Sterile insect technique (SIT) is a promising, environmentally friendly alternative to the use of pesticides for insect pest control. However, implementing SIT with Aedes aegypti (Linnaeus) mosquitoes presents unique challenges. For example, during transport from the rearing facility to the release site and during the actual release in the field, damage to male mosquitoes should be minimized to preserve their reproductive competitiveness. The short flight range of male Ae. aegypti requires elaborate release strategies such as release via Unmanned Aircraft Systems, more commonly referred to as drones. Two key parameters during transport and release are storage temperature and compaction rate. We performed a set of laboratory experiments to identify the optimal temperatures and compaction rates for storage and transport of male Ae. aegypti. We then conducted shipping experiments to test our laboratory-derived results in a ‘real-life’ setting. The laboratory results indicate that male Ae. aegypti can survive at a broad range of storage temperatures ranging from 7 to 28°C, but storage time should not exceed 24 h. Male survival was high at all compaction rates we tested with a low at 40 males/cm3. Interestingly, results from our ‘real-life’ shipping experiment showed that high compaction rates were beneficial to survival. This study advances key understudied aspects of the practicalities of moving lab-reared insects into the field and lies the foundation for further studies on the effect of transport conditions on male reproductive fitness.

Background: the mosquito Aedes aegypti (Ae. aegypti) is an important vector of arboviruses, including Zika, Dengue, and Chikungunya. The dietary requirements of larval Ae. aegypti are not well understood and likely impact developmental and physiological parameters knowledge of which could be important for vector control. This study examines the effects nutrition has on growth and development of larval Ae. aegypti of laboratory-reared Rockefeller strain mosquitoes. Methods: mosquito larvae were split into five feeding groups with diets providing different ratios of protein and carbohydrates. Each group received autolyzed Brewer’s yeast (AY - high-protein), and/or rice flour (RF—high-carbohydrate). The groups were monitored to record larval developmental times, adult sizes and nutritional stores. Results: the 100% AY group failed to pupate, suggesting the AY alone is either lacking in critical nutrients or is toxic at higher concentrations. The 100% RF group resulted in the smallest adults that took the longest time to reach pupation. Of the remaining groups, the 25% AY/75% RF (Med–low) diet yielded adult mosquitoes with highest average weight, wing length, and lipid stores relative to the other diets. Conclusions: the dietary requirements for development, body size, and nutrient stores of Ae. aegypti mosquitoes appear to be dependent on a relatively low but essential proportion of dietary protein to carbohydrates to achieve optimal developmental outcomes.

Tsetse flies (genus Glossina), the sole vectors of African trypanosomiasis, are distinct from most other insects, due to dramatic morphological and physiological adaptations required to support their unique biology. These adaptations are driven by demands associated with obligate hematophagy and viviparous reproduction. Obligate viviparity entails intrauterine larval development and the provision of maternal nutrients for the developing larvae. The reduced reproductive capacity/rate associated with this biology results in increased inter- and intra-sexual competition. Here, we use phase contrast microcomputed tomography (pcMicroCT) to analyze morphological adaptations associated with viviparous biology. These include (1) modifications facilitating abdominal distention required during blood feeding and pregnancy, (2) abdominal and uterine musculature adaptations for gestation and parturition of developed larvae, (3) reduced ovarian structure and capacity, (4) structural features of the male-derived spermatophore optimizing semen/sperm delivery and inhibition of insemination by competing males and (5) structural features of the milk gland facilitating nutrient incorporation and transfer into the uterus. Three-dimensional analysis of these features provides unprecedented opportunities for examination and discovery of internal morphological features not possible with traditional microscopy techniques and provides new opportunities for comparative morphological analyses over time and between species.

Tsetse flies (genus Glossina), the sole vectors of African trypanosomiasis, are distinct from other disease vectors, and most other insects, due to dramatic evolutionary adaptations required to support their unique life history. These morphological and physiological adaptations are driven by demands associated with their strict dietary and reproductive requirements. Tsetse reproduce by obligate viviparity which entails obligate intrauterine larval development and provisioning of nutrients for the developing larvae. Viviparous reproduction reduces reproductive capacity/rate which also drives increased inter-and intra-sexual competition. This work describes three-dimensional (3D) analysis of viviparity associated morphological adaptations of tsetse female reproductive tract as well as that of male seminal secretions by phase contrast microcomputed tomography (pcMicroCT). Structural features of note include abdominal modifications facilitating the extreme abdominal distention required during blood feeding and pregnancy; abdominal and uterine musculature required for parturition of developed larvae; reduction of ovarian structure and capacity; structural features of the male seminal spermatophore that enhance sperm delivery and inhibition of insemination by competing males; uterine morphological features facilitating expansion and contraction before, during and after pregnancy; analysis of structural optimizations of the milk gland facilitating nutrient incorporation and transfer into the uterus. The use of pcMicroCT provides unprecedented opportunities for examination and discovery of internal morphological features not possible with traditional microscopy techniques and new opportunities for comparative morphological analyses over time and between species.3