Sterile Insect Technique Component Research Articles

Two invasions at once: update on the introduction of the invasive species Aedes aegypti and Aedes albopictus in Cyprus - a call for action in Europe.

Aedes aegypti, the yellow fever mosquito and Aedes albopictus, the tiger mosquito, continue to expand their geographical distribution, reshaping the European epidemiological risks for mosquito-borne diseases. The reintroduction of Aedes aegypti near the airport and port in Larnaka and the detection of Aedes albopictus near the marina and old port of the Limassol area in Cyprus are reported herein. The measures taken to investigate these events included (i) communication to health authorities, (ii) expert on-site visits and verification of findings, (iii) enhanced active surveillance, and (iv) development of an Emergency Action Plan followed by a Contingency Plan. These emergency action plans were developed to delimitate the infested areas and to prevent the spreading of the mosquito populations into new areas. The general principles are presented along with their rationale to serve as guidelines for other geographical regions targeting suppression/eradication with a sterile insect technique component. In parallel, this manuscript serves as a call for action at the European level to impede the further spread of these species and support the activities being undertaken in Cyprus to combat the incursions of Aedes invasive species.

Near-infrared imaging for automated tsetse pupae sex sorting in support of the sterile insect technique.

Tsetse flies are the cyclical vectors of African trypanosomes and one of several methods to manage this vector is the sterile insect technique (SIT). The ability to determine the sex of tsetse pupae with the objective to separate the sexes before adult emergence has been a major goal for decades for tsetse management programmes with an SIT component. Tsetse females develop faster and pharate females inside the pupae melanise 1-2 days before males. This earlier melanisation can be detected by infrared cameras through the pupal shell, and the newly developed Near InfraRed Pupae Sex Sorter (NIRPSS) takes advantage of this. The melanisation process is not homogeneous for all fly organs and the pupa needs to be examined ventrally, dorsally and laterally to ensure accurate classification by an image analysis algorithm. When the pupae are maturing at a constant temperature of 24°C and sorted at the appropriate age, 24 days post-larviposition for Glossina palpalis gambiensis, the sorting machine can efficiently separate the sexes. The recovered male pupae can then be sterilised for field releases of males, while the rest of the pupae can be used to maintain the laboratory colony. The sorting process with the new NIRPSS had no negative impact on adult emergence and flight ability. A mean male recovery of 62.82±3.61% was enough to provide sterile males to an operational SIT programme, while mean contamination with females (4.69±3.02%) was low enough to have no impact on the maintenance of a laboratory colony.

Gamma-radiation of Glossina palpalis gambiensis revisited: effect on fertility and mating competitiveness

African animal trypanosomoses are vector-borne diseases that cause enormous livestock losses in sub-Saharan Africa, with drastic socio-economic impacts. Vector control in the context of an area-wide integrated pest management program with a sterile insect technique component requires the production of high-quality sterile male tsetse flies. In our study, we evaluated the effect of irradiation on the fecundity of Glossina palpalis gambiensis to identify the optimal dose that will induce maximum sterility while maintaining biological performance as much as possible. In addition, male mating performance was evaluated in semi-field cages. The irradiation doses used were 90, 100, 110, 120, 130, 140, and 150 Gy, and untreated males were used as the control. The results showed that pupal production and emergence rates were higher in batches of females that had mated with fertile males than in those that had mated with irradiated males with any experimental dose. A dose of 120 Gy administered to male flies induced 97-99% sterility after mating with virgin females. For the semi-field cage experiments, males irradiated with 120 Gy showed good sexual competitiveness as compared to fertile males and those irradiated with 140 Gy, considering the level of filling of spermatheca and the number of pairs formed. The optimal radiation dose of 120 Gy found in this study is slightly different from the traditional dose of 110 Gy that has been used in several eradication programmes in the past. The potential reasons for this difference are discussed, and an argument is made for the inclusion of reliable dosimetry systems in these types of studies.

Temperature Sensitivity of Wild-Type, Mutant and Genetic Sexing Strains of Ceratitis capitata.

Simple SummaryThe Mediterranean fruit fly (medfly), Ceratitis capitata, is a major agricultural insect pest species worldwide. The sterile insect technique (SIT), as a component of area-wide integrated management (AW-IPM) programmes, is currently used to control populations of this pest. SIT is based on the mass production and release of sexually sterile insects, ideally males, over a target area. Male-only releases can be achieved by using genetic sexing strains (GSS) such as the medfly VIENNA 8 GSS. Females of this strain are homozygous for a mutation in the temperature-sensitive lethal (tsl) gene, which kills them when exposed to high temperatures during the embryonic stage. In the present study, we employed a temperature-sensitive lethal test (TSLT) to determine the temperature sensitivity or tolerance of twenty-seven Ceratitis capitata wild-type, genetic sexing, and tsl mutant strains. Significant differences were detected among the strains studied with respect to egg hatching, pupal, and adult recovery rates. We discussed our findings in the context of SIT applications and climate change. Area-wide integrated pest management (AW-IPM) programmes with a sterile insect technique component (SIT) are used to control populations of insect pests worldwide, including the Mediterranean fruit fly, Ceratitis capitata. SIT consists of the mass rearing, radiation-induced sterilization, handling, and release of sterile insects over the target area. Although SIT can be performed by using both sterile males and females, male-only releases significantly increase the efficiency and cost-effectiveness of SIT applications. Male-only releases can be achieved by using genetic sexing strains (GSS). The medfly VIENNA 8 GSS is based on two selectable markers, the white pupae (wp) gene, and the temperature-sensitive lethal (tsl) genes. The latter allows the elimination of females by exposing embryos to elevated temperatures. This study assessed the temperature sensitivity of twenty-seven medfly strains through a TSLT. Our results indicated significant differences among the strains regarding egg hatching as well as pupal and adult recovery rates due to the presence or absence of the tsl mutation and/or the genetic background of the strains. Our findings are discussed in the context of SIT applications, the importance of the tsl gene for developing genetic sexing strains, and climate change.

Genetic structure and symbiotic profile of worldwide natural populations of the Mediterranean fruit fly, Ceratitis capitata

BackgroundThe Mediterranean fruit fly, Ceratitis capitata, is a cosmopolitan agricultural pest of worldwide economic importance and a model for the development of the Sterile Insect Technique (SIT) for fruit flies of the Tephritidae family (Diptera). SIT relies on the effective mating of laboratory-reared strains and natural populations, and therefore requires an efficient mass-rearing system that will allow for the production of high-quality males. Adaptation of wild flies to an artificial laboratory environment can be accompanied by negative effects on several life history traits through changes in their genetic diversity and symbiotic communities. Such changes may lead to reduced biological quality and mating competitiveness in respect to the wild populations. Profiling wild populations can help understand, and maybe reverse, deleterious effects accompanying laboratory domestication thus providing insects that can efficiently and effectively support SIT application.ResultsIn the present study, we analyzed both the genetic structure and gut symbiotic communities of natural medfly populations of worldwide distribution, including Europe, Africa, Australia, and the Americas. The genetic structure of 408 individuals from 15 distinct populations was analyzed with a set of commonly used microsatellite markers. The symbiotic communities of a subset of 265 individuals from 11 populations were analyzed using the 16S rRNA gene-based amplicon sequencing of single individuals (adults). Genetic differentiation was detected among geographically distant populations while adults originated from neighboring areas were genetically closer. Alpha and beta diversity of bacterial communities pointed to an overall reduced symbiotic diversity and the influence of the geographic location on the bacterial profile.ConclusionsOur analysis revealed differences both in the genetic profile and the structure of gut symbiotic communities of medfly natural populations. The genetic analysis expanded our knowledge to populations not analyzed before and our results were in accordance with the existing scenarios regarding this species expansion and colonization pathways. At the same time, the bacterial communities from different natural medfly populations have been characterized, thus broadening our knowledge on the microbiota of the species across its range. Genetic and symbiotic differences between natural and laboratory populations must be considered when designing AW-IPM approaches with a SIT component, since they may impact mating compatibility and mating competitiveness of the laboratory-reared males. In parallel, enrichment from wild populations and/or symbiotic supplementation could increase rearing productivity, biological quality, and mating competitiveness of SIT-important laboratory strains.

High sensitivity of one-step real-time reverse transcription quantitative PCR to detect low virus titers in large mosquito pools

BackgroundMosquitoes are the deadliest animals in the world. Their ability to carry and spread diseases to humans causes millions of deaths every year. Due to the lack of efficient vaccines, the control of mosquito-borne diseases primarily relies on the management of the vector. Traditional control methods are insufficient to control mosquito populations. The sterile insect technique (SIT) is an additional control method that can be combined with other control tactics to suppress specific mosquito populations. The SIT requires the mass-rearing and release of sterile males with the aim to induce sterility in the wild female population. Samples collected from the environment for laboratory colonization, as well as the released males, should be free from mosquito-borne viruses (MBV). Therefore, efficient detection methods with defined detection limits for MBV are required. Although a one-step reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) method was developed to detect arboviruses in human and mosquito samples, its detection limit in mosquito samples has yet to be defined.MethodsWe evaluated the detection sensitivity of one step RT-qPCR for targeted arboviruses in large mosquito pools, using pools of non-infected mosquitoes of various sizes (165, 320 and 1600 mosquitoes) containing one infected mosquito body with defined virus titers of chikungunya virus (CHIKV), usutu virus (USUV), West Nile virus (WNV) and Zika virus (ZIKV).ResultsCHIK, USUV, ZIKV, and WNV virus were detected in all tested pools using the RT-qPCR assay. Moreover, in the largest mosquito pools (1600 mosquitoes), RT-qPCR was able to detect the targeted viruses using different total RNA quantities (10, 1 and 0.1 ng per reaction) as a template. Correlating the virus titer with the total RNA quantity allowed the prediction of the maximum number of mosquitoes per pool in which the RT-qPCR can theoretically detect the virus infection.ConclusionsMosquito-borne viruses can be reliably detected by RT-qPCR assay in pools of mosquitoes exceeding 1000 specimens. This will represent an important step to expand pathogen-free colonies for mass-rearing sterile males for programmes that have a SIT component by reducing the time and the manpower needed to conduct this quality control process.

Demonstration of resistance to satyrization behavior in Aedes aegypti from La Réunion island.

Aedes aegypti and Aedes albopictus are competent vectors of arboviruses such as dengue and chikungunya viruses which co-exist in some areas, including La Réunion island, Indian Ocean. A type of reproductive interference called satyrization has been described in sympatric species where dominant species mating fails to produce hybrids and thus reduces the fitness and tends to control the spread of the other species. Here, we investigated satyrization in laboratory experiments to provide insights on the potential impact on Ae. aegypti of a control campaign including a sterile insect technique component against Ae. albopictus. Different mating crosses were used to test sympatric, conspecific-interspecific and allopatric effects of irradiated and non-irradiated male Ae. albopictus on female Ae. aegypti, including in a situation of skewed male ratio. Our results suggest that there was only a low level of satyrization between sympatric populations of Ae. aegypti and Ae. albopictus colonized from La Réunion island. A male Ae. albopictus to female Ae. aegypti ratio of 3:1 did not increase the level of satyrization. Female Ae. aegypti previously mated to male Ae. albopictus were not prevented from being inseminated by conspecific males. A satyrization effect was not seen between allopatric Ae. albopictus and Ae. aegypti strains from La Réunion Island either. The tested Ae. aegypti strain from La Réunion island has therefore developed full resistance to satyrization and so releasing sterile male Ae. albopictus may not suppress Ae. aegypti populations if an overflooding of irradiated male Ae. albopictus leads to similar results. The management strategy of two competent species in a sympatric area is discussed.

Cryopreservation of the Mediterranean fruit fly (Diptera: Tephritidae) VIENNA 8 genetic sexing strain: No effect on large scale production of high quality sterile males for SIT applications.

The sterile insect technique (SIT) integrated in area-wide integrated pest management (AW-IPM) programmes is being used for the successful management of the Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) which is a horticultural pest of economic importance in tropical and subtropical countries. All programmes with an SIT component are using the VIENNA genetic sexing strains (GSS), mainly the VIENNA 8 GSS, which have been developed by applying classical genetic approaches. The VIENNA 8 GSS carries two selectable markers, the white pupae and the temperature sensitive lethal genes, which allows the production and release of only males thus increasing the biological efficiency and cost effectiveness of SIT applications. However, mass rearing may affect quality traits of the GSS, in which case replenishment of the colony with wild flies is recommended, a process which is tedious and time consuming. We previously reported the development of a cryopreservation protocol for the VIENNA 8D53+ strain. In the present study, we report on the evaluation of the cryopreserved strain VIENNA 8D53+/Cryo-228L, reared under semi mass rearing conditions, for production parameters, quality control indices and mating competitiveness of males, in a comparative way with the non-cryopreserved VIENNA 8D53+ strain, against wild type males. The VIENNA 8D53+ and VIENNA 8D53+/Cryo-228L strains were similar for production parameters viz. egg production, pupal production, pupal recovery, and quality control indices like fly emergence, sex ratio and flight ability. Males from both strains were equally competitive with males of the wild type strain in achieving mating with wild type females under field cage conditions. Results are discussed in the context of cryopreservation as a potential backup strategy for refreshing the mass rearing colony with biological material from a cryopreserved stock.

Longevity of mass-reared, irradiated and packed male Anopheles arabiensis and Aedes aegypti under simulated environmental field conditions

BackgroundTo ensure the success of a mosquito control programme that integrates the sterile insect technique (SIT), it is highly desirable to release sterile males with a maximal lifespan to increase release effectiveness. Understanding sterile male survival under field conditions is thus critical for determining the number of males to be released. Our study aimed to investigate the effect of mass rearing, irradiation, chilling, packing and release time on irradiated male mosquito longevity.MethodsAnopheles arabiensis and Aedes aegypti immature stages were mass-reared using a rack and tray system. Batches of 50 males irradiated at the pupal stage were immobilised, packed into canisters and chilled for 6 hours at 6 °C. Mosquitoes were then transferred either in the early morning or early evening into climate chambers set to simulate the weather conditions, typical of the beginning of the rainy season in Khartoum, Sudan and Juazeiro, Brazil for An. arabiensis and Ae. aegypti, respectively. The longevity of experimental males was assessed and compared to mass-reared control males subjected either to simulated field or laboratory conditions.ResultsThe combined irradiation, chilling and packing treatments significantly reduced the longevity of both An. arabiensis and Ae. aegypti under simulated field conditions (P < 0.001). However, packing alone did not significantly reduce longevity of Ae. aegypti (P = 0.38) but did in An. arabiensis (P < 0.001). Overall, the longevity of mass reared, irradiated and packed males was significantly reduced, with the median survival time (days) lower following an early morning introduction (4.62 ± 0.20) compared to an evening (7.34 ± 0.35) in An. arabiensis (P < 0.001). However, there was no significant difference in longevity between morning (9.07 ± 0.54) and evening (7.76 ± 0.50) in Ae. aegypti (P = 0.14).ConclusionsOur study showed that sterile mass-reared males have a reduced lifespan in comparison to laboratory-maintained controls under simulated field conditions, and that An. arabiensis appeared to be more sensitive to the handling process and release time than Ae. aegypti. Longevity and release time are important parameters to be considered for a successful area-wide integrated vector control programme with a SIT component.

Impact of Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) on a heterologous tsetse fly host, Glossina fuscipes fuscipes

BackgroundTsetse flies (Diptera: Glossinidae) are the vectors of African trypanosomosis, the causal agent of sleeping sickness in humans and nagana in animals. Glossina fuscipes fuscipes is one of the most important tsetse vectors of sleeping sickness, particularly in Central Africa. Due to the development of resistance of the trypanosomes to the commonly used trypanocidal drugs and the lack of effective vaccines, vector control approaches remain the most effective strategies for sustainable management of those diseases. The Sterile Insect Technique (SIT) is an effective, environment-friendly method for the management of tsetse flies in the context of area-wide integrated pest management programs (AW-IPM). This technique relies on the mass-production of the target insect, its sterilization with ionizing radiation and the release of sterile males in the target area where they will mate with wild females and induce sterility in the native population. It has been shown that Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) infection causes a decrease in fecundity and fertility hampering the maintenance of colonies of the tsetse fly G. pallidipes. This virus has also been detected in different species of tsetse files. In this study, we evaluated the impact of GpSGHV on the performance of a colony of the heterologous host G. f. fuscipes, including the flies’ productivity, mortality, survival, flight propensity and mating ability and insemination rates.ResultsEven though GpSGHV infection did not induce SGH symptoms, it significantly reduced all examined parameters, except adult flight propensity and insemination rate.ConclusionThese results emphasize the important role of GpSGHV management strategy in the maintenance of G. f. fuscipes colonies and the urgent need to implement measures to avoid virus infection, to ensure the optimal mass production of this tsetse species for use in AW-IPM programs with an SIT component.

Competitiveness and survival of two strains of Glossina palpalis gambiensis in an urban area of Senegal

BackgroundIn the Niayes area, located in the west of Senegal, only one tsetse species, Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) was present. The Government of Senegal initiated and implemented an elimination programme in this area that included a sterile insect technique (SIT) component. The G. p. gambiensis strain (BKF) mass-reared at the Centre International de Recherche-Développement sur l'Elevage en zone Subhumide (CIRDES) in Burkina Faso was used for the SIT component.Methodology/principal findingsStudies conducted in 2011 in four localities in the Niayes area (Pout, Sébikotane, Diacksao Peul and the Parc de Hann) showed that the BKF strain demonstrated inferior survival in the ecosystem of the Parc de Hann, a forested area in the city centre of the capital Dakar. Therefore, G. p. gambiensis flies from the Niayes area (SEN strain) were colonized. Here we compared the competitiveness and survival of the two strains (BKF and SEN) in the Parc de Hann. Released sterile males of the SEN colony showed a daily mortality rate of 0.08 (SD 0.08) as compared with 0.14 (SD 0.08) for the BKF flies but the difference was not significant (p-value = 0.14). However, the competitiveness of the SEN males was lower (0.14 (SD 0.10)) as compared with that of the BKF males (0.76 (SD 0.11)) (p-value < 10−3).Conclusions/significanceBased on the results of this study, it can be concluded that the BKF strain will remain the main strain to be used in the elimination programme. Despite the slightly longer survival of the SEN males in the Parc de Hann, the superior competitiveness of the BKF males is deemed more important for the SIT component, as their shorter survival rates can be easily compensated for by more frequent fly releases.

Evaluation of radiation sensitivity and mating performance of Glossina brevipalpis males.

BackgroundArea-wide integrated pest management strategies that include a sterile insect technique component have been successfully used to eradicate tsetse fly populations in the past. To ensure the success of the sterile insect technique, the released males must be adequately sterile and be able to compete with their native counterparts in the wild.Methodology/Principal findingsIn the present study the radiation sensitivity of colonised Glossina brevipalpis Newstead (Diptera; Glossinidae) males, treated either as adults or pupae, was assessed. The mating performance of the irradiated G. brevipalpis males was assessed in walk-in field cages. Glossina brevipalpis adults and pupae were highly sensitive to irradiation, and a dose of 40 Gy and 80 Gy induced 93% and 99% sterility respectively in untreated females that mated with males irradiated as adults. When 37 to 41 day old pupae were exposed to a dose of 40 Gy, more than 97% sterility was induced in untreated females that mated with males derived from irradiated pupae. Males treated as adults with a dose up to 80 Gy were able to compete successfully with untreated fertile males for untreated females in walk-in field cages.Conclusions/SignificanceThe data emanating from this field cage study indicates that, sterile male flies derived from the colony of G. brevipalpis maintained at the Agricultural Research Council-Onderstepoort Veterinary Institute in South Africa are potential good candidates for a campaign that includes a sterile insect technique component. This would need to be confirmed by open field studies.

A sound trap for Aedes albopictus (Skuse) male surveillance:Response analysis to acoustic and visual stimuli

Aedes albopictus (Skuse) is one of the most invasive mosquito species capable of transmitting various harmful pathogens to humans. Failure of vector control strategies against this species requires the development of new effective vector control methods. Among the alternative genetic control measures under development, the sterile insect technique (SIT) is today receiving a renewed interest as a possible effective tool to be integrated in an area-wide pest management approach. The monitoring of the abundance, distribution, movements and ratio of released sterile and wild fertile males is a fundamental requirement for the successful management of any pest control activities integrating an SIT component. Although several traps exist for adult female mosquito surveillance and population control, effective methods to monitor large quantities of non-biting males were less researched in the past and more difficult to obtain. In this study we analyzed the response of A. albopictus males to various sound stimuli produced with different volumes and frequencies in association with visual cues for the optimization of male catches. The production of modulated frequencies continuously varying within the typical female sound emission range (500–650Hz) showed the best results for a sound pressure level between 75 and 79dB measured at the speaker level. The black color of the trap, however, seems decisive to attract males in the vicinity of sound traps and the combined effect produced by these stimuli is additive. We also observed that males increase their response to acoustic stimulation up to 4days of age and then show a continuous and progressive decline of their sound responsiveness. A plastic sound trap prototype capable of producing the most effective sound stimuli tested at laboratory conditions showed a good A. albopictus males’ attractiveness in the field despite a reduced use of color stimuli. The use of sound stimuli therefore appears to be a promising prospect to increase the capture rate of A. albopictus males in new or in already existing mosquito traps.

Effect of Genetically Modified Bt Maize in an Artificial Diet on the Survival ofCydia pomonella(Lepidoptera: Tortricidae)

Abstract Genetically modified maize contains an insecticidal gene from the soil bacterium Bacillus thuringiensis (Bt), which is an important component in integrated pest management strategies against lepidopteran pests of maize. A project is being implemented in the Western Cape of South Africa against the codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), a pome fruit pest, using an area-wide integrated pest management approach with a sterile insect technique component. The project requires rearing of large numbers of the target pest for which an artificial diet that contains maize meal as the main ingredient is used. Most of the maize produced in South Africa is Bt maize, which is known to be toxic to codling moth. The aim of this study was to assess the impact of Bt maize in the diet of codling moth on its production parameters. Codling moths were reared for a period of 44 d on artificial diets that contained 5 different concentrations of Bt maize meal and a control using non-Bt maize. The ...

Mating Compatibility and Competitiveness between Wild and Laboratory Strains ofEldana saccharina(Lepidoptera: Pyralidae) after Radiation Treatment

Abstract The efficacy of the sterile insect technique (SIT) applied as part of area-wide integrated pest management (AW-IPM) depends on efficient transfer of sperm carrying dominant lethal mutations from sterile males to wild females. Success or failure of this strategy is therefore critically dependent on quality and ability of sterile males to search for and copulate with wild females. The African sugarcane borer, Eldana saccharina Walker (Lepidoptera: Pyralidae) is an economic pest of sugarcane targeted for control in South Africa using an AW-IPM approach with a SIT component. As part of further steps towards development of the technique, levels of mating competitiveness and compatibility were assessed by observing the extent to which individuals from different populations interbreed when confined together under both laboratory and semi-field conditions. Three types of pair-wise competition experiments were conducted: non-irradiated laboratory adults vs. non-irradiated wild adults, irradiated (200 Gy) ...

A Molecular Method to Discriminate between Mass-Reared Sterile and Wild Tsetse Flies during Eradication Programmes That Have a Sterile Insect Technique Component.

BackgroundThe Government of Senegal has embarked several years ago on a project that aims to eradicate Glossina palpalis gambiensis from the Niayes area. The removal of the animal trypanosomosis would allow the development more efficient livestock production systems. The project was implemented using an area-wide integrated pest management strategy including a sterile insect technique (SIT) component. The released sterile male flies originated from a colony from Burkina Faso.Methodology/Principal FindingsMonitoring the efficacy of the sterile male releases requires the discrimination between wild and sterile male G. p. gambiensis that are sampled in monitoring traps. Before being released, sterile male flies were marked with a fluorescent dye powder. The marking was however not infallible with some sterile flies only slightly marked or some wild flies contaminated with a few dye particles in the monitoring traps. Trapped flies can also be damaged due to predation by ants, making it difficult to discriminate between wild and sterile males using a fluorescence camera and / or a fluorescence microscope. We developed a molecular technique based on the determination of cytochrome oxidase haplotypes of G. p. gambiensis to discriminate between wild and sterile males. DNA was isolated from the head of flies and a portion of the 5’ end of the mitochondrial gene cytochrome oxidase I was amplified to be finally sequenced. Our results indicated that all the sterile males from the Burkina Faso colony displayed the same haplotype and systematically differed from wild male flies trapped in Senegal and Burkina Faso. This allowed 100% discrimination between sterile and wild male G. p. gambiensis.Conclusions/SignificanceThis tool might be useful for other tsetse control campaigns with a SIT component in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign (PATTEC) and, more generally, for other vector or insect pest control programs.

Effects of laboratory colonization on Bactrocera dorsalis (Diptera, Tephritidae) mating behaviour: 'what a difference a year makes'.

Laboratory-reared insects are widely known to have significantly reduced genetic diversity in comparison to wild populations; however, subtle behavioural changes between laboratory-adapted and wild or ‘wildish’ (i.e., within one or very few generations of field collected material) populations are less well understood. Quantifying alterations in behaviour, particularly sexual, in laboratory-adapted insects is important for mass-reared insects for use in pest management strategies, especially those that have a sterile insect technique component. We report subtle changes in sexual behaviour between ‘wildish’ Bactrocera dorsalis flies (F1 and F2) from central and southern Thailand and the same colonies 12 months later when at six generations from wild. Mating compatibility tests were undertaken under standardised semi-natural conditions, with number of homo/heterotypic couples and mating location in field cages analysed via compatibility indices. Central and southern populations of Bactrocera dorsalis displayed positive assortative mating in the 2010 trials but mated randomly in the 2011 trials. ‘Wildish’ southern Thailand males mated significantly earlier than central Thailand males in 2010; this difference was considerably reduced in 2011, yet homotypic couples from southern Thailand still formed significantly earlier than all other couple combinations. There was no significant difference in couple location in 2010; however, couple location significantly differed among pair types in 2011 with those involving southern Thailand females occurring significantly more often on the tree relative to those with central Thailand females. Relative participation also changed with time, with more southern Thailand females forming couples relative to central Thailand females in 2010; this difference was considerably decreased by 2011. These results reveal how subtle changes in sexual behaviour, as driven by laboratory rearing conditions, may significantly influence mating behaviour between laboratory-adapted and recently colonised tephritid fruit flies over a relatively short period of time.

Determination of the optimal mating age of colonised Glossina brevipalpis and Glossina austeni using walk-in field cages in South Africa.

BackgroundFor the control of Glossina brevipalpis and Glossina austeni that occur in South Africa an area-wide integrated pest management (AW-IPM) program with a sterile insect technique (SIT) component has been proposed. The quality of the released sterile male tsetse flies will greatly determine the success of the SIT component of the programme. Sterile males need to be able to compete with wild males immediately after their release in the affected area. The mating competitiveness can be affected by many factors including the optimal mating age of the fly which can have an impact on the timing of the release.MethodsTo assess the optimal mating age for G. brevipalpis and G. austeni, mating competitiveness studies were carried out in a walk-in field cage. First, the time of peak fly activity was determined by performing the experiment in the morning and then again in the afternoon. Thereafter, 3, 6 and 9-day-old male flies competed for 3-day-old virgin females.ResultsThere were no significant differences in mating performance when the field cage experiments were done in the morning or in the afternoon. However, the mating latency was shorter in the afternoon than in the morning. For both species 9-day-old males mated significantly more often than 6 or 3-day-old males. Age did not affect the males’ ability to transfer sperm, mating duration or the mating latency. All females that mated were inseminated.ConclusionsAge did influence the mating competitiveness of G. brevipalpis and G. austeni and it is recommended that sterile males are not released before the age of 9 days. Keeping the male flies in the rearing facility for 8 days will have economic and logistic consequences for AW-IPM programmes that have a SIT component.

Standard operating procedures for standardized mass rearing of the dengue and chikungunya vectors Aedes aegypti and Aedes albopictus (Diptera: Culicidae) - II - Egg storage and hatching.

BackgroundManagement of large quantities of eggs will be a crucial aspect of the efficient and sustainable mass production of mosquitoes for programmes with a Sterile Insect Technique component. The efficiency of different hatching media and effectiveness of long term storage methods are presented here.MethodsThe effect on hatch rate of storage duration and three hatching media was analysed: deionized water, boiled deionized water and a bacterial broth, using Two-way ANOVA and Post hoc Tukey tests, and the Pearson correlation coefficient was used to find the effect on the proportion of collapsed eggs. Two long term storage methods were also tested: conventional storage (egg paper strips stored in zip lock bags within a sealed plastic box), and water storage (egg papers in a covered plastic cup with deionized water). Regression analyses were used to find the effect of water storage and storage duration on hatch rate.ResultsBoth species hatched most efficiently in bacterial broth. Few eggs hatched in deionized water, and pre-boiling the water increased the hatch rate of Ae. aegypti, but not Ae. albopictus. A hatch rate greater than 80 % was obtained after 10 weeks of conventional storage in Ae. aegypti and 11 weeks in Ae. albopictus. After this period, hatching decreased dramatically; no eggs hatched after 24 weeks. Storing eggs in water produced an 85 % hatch rate after 5 months in both species. A small but significant proportion of eggs hatched in the water, probably due to combined effects of natural deoxygenation of the water over time and the natural instalment hatching typical of the species.ConclusionsThe demonstrated efficiency of the bacterial broth hatching medium for both Ae. albopictus and Ae. aegypti facilitates mass production of these two important vector species in the same facility, with use of a common hatching medium reducing cost and operational complexity. Similarly the increased hatch rate of eggs stored in water would allow greater flexibility of egg management in a large programme over the medium term, particularly if oxygenation of the water by bubbling oxygen through the storage tray could be applied to prevent hatching during storage.

Anopheles arabiensis egg treatment with dieldrin for sex separation leaves residues in male adult mosquitoes that can bioaccumulate in goldfish (Carassius auratus auratus).

The sterile insect technique (SIT) is a biological control tactic that is used as a component of area-wide integrated pest management (AW-IPM) programs. The SIT can only be applied against disease-transmitting mosquitoes when only sterile male mosquitoes are released, and the blood-sucking and potentially disease-transmitting females are eliminated from the production line. For Anopheles arabiensis, a potent vector of malaria, a genetic sexing strain was developed whereby females can be eliminated by treating the eggs or larvae with the insecticide dieldrin. To evaluate the presence of dieldrin residues in male mosquitoes designated for SIT releases, a simple, sensitive, and accurate gas chromatography–electron capture detector (GC–ECD) method was developed. In addition, bioaccumulation and food chain transfer of these residues to fish after feeding with treated mosquitoes was demonstrated. The overall recovery from method validation studies was 77.3 ± 2.2% (mean ± relative standard deviation [RSD]) for the mosquitoes, and 99.1 ± 4.4% (mean ± RSD) for the fish. The average dieldrin concentration found in adult male An. arabiensis was 28.1 ± 2.9 µg/kg (mean ± standard deviation [SD]). A range of 23.9 ± 1.1 µg/kg to 73.9 ± 5.2 µg/kg (mean ± SD) of dieldrin was found in the fish samples. These findings indicate the need to reassess the environmental and health implications of control operations with a SIT component against An. arabiensis that involves using persistent organochlorines in the sexing process.