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The sterile insect technique is based on the overflooding of a target population with released sterile males inducing sterility in the wild female population. It has proven to be effective against several insect pest species of agricultural and veterinary importance and is under development for Aedes mosquitoes. Here, we show that the release of sterile males at high sterile male to wild female ratios may also impact the target female population through mating harassment. Under laboratory conditions, male to female ratios above 50 to 1 reduce the longevity of female Aedes mosquitoes by reducing their feeding success. Under controlled conditions, blood uptake of females from an artificial host or from a mouse and biting rates on humans are also reduced. Finally, in a field trial conducted in a 1.17 ha area in China, the female biting rate is reduced by 80%, concurrent to a reduction of female mosquito density of 40% due to the swarming of males around humans attempting to mate with the female mosquitoes. This suggests that the sterile insect technique does not only suppress mosquito vector populations through the induction of sterility, but may also reduce disease transmission due to increased female mortality and lower host contact. The sterile insect technique involves the introduction of sterile males to induce sterility in females and reduce population size. Here, the authors show that sterile males may also reduce female density and inhibit female blood feeding through mating harassment.

The sterile insect technique is a promising environmentally friendly method for mosquito control. This technique involves releasing laboratory-produced sterile males into a target field site, and its effectiveness may be affected by the extent of adult mosquito predation. Sterile males undergo several treatments. Therefore, it is vital to understand which treatments are essential in minimizing risks to predation once released. The present study investigates the predation propensity of four mantis species ( Phyllocrania paradoxa, Hymenopus coronatus, Blepharopsis mendica, Deroplatys desiccata ) and two gecko species ( Phelsuma standingi, P. laticauda) on adult Aedes aegypti , Ae. albopictus and Anopheles arabiensis mosquitoes in a laboratory setting. First, any inherent predation preferences regarding mosquito species and sex were evaluated. Subsequently, the effects of chilling, marking, and irradiation, on predation rates were assessed. The selected predators effectively preyed on all mosquito species regardless of the treatment. Predation propensity varied over days for the same individuals and between predator individuals. Overall, there was no impact of laboratory treatments of sterile males on the relative risk of predation by the test predators, unless purposely exposed to double the required sterilizing irradiation dose. Further investigations on standardized predation trials may lead to additional quality control tools for irradiated mosquitoes.

Reproductive sterility is the basis of the sterile insect technique (SIT) and essential for its success in the field. Numerous factors that influence dose–response in insects have been identified. However, historically the radiation dose administered has been considered a constant. Efforts aiming to standardize protocols for mosquito irradiation found that, despite carefully controlling many variable factors, there was still an unknown element responsible for differences in expected sterility levels of insects irradiated with the same dose and handling protocols. Thus, together with previous inconclusive investigations, the question arose whether dose really equals dose in terms of biological response, no matter the rate at which the dose is administered. Interestingly, the dose rate effects studied in human nuclear medicine indicated that dose rate could alter dose–response in mammalian cells. Here, we conducted experiments to better understand the interaction of dose and dose rate to assess the effects in irradiated mosquitoes. Our findings suggest that not only does dose rate alter irradiation-induced effects, but that the interaction is not linear and may change with dose. We speculate that the recombination of reactive oxygen species (ROS) in treatments with moderate to high dose rates might minimize indirect radiation-induced effects in mosquitoes and decrease sterility levels, unless dose along with its direct effects is increased. Together with further studies to identify an optimum match of dose and dose rate, these results could assist in the development of improved methods for the production of high-quality sterile mosquitoes to enhance the efficiency of SIT programs.

This study evaluated the performance of three packing methods for long distance transport of sterile adult Aedes albopictus males, considering the specific effects of pupal vs. adult irradiation and two compaction methods. The results indicated that while transportation significantly impacted the quality of sterile males, all the three packing methods (IAEA V.1.0, IAEA V.2.0, and CAA) tested under simulated transportation durations of 25–48 h resulted in average mortality rates below 12%, with male escape rates ranging between 30 and 50% after a 24-h recovery period. The transported males were able to survive for about 2 days under stress conditions. Additionally, adult irradiation led to higher male survival and escape rates than pupal irradiation, for both before and after transportation. The study also found that stacked cups were more effective as compaction method than the square plastic boxes, despite having a reduced maximum transport capacity. Providing sterile males with 10% sugar for a 24-h recovery period after transportation, also played a role in enhancing escape rate and survival. Although Ae. albopictus prove to be more sensitive to packaging and shipping procedures compared to Aedes aegypti , this study provides valuable insights for developing and improving handling packing and shipping guidelines.

Balancing process efficiency and adult sterile male biological quality is one of the challenges in the success of the sterile insect technique (SIT) against insect pest populations. For the SIT against mosquitoes, many stress factors need to be taken into consideration when producing sterile males that require high biological quality to remain competitive once released in the field. Pressures of mass rearing, sex sorting, irradiation treatments, packing, transport and release including handling procedures for each step, add to the overall stress budget of the sterile male post-release. Optimizing the irradiation step to achieve maximum sterility while keeping off-target somatic damage to a minimum can significantly improve male mating competitiveness. It is therefore worth examining various protocols that have been found to be effective in other insect species, such as dose fractionation. A fully sterilizing dose of 70 Gy was administered to Aedes aegypti males as one acute dose or fractionated into either two equal doses of 35 Gy, or one low dose of 10 Gy followed by a second dose of 60 Gy. The two doses were separated by either 1- or 2-day intervals. Longevity, flight ability, and mating competitiveness tests were performed to identify beneficial effects of the various treatments. Positive effects of fractionating dose were seen in terms of male longevity and mating competitiveness. Although applying split doses generally improved male quality parameters, the benefits may not outweigh the added labor in SIT programmes for the management of mosquito vectors. Équilibrer l’efficacité du processus et la qualité biologique des mâles adultes stériles est l’un des défis du succès de la technique des insectes stériles (TIS) contre les populations d’insectes nuisibles. Pour la TIS contre les moustiques, de nombreux facteurs de stress sont à prendre en compte lors de la production de mâles stériles qui nécessitent une haute qualité biologique pour rester compétitifs une fois relâchés au champ. Les pressions de l’élevage en masse, du triage par sexe, des traitements d’irradiation, de l’emballage, du transport et de la libération, y compris les procédures de manipulation pour chaque étape, s’ajoutent au budget de stress global du mâle stérile après la libération. L’optimisation de l’étape d’irradiation pour atteindre une stérilité maximale tout en minimisant les dommages somatiques hors cible peut améliorer considérablement la compétitivité de l’accouplement des mâles et il est donc important d’examiner divers protocoles qui se sont révélés efficaces chez d’autres espèces d’insectes, comme le fractionnement de dose. Une dose entièrement stérilisante de 70 Gy a été administrée aux mâles Aedes aegypti en une dose unique ou fractionnée en deux doses égales de 35 Gy, ou une faible dose de 10 Gy suivie d’une seconde dose de 60 Gy. Les deux doses étaient séparées par des intervalles de 1 ou 2 jours. Des tests de longévité, d’aptitude au vol et de compétitivité à l’accouplement ont été réalisés pour identifier les effets bénéfiques des différents traitements. Des effets positifs de la dose de fractionnement ont été observés en termes de longévité des mâles et de compétitivité à l’accouplement. Bien que l’application de doses fractionnées améliore généralement les paramètres de qualité des mâles, les avantages peuvent ne pas compenser le travail supplémentaire dans les programmes TIS pour la gestion des moustiques vecteurs.

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 .

Background The sterile insect technique (SIT) for use against mosquitoes consists of several steps including the production of the target species in large numbers, the separation of males and females, the sterilization of the males, and the packing, transport and release of the sterile males at the target site. The sterility of the males is the basis of the technique; for this, efficient and standardized irradiation methods are needed to ensure that the required level of sterility is reliably and reproducibly achieved. While several reports have found that certain biological factors, handling methods and varying irradiation procedures can alter the level of induced sterility in insects, few studies exist in which the methodologies are adequately described and discussed for the reproductive sterilization of mosquitoes. Numerous irradiation studies on mosquito pupae have resulted in varying levels of sterility. Therefore, we initiated a series of small-scale experiments to first investigate variable parameters that may influence dose-response in mosquito pupae, and secondly, identify those factors that potentially have a significantly large effect and need further attention. Methods In this study, we compiled the results of a series of experiments investigating variable parameters such as pupal age ( Aedes aegypti ), pupal size ( Ae. aegypti ), geographical origin of mosquito strains ( Ae. aegypti and Ae. albopictus ), exposure methods (in wet versus dry conditions, Ae. albopictus ) and subsequently in low versus high oxygen environments [submerged in water (low O 2 (< 5 %)] and in air [high O 2 (~ 21 %)] on the radiosensitivity of male pupae ( Ae. aegypti , Ae. albopictus and Anopheles arabiensis ). Results Results indicate that radiosensitvity of Ae. aegypti decreases with increasing pupal age (99% induced sterility in youngest pupae, compared to 93% in oldest pupae), but does not change with differences in pupal size ( P  = 0.94). Differing geographical origin of the same mosquito species did not result in variations in radiosensitivity in Ae. aegypti pupae [Brazil, Indonesia, France (La Reunion), Thailand] or Ae. albopictus [Italy, France (La Reunion)]. Differences in induced sterility were seen following irradiation of pupae that were in wet versus dry conditions, which led to further tests showing significant radioprotective effects of oxygen depletion during irradiation procedures in three tested mosquito species, as seen in other insects. Conclusions These findings infer the necessity to further evaluate significant factors and reassess dose-response for mosquitoes with controlled variables to be able to formulate protocols to achieve reliable and reproducible levels of sterility for application in the frame of the SIT.

The Joint FAO/IAEA Centre (formerly called Division) of Nuclear Techniques in Food and Agriculture was established in 1964 and its accompanying laboratories in 1961. One of its subprograms deals with insect pest control, and has the mandate to develop and implement the sterile insect technique (SIT) for selected key insect pests, with the goal of reducing the use of insecticides, reducing animal and crop losses, protecting the environment, facilitating international trade in agricultural commodities and improving human health. Since its inception, the Insect Pest Control Laboratory (IPCL) (formerly named Entomology Unit) has been implementing research in relation to the development of the SIT package for insect pests of crops, livestock and human health. This paper provides a review of research carried out between 2010 and 2020 at the IPCL. Research on plant pests has focused on the development of genetic sexing strains, characterizing and assessing the performance of these strains (e.g., Ceratitis capitata), elucidation of the taxonomic status of several members of the Bactrocera dorsalis and Anastrepha fraterculus complexes, the use of microbiota as probiotics, genomics, supplements to improve the performance of the reared insects, and the development of the SIT package for fruit fly species such as Bactrocera oleae and Drosophila suzukii. Research on livestock pests has focused on colony maintenance and establishment, tsetse symbionts and pathogens, sex separation, morphology, sterile male quality, radiation biology, mating behavior and transportation and release systems. Research with human disease vectors has focused on the development of genetic sexing strains (Anopheles arabiensis, Aedes aegypti and Aedes albopictus), the development of a more cost-effective larvae and adult rearing system, assessing various aspects of radiation biology, characterizing symbionts and pathogens, studying mating behavior and the development of quality control procedures, and handling and release methods. During the review period, 13 coordinated research projects (CRPs) were completed and six are still being implemented. At the end of each CRP, the results were published in a special issue of a peer-reviewed journal. The review concludes with an overview of future challenges, such as the need to adhere to a phased conditional approach for the implementation of operational SIT programs, the need to make the SIT more cost effective, to respond with demand driven research to solve the problems faced by the operational SIT programs and the use of the SIT to address a multitude of exotic species that are being introduced, due to globalization, and established in areas where they could not survive before, due to climate change.

The production of a large number of mosquitoes of high biological qualities and reliable sex sorting before release are key challenges when applying the sterile insect technique as part of an area-wide integrated pest management approach. There is a need to fully evaluate the production capacity of the equipment developed in order to plan and maintain a daily production level for large-scale operational release activities. This study aimed to evaluate the potential use of the FAO/IAEA larval rearing unit for Aedes aegypti and the subsequent female contamination rate after sex sorting with a Fay–Morlan glass separator. Trays from each rack were tilted and their contents sorted either for each individual tray or after mixing the content of all trays from the rack. The pupal production and the female contamination rate were estimated with respect to day of collection, position of the tray, type of pupae collection, and sorting operator. Results showed significant daily variability of pupal production and female contamination rate, with a high male pupal production level achieved on the second day of collection and estimated female contamination of male pupae reached around 1%. Neither tray position nor type of pupae collection affected the pupal production and female contamination rate. However, the operator had a significant effect on the female contamination rate. These results highlight the need to optimize pupal production at early days of collection and to develop a more effective and automated method of sex separation. La production d’un grand nombre de moustiques de haute qualité biologique et le tri des sexes avant les lâchers sont des défis clés lors de l’application de la technique des insectes stériles, dans le cadre d’une approche de lutte intégrée contre les ravageurs à l’échelle d’une zone. Il est nécessaire d’évaluer pleinement la capacité de production des équipements développés afin de planifier et de maintenir un niveau de production quotidien pour les activités de libération opérationnelle à grande échelle. Cette étude visait à évaluer l’utilisation potentielle de l’unité d’élevage larvaire FAO/AIEA pour Aedes aegypti et le taux de contamination par des femelles après le tri sexuel avec un séparateur en verre Fay–Morlan. Les plateaux de chaque rack ont été inclinés et leur contenu trié soit pour chaque plateau, soit après avoir mélangé le contenu de tous les plateaux du rack. La production de pupes et le taux de contamination par des femelles ont été estimés en fonction du jour de collecte, de la position du bac, du type de collecte des pupes et de l’opérateur du tri. Les résultats ont montré une variabilité quotidienne significative de la production de pupes et du taux de contamination par des femelles, avec un niveau élevé de production de pupes mâles atteint le deuxième jour de collecte et la contamination estimée des pupes mâles par des femelles a atteint environ 1 %. Ni la position du plateau ni le type de collecte des pupes n’ont affecté la production de pupes et le taux de contamination par des femelles. Cependant, l’opérateur avait un effet significatif sur le taux de contamination par les femelles. Ces résultats mettent en évidence la nécessité d’optimiser la production des pupes dès les premiers jours de la collecte et de développer une méthode de séparation des sexes plus efficace et automatisée.

Background South Africa has set a mandate to eliminate local malaria transmission by 2023. In pursuit of this objective a Sterile Insect Technique programme targeting the main vector Anopheles arabiensis is currently under development. Significant progress has been made towards operationalizing the technology. However, one of the main limitations being faced is the absence of an efficient genetic sexing system. This study is an assessment of an An. arabiensis (AY-2) strain carrying the full Y chromosome from Anopheles gambiae , including a transgenic red fluorescent marker, being introgressed into a South African genetic background as a potential tool for a reliable sexing system. Methods Adult, virgin males from the An. arabiensis AY-2 strain were outcrossed to virgin females from the South African, Kwazulu-Natal An. arabiensis (KWAG strain) over three generations. Anopheles arabiensis AY-2 fluorescent males were sorted as first instar larvae (L1) using the Complex Object Parametric Analyzer and Sorter (COPAS) and later screened as pupae to verify the sex. Life history traits of the novel hybrid KWAG-AY2 strain were compared to the original fluorescent AY-2 strain, the South African wild-type KWAG strain and a standard laboratory An. arabiensis (Dongola reference strain). Results The genetic stability of the sex-linked fluorescent marker and the integrity and high level of sexing efficiency of the system were confirmed. No recombination events in respect to the fluorescent marker were detected over three rounds of introgression crosses. KWAG-AY2 had higher hatch rates and survival of L1 to pupae and L1 to adult than the founding strains. AY-2 showed faster development time of immature stages and larger adult body size, but lower larval survival rates. Adult KWAG males had significantly higher survival rates. There was no significant difference between the strains in fecundity and proportion of males. KWAG-AY2 males performed better than reference strains in flight ability tests. Conclusion The life history traits of KWAG-AY2, its rearing efficiency under laboratory conditions, the preservation of the sex-linked fluorescence and perfect sexing efficiency after three rounds of introgression crosses, indicate that it has potential for mass rearing. The potential risks and benefits associated to the use of this strain within the Sterile Insect Technique programme in South Africa are discussed.