin embargo, el análisis de ADN mitocondrial (región código de barras, 658 pb) mostró una divergencia genética entre An. oswaldoi B y An. oswaldoi s.s. de 3,2%, mientras entre An. albirtarsis F y An. marajoara s.s. esta fue de 5,2%, valores que están por encima de los limites propuestos para delimitación de especies.

Anopheles albitarsis F and Anopheles oswaldoi B were first recorded from collections in Sucre and Cedeño Municipalities, Bolivar State. Morphometric analysis of dark and pale spots on wing Costa and hind tarsomeres of adult females, showed that the range of variation for An. albitarsis F are within the reported ranges for An. marajoara sensu latu For An. oswaldoi B the range of variation are within the ranges reported for An. oswaldoi s.l., An. oswaldoi sensu stricto from Brazil and An. konderi. DNA mitochondrial analysis (Barcode region 658 bp) showed 3.2% of genetic divergence between An. oswaldoi B and An. oswaldoi s.s., while this divergence was 5.2% between An. albirtarsis F and An. marajoara s.s. Both these values are above the proposed limits for species delimitation.

Systematics of the oswaldoi complex (Anopheles, Nyssorhynchus) in South America.

BACKGROUND: Effective malaria control relies on accurate identification of those Anopheles mosquitoes responsible for the transmission of Plasmodium parasites. Anopheles oswaldoi s.l. has been incriminated as a malaria vector in Colombia and some localities in Brazil, but not ubiquitously throughout its Neotropical range. This evidence together with variable morphological characters and genetic differences supports that An. oswaldoi s.l. compromises a species complex. The recent fully integrated redescription of An. oswaldoi s.s. provides a solid taxonomic foundation from which to molecularly determine other members of the complex. METHODS: DNA sequences of the Second Internal Transcribed Spacer (ITS2 - rDNA) (n = 192) and the barcoding region of the Cytochrome Oxidase I gene (COI - mtDNA) (n = 110) were generated from 255 specimens of An. oswaldoi s.l. from 33 localities: Brazil (8 localities, including the lectotype series of An. oswaldoi), Ecuador (4), Colombia (17), Trinidad and Tobago (1), and Peru (3). COI sequences were analyzed employing the Kimura-two-parameter model (K2P), Bayesian analysis (MrBayes), Mixed Yule-Coalescent model (MYC, for delimitation of clusters) and TCS genealogies. RESULTS: Separate and combined analysis of the COI and ITS2 data sets unequivocally supported four separate species: two previously determined (An. oswaldoi s.s. and An. oswaldoi B) and two newly designated species in the Oswaldoi Complex (An. oswaldoi A and An. sp. nr. konderi). The COI intra- and inter-specific genetic distances for the four taxa were non-overlapping, averaging 0.012 (0.007 to 0.020) and 0.052 (0.038 to 0.064), respectively. The concurring four clusters delineated by MrBayes and MYC, and four independent TCS networks, strongly confirmed their separate species status. In addition, An. konderi of Sallum should be regarded as unique with respect to the above. Despite initially being included as an outgroup taxon, this species falls well within the examined taxa, suggesting a combined analysis of these taxa would be most appropriate. CONCLUSIONS: Through novel data and retrospective comparison of available COI and ITS2 DNA sequences, evidence is shown to support the separate species status of An. oswaldoi s.s., An. oswaldoi A and An. oswaldoi B, and at least two species in the closely related An. konderi complex (An. sp. nr. konderi, An. konderi of Sallum). Although An. oswaldoi s.s. has never been implicated in malaria transmission, An. oswaldoi B is a confirmed vector and the new species An. oswaldoi A and An. sp. nr. konderi are circumstantially implicated, most likely acting as secondary vectors.

DNA barcoding reveals both known and novel taxa in the Albitarsis Group (Anopheles: Nyssorhynchus) of Neotropical malaria vectors.

BACKGROUND: Mosquitoes belonging to the Albitarsis Group (Anopheles: Nyssorhynchus) are of importance as malaria vectors across the Neotropics. The Group currently comprises six known species, and recent studies have indicated further hidden biodiversity within the Group. DNA barcoding has been proposed as a highly useful tool for species recognition, although its discriminatory utility has not been verified in closely related taxa across a wide geographic distribution. METHODS: DNA barcodes (658 bp of the mtDNA Cytochrome c Oxidase--COI) were generated for 565 An. albitarsis s.l. collected in Argentina, Brazil, Colombia, Paraguay, Trinidad and Venezuela over the past twenty years, including specimens from type series and type localities. Here we test the utility of currently advocated barcoding methodologies, including the Kimura-two-parameter distance model (K2P) and Neighbor-joining analysis (NJ), for determining species delineation within mosquitoes of the Neotropical Albitarsis Group of malaria vectors (Anopheles: Nyssorhynchus), and compare results with Bayesian analysis. RESULTS: Species delineation through barcoding analysis and Bayesian phylogenetic analysis, fully concur. Analysis of 565 sequences (302 unique haplotypes) resolved nine NJ tree clusters, with less than 2% intra-node variation. Mean intra-specific variation (K2P) was 0.009 (range 0.002-0.014), whereas mean inter-specific divergence were several-fold higher at 0.041 (0.020-0.056), supporting the reported "barcoding gap". These results show full support for separate species status of the six known species in the Albitarsis Group (An. albitarsis s.s., An. albitarsis F, An. deaneorum, An. janconnae, An. marajoara and An. oryzalimnetes), and also support species level status for two previously detected lineages--An. albitarsis G &An. albitarsis I (designated herein). In addition, we highlight the presence of a unique mitochondrial lineage close to An. deaneorum and An. marajoara (An. albitarsis H) from Rondônia and Mato Grosso in southwestern Brazil. Further integrated studies are required to confirm the status of this lineage. CONCLUSIONS: DNA barcoding provides a reliable means of identifying both known and undiscovered biodiversity within the closely related taxa of the Albitarsis Group. We advocate its usage in future studies to elucidate the vector competence and respective distributions of all eight species in the Albitarsis Group and the novel mitochondrial lineage (An. albitarsis H) recovered in this study.

Anopheles (Kerteszia) lepidotus (Diptera: Culicidae), not the malaria vector we thought it was: Revised male and female morphology; larva, pupa, and male genitalia characters; and molecular verification.

The name Anopheles (Kerteszia) lepidotus Zavortink, commonly used for an important malaria vector in the eastern cordillera of the Andes, is here corrected to An. pholidotus Zavortink. We discovered that An. (Ker.) specimens from Peru, and reared-associated specimens from Ecuador, had unambiguous habitus characters that matched those on the male holotype of An. lepidotus. However, the specimens do not exhibit characters of the female allotype and female paratypes of An. lepidotus, which are actually An. pholidotus. Our specimens are the first correctly associated females of An. lepidotus, which allow us to provide a new morphological interpretation for the adult habitus of this species. This finding is also corroborated by molecular data from a portion of the Cytochrome Oxidase I (COI) gene and ribosomal DNA Internal Transcribed Spacer 2 (rDNA ITS2). The pupal stage of An. lepidotus is described for the first time, and additional larval characters are also noted. Diagnostic morphological characters for the adult, pupal, and larval stages of An. pholidotus are provided to separate the two species. All stages of An. lepidotus are easily separated from other currently known species in subgenus Kerteszia and a new key to the females of An. (Kerteszia) is given. Previously published distribution, bionomics, and medical significance data are corrected and enhanced.

Malaria vector species in Colombia: a review.

Here we present a comprehensive review of the literature on the vectorial importance of the major Anopheles malaria vectors in Colombia. We provide basic information on the geographical distribution, altitudinal range, immature habitats, adult behaviour, feeding preferences and anthropophily, endophily and infectivity rates. We additionally review information on the life cycle, longevity and population fluctuation of Colombian Anopheles species. Emphasis was placed on the primary vectors that have been epidemiologically incriminated in malaria transmission: Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. The role of a selection of local, regional or secondary vectors (e.g., Anopheles pseudopunctipennis and Anopheles neivai) is also discussed. We highlight the importance of combining biological, morphological and molecular data for the correct taxonomical determination of a given species, particularly for members of the species complexes. We likewise emphasise the importance of studying the bionomics of primary and secondary vectors along with an examination of the local conditions affecting the transmission of malaria. The presence and spread of the major vectors and the emergence of secondary species capable of transmitting human Plasmodia are of great interest. When selecting control measures, the anopheline diversity in the region must be considered. Variation in macroclimate conditions over a species' geographical range must be well understood and targeted to plan effective control measures based on the population dynamics of the local Anopheles species.

Malaria vector species in Colombia: a review

Here we present a comprehensive review of the literature on the vectorial importance of the major Anopheles malaria vectors in Colombia. We provide basic information on the geographical distribution, altitudinal range, immature habitats, adult behaviour, feeding preferences and anthropophily, endophily and infectivity rates. We additionally review information on the life cycle, longevity and population fluctuation of Colombian Anopheles species. Emphasis was placed on the primary vectors that have been epidemiologically incriminated in malaria transmission: Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. The role of a selection of local, regional or secondary vectors (e.g., Anopheles pseudopunctipennis and Anopheles neivai) is also discussed. We highlight the importance of combining biological, morphological and molecular data for the correct taxonomical determination of a given species, particularly for members of the species complexes. We likewise emphasise the importance of studying the bionomics of primary and secondary vectors along with an examination of the local conditions affecting the transmission of malaria. The presence and spread of the major vectors and the emergence of secondary species capable of transmitting human Plasmodia are of great interest. When selecting control measures, the anopheline diversity in the region must be considered. Variation in macroclimate conditions over a species' geographical range must be well understood and targeted to plan effective control measures based on the population dynamics of the local Anopheles species.