FLAVIVIRUS SUSCPETIBILITY IN Aedes aegypti

Aedes aegypti is the primary vector of yellow fever (YF) and dengue fever (DF) flaviviruses worldwide. In this review we focus on past and present research on genetic components and environmental factors in Aedes aegypti that appear to control flavivirus transmission. We review genetic relationships among Ae. Aegypti populations throughout the world and discuss how variation in vector competence is correlated with overall genetic difference among populations. We describe current research into how genetic and environmental factors jointly affect distribution of vector competence in natural populations. Based on this information, we propose a population genetic model for vector competence and discuss our recent progress in testing this model. We end with a discussion of approaches being taken to identify the genes that may control flavivirus susceptibility in Ae. Aegypti

Universal primers for the amplification and sequence analysis pf actin-1 from diverse mosquito species

We report the development of universal primers for the reverse-transcription polymerase chain reaction (RT-PCR) amplification and nucleotide sequence analysis of actin cDNAs from taxonomically diverse mosquito species. Primers specific to conserved regions of the invertebrate actin-1 gene were designed after actin cDNA sequences of Anopheles gambiae, Bombyx mori, Drosophila melanogaster, and Caenorhabditis elegans. The efficacy of these primers was determined by RT-PCR with the use of total RNA from mosquitoes belonging to 30 species and 8 genera (Aedes, Anopheles, Culex, Deinocerites, Mansonia, Psorophora, Toxorhynchites, and Wyeomyia). The RT-PCR products were sequenced, and sequence data were used to design additional primers. One primer pair, denoted as Act-2F (5?-ATGGTCGGYATGGGNCAGAAGGACTC-3?) and Act-8R (5?-GATTCCATACCCAGGAAG-GADGG-3?), successfully amplified an RT-PCR product of the expected size (683-nt) in all mosquito spp. tested. We propose that this primer pair can be used as an internal control to test the quality of RNA from mosquitoes collected in vector surveillance studies. These primers can also be used in molecular experiments in which the detection, amplification or silencing of a ubiquitously expressed mosquito housekeeping gene is necessary. Sequence and phylogenetic data are also presented in this report

The neovolcanic axis is a barrier to gene flow among Aedes aegypti populations in México that differ in vector competence for Dengue 2 virus.

The Neovolcanic axis (NVA) traverses Mexico at the 19th parallel and is considered to be a geographic barrier to many species. We have demonstrated that the intersection of the NVA with the coast in Veracruz state is a barrier to gene flow in Ae. aegypti. This was unexpected because the intersection of the NVA with the Pacific Coast is not a barrier to gene flow. Further studies to identify the actual mechanism(s) that is(are) contributing to the lack of gene flow will provide important information on the trafficking potential of Ae. aegypti, which will be of great value to Ae. aegypti control programs. There are significant differences in vector competence for dengue virus between mosquitoes north and south of the NVA, but the epidemiological significance of these finding remains to be determined. Future studies will determine if, for example, the genes that condition midgut infection and vector competence of Ae. aegypti populations provide biomarkers for risk of dengue transmission. Such biomarkers could be of great value to control programs in resource limited environments by allowing targeting of vector control efforts to areas at most risk for epidemic dengue and dengue hemorrhagic fever

Gonotrophic cycle estimate for Culex quinquefasciatus in Mérida, Yucatán, México

West Nile virus (WNV) has been present in the Yucatán State, México, since 2002. Culex quinquefasciatus, one of the main vectors of WNV transmission in the United States, is also common in Mexico and may be a key vector of WNV transmission to humans in the Yucatán. The aim of this study was to determine the length of the gonotrophic cycle and the survival rates of Cx. quinquefasciatus from Mérida, Yucatán, during the rainy versus the dry season. Mosquitoes were collected during 25-day periods in October (rainy season) and in April (dry season), and captured females were classified by abdominal appearance (freshly fed, late-stage fed, half gravid, and subgravid). To determine the age structure as nulliparous and parous females and to calculate the gonotrophic cycle through a time series and the mosquito survival, we used Davidson formulae. Also, vitellogenesis analysis to monitor egg maturity was conducted during both seasons. Cross-correlation data suggested a similar length of the gonotrophic cycle (4 days) in both seasons. Oogenic development required a minimum of 72 h in each season. However, survival of the mosquito population collected in the rainy season was significantly higher (0.91) with a mean temperature of 28 ± 1.57°C than was survival in the dry season (0.78) with a mean temperature of 29 ± 1.10°C. Survival, although higher during the rainy season, did not influence the length of the gonotrophic cycle of Cx. quinquefasciatus in Yucatán

VARIATION IN VECTOR COMPETENCE FOR DENGUE 2 VIRUS AMONG 24 COLLECTIONS OF AEDES AEGYPTI FROM MEXICO AND THE UNITED STATES

Aedes aegypti from 24 collections in Mexico and the United States were challenged orally with dengue 2 virus JAM1409 (DEN-2-JAM 1409). The vector competence (VC) of the populations ranged from 24% to 83%. Mosquito populations from the Yucatan exhibited greater VC than those from other areas of Mexico. The presence of absence of a midgut infection barrier (MIB) and a midgut escape barrier(MEB) was determined for mosquitoes in each ranged from 4% to 43% in the collections. The MIB and MEB were not completely independent as determined by regression analysis. Midgut infection rates were dose dependent