Automated differentiation of mixed populations of free-flying female mosquitoes under semi-field conditions.

Great advances in automated identification systems, or 'smart traps', that differentiate insect species have been made in recent years, yet demonstrations of field-ready devices under free-flight conditions remain rare. Here, we describe the results of mixed-species identification of female mosquitoes using an advanced optoacoustic smart trap design under free-flying conditions. Point-of-capture classification was assessed using mixed populations of congeneric (Aedes albopictus and Aedes aegypti) and non-congeneric (Ae. aegypti and Anopheles stephensi) container-inhabiting species of medical importance. Culex quinquefasciatus, also common in container habitats, was included as a third species in all assessments. At the aggregate level, mixed collections of non-congeneric species (Ae. aegypti, Cx. quinquefasciatus, and An. stephensi) could be classified at accuracies exceeding 90% (% error = 3.7-7.1%). Conversely, error rates increased when analysing individual replicates (mean % error = 48.6; 95% CI 8.1-68.6) representative of daily trap captures and at the aggregate level when Ae. albopictus was released in the presence of Ae. aegypti and Cx. quinquefasciatus (% error = 7.8-31.2%). These findings highlight the many challenges yet to be overcome but also the potential operational utility of optoacoustic surveillance in low diversity settings typical of urban environments.