Optimizing larval mass-rearing techniques for

The quantity and quality of laboratory-reared insects are pivotal for the success of any sterile male-release program. Optimizing larval mass-rearing methods to enhance both production and quality in Aedes mosquitoes is essential to meet the growing demand from FAO/IAEA Member States for the sterile insect technique (SIT) as a component of area-wide integrated pest management to control or suppress disease vectors. This study was designed to identify the most effective feeding regime and schedule that maximize pupae production with a single tilt/sorting event and to evaluate an alternative larval-rearing unit. The results demonstrated that ingredient particle size, mosquito strain and feeding regime significantly influenced insect production and quality, underscoring the critical need to account for these factors in mass-rearing operations. A daily feeding regime of 0.17, 0.33, 0.67, 0.67 and 0.5 mg per larva was identified as optimal for both species (Ae. aegypti and Ae. albopictus) achieving up to 80 ± 2.5% male pupae recovery rate when sorted 48 h after the onset of pupation. Production outcomes were not compromised with the exclusion of feeding on Days 2 and 3. Furthermore, under the conditions of this study, the Wolbaki rack (Model WBK-P0003-V2) was shown to be sufficient for mass-rearing Aedes mosquitoes. Finally, a 4-day feeding regime was implemented in a field program on Reunion island, yielding similar pupae recovery rates and contamination as the reference regime, a significant step toward improving cost-efficiency and scaling-up the program. These findings provide valuable information for refining standard operating procedures (SOPs) for mass-rearing, thereby enhancing the efficiency and scalability of SIT programs.