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Aedes aegypti, the primary vector of dengue virus, is well established throughout urban areas of the Southwestern US, including Tucson, AZ. Local transmission of the dengue virus, however, has not been reported in this area. Although many factors influence the distribution of the dengue virus, we hypothesize that one contributing factor is that the lifespan of female Ae. aegypti mosquitoes in the Southwestern US is too short for the virus to complete development and be transmitted to a new host. To test this we utilized two age grading techniques. First, we determined parity by analyzing ovarian tracheation and found that only 40% of Ae. aegypti females collected in Tucson, AZ were parous. The second technique determined transcript levels of an age-associated gene, Sarcoplasmic calcium-binding protein 1 (SCP-1). SCP-1 expression decreased in a predictable manner as the age of mosquitoes increased regardless of rearing conditions and reproductive status. We developed statistical models based on parity and SCP-1 expression to determine the age of individual, field collected mosquitoes within three age brackets: nonvectors (0-5 days post-emergence), unlikely vectors (6-14 days post-emergence), and potential vectors (15+ days post-emergence). The statistical models allowed us to accurately group individual wild mosquitoes into the three age brackets with high confidence. SCP-1 expression levels of individual, field collected mosquitoes were analyzed in conjunction with parity status. Based on SCP-1 transcript levels and parity data, 9% of collected mosquitoes survived more than 15 days post emergence.

Abstract Background The Aedes aegypti mosquito is a vector of several viruses including dengue, chikungunya, zika, and yellow fever. Vector surveillance and control are the primary methods used for the control and prevention of disease transmission; however, public health institutions largely rely on measures of population abundance as a trigger for initiating control activities. Previous research found evidence that at the northern edge of Ae. aegypti’s geographic range, survival, rather than abundance, is likely to be the factor limiting disease transmission. In this study, we sought to test the utility of using body size as an entomological index to surveil changes in the age structure of field-collected female Aedes aegypti. Methods We collected female Ae. aegypti mosquitoes using BG sentinel traps in three cities at the northern edge of their geographic range. Collections took place during their active season over the course of 3 years. Female wing size was measured as an estimate of body size, and reproductive status was characterized by examining ovary tracheation. Chronological age was determined by measuring transcript abundance of an age-dependent gene. These data were then tested with female abundance at each site and weather data from the estimated larval development period and adulthood (1 week prior to capture). Two sources of weather data were tested to determine which was more appropriate for evaluating impacts on mosquito physiology. All variables were then used to parameterize structural equation models to predict age. Results In comparing city-specific NOAA weather data and site-specific data from HOBO remote temperature and humidity loggers, we found that HOBO data were more tightly associated with body size. This information is useful for justifying the cost of more precise weather monitoring when studying intra-population heterogeneity of eco-physiological factors. We found that body size itself was not significantly associated with age. Of all the variables measured, we found that best fitting model for age included temperature during development, body size, female abundance, and relative humidity in the 1 week prior to capture . The strength of models improved drastically when testing one city at a time, with Hermosillo (the only study city with seasonal dengue transmission) having the best fitting model for age. Despite our finding that there was a bias in the body size of mosquitoes collected alive from the BG sentinel traps that favored large females, there was still sufficient variation in the size of females collected alive to show that inclusion of this entomological indicator improved the predictive capacity of our models. Conclusions Inclusion of body size data increased the strength of weather-based models for age. Importantly, we found that variation in age was greater within cities than between cities, suggesting that modeling of age must be made on a city-by-city basis. These results contribute to efforts to use weather forecasts to predict changes in the probability of disease transmission by mosquito vectors. Graphical abstract

As gene drive mosquito projects advance from contained laboratory testing to semi-field testing and small-scale field trials, there is a need to assess monitoring requirements to: i) assist with the effective introduction of the gene drive system at field sites, and ii) detect unintended spread of gene drive mosquitoes beyond trial sites, or resistance mechanisms and non-functional effector genes that spread within trial and intervention sites. This is of particular importance for non-localized gene drive projects, as the potential scale of intervention means that monitoring is expected to be more costly than research, development and deployment. Regarding monitoring needs for population replacement systems, lessons may be learned from experiences with Wolbachia-infected mosquitoes, and for population suppression systems, from experiences with releases of genetically sterile male mosquitoes. For population suppression systems, assessing monitoring requirements for tracking population size and detecting rare resistant alleles are priorities, while for population replacement systems, allele frequencies must be tracked, and pressing concerns include detection of gene drive alleles with non-functional effector genes, and resistance of pathogens to functional effector genes. For spread to unintended areas, open questions relate to the optimal density and placement of traps and frequency of sampling in order to detect gene drive alleles, drive-resistant alleles or non-functional effector genes while they can still be effectively managed. Invasive species management programs face similar questions, and lessons may be learned from these experiences. We explore these monitoring needs for gene drive mosquito projects progressing through the phases of pre-release, release and post-release.