Phylogeography of the neotropical Anopheles triannulatus complex (Diptera: Culicidae) supports deep structure and complex patterns.

BACKGROUND: The molecular phylogenetic relationships and population structure of the species of the Anopheles triannulatus complex: Anopheles triannulatus s.s., Anopheles halophylus and the putative species Anopheles triannulatus C were investigated. METHODS: The mitochondrial COI gene, the nuclear white gene and rDNA ITS2 of samples that include the known geographic distribution of these taxa were analyzed. Phylogenetic analyses were performed using Bayesian inference, Maximum parsimony and Maximum likelihood approaches. RESULTS: Each data set analyzed septely yielded a different topology but none provided evidence for the seption of An. halophylus and An. triannulatus C, consistent with the hypothesis that the two are undergoing incipient speciation. The phylogenetic analyses of the white gene found three main clades, whereas the statistical parsimony network detected only a single metapopulation of Anopheles triannulatus s.l. Seven COI lineages were detected by phylogenetic and network analysis. In contrast, the network, but not the phylogenetic analyses, strongly supported three ITS2 groups. Combined data analyses provided the best resolution of the trees, with two major clades, Amazonian (clade I) and trans-Andean + Amazon Delta (clade II). Clade I consists of multiple subclades: An. halophylus + An. triannulatus C; trans-Andean Venezuela; central Amazonia + central Bolivia; Atlantic coastal lowland; and Amazon delta. Clade II includes three subclades: Panama; cis-Andean Colombia; and cis-Venezuela. The Amazon delta specimens are in both clades, likely indicating local sympatry. Spatial and molecular variance analyses detected nine groups, corroborating some of subclades obtained in the combined data analysis. CONCLUSION: Combination of the three molecular markers provided the best resolution for differentiation within An. triannulatus s.s. and An. halophylus and C. The latest two species seem to be very closely related and the analyses performed were not conclusive regarding species differentiation. Further studies including new molecular markers would be desirable to solve this species status question. Besides, results of the study indicate a trans-Andean origin for An. triannulatus s.l. The potential implications for malaria epidemiology remain to be investigated.

Invalidación de autapomorfías putativas mediante análisis de filogenia molecular en Anopheles subgénero Nyssorhynchus / Invalidation of putative autapomorphic characters using molecular phylogenetic analyses in Anopheles subgenus Nyssorhynchus

Colectas e identificaciones realizadas en un área de transmisión malárica en el estado Sucre, Venezuela, mostraron mediante el uso de las claves ad hoc, la posible presencia en simpatría de poblaciones de Anopheles aquasalis Curry y An. benarrochi Gabaldón, Cova-García & López. Adicionalmente, se detectaron poblaciones de individuos que mostraron mezcla de los caracteres diagnósticos señalados en las claves. Debido a la importancia epidemiológica que representan ambas especies en dicha zona malárica, este estudio se orientó hacia la determinación de las tres entidades señaladas, mediante filogenia molecular con las secuencias de ADN mitocondrial (region Citocromo Oxidasa 1). En este sentido, el concepto de especie filogenética fue evaluada utilizando análisis de parsimonia máxima que mostraron politomías (nodos no resueltos) fuertemente apoyadas entre las secuencias inclusive la del haplotipo proveniente de un macho identificado como aquasalis, mostrando hipótesis (árboles) totales sin resolución ni apoyo de grupos internos con los tres morfotipos como un grupo natural único. Las distancias genéticas (Kimura-2P) mostraron que la variablidad inter-morfotipos están dentro de la varianza intra sugiriendo que los haplotipos problema son una única entidad polimórfica y con plasticidad fenotípica. Se concluye que An. aquasalis es la entidad única de las poblaciones estudiadas y que los caracteres morfológicos como: 1) vena Medial con escamas negras y 2) palpos maxilares con superficie ventral cubierta de escamas blancas, que son propuestas en la mayoría de las claves no son caracteres válidos para la identificación de hembras de An. benarrochi y que tales caracteres forman parte de la variación intraespecífica en An. aquasalis. Esta conclusión apoya la revisión de Faran (1980), quién no propone caracteres diferenciales para hembras de ambas especies, siendo indistinguibles por medio de los caracteres morfológicos tradicionalmente utilizados.

Collected specimens from a malaric area in Sucre State, Venezuela, using the ad hoc Keys, were identified as Anopheles aquasalis Curry and An. benarrochi Gabaldón, Cova-García & López populations. Additionally, other individuals have shown a mixture of diagnostic characters of both species. As a consequence of the epidemiological importance that both species represent in this malaric area and its correct identification for vector control aims, we addressed the research to the determination of both entities by mean of phylogenetic methods using mitochondrial DNA (Cytochrome Oxidase 1) from these three entities. In order to evaluate the phylogenetic species concept, the Maximum Parsimony analyses showed strong supported politomies (unresolved nodes), yielded solutions with no supported groups correlated with two or three morphological entities including the male haplotype identified as An. aquasalis. The genetic distances (K-2P) showed that the variability inter-morphotype is in the variance intra, suggesting that the haplotypes belong to the three morphotypes and represent a unique polymorphic species with phenotypic plasticity. We conclude that An. aquasalis is a unique taxonomic entity in the populations studied and also that the morphological characters such as: 1) medial vein with black scales and 2) maxillary palpus with ventral surface covered with white scales, proposed in most of the keys used are not valid characters to distinguish females of An. benarrochi from An. aquasalis as these characters belong to the intra-specific variation of An. aquasalis. This conclusion supports the Faran (1980) key, which did not report any morphological characters, showing undistinguished females under classical characters used

[Asymmetric reproductive isolation between Lutzomyia pseudolongipalpis and Lutzomyia longipalpis (species C2), neotropical vectors of visceral leishmaniasis (Diptera: Pshychodidae)]. / Aislamiento reproductive asimétrico entre Lutzomyia pseudolongipalpis y Lutzomyia longipalpis (especie C2), vectores neotropicales de leishmaniasis visceral (Diptera: Pshychodidae).

Lutzomyia pseudolongipalpis and Lutzomyia longipalpis (species C2, L. sp n.) are two endemic species of Phlebotominae sand fly vectors from Venezuela. The two insects are sympatric and monophyletic but have deeply diverging, morphological differences. They belong to the L. longipalpis complex. A study of their reproductive isolation is necessary to understand the process of speciation and maintenance of the two sister species as two discrete taxonomic and biological entities. Cross-mating tests were conducted (homo and hetero-specific) and monitored under two criteria: biological (presence of copulation and offspring) and genetic (using two isozymic markers diagnostic for the L. longipalpis complex; enzyme diagnosis AK and HK). Results indicate reproductive isolation, with an asymmetrical genetic exchange in a direction of hybridization between the two species under experimental conditions, and production of a low number of heterozygotes. These findings support the existence of negative selection on hybrids, and explain the absence of hybrids under natural conditions, in the sympatric locality.

Aislamiento reproductivo asimétrico entre Lutzomyia pseudolongipalpis y Lutzomyia longipalpis (especie C2), vectores neotropicales de leishmaniasis visceral (Diptera: Pshychodidae)

Asymmetric reproductive isolation between Lutzomyia pseudolongipalpis and Lutzomyia longipalpis (species C2), Neotropical vectors of visceral leishmaniasis (Diptera: Pshychodidae). Lutzomyia pseudolongipalpis and Lutzomyia longipalpis (species C2, L. sp n.) are two endemic species of Phlebotominae sand fly vectors from Venezuela. The two insects are sympatric and monophyletic but have deeply diverging, morphological differences. They belong to the L. longipalpis complex. A study of their reproductive isolation is necessary to understand the process of speciation and maintenance of the two sister species as two discrete taxonomic and biological entities. Cross-mating tests were conducted (homo and hetero-specific) and monitored under two criteria: biological (presence of copulation and offspring) and genetic (using two isozymic markers diagnostic for the L. longipalpis complex; enzyme diagnosis AK and HK). Results indicate reproductive isolation, with an asymmetrical genetic exchange in a direction of hybridization between the two species under experimental conditions, and production of a low number of heterozygotes. These findings support the existence of negative selection on hybrids, and explain the absence of hybrids under natural conditions, in the sympatric locality. Rev. Biol. Trop. 57 (1-2): 23-31. Epub 2009 June 30.

Lutzomyia pseudolongipalpis y Lutzomyia longipalpis (especie C2, L. n. sp) son dos especies de flebotominos vectores endémicos de Venezuela, simpátricas, monofiléticas, con profundas divergencias, morfológicamente diferentes y pertenecientes al complejo de especies L. longipalpis. El estudio de su aislamiento reproductivo es clave para entender el proceso de especiación y el mantenimiento de estas dos especies hermanas como entidades taxonómicas y biológicas discretas. Por tal motivo, se realizaron ensayos de entrecruzamiento homo y heteroespecíficos, los cuales fueron monitoreados por dos criterios: biológico (presencia de cópula y progenie) y genético (utilizando dos marcadores isoenzimáticos diagnósticos para el complejo L. longipalpis, las enzimas AK y HK). Los resultados indican aislamiento reproductivo, con un intercambio genético asimétrico hacia la hibridización en condiciones experimentales, y producción de un bajo número de heterocigotos, lo que apoya la existencia de selección negativa sobre los híbridos y explica su ausencia en condiciones naturales en la localidad simpátrica.

Monitoring of argatroban in ARG310 study: potential recommendations for its use in interventional cardiology.

Argatroban is a peptidomimimetic synthetic direct thrombin inhibitor with reversible and specific properties resulting in predictable anticoagulant effects. Usually, argatroban therapy is monitored by the activated dotting time (ACT) or activated partial thromboplastin time (aPTT). While other global dotting tests for the monitoring of anticoagulants are useful, their applicability to antithrombin agents (particularly of argatroban at higher dosages) is rather questionable. In this study, we sought to compare the argatroban anticoagulant levels in patients undergoing percutaneous transluminal angioplasty (PTCA) and stenting procedures, utilizing both functional and absolute quantitation methods. Argatroban produced a comparable increase of ACT and HMT, 5 to 10 minutes after administration. The level of anticoagulation achieved (400-450 seconds with ACT and HMT) following a slow bolus of argatroban (350 microg/kg) was maintained throughout the procedure using 25 microg/kg/min infusion. Following discontinuation of argatroban at the end of the procedure, the ACTs and HMTs showed a comparable progressive reduction in the anticoagulant response, which reflected the elimination of argatroban 2 to 3 hours after the procedure. No significant differences between the three methods (Hemotec, Hemochron, and HMT) were noted at any given sampling time. Argatroban dosage at 350 microg/kg intravenous slow bolus followed by 25 microg/kg/min infusion was adequate to perform PTCA and stenting procedures. There was no incidence of bleeding complications. Absolute quantitation of argatroban levels in patient plasmas by a newly developed HPLC method was found to be quite comparable with the ecarin dotting time (ECT) results. The ECT system was found to be less sensitive when compared to other tests, and therefore, could be used as a point-of-care test during the PTCA/stenting procedures to monitor argatroban.

Estudio eco-epidemiológico de la reintroducción de la malaria en el nororiente de Venezuela mediante Sistemas de Información geográfica y sensores remotos / Study of eco-epidemiological of the reintroduction of the malaria in the northeastern of Venezuela geographical information systems and remote sensingg

El principal objetivo de este estudio es el análisis espacio-temporal del proceso de la reintroducción de la malaria en la población humana en la zona de Santa Fe, estado Sucre, mediante un enfoque eco-epidemiológico y el empleo de Sistemas de Información Geográfica (SIG) y Sensores Remotos (imagen Landsat TM). El SIG integra datos epidemiológicos (casos de malaria), meteorológicos (precipitación), geográficos (relieve, hidrografía, vialidad, población humana, uso de la tierra, etc.) y entomológicos (criaderos de anofelinos vectores). Los resultados del presente estudio señalan los siguientes factores de riesgo malárico para la zona de Santa Fe: Altitudes menores a 50 m sobre el nivel del mar, pendiente del terreno inferior al 10 por ciento, viviendas ubicadas a menos de 1 Km de criaderos de Anopheles aquasalis, especialemente aquellas cercanas a ríos y próximas a canales, presencia de vegetación boscosa o de manglar, concentraciones urbanas y presencia de herbazales altos (caña amarga). Mediante el SIG podemos delimitar las áreas de mayor riesgo malárico, por lo cual se recomienda la aplicación de esta tecnología para reducir los costos operacionales de control en zonas que están actualmente infectadas. Se hacen recomendaciones para el control de An.aquasalis mediante técnicas de manejo ambiental de los criaderos para proteger a la población. Se recomienda una adecuada vigilancia epidemiológica dado que existen las condiciones necesarias para una eventual reinfección de la zona

Estudio biosistemático y ecológico de anopleles aquasalis y sus implicaciones para el control de la malaria en el estado Sucre: I genética de poblaciones / Study biosystematic and ecologic of anopheles aquasalis and it's implications for the malaria control in the Sucre state: I genetic populations

Se realizaron sub-poblaciones del vector de malaria Anopheles aquasalis de Venezuela y Trinidad, mediante 18 loci enzimáticos. En Venezuela, los estimados de variabilidad genética fueron relativamente mayores que los obtenidos en Trinidad. Los valores de FST estuvieron en un intervalo de poca a gran diferenciación en Venezuela y de baja estructura entre Capure, Venezuela y Trinidad. Los valores de distancia genética fueron pequeños y soportan la hipotesis que Anopheles aquasalis es una especie. Los resultados indican la necesidad de un control local del vector para reducir la incidencia de la enfermedad