Rhodamine-B for the mark, release, and recapture experiments in gamma-irradiated male Aedes aegypti (Diptera: Culicidae): Persistence, dispersal, and its effect on survival.

Background and Aim: Rhodamine-B (Rh-B) marking shows a great potential for use in mark-release-recapture (MRR) studies for rear-and-release mosquito control strategies, including the radiation-based sterile insect technique. However, its applicability and evaluation in body-stain-irradiated males of Aedes aegypti have received little attention. The present study evaluated the use of Rh-B to mark gamma-irradiated male A. aegypti. Materials and Methods: Male A. aegypti were irradiated at the pupal stage at a dose of 70 Gy. After emergence, males were fed 0.1, 0.2, 0.3, or 0.4% Rh-B in 10% glucose solution for 4 days. Groups of unirradiated males that received the same feeding treatments were used as control groups. We evaluated the persistence of Rh-B and the longevity of males after Rh-B feeding. Furthermore, the use of Rh-B in irradiated A. aegypti for MRR experiments was evaluated at an urban site. Results: No difference was observed in the Rh-B persistence among all concentrations at the 24-h postmarking period ranging from 91.25 ± 1.61% to 96.25 ± 1.61% and from 90.00 ± 2.28% to 93.13 ± 2.77% for the unirradiated and irradiated groups, respectively. Rh-B persistence significantly decreased over time, and persistence was significantly longer with increased concentrations in both the unirradiated and irradiated groups. Longevity was considerably decreased by Rh-B feeding and irradiation. However, no significant difference in longevity was found among males fed various concentrations of Rh-B. Through MRR experiments, irradiated-Rh-B marked males were mostly detected within a radius of 20 m and 40 m from the center-release point. The mean distance traveled of the released males from the three MRR events was calculated to be 42.6 m. Conclusion: This study confirms that Rh-B body marking through sugar feeding is applicable for irradiated male A. aegypti, with only a slight effect on longevity. Furthermore, considering the significant reduction in persistence over time, further study is needed to assess the impact of this reduction on the calculation of field biological parameters resulting from MRR experiments.

Assessment of Compaction, Temperature, and Duration Factors for Packaging and Transporting of Sterile Male Aedes aegypti (Diptera: Culicidae) under Laboratory Conditions.

Optimized conditions for the packaging and transportation of sterile males are crucial factors in successful SIT programs against mosquito vector-borne diseases. The factors influencing the quality of sterile males in packages during transportation need to be assessed to develop standard protocols. This study was aimed to investigate the impact of compaction, temperature, and duration factors during packaging and transportation on the quality of gamma-sterilized male Ae. aegypti. Aedes aegypti males were sterilized at a dose of 70 Gy, compacted into Falcon tubes with densities of 40, 80, and 120 males/2 mL; and then exposed to temperatures of 7, 14, 21, and 28 °C. Each temperature setup was held for a duration of 3, 6, 12, 24, and 48 h at a 60 rpm constant vibration to simulate transportation. The parameters of mortality, flight ability, induced sterility, and longevity were investigated. Results showed that increases in density, temperature, and duration significantly increased mortality and reduced flight ability and longevity, but none of the factors significantly affected induced sterility. With a mortality rate of less than 20%, an escaping rate of more than 70%, considerable longevity, and the most negligible effect on induced sterility (approximately 98%), a temperature of 7 °C and a compaction density of 80 males/2 mL were shown to be optimized conditions for short-term transportation (no more than 24 h) with the minimum adverse effects compared with other condition setups.

Assessment of packing density and transportation effect on sterilized pupae and adult Aedes aegypti (Diptera: Culicidae) in non-chilled conditions.

Long-distance transportation from a radiation facility to a target site potentially affects the quantity and quality of sterile male mosquitoes. This study tested the effects of multi-hour land transportation on the survival, longevity, and mating performance of gamma-rays sterilized adult and pupal male mosquitoes at different densities in non-chilled condition. The results demonstrated that mortality rate, longevity, induced sterility (IS) level, and mating competitiveness (C index) were significantly affected by life stage, transportation treatment, and density. Transportation was detrimental to the survival and longevity of the adults, and transporting pupae was restricted by the overcrowding effect; particularly, those packing density of 200 pupae. The longevity of transported mosquitoes were 1-5 days shorter than that of non-transported mosquitoes regardless of packing density. The irradiated transported adult males exhibited an equal IS and C index to their non-transported counterparts. Although there was no evidence suggested an association between low mating competitiveness and packing density in the transported adults, the mating competitiveness of adult mosquitoes decreased with increased packing density. Additionally, the effects of transportation and packing density on the mating ability of transported pupal males were also notable. The results indicate the factors of packing density and life stages in transporting sterile males under non-chilled conditions should be taken into account in formulating the procedure in SIT operation.

Ovitrap surveillance of dengue vector mosquitoes in Bandung City, West Java Province, Indonesia.

Larval surveillance is the central approach for monitoring dengue vector populations in Indonesia. However, traditional larval indices are ineffective for measuring mosquito population dynamics and predicting the dengue transmission risk. We conducted a 14-month ovitrap surveillance. Eggs and immature mosquitoes were collected on a weekly basis from an urban village of Bandung, namely Sekejati. Ovitrap-related indices, namely positive house index (PHI), ovitrap index (OI), and ovitrap density index (ODI), were generated and correlated with environmental variables, housing type (terraced or high-density housing), ovitrap placement location (indoor or outdoor; household or public place), and local dengue cases. Our results demonstrated that Aedes aegypti was significantly predominant compared with Aedes albopictus at each housing type and ovitrap placement location. Ovitrap placement locations and rainfall were the major factors contributing to variations in PHI, OI, and ODI, whereas the influences of housing type and temperature were subtle. Indoor site values were significantly positively correlated to outdoor sites' values for both OI and ODI. OI and ODI values from households were best predicted with those from public places at 1- and 0-week lags, respectively. Weekly rainfall values at 4- and 3-week lags were the best predictors of OI and ODI for households and public places, respectively. Monthly mean PHI, OI, and ODI were significantly associated with local dengue cases. In conclusion, ovitrap may be an effective tool for monitoring the population dynamics of Aedes mosquitoes, predicting dengue outbreaks, and serving as an early indicator to initiate environmental clean-up. Ovitrap surveillance is easy for surveyors if they are tasked with a certain number of ovitraps at a designated area, unlike the existing larval surveillance methodology, which entails identifying potential breeding sites largely at the surveyors' discretion. Ovitrap surveillance may reduce the influence of individual effort in larval surveillance that likely causes inconsistency in results.

Effects of larval diets and temperature regimes on life history traits, energy reserves and temperature tolerance of male Aedes aegypti (Diptera: Culicidae): optimizing rearing techniques for the sterile insect programmes.

BACKGROUND: Producing high quality sterile males is vital in Aedes aegypti rear-and-release birth control strategies. Larval diets, rearing temperatures, and their interactions determine the accumulation rates of essential nutrients in larvae, but these factors have been understudied in relation to mass-rearing techniques for producing eminent males. METHODS: We compared the effects of two larval diets, a cereal-legume-based diet (Khan's diet) and a standard larval diet developed in the FAO/IAEA Insect Pest Control Laboratory (IAEA 2 diet). Diets were tested at selected temperatures for both larval and male adult life history traits, adult extreme temperature tolerance, and mating capacity relative to energy reserves of reared male adult Ae. aegypti. RESULTS: Khan's diet resulted in shorter immature development time at each test temperature (except for 25 °C) than an IAEA 2 diet. Larvae reared at 28 °C and 32 °C with Khan's diet demonstrated low pupation rates (c.80%). We accounted for these phenomena as secondary sex ratio manipulation, because a higher proportion of male adults emerged at 28 °C and 32 °C than that for the IAEA 2 diet. In general, the pupal development time shortened as temperature increased, resulting in higher teneral energy reserves in male mosquitoes. High energy reserves allowed male mosquitoes reared with Khan's diet to have higher adult longevity (5-6 days longer when sugar-fed and 2-3 days longer when water-fed) and tolerance of heat stress than those fed on the IAEA 2 diet. The IAEA 2 diet produced larger male mosquitoes than Khan's diet did: mosquitoes fed on Khan's diet were 1.03-1.05 times smaller than those fed on the IAEA 2 diet at 28 °C and 32 °C. No evidence indicated reduced mating capacity for small mosquitoes fed on Khan's diet. CONCLUSIONS: Larvae reared at 28 °C and 32 °C with Khan's diet were characterized by shorter immature development time compared with those fed on the IAEA 2 diet. Adult mosquitoes produced from that larval rearing condition exhibited a significant male bias, long lifespan, and better endurance against extreme temperatures relative to energy reserves. Thus, the larval diet at rearing temperature of 28 °C and 32 °C optimized rearing techniques for the sterile insect programmes. However, mating competitiveness and flight performance of adult males require further investigation.