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The sterile insect technique (SIT) may offer a means to control the transmission of mosquito borne diseases. SIT involves the release of male insects that have been sterilized by exposure to ionizing radiation. We determined the effects of different doses of radiation on the survival and reproductive capacity of local strains of Aedes aegypti and Ae. albopictus in southern Mexico. The survival of irradiated pupae was invariably greater than 90% and did not differ significantly in either sex for either species. Irradiation had no significant adverse effects on the flight ability (capacity to fly out of a test device) of male mosquitoes, which consistently exceeded 91% in Ae. aegypti and 96% in Ae. albopictus. The average number of eggs laid per female was significantly reduced in Ae. aegypti at doses of 15 and 30 Gy and no eggs were laid by females that had been exposed to 50 Gy. Similarly, in Ae. albopictus, egg production was reduced at doses of 15 and 25 Gy and was eliminated at 35 Gy. In Ae. aegypti, fertility in males was eliminated at 70 Gy and was eliminated at 30 Gy in females, whereas in Ae. albopictus, the fertility of males that mated with untreated females was almost zero (0.1%) in the 50 Gy treatment and female fertility was eliminated at 35 Gy. Irradiation treatments resulted in reduced ovary length and fewer follicles in both species. The adult median survival time of both species was reduced by irradiation in a dose-dependent manner. However, sterilizing doses of 35 Gy and 50 Gy resulted in little reduction in survival times of males of Ae. albopictus and Ae. aegypti, respectively, indicating that these doses should be suitable for future evaluations of SIT-based control of these species. The results of the present study will be applied to studies of male sexual competitiveness and to stepwise evaluations of the sterile insect technique for population suppression of these vectors in Mexico.

The spotted-wing drosophila ( Drosophila suzukii ), a highly invasive agricultural pest, poses significant challenges to fruit production worldwide. Traditional chemical control methods are costly and raise concerns about resistance and environmental sustainability. The Heterospecific Sterile Insect Technique (h-SIT) has emerged as a promising alternative. Sterile heterospecific males ( Drosophila melanogaster ) can be used to suppress D. suzukii populations through reproductive interference, primarily mediated by post-zygotic isolation mechanisms. Although this approach ensures the absence of viable offspring from heterospecific matings, male sterilization through irradiation remains essential. It prevents unintended ecological effects from D. melanogaster proliferation in the release area and allows for safe large-scale implementation. Therefore, determining an optimal irradiation dose is critical for achieving high levels of male sterility and maintaining biological quality and mating performance. This study aimed to determine the optimal irradiation dose by assessing induced sterility in D. melanogaster males exposed to gamma ray doses ranging from 80–180 Gy. Subsequently, the longevity and the time spent by irradiated D. melanogaster males courting D. suzukii females were also assessed. Results showed a significant dose-dependent increase in induced sterility, with near-complete sterility at 180 Gy. However, longevity decreased with increasing doses, with males irradiated at 160–180 Gy showing a lifespan reduction of up to 50 days compared to controls. Regardless of the irradiation dose received, D. melanogaster males retained their courtship ability toward D. suzukii females, although males exposed to 160 Gy exhibited reduced courtship activity. These findings showed that, among the tested doses, 80 Gy was the most effective in preserving male longevity and mating performance, significantly reducing fertility, while 180 Gy induced the highest sterility. The potential lifespan and courtship behavior trade-offs warrant further evaluation. Future studies should evaluate field performance to refine the balance between sterility, longevity, and mating performance for effective D. suzukii population suppression.

Sri Lanka has experienced severe dengue epidemics in recent years, despite the extensive vector control measures taken. Therefore, it is necessary to find sustainable vector control strategies against dengue. Novel vector control tools need to be tested for the feasibility of applying them against local vectors. Sterile Insect Technique (SIT) is an increasingly popular vector control technique which has been adopted by many countries to suppress insect pest populations and is being tested for dengue vectors. In this study, SIT was developed for Aedes albopictus (Skuse, 1895), one of the 2 dengue vectors present in Sri Lanka. The optimum radiation dose for sterilizing male pupae (age 24-48 hours) using a Co 60 source was determined based on the post-irradiation pupal and adult survival in males and induced sterility in females at different doses. Further, the effect of irradiation on mating competitiveness of the selected mosquito strain was assessed under laboratory and semi-field conditions. The optimum release ratio of irradiated males to wild males was assessed in laboratory and semi-field settings. The optimum radiation dose was 50 Gy among the series of doses (25, 30, 40, 50, 60, and 70 Gy) tested. When pupae were exposed to the optimal radiation dose, 100% pupal survival, 19-day median adult survival time and 99% induced sterility resulted. A 5:1 ratio of irradiated males to non-irradiated laboratory-reared or wild males in laboratory cages resulted in induced sterility of 75% and 62%, respectively. The respective values were 74% and 61% in large semi-field cages. Fried Competitiveness Index (FCI) of irradiated males against wild males of laboratory and wild origin were 0.63 and 0.43 in laboratory cages and 0.57 and 0.55 in large semi-field cages. The males of Ae. albopictus irradiated at 50 Gy are adequately sterile and are competitive against the wild males. The release ratio of 5:1 irradiated males to wild males is a suitable ratio for the field application of SIT. The findings of the study will be important for the development of a protocol for future application of SIT for Ae. albopictus in Sri Lanka.

Novel and alternative vector control approaches using a sterile male-based release to suppress Aedes aegypti mosquito vectors have recently been tested in the field in many countries. These approaches included the sterile insect technique (SIT), incompatible insect technique (IIT), and a combination of both techniques. In this study, we conducted a series of experiments to compare the quality between radiation-based and Wolbachia-induced sterile males in terms of flight ability, sterility, mating competitiveness, survival rate, and longevity. Aedes aegypti mosquitoes irradiated at 50 Gy (SIT) and those trans-infected with wAlbB Wolbachia (IIT) were used for quality comparison. Our results showed that irradiated and Wolbachia trans-infected males were not significantly different in flight ability (p > 0.05) and both could induce sterility in wild-type females. In addition, although irradiation at 50 Gy or Wolbachia trans-infection reduced male mating competitiveness, combined irradiation and Wolbachia wAlbB trans-infection increased male competitiveness at the one-to-one ratio. Increasing the number of sterile males released could compensate for reduced competitiveness but it does not make them more competitive. Irradiation did not affect the survival and longevity of irradiated males, but it showed significant negative impacts on females (p < 0.05); while the opposite was observed in the case of Wolbachia infection, i.e., with significant increase in the survival rate of Wolbachia trans-infected males (p < 0.05), but both survival and longevity were reduced in Wolbachia trans-infected females with no significant impacts (p > 0.05). In conclusion, neither irradiation nor Wolbachia trans-infection significantly affected the quality of sterile males except their mating competitiveness; but this could compensate by increasing the number of sterile males released. Sterility could be induced by either 50 Gy irradiation or wAlbB trans-infection. Mating competitiveness results showed that a higher number of sterile males produced by irradiation need to be released in comparison to those produced by Wolbachia trans-infection. Our results should be useful for planning SIT, IIT, or a combination for Ae. aegypti vector control.

The Sterile Insect Technique (SIT) is a successful autocidal control method that uses ionizing radiation to sterilize insects. However, irradiation in normal atmospheric conditions can be damaging for males, because irradiation generates substantial biological oxidative stress that, combined with domestication and mass-rearing conditions, may reduce sterile male sexual competitiveness and quality. In this study, biological oxidative stress and antioxidant capacity were experimentally manipulated in Anastrepha suspensa using a combination of low-oxygen conditions and transgenic overexpression of mitochondrial superoxide dismutase (SOD2) to evaluate their role in the sexual behavior and quality of irradiated males. Our results showed that SOD2 overexpression enhances irradiated insect quality and improves male competitiveness in leks. However, the improvements in mating performance were modest, as normoxia-irradiated SOD2 males exhibited only a 22% improvement in mating success compared to normoxia-irradiated wild type males. Additionally, SOD2 overexpression did not synergistically improve the mating success of males irradiated in either hypoxia or severe hypoxia. Short-term hypoxic and severe-hypoxic conditioning hormesis, per se, increased antioxidant capacity and enhanced sexual competitiveness of irradiated males relative to non-irradiated males in leks. Our study provides valuable new information that antioxidant enzymes, particularly SOD2, have potential to improve the quality and lekking performance of sterile males used in SIT programs.

Background Symbiotic microbes represent a driving force of evolutionary innovation by conferring novel ecological traits to their hosts. Many insects are associated with microbial symbionts that contribute to their host’s nutrition, digestion, detoxification, reproduction, immune homeostasis, and defense. In addition, recent studies suggest a microbial involvement in chemical communication and mating behavior, which can ultimately impact reproductive isolation and, hence, speciation. Here we investigated whether a disruption of the microbiota through antibiotic treatment or irradiation affects cuticular hydrocarbon profiles, and possibly mate choice behavior in the tsetse fly, Glossina morsitans morsitans . Four independent experiments that differentially knock down the multiple bacterial symbionts of tsetse flies were conducted by subjecting tsetse flies to ampicillin, tetracycline, or gamma-irradiation and analyzing their cuticular hydrocarbon profiles in comparison to untreated controls by gas chromatography – mass spectrometry. In two of the antibiotic experiments, flies were mass-reared, while individual rearing was done for the third experiment to avoid possible chemical cross-contamination between individual flies. Results All three antibiotic experiments yielded significant effects of antibiotic treatment (particularly tetracycline) on cuticular hydrocarbon profiles in both female and male G. m. morsitans , while irradiation itself had no effect on the CHC profiles. Importantly, tetracycline treatment reduced relative amounts of 15,19,23-trimethyl-heptatriacontane, a known compound of the female contact sex pheromone, in two of the three experiments, suggesting a possible implication of microbiota disturbance on mate choice decisions. Concordantly, both female and male flies preferred non-treated over tetracycline-treated flies in direct choice assays. Conclusions While we cannot exclude the possibility that antibiotic treatment had a directly detrimental effect on fly vigor as we are unable to recolonize antibiotic treated flies with individual symbiont taxa, our results are consistent with an effect of the microbiota, particularly the obligate nutritional endosymbiont Wigglesworthia , on CHC profiles and mate choice behavior. These findings highlight the importance of considering host-microbiota interactions when studying chemical communication and mate choice in insects.

Background The success of a sterile insect technique programme to control target mosquito populations depends on producing high-quality sterile males that can compete with wild males to inseminate wild females, to consequently inhibit procreation. To succeed, the released males require high biological quality. The quality of laboratory-reared, sterile males can be affected by the rearing conditions, handling, transportation, and irradiation. Limiting the cumulative damage caused by all these processes is key to maintain sterile male performance in the field once released. The ideal life stage to irradiate Anopheles arabiensis males is still an important question for this species. This study aims to assess the dose response of irradiated males at pupal versus adult stages and use quality control (QC) parameters to assess the feasibility and possible advantages of shifting to adult irradiation in this species. Methods Laboratory reared An. arabiensis males were irradiated as late-stage pupae or as adults at varying doses to compare the radiosensitivity in both developmental stages. Males irradiated at fully sterilizing doses were further subjected to quality control tests to compare the quality of males irradiated at adult versus pupae stages. The QC parameters tested included flight ability, longevity and mating competitiveness. Results The results show that adults do not require more radiation dose to achieve the target sterility, and females are more radiosensitive than males. Longevity, flight ability and mating competitiveness were improved significantly in males irradiated as adults compared to that of males irradiated as pupae. Conclusions Given the challenges in synchronizing pupation in a mass rearing setting and achieving a homogenous age (i.e. late-stage pupae) of collected pupae in this species, as well as these findings on the overall improvement of sterile male quality in the males irradiated as adults, this report provides a strong argument for irradiation protocols to shift towards adult stage irradiation in SIT programmes against An. arabiensis .

Many reef-building corals participate in a mass-spawning event that occurs yearly on the Great Barrier Reef. This coral reproductive event is one of earth's most prominent examples of synchronised behavior, and coral reproductive success is vital to the persistence of coral reef ecosystems. Although several environmental cues have been implicated in the timing of mass spawning, the specific sensory cues that function together with endogenous clock mechanisms to ensure accurate timing of gamete release are largely unknown. Here, we show that moonlight is an important external stimulus for mass spawning synchrony and describe the potential mechanisms underlying the ability of corals to detect environmental triggers for the signaling cascades that ultimately result in gamete release. Our study increases the understanding of reproductive chronobiology in corals and strongly supports the hypothesis that coral gamete release is achieved by a complex array of potential neurohormones and light-sensing molecules. Sexual reproduction in corals is possibly the most important process for replenishing degraded coral reefs. Most corals are “broadcast spawners” that reproduce by releasing their egg cells and sperm cells into the sea water surface. To maximize their chances of reproductive success, most coral in the Great Barrier Reef – over 130 species – spawn on the same night, during a time window that is approximately 30-60 minutes long. This is the largest-scale mass spawning event of coral in the world, and is triggered by changes in sea water temperature, tides, sunrise and sunset and by the intensity of the moonlight. How corals tune their spawning behavior with the phases of the moonlight was an unanswered question for decades. Now, Kaniewska, Alon et al. have exposed the coral Acropora millepora – which makes up part of the Great Barrier Reef – to different light treatments and sampled the corals before, during and after their spawning periods. This revealed that light causes changes to gene expression and signaling processes inside cells. These changes are specifically related to the release of egg and sperm cells, and occur only on the night of spawning. Furthermore, by exposing corals to light conditions that mimic artificial urban “light pollution”, Kaniewska, Alon et al. caused a mismatch in certain cellular signaling processes that prevented the corals from spawning. Reducing the exposure of corals to artificial lighting could therefore help to protect and regenerate coral reefs. Future work will involve comparing these results with information about a coral species from another part of the world to investigate whether there is a universal mechanism used by corals to control when they spawn.

Background Aedes aegypti is a vector that transmits various viral diseases, including dengue and Zika. The radiation-based sterile insect technique (SIT) has a limited effect on mosquito control because of the difficulty in irradiating males without reducing their mating competitiveness. In this study, the insect sex pheromone heptacosane was applied to Ae. aegypti males to investigate whether it could enhance the mating competitiveness of irradiated males. Methods Heptacosane was smeared on the abdomens of Ae. aegypti males that were allowed to mate with untreated virgin females. The insemination rate was used to assess the attractiveness of heptacosane-treated males to females. The pupae were irradiated with different doses of X-rays and γ-rays, and the emergence, survival time, egg number, and hatch rate were detected to find the optimal dose of X-ray and γ-ray radiation. The males irradiated at the optimal dose were smeared with heptacosane, released in different ratios with untreated males, and mated with females. The effect of heptacosane on the mating competitiveness of irradiated mosquitoes was then evaluated by the hatch rate, induced sterility, and mating competitiveness index. Results Applying heptacosane to Ae. aegypti males significantly increased the insemination rate of females by 20%. Pupal radiation did not affect egg number but significantly reduced survival time and hatch rate. The emergence of the pupae was not affected by X-ray radiation but was affected by γ-ray radiation. Pupae exposed to 60 Gy X-rays and 40 Gy γ-rays were selected for subsequent experiments. After 60 Gy X-ray irradiation or 40 Gy γ-ray irradiation, the average hatch rate was less than 0.1%, and the average survival time was more than 15 days. Moreover, at the same release ratio, the hatch rate of the irradiated group perfumed with heptacosane was lower than that of the group without heptacosane. Conversely, the male sterility and male mating competitiveness index were significantly increased due to the use of heptacosane. Conclusions The sex pheromone heptacosane enhanced the interaction between Ae. aegypti males and females. Perfuming males irradiated by X-rays or γ-rays with heptacosane led to a significant increase in mating competitiveness. This study provided a new idea for improving the application effect of SIT. Graphical Abstract

There is currently renewed interest in assessing the feasibility of the sterile insect technique (SIT) to control African malaria vectors in designated areas. The SIT relies on the sterilization of males before mass release, with sterilization currently being achieved through the use of ionizing radiation. This paper reviews previous work on radiation sterilization of Anopheles mosquitoes. In general, the pupal stage was irradiated due to ease of handling compared to the adult stage. The dose-response curve between the induced sterility and log (dose) was shown to be sigmoid, and there was a marked species difference in radiation sensitivity. Mating competitiveness studies have generally been performed under laboratory conditions. The competitiveness of males irradiated at high doses was relatively poor, but with increasing ratios of sterile males, egg hatch could be lowered effectively. Males irradiated as pupae had a lower competitiveness compared to males irradiated as adults, but the use of partially-sterilizing doses has not been studied extensively. Methods to reduce somatic damage during the irradiation process as well as the use of other agents or techniques to induce sterility are discussed. It is concluded that the optimal radiation dose chosen for insects that are to be released during an SIT programme should ensure a balance between induced sterility of males and their field competitiveness, with competitiveness being determined under (semi-) field conditions. Self-contained 60 Co research irradiators remain the most practical irradiators but these are likely to be replaced in the future by a new generation of high output X ray irradiators.