A Three-Pronged Approach to Studying Sublethal Insecticide Doses: Characterising Mosquito Fitness, Mosquito Biting Behaviour, and Human/Environmental Health Risks.

Worldwide, pyrethroids are one of the most widely used insecticide classes. In addition to serving as personal protection products, they are also a key line of defence in integrated vector management programmes. Many studies have assessed the effects of sublethal pyrethroid doses on mosquito fitness and behaviour. However, much remains unknown about the biological, physiological, demographic, and behavioural effects on individual mosquitoes or mosquito populations when exposure occurs via spatial treatments. Here, females and males of two laboratory-reared mosquito species, Culex pipiens and Aedes albopictus, were exposed to five different treatments: three doses of the pyrethroid prallethrin, as well as an untreated and a negative control. The effects of each treatment on mosquito species, sex, adult mortality, fertility, F1 population size, and biting behaviour were also evaluated. To compare knockdown and mortality among treatments, Mantel-Cox log-rank tests were used. The results showed that sublethal doses reduced mosquito survival, influencing population size in the next generation. They also provided 100% protection to human hosts and presented relatively low risks to human and environmental health. These findings emphasise the need for additional studies that assess the benefits of using sublethal doses as part of mosquito management strategies.

Two New Alternatives to the Conventional Arm-in-Cage Test for Assessing Topical Repellents.

European guidelines for testing attractant and repellent efficacy (i.e., Product type 19 [PT19]) have been in revision since 2017. A key topic of discussion is the current approach to evaluating topical repellents. The European Chemical Agency has stated field testing should be avoided because of mosquito-borne disease risks. However, the most common laboratory method, the arm-in-cage (AIC) test, may limit the reliable extrapolation of lab results to field conditions. This study's main goal was to assess alternative laboratory methods for evaluating topical mosquito repellents that use mosquito landing rates more representative of those in the field. The study took place at three European testing labs using 30 study participants per test and the mosquito, Aedes albopictus (Skuse, 1894, Diptera: Culicidae). In phase 1, a conventional AIC test and a sleeved AIC test were performed. Respectively, the arm area exposed was 600 and 100 cm2, and cage volume was 0.040 and 0.064 m3. Mosquito density was the same for both: 1 female/840 cm3. In phase 2, room-based testing (40 ± 5 mosquitoes in 25-30 m3) was used as a proxy for field testing. The mosquito repellent employed was 15% N,N-diethyl-m-toluamide in ethanol at two doses: 1 and 0.5 g/600 cm2. The protection times measured at each laboratory were analyzed both separately and together using nonparametric (Kruskal-Wallis) test. The two alternatives methods showed to be potential alternatives to the current AIC method recreated field mosquito landing rates and achieved reproducible protection times across laboratories.

From the Field to the Laboratory: Quantifying Outdoor Mosquito Landing Rate to Better Evaluate Topical Repellents.

Vector-borne diseases are a worldwide threat to human health. Often, no vaccines or treatments exist. Thus, personal protection products play an essential role in limiting transmission. The World Health Organization (WHO) arm-in-cage (AIC) test is the most common method for evaluating the efficacy of topical repellents, but it remains unclear whether AIC testing conditions recreate the mosquito landing rates in the field. This study aimed to estimate the landing rate outdoors, in an area of Europe highly infested with the Asian tiger mosquito (Aedes albopictus (Skuse, 1894, Diptera: Culididae)), and to determine how to replicate this rate in the laboratory. To assess the landing rate in the field, 16 individuals were exposed to mosquitoes in a highly infested region of Italy. These field results were then compared to results obtained in the laboratory: 1) in a 30 m3 room where nine volunteers were exposed to different mosquito abundances (ranges: 15-20, 25-30, and 45-50) and 2) in a 0.064 m3 AIC test cage where 10 individuals exposed their arms to 200 mosquitoes (as per WHO requirements). The highest mosquito landing rate in the field was 26.8 landings/min. In the room test, a similar landing rate was achieved using 15-20 mosquitoes (density: 0.50-0.66 mosquitoes/m3) and an exposure time of 3 min. In the AIC test using 200 mosquitoes (density: 3,125 mosquitoes/m3), the landing rate was 229 ± 48 landings/min. This study provides useful reference values that can be employed to design new evaluation standards for topical repellents that better simulate field conditions.