Host Usage in Aedes aegypti from Houston, Texas, and Phoenix, Arizona, Using Third-Generation Sequencing Blood Meal Analysis

The ecology and establishment of <i>Aedes aegypti</i> in tropical, sub-tropical, and temperate areas makes them one of the most medically relevant mosquito species. While they have been reported to be highly anthropophilic, several studies indicate a broader host range. They also reportedly take multiple blood meals between gonotrophic cycles, which makes determination of host usage difficult when using common blood meal analysis methods. In this study, we examined host usage of <i>Ae. aegypti</i> in Harris County, Texas (Houston), and Maricopa County, Arizona (Phoenix), using a nanopore-based third-generation sequencing protocol to resolve host usage and multiplicity. Using this method, approximately 80% of samples from each location with evidence of blood feeding resulted in blood meal identification, with a single host blood meal identified in about 80% of samples and approximately 20% containing evidence of multiple blood meals. Overall, we observed a wide host range with human DNA being the most prevalent followed by feline (<i>Felis catus</i>) and canine (<i>Canis lupus</i>). We also identified avian, rodent, ungulate and even ectotherm usage by <i>Ae. aegypti</i> from Maricopa County. This study demonstrates the utility of a third-generation sequencing method to identify less common hosts, like ectotherms in Arizona, and to resolve multiple feedings which remain difficult to resolve by current Sanger-based methods.