Background: Dengue virus is spread in tropical areas of the world and is the causative agent of dengue fever and dengue hemorrhagic fever. It is horizontally transmitted to humans by infected Aedes mosquitoes, but it is also able to be vertically or transovarially transmitted to insect progeny. Objective: In this work, we analyzed the vertical transmission of dengue virus in Aedes aegypti mosquitoes collected in two endemic localities in the state of Oaxaca, Mexico. Methods: The collected larvae were grown in the laboratory and transovarial transmission of dengue virus, either in larvae or newly emerged mosquitoes, was investigated using a semi-nested reverse transcription-polymerase chain reaction method. Results: Although the presence of dengue virus in larvae could not be demonstrated, the viral genome was amplified in 4 out of 43 pools of in-cage born mosquitoes: DEN 2, 3 and 4 serotypes were detected in 2 pools from Tuxtepec and two from Juchitán. Conclusion: The results presented here strongly suggest that dengue virus can be vertically transmitted in mosquitoes from Oaxaca, but more studies will be necessary to analyze the epidemiological impact of this mechanism of transmission.

BackgroundCulex spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, Culex pipiens pipiens, Culex pipiens quinquefasciatus, and Culex tarsalis are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of Culex pipiens complex mosquitoes and hybrids in Mexico City. Culex pipiens pipiens preferentially feed on avian hosts; Culex pipiens quinquefasciatus have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of Culex pipiens complex mosquitoes and their hybrids.ResultsMosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as Culex, Culiseta or Ochlerotatus by morphological analysis. Within the genus Culex, specimens were further identified as Culex tarsalis or as belonging to the Culex pipiens complex. Members of the Culex pipiens complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the Ace.2 gene. Culex pipiens quinquefasciatus was the most abundant form collected.ConclusionsImportant WNV vectors species, Cx. p. pipiens, Cx. p. quinquefasciatus and Cx. tarsalis, are all present in Mexico City. Hybrids of Cx. p. pipiens and Cx. p. quinquefasciatus were also collected and identified. The presence and abundance of these WNV competent vectors is a cause for concern. Understanding the distribution of these vectors can help improve viral surveillance activities and mosquito control efforts in Mexico City.

Here; we have described and tested a microarray based-method for the screening of dengue virus (DENV) serotypes. This DNA microarray assay is specific and sensitive and can detect dual infections with two dengue virus serotypes and single-serotype infections. Other methodologies may underestimate samples containing more than one serotype. This technology can be used to discriminate between the four DENV serotypes. Single-stranded DNA targets were covalently attached to glass slides and hybridised with specific labelled probes. DENV isolates and dengue samples were used to evaluate microarray performance. Our results demonstrate that the probes hybridized specifically to DENV serotypes; with no detection of unspecific signals. This finding provides evidence that specific probes can effectively identify single and double infections in DENV samples.