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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.

Simple Summary The Insect Pest Control (IPC) Section and its associated laboratory (IPCL) is part of the Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture. Its mandate is to develop and implement the sterile insect technique (SIT) for selected key insect pests, and thereby to reduce the use of insecticides, to reduce animal and crop losses, to protect the environment, to facilitate international trade in agricultural commodities and to improve human health. With this aim, the IPCL 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 the research carried out between 2010 and 2020 at the IPCL. Abstract 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 sterile insect technique may prove useful for the suppression of mosquito vectors of medical importance in regions where arboviruses pose a serious public health threat. In the present study, we examined the effects of sterilizing irradiation doses across different ratios of fertile:irradiated males on the mating competitiveness of Ae. aegypti and Ae. albopictus under laboratory and field-cage conditions. For both species, the percentage of females inseminated and the number of eggs laid over two gonotrophic cycles varied significantly in mating treatments involving 1:1, 1:5, and 1:10 fertile:irradiated males compared to controls of entirely fertile or entirely irradiated males but was not generally affected by the irradiation dose. Egg hatching was negatively affected in females exposed to increasing proportions of irradiated males in both laboratory and field cages. Male competitiveness (Fried’s index) values varied from 0.19 to 0.58 in the laboratory and were between 0.09 and 1.0 in field cages, depending on th species. Competitiveness values were negatively affected by th eirradiation dose in both species under field-cage conditions, whereas in the laboratory, Ae. albopictus was sensitive to the dose but Ae. aegypti was not. In general, male competitiveness was similar across all mating regimes. Most importantly, induced egg sterility was positively correlated with the proportion of irradiated males present in the mating treatments, reaching a maximum of 88% under field-cage conditions for both Ae. aegypti and Ae. albopictus males treated with 50 and 40 Gy irradiation, respectively. These results indicate that sterile males produced at our facility are suitable and competitive enough for field pilot SIT projects and provide guidance to decide the optimal sterile:fertile ratios.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://royalsocietypublishing.org/doi/epdf/10.1098/rstb.2019.0808 En este artículo también participan: Rafael Argilés Herrero y Kostas Bourtzis. Este artículo pertenece al número que lleva por título "Novel control strategies for mosquito-borne diseases". Aedes aegypti is the primary vector of arthropod-borne viruses including dengue, chikungunya and Zika. Vector population control methods are reviving to impede disease transmission. An efficient sex separation for male-only releases is crucial for area-wide mosquito population suppression strategies. Here, we report on the construction of two genetic sexing strains using red- and white-eye colour mutations as selectable markers. Quality control analysis showed that the Red-eye genetic sexing strains (GSS) is better and more genetically stable than the White-eye GSS. The introduction of an irradiation-induced inversion (Inv35) increases genetic stability and reduces the probability of female contamination of the male release batches. Bi-weekly releases of irradiated males of both the Red-eye GSS and the Red-eye GSS/Inv35 fully suppressed target laboratory cage populations within six and nine weeks, respectively. An image analysis algorithm allowing sex determination based on eye colour identification at the pupal stage was developed. The next step is to automate the Red-eye-based genetic sexing and validate it in pilot trials prior to its integration in large-scale population suppression programmes.

Simple Summary The sterile insect technique (SIT) involves the release of massive numbers of male insects that have been sterilized by irradiation treatment during their development. Wild females that mate with sterilized males are not able to produce offspring, resulting in rapid decline in the target insect population over a large area. The success of this technique depends on the ratio of wild:sterile males achieved following male releases and the ability of sterile males to mate with wild females, i.e., their sexual competitiveness compared to fertile wild male insects. There is growing interest in applying SIT to the area-wide control of mosquitoes, such as Aedes aegypti and Aedes albopictus, that transmit important human diseases caused by dengue, chikungunya, and Zika viruses. In the present study, the sexual competitiveness of both mosquito species was affected by irradiation treatments but did not vary greatly with different ratios of fertile:sterile males in mating cages. Most importantly, the fertility of eggs was greatly reduced when more sterile males were present in mating cages, resulting in an 88% decrease in the production of fertile eggs by both species of mosquitoes in some experiments. We will use these results to perform small-scale trials in rural villages frequently affected by outbreaks of mosquito-borne diseases in southern Mexico. Abstract The sterile insect technique may prove useful for the suppression of mosquito vectors of medical importance in regions where arboviruses pose a serious public health threat. In the present study, we examined the effects of sterilizing irradiation doses across different ratios of fertile:irradiated males on the mating competitiveness of Ae. aegypti and Ae. albopictus under laboratory and field-cage conditions. For both species, the percentage of females inseminated and the number of eggs laid over two gonotrophic cycles varied significantly in mating treatments involving 1:1, 1:5, and 1:10 fertile:irradiated males compared to controls of entirely fertile or entirely irradiated males but was not generally affected by the irradiation dose. Egg hatching was negatively affected in females exposed to increasing proportions of irradiated males in both laboratory and field cages. Male competitiveness (Fried’s index) values varied from 0.19 to 0.58 in the laboratory and were between 0.09 and 1.0 in field cages, depending on th species. Competitiveness values were negatively affected by th eirradiation dose in both species under field-cage conditions, whereas in the laboratory, Ae. albopictus was sensitive to the dose but Ae. aegypti was not. In general, male competitiveness was similar across all mating regimes. Most importantly, induced egg sterility was positively correlated with the proportion of irradiated males present in the mating treatments, reaching a maximum of 88% under field-cage conditions for both Ae. aegypti and Ae. albopictus males treated with 50 and 40 Gy irradiation, respectively. These results indicate that sterile males produced at our facility are suitable and competitive enough for field pilot SIT projects and provide guidance to decide the optimal sterile:fertile ratios.

Aedes aegypti is the primary vector of arthropod-borne viruses including dengue, chikungunya and Zika. Vector population control methods are reviving to impede disease transmission. An efficient sex separation for male-only releases is crucial for area-wide mosquito population suppression strategies. Here, we report on the construction of two genetic sexing strains using red- and white-eye colour mutations as selectable markers. Quality control analysis showed that the Red-eye genetic sexing strains (GSS) is better and more genetically stable than the White-eye GSS. The introduction of an irradiation-induced inversion (Inv35) increases genetic stability and reduces the probability of female contamination of the male release batches. Bi-weekly releases of irradiated males of both the Red-eye GSS and the Red-eye GSS/Inv35 fully suppressed target laboratory cage populations within six and nine weeks, respectively. An image analysis algorithm allowing sex determination based on eye colour identification at the pupal stage was developed. The next step is to automate the Red-eye-based genetic sexing and validate it in pilot trials prior to its integration in large-scale population suppression programmes. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.

Simple Summary Aedes aegypti and Ae. albopictus are the most important mosquito vectors of dengue, chikungunya and Zika virus. There is growing interest in the control of these vectors using the sterile insect technique in which large numbers of sterilized males are released and compete with wild fertile males for mates. Females that mate with sterile males do not produce viable offspring. A study was performed on the population fluctuations of Ae. aegypti and Ae. albopictus using egg traps in two rural villages with a history of dengue in Chiapas, southern Mexico. Higher numbers of Aedes eggs were recorded in Hidalgo village compared with the village of Río Florido. In contrast, higher number of eggs were collected in areas surrounding Río Florido, compared with those around Hidalgo. Aedes aegypti was the dominant species during the dry season and at the start of the rainy season in both villages. Aedes albopictus populations were lower for most of the dry season, but increased during the rainy season and became dominant at the end of the rainy season in both villages. Aedes albopictus was also the dominant species in the zones of natural vegetation surrounding both villages. We conclude that the efficacy of a program of vector control involving the sterile insect technique could be evaluated in future studies on the isolated mosquito populations in these rural villages, in combination with habitat elimination and appropriate treatment of water sources. Abstract Indoor and outdoor ovitraps were placed in 15 randomly selected houses in two rural villages in Chiapas, southern Mexico. In addition, ovitraps were placed in five transects surrounding each village, with three traps per transect, one at the edge, one at 50 m, and another at 100 m from the edge of the village. All traps were inspected weekly. A transect with eight traps along a road between the two villages was also included. Population fluctuations of Aedes aegypti and Ae. albopictus were examined during 2016–2018 by counting egg numbers. A higher number of Aedes spp. eggs was recorded at Hidalgo village with 257,712 eggs (60.9%), of which 58.1% were present in outdoor ovitraps and 41.9% in indoor ovitraps, compared with 165,623 eggs (39.1%) collected in the village of Río Florido, 49.0% in outdoor and 51.0% in indoor ovitraps. A total of 84,047 eggs was collected from ovitraps placed along transects around Río Florido, compared to 67,542 eggs recorded from transects around Hidalgo. Fluctuations in egg counts were associated with annual variation in precipitation, with 2.3 to 3.2-fold more eggs collected from ovitraps placed in houses and 4.8 to 5.1-fold more eggs in ovitraps from the surrounding transects during the rainy season than in the dry season, respectively. Aedes aegypti was the dominant species during the dry season and at the start of the rainy season in both villages. Aedes albopictus populations were lower for most of the dry season, but increased during the rainy season and predominated at the end of the rainy season in both villages. Aedes albopictus was also the dominant species in the zones surrounding both villages. The numbers of eggs collected from intradomiciliary ovitraps were strongly correlated with the numbers of eggs in peridomiciliary ovitraps in both Río Florido (R2adj = 0.92) and Hidalgo (R2adj = 0.94), suggesting that peridomiciliary sampling could provide an accurate estimate of intradomiciliary oviposition by Aedes spp. in future studies in these villages. We conclude that the feasibility of sterile insect technique (SIT)-based program of vector control could be evaluated in the isolated Ae. aegypti populations in the rural villages of our baseline study.

BackgroundRadiation induced sterility is the basis of the Sterile Insect Technique, by which a target insect pest population is suppressed by releasing artificially reared sterile males of the pest species in overflooding numbers over a target site. In order for the sterile males to be of high biological quality, effective standard irradiation protocols are required. Following studies investigating the effects of mosquito pupae irradiation in waterversusin air, there is a need to investigate the oxy-regulatory behavior of mosquito pupae in water to better understand the consequences of irradiation in hypoxic versus normoxic conditions.MethodsPupae ofAedes aegypti,Ae. albopictus, andAnopheles arabiensiswere submerged in water inside air-tight 2 ml glass vials at a density of 100 pupae/ml and the dissolved oxygen (DO) levels in the water were measured and plotted over time. In addition, male pupae ofAe. aegypti(aged 40–44 h),Ae. albopictus(aged 40–44 h) andAn. arabiensis(aged 20–24 h) were irradiated in a gammacell220 at increasing doses in either hypoxic (water with 2content) or normoxic (in air) conditions. The males were then mated to virgin females and resulting eggs were checked for induced sterility.ResultsAll three species depleted the water of DO to levels under 0.5% within 30 minutes, withAn. arabiensisconsuming oxygen the fastest at under 10 minutes. Following irradiation, the protective effect of hypoxia was observed across species and doses (P An. arabiensis.ConclusionsThe consumption of dissolved oxygen by pupae submerged in water was significantly different between species, indicating that their oxy-regulatory capacity seems to have possibly evolved according to their preferred breeding site characteristics. This needs to be considered when sterilizing male mosquitoes at pupal stage in water. Depending on species, their DO consumption rates and their density, irradiation doses needed to achieve full sterility may vary significantly. Further assessments are required to ascertain optimal conditions in terms of ambient atmosphere during pupal irradiation to produce competitive sterile males, and temperature and density dependent effects are expected.

BackgroundThe 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.MethodsIn 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. aegyptiandAe. albopictus), exposure methods (in wetversusdry conditions,Ae. albopictus) and subsequently in lowversushigh oxygen environments [submerged in water (low O2(2(~ 21 %)] on the radiosensitivity of male pupae (Ae. aegypti,Ae. albopictusandAnopheles arabiensis).ResultsResults indicate that radiosensitvity ofAe. aegyptidecreases 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 inAe. aegyptipupae [Brazil, Indonesia, France (La Reunion), Thailand] orAe. albopictus[Italy, France (La Reunion)]. Differences in induced sterility were seen following irradiation of pupae that were in wetversusdry 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.ConclusionsThese 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.

Aedes aegypti is the principal vector of the urban arboviruses and the blood ingestion is important to produce the eggs in this species. To analyze the egg production in Ae. aegypti, researchers frequently use small cages or Drosophila vials to collect eggs from gravid females. Although it is affordable, the setup is time- and space-consuming, mainly when many mosquitoes need to be individually analyzed. This study presents an easy, cheap, and space-saving method to perform individual oviposition assays in Ae. aegypti using cell culture plates. This new method to access fecundity rate was named &ldquo, oviplate&rdquo, The oviplates are setup with 12- or 24-well plates, distilled water and filter paper and they are 78 to 88% cheaper than the traditional Drosophila vial assay, respectively. Furthermore, to allocate 72 vitellogenic females in an insectary using Drosophila vial is necessary 4100 cm3 against 1400 cm3 and 700 cm3 when using 12- and 24-well plates, respectively. No statistical differences were found between the number of eggs laid in Drosophila vials and the oviplates, validating the method. The oviplate method is an affordable, and time- and space-efficient device, and it is simpler to perform individual fecundity analyses in Ae. aegypti.

Ades aegypti is the most important arbovirus vector in the world, and new strategies are under evaluation. Biological studies mentioning the occurrence of a second mate in Aedes aegypti can interfere with vector control program planning, which involves male mosquito release technique. This study presents different experiments to show the occurrence of mixed progeny. Mixed male crosses (using a combination of different type of males in confinement with virgin females) showed no polyandric female. Individual crosses with male substitution in every gonotrophic cycle also did not show any polyandric female. Individual crosses with a 20 minutes interval, with subsequent male change, showed that only a few females presented mixed offspring. The copulation breach in three different moments, group A with full coitus length, group B the coitus was interrupted in 5–7 seconds after the start; and group C, which the copulation was interrupted 3 seconds after started. In summary, group A showed a majority of unique progeny from the first male; group B showed the higher frequency of mixed offspring and group C with the majority of the crosses belonging to the second male. To conclude, the occurrence of a viable second mate and mixed offspring is only possible when the copulation is interrupted; otherwise, the first mate is responsible for mixed progeny.

Ades aegypti is the most important arbovirus vector in the world, and new strategies are under evaluation. Biological studies mentioning the occurrence of a second mate in Aedes aegypti can interfere with vector control program planning, which involves male mosquito release technique. This study presents different experiments to show the occurrence of mixed progeny. Mixed male crosses (using a combination of different type of males in confinement with virgin females) showed no polyandric female. Individual crosses with male substitution in every gonotrophic cycle also did not show any polyandric female. Individual crosses with a 20 minutes interval, with subsequent male change, showed that only a few females presented mixed offspring. The copulation breach in three different moments, group A with full coitus length, group B the coitus was interrupted in 5-7 seconds after the start; and group C, which the copulation was interrupted 3 seconds after started. In summary, group A showed a majority of unique progeny from the first male; group B showed the higher frequency of mixed offspring and group C with the majority of the crosses belonging to the second male. To conclude, the occurrence of a viable second mate and mixed offspring is only possible when the copulation is interrupted; otherwise, the first mate is responsible for mixed progeny.

The number of mosquito populations resistant to insecticides is increasing along with the reemerging of vector-borne diseases. New technologies are under evaluation to complement the strategies used against these mosquitoes. Transgenic mosquitoes are one approach that some countries are considering and they are being evaluated to control the wild population. Although they have achieved success in population suppression of Aedes aegypti (L.) (Diptera: Culicidae), these studies have not demonstrated what the outcomes are when releases are interrupted (ceased). In this study, after demonstrating suppression of Ae. aegypti using transgenic technology, changes in the spatial distribution of the infestation and the abundance of the vector Ae. aegypti were assessed in the post-release period, along with fluctuation of transgenic mosquitoes in two areas of Brazil. In both pilot trials, there was an average suppression of ca. 70% of the wild population due to the release of transgenic males compared to the pre-release period. In Juazeiro (Mandacaru), in the post-release phase, the number of eggs per trap ranged between 0.06 and 14.41 (mean ± SE = 4.44 ± 0.44), and the ovitrap index (OI = number of ovitraps with eggs/total number of ovitraps recovered) ranged from 0.01 to 0.43 (0.13 ± 0.01). In Jacobina (Pedra Branca), during the post-release phase, the number of eggs per trap ranged between 1 and 7.2 (1.72 ± 0.72), and the OI ranged from 1 to 0.83 (0.095 ± 0.032). The mosquito population in Juazeiro (Mandacaru) remained suppressed for 17 weeks after the release interruption, whereas in Jacobina (Pedra Branca) suppression lasted 32 weeks. In Juazeiro, transgenic larvae were detected up to 5 months after the interruption of the over-flooding releases of transgenic males. In Jacobina, they were found up to 2 months after the release interruption. The number of eggs collected increased 4–5 months after the release interruption, which indicated that the Ae. aegypti population had been re-established after the interruption of releases. The results demonstrate that the technique requires a continuous release in the treated areas, and after suppression, the release rate can be decreased and used as a barrier against external migration.

Mosquitoes are responsible for the transmission of pathogens that cause devastating human diseases such as malaria and dengue. The current increase in mean global temperature and changing sea level interfere with precipitation frequency and some other climatic conditions which, in general, influence the rate of development of insects and etiologic agents causing acceleration as the temperature rises. The most common strategy employed to combat target mosquito species is the Integrated Vector Management (IVM), which comprises the use of multiple activities and various approaches to preventing the spread of a vector in infested areas. IVM programmes are becoming ineffective; and the global scenario is threatening, requiring new interventions for vector control and surveillance. Not surprisingly, there is a growing need to find alternative methods to combat the mosquito vectors. The possibility of using transgenic mosquitoes to fight against those diseases has been discussed over the last two decades and this use of transgenic lines to suppress populations or to replace them is still under investigation through field and laboratory trials. As an alternative, the available transgenic strategies could be improved by coupling suppression and substitution strategies. The idea is to first release a suppression line to significantly reduce the wild population, and once the first objective is reached a second release using a substitution line could be then performed. Examples of targeting this approach against vectors of malaria and dengue are discussed.

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.

For the sterile insect technique, and other related biological control methods where large numbers of the target mosquito are reared artificially, production efficiency is key for the economic viability of the technique. Rearing success begins with high quality eggs. Excess eggs are often stockpiled and stored for longer periods of time. Any pests that prey on these eggs are detrimental to stockpiles and need to be avoided. Psocids of the genus Liposcelis (Psocoptera, Liposcelididae) are common scavengers consuming various types of organic material that are distributed globally and thrive in warm damp environments, making insectaries ideal habitats. In this short report, we investigated the species that has been found scavenging stored mosquito eggs in our insectary and identified it to be Liposcelis bostrychophila Badonnel, 1931. Additional observations were made to determine whether these predators indeed feed on mosquito eggs, and to suggest simple, effective ways of avoiding infestation.Élevage de masse de moustiques : mais qui mange les œufs ?Pour la technique des insectes stériles et les autres méthodes de lutte biologique associées, dans lesquelles un grand nombre de moustiques cibles sont élevés artificiellement, l’efficacité de la production est essentielle pour la viabilité économique de la technique. Le succès de l’élevage commence par des œufs de bonne qualité. Les œufs excédentaires sont souvent stockés pendant de longues périodes. Tous les organismes nuisibles qui exploitent ces œufs nuisent à ces stocks et doivent être évités. Les psoques du genre Liposcelis (Psocoptera, Liposcelididae) sont des charognards répandus qui consomment diverses matières organiques, sont répartis dans le monde entier et prospèrent dans des environnements chauds et humides, ce qui rend les insectariums des habitats idéaux pour eux. Dans ce court rapport, nous avons étudié l’espèce qui mangeait des œufs de moustiques stockés dans notre insectarium et nous avons déterminé qu’il s’agissait de Liposcelis bostrychophila Badonnel, 1931. D’autres observations ont été faites pour déterminer si ces prédateurs se nourrissent effectivement des œufs de moustiques et suggérer des moyens simples et efficaces pour éviter l’infestation.

New studies and strategies are under development for mosquito control. The resulting new information might be evaluated and shared with the local population. They must face the possibility that those strategies will result in increased numbers of mosquitoes in local houses, gardens, and streets. Here is presented a brief collection of studies using this kind of technology and the outcomes from the public engagement used by them, from failures as in India to the one demonstrating a lot of success in Brazil. We describe the set of activities and discuss about the need to adapt any action plan to every target population. Based on the Brazilian experience, contrasted against Cayman Island, the most important characteristic that shifts from a good community engagement is the lack of transparency and distance from local population.

Vector-borne diseases are currently the worst challenge of public health in Brazil and worldwide, being the current control measures inefficient. Innovations concerning vector control point to a new perspective to handling with gene engineering, as they may interfere with disease transmission, preventing the pathogen to complete its cycle in the vector, ie, reducing mosquito population. We present some of the mean genetic tools available, SIT and RIDL for example. We discuss how these tools work, some risks and benefits

Background Aedes aegypti, the principal vector of dengue fever, have been genetically engineered for use in a sterile insect control programme. To improve our understanding of the dispersal ecology of mosquitoes and to inform appropriate release strategies of ‘genetically sterile’ male Aedes aegypti detailed knowledge of the dispersal ability of the released insects is needed. Methodology/Principal Findings The dispersal ability of released ‘genetically sterile’ male Aedes aegypti at a field site in Brazil has been estimated. Dispersal kernels embedded within a generalized linear model framework were used to analyse data collected from three large scale mark release recapture studies. The methodology has been applied to previously published dispersal data to compare the dispersal ability of ‘genetically sterile’ male Aedes aegypti in contrasting environments. We parameterised dispersal kernels and estimated the mean distance travelled for insects in Brazil: 52.8m (95% CI: 49.9m, 56.8m) and Malaysia: 58.0m (95% CI: 51.1m, 71.0m). Conclusions/Significance Our results provide specific, detailed estimates of the dispersal characteristics of released ‘genetically sterile’ male Aedes aegypti in the field. The comparative analysis indicates that despite differing environments and recapture rates, key features of the insects’ dispersal kernels are conserved across the two studies. The results can be used to inform both risk assessments and release programmes using ‘genetically sterile’ male Aedes aegypti., Author Summary Vector control using releases of sterile insects is a well-known approach. ‘Genetically sterile’ male Aedes aegypti have been developed and released in a modern realisation of the sterile insect technique. Released engineered males seek out and mate with wild females, with the resultant offspring dying before they reach maturity. Control of a wild vector population can therefore be achieved by maintaining sustained releases of sterile males whilst ensuring sufficient distribution and coverage of released males across the target area. In order to efficiently plan releases of these, individuals’ detailed knowledge of how they disperse in the field is required. We present an analysis of the dispersal of these engineered male Aedes aegypti using data from field experiments in Brazil. Our results provide detailed information on how the mosquitoes disperse over their potential flight range.

The increasing burden of dengue, and the relative failure of traditional vector control programs highlight the need to develop new control methods. SIT using self-limiting genetic technology is one such promising method. A self-limiting strain of Aedes aegypti, OX513A, has already reached the stage of field evaluation. Sustained releases of OX513A Ae. aegypti males led to 80% suppression of a target wild Ae. aegypti population in the Cayman Islands in 2010. Here we describe sustained series of field releases of OX513A Ae. aegypti males in a suburb of Juazeiro, Bahia, Brazil. This study spanned over a year and reduced the local Ae. aegypti population by 95% (95% CI: 92.2%-97.5%) based on adult trap data and 81% (95% CI: 74.9-85.2%) based on ovitrap indices compared to the adjacent no-release control area. The mating competitiveness of the released males (0.031; 95% CI: 0.025-0.036) was similar to that estimated in the Cayman trials (0.059; 95% CI: 0.011 – 0.210), indicating that environmental and target-strain differences had little impact on the mating success of the OX513A males. We conclude that sustained release of OX513A males may be an effective and widely useful method for suppression of the key dengue vector Ae. aegypti. The observed level of suppression would likely be sufficient to prevent dengue epidemics in the locality tested and other areas with similar or lower transmission., Author Summary Dengue is a major mosquito-borne disease, increasing in prevalence and severity; there are no specific drugs or licensed vaccine. It is primarily transmitted by one mosquito species, Aedes aegypti. We released transgenic ‘sterile’ male mosquitoes in Itaberaba, a suburb of Juazeiro, a Brazilian city. Sustained release of these males, whose offspring typically die before adulthood as a consequence of the transgenic modification, strongly suppressed the target wild population—by 80–95% according to different measures. These data are consistent with previous releases in the Cayman Islands, suggesting that differences between the two locations, including the environment or wild mosquito strain, made little difference. Mathematical models suggest that this degree of suppression would be highly effective in preventing epidemic dengue.

New techniques and methods are being sought to try to win the battle against mosquitoes. Recent advances in molecular techniques have led to the development of new and innovative methods of mosquito control based around the Sterile Insect Technique (SIT)(1-3). A control method known as RIDL (Release of Insects carrying a Dominant Lethal)(4), is based around SIT, but uses genetic methods to remove the need for radiation-sterilization(5-8). A RIDL strain of Ae. aegypti was successfully tested in the field in Grand Cayman(9,10); further field use is planned or in progress in other countries around the world. Mass rearing of insects has been established in several insect species and to levels of billions a week. However, in mosquitoes, rearing has generally been performed on a much smaller scale, with most large scale rearing being performed in the 1970s and 80s. For a RIDL program it is desirable to release as few females as possible as they bite and transmit disease. In a mass rearing program there are several stages to produce the males to be released: egg production, rearing eggs until pupation, and then sorting males from females before release. These males are then used for a RIDL control program, released as either pupae or adults(11,12). To suppress a mosquito population using RIDL a large number of high quality male adults need to be reared(13,14). The following describes the methods for the mass rearing of OX513A, a RIDL strain of Ae. aegypti (8), for release and covers the techniques required for the production of eggs and mass rearing RIDL males for a control program.

New techniques and methods are being sought to try to win the battle against mosquitoes. Recent advances in molecular techniques have led to the development of new and innovative methods of mosquito control based around the Sterile Insect Technique (SIT)(1-3). A control method known as RIDL (Release of Insects carrying a Dominant Lethal)(4), is based around SIT, but uses genetic methods to remove the need for radiation-sterilization(5-8). A RIDL strain of Ae. aegypti was successfully tested in the field in Grand Cayman(9,10); further field use is planned or in progress in other countries around the world. Mass rearing of insects has been established in several insect species and to levels of billions a week. However, in mosquitoes, rearing has generally been performed on a much smaller scale, with most large scale rearing being performed in the 1970s and 80s. For a RIDL program it is desirable to release as few females as possible as they bite and transmit disease. In a mass rearing program there are several stages to produce the males to be released: egg production, rearing eggs until pupation, and then sorting males from females before release. These males are then used for a RIDL control program, released as either pupae or adults(11,12). To suppress a mosquito population using RIDL a large number of high quality male adults need to be reared(13,14). The following describes the methods for the mass rearing of OX513A, a RIDL strain of Ae. aegypti (8), for release and covers the techniques required for the production of eggs and mass rearing RIDL males for a control program.

Vector-borne diseases are transmitted to humans by hematophagous arthropods and these blood-sucking organisms are target to researches worldwide. The laboratory colonization of these species is an important factor in the development of innovative strategies to control these vectors. However, this maintenance requires blood to make these invertebrates able to complete their life cycle. Although live vertebrate animals are frequently used for this feeding procedure, artificial feeders are available as potential alternatives to replace the use of live animals in some situations, especially in vector colony maintenance. The aim of this commentary is to discuss the use of artificial feeding methods concerning the 3Rs principle application. The scientific community focused on vector-borne diseases studies needs to strongly consider these artificial feeding options as a bioethical alternative to maintain blood-feeding arthropods in laboratory.

Background The circumsporozoite protein is the most abundant polypeptide expressed by sporozoites, the malaria parasite stage capable of infecting humans. Sporozoite invasion of mosquito salivary glands prior to transmission is likely mediated by a receptor/ligand-like interaction of the parasites with the target tissues, and the amino (NH2)-terminal portion of CSP is involved in this interaction but not the TSR region on the carboxyl (C)-terminus. Peptides based on the NH2-terminal domain could compete with the parasites for the salivary gland receptors and thus inhibit penetration. Methods Peptides based on the NH2-terminus and TSR domains of the CSP from avian or human malaria parasites, Plasmodium gallinaceum and Plasmodium falciparum, respectively, were expressed endogenously in mosquito haemolymph using a transient (Sindbis virus-mediated) or stable (piggyBac-mediated transgenesis) system. Results Transient endogenous expression of partial NH2-terminus peptide from P. falciparum CSP in P. gallinaceum-infected Aedes aegypti resulted in a reduced number of sporozoites in the salivary glands. When a transgenic approach was used to express a partial CSP NH2-terminal domain from P. gallinaceum the number of sporozoites in the salivary glands did not show a difference when compared to controls. However, a significant difference could be observed when mosquitoes with a lower infection were analysed. The same result could not be observed with mosquitoes endogenously expressing peptides based on the TSR domain from either P. gallinaceum or P. falciparum. Conclusion These results support the conclusion that CSP partial NH2-terminal domain can be endogenously expressed to promote a competition for the receptor used by sporozoites to invade salivary glands, and they could be used to block this interaction and reduce parasite transmission. The same effect cannot be obtained with peptides based on the TSR domain. Electronic supplementary material The online version of this article (doi:10.1186/s12936-016-1207-8) contains supplementary material, which is available to authorized users.