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Background  Sex separation of mosquitoes at different stages is currently being attempted to ensure the successful release of male mosquitoes in novel vector control approaches. Mechanical and behavioral techniques have been tried most frequently.    Methods  Batches of ( n  = 300)  Aedes aegypti  and  Ae. albopictus  pupae were used for standard sieving (using sieves with 1.12, 1.25, 1.40 and 1.60 mm mesh sizes) and the Fay-Morlan glass plate separation methods. Male and female separation by each method was calculated. For behavioral separation, a spiked blood meal with different concentrations (0, 2, 4, 6, 8 and 10 ppm) of ivermectin and spinosad (spinosyn, 12% w/v), were provided to a batch ( n  = 300) of adult  Ae. aegypti  and  Ae. albopictus  (1:1 sex ratio) followed by observation of mortality. An additional “double feeding method” involved provision of a further blood meal after 24 h, with the same concentrations of ivermectin and spinosad as the initial feeding, followed by a 48-h observation of mortality. All experiments were repeated five times.    Results  In the standard sieving method, the percentage of males and females separated at different pore sizes differed significantly ( P  < 0.05). The majority of the male pupae were collected in the 1.12 mm pore sized sieve for both  Ae. aegypti  (73%) and  Ae. albopictus  (69%) while females were retained mainly in the sieve with the pore size of 1.25 mm. In the Fay-Morlan glass plate separation, 99.0% of the  Ae. aegypti  and 99.2% of the  Ae. albopictus  introduced male pupae could be separated, but with female contaminations of 16 and 12%, respectively. Provision of a blood meal spiked with 8 ppm of ivermectin under the “double feeding” was identified as the most effective way of achieving 100% female elimination for both  Aedes  species.    Conclusions  With 100% separation, use of a spiked blood meal is a more effective method of sex separation than the mechanical methods. Application of the spiked blood meal approach as a second separation level for sexes, after applying the Fay-Morlan glass plate method, could achieve 100% sex separation of sexes whilst allowing a reduction in the amount of toxicants required.

parasites and vectors

Use of mechanical and behavioural methods to eliminate female Aedes aegypti and Aedes albopictus for sterile insect technique and incompatible insect technique applications