Mejoramiento de un método no comercial para la extracción de ADN de moscas de interés médico-legal

La obtención de ADN de moscas de interés médico-legal es de relevancia para una variedad de aplicaciones. Aunque existen métodos de extracción comerciales de ADN, su uso rutinario es limitado, en algunos escenarios. En este contexto, el uso de métodos no comerciales constituye una alternativa; sin embargo, su optimización es clave para mejorar el flujo de trabajo y los resultados. Este trabajo evaluó el impacto de variaciones a un método de precipitación salina sobre la concentración y la pureza del ADN recuperado. No se encontraron diferencias significativas en la concentración de ADN extraído entre los diferentes tiempos de incubación, probados durante la fase de extracción, mientras que el incremento en el volumen de etanol absoluto, en la fase de precipitación de ADN, mejoró significativamente la concentración de ADN obtenido. Las modificaciones propuestas reducen el tiempo de ejecución y la concentración de ADN obtenido comparado con el protocolo original.

Obtaining DNA from flies of medico-legal interest is relevant for a variety of applications. Although commercial extraction methods offer optimal DNA, their routine use is limited in some settings. In this context, the use of non-commercial methods constitutes an alternative in laboratories with limited resources however, its optimization is key to improving the workflow and the results. This work evaluated the impact of variations to a saline precipitation method on the concentration and purity of the recovered DNA. No significant differences were found in the concentration of extracted DNA between the different incubation times tested during the extraction phase. In contrast, the increased volume of absolute ethanol in the DNA precipitation phase significantly improved the concentration of DNA obtained. The proposed modifications reduce the runtime and DNA concentration obtained compared with the original protocol.

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 .

A new mtDNA COI gene lineage closely related to Anopheles janconnae of the Albitarsis complex in the Caribbean region of Colombia

An understanding of the taxonomic status and vector distribution of anophelines is crucial in controlling malaria. Previous phylogenetic analyses have supported the description of six species of the Neotropical malaria vector Anopheles (Nyssorhynchus) albitarsis s.l. (Diptera: Culicidae): An. albitarsis, Anopheles deaneorum, Anopheles marajoara, Anopheles oryzalimnetes, Anopheles janconnae and An. albitarsis F. To evaluate the taxonomic status of An. albitarsis s.l. mosquitoes collected in various localities in the Colombian Caribbean region, specimens were analyzed using the complete mitochondrial DNA cytochrome oxidase I (COI) gene, the ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2) region and partial nuclear DNA white gene sequences. Phylogenetic analyses of the COI gene sequences detected a new lineage closely related to An. janconnae in the Caribbean region of Colombia and determined its position relative to the other members of the complex. However, the ITS2 and white gene sequences lacked sufficient resolution to support a new lineage closely related to An. janconnae or the An. janconnae clade. The possible involvement of this new lineage in malaria transmission in Colombia remains unknown, but its phylogenetic closeness to An. janconnae, which has been implicated in local malaria transmission in Brazil, is intriguing.

Species composition and natural infectivity of anthropophilic Anopheles (Diptera: Culicidae) in the states of Córdoba and Antioquia, Northwestern Colombia

Malaria is a serious health problem in the states of Córdoba and Antioquia, Northwestern Colombia, where 64.4 percent of total Colombian cases were reported in 2007. Because little entomological information is available in this region, the aim of this work was to identify the Anopheles species composition and natural infectivity of mosquitoes distributed in seven localities with highest malaria transmission. A total of 1,768 Anopheles mosquitoes were collected using human landing catches from March 2007-July 2008. Ten species were identified; overall, Anopheles nuneztovari s.l. was the most widespread (62 percent) and showed the highest average human biting rates. There were six other species of the Nyssorhynchus subgenus: Anopheles albimanus (11.6 percent), Anopheles darlingi (9.8 percent), Anopheles braziliensis (6.6 percent), Anopheles triannulatus s.l. (3.5 percent), Anopheles albitarsis s.l. and Anopheles oswaldoi s.l. at < 1 percent; and three of the Anopheles subgenus: Anopheles punctimacula, Anopheles pseudopunctipennis s.l. and Anopheles neomaculipalpusat < 1 percent each. Two species from Córdoba, An. nuneztovari and An. darlingi, were found to be naturally infected by Plasmodium vivax VK247, as determined by ELISA and confirmed by nested PCR. All species were active indoors and outdoors. These results provide basic information for targeted vector control strategies in these localities.