Clelia F. Oliva

The sterile insect technique for controlling populations of Aedes albopictus (Diptera: Culicidae) on Reunion Island: mating vigour of sterilized males.

Reunion Island suffers from high densities of the chikungunya and dengue vector Aedes albopictus. The sterile insect technique (SIT) offers a promising strategy for mosquito-borne diseases prevention and control. For such a strategy to be effective, sterile males need to be competitive enough to fulfil their intended function by reducing wild mosquito populations in natura. We studied the effect of irradiation on sexual maturation and mating success of males, and compared the sexual competitiveness of sterile versus wild males in the presence of wild females in semi-field conditions. For all untreated or sterile males, sexual maturation was completed within 13 to 20 h post-emergence and some males were able to inseminate females when 15 h old. In the absence of competition, untreated and sterile males were able to inseminate the same number of virgin females during 48 h, in small laboratory cages: an average of 93% of females was inseminated no matter the treatment, the age of males, and the sex ratio. Daily mating success of single sterile males followed the same pattern as for untreated ones, although they inseminated significantly fewer females after the ninth day. The competitiveness index of sterile males in semi-field conditions was only 0.14 when they were released at 1-day old, but improved to 0.53 when the release occurred after a 5-day period in laboratory conditions. In SIT simulation experiments, a 5∶1 sterile to wild male ratio allowed a two-fold reduction of the wild population’s fertility. This suggests that sterile males could be sufficiently competitive to mate with wild females within the framework of an SIT component as part of an AW-IPM programme for suppressing a wild population of Ae. albopictus in Reunion Island. It will be of interest to minimise the pre-release period in controlled conditions to ensure a good competitiveness without increasing mass rearing costs.

Reproductive Strategies of Aedes albopictus (Diptera: Culicidae) and Implications for the Sterile Insect Technique

Male insects are expected to optimize their reproductive strategy according to the availability of sperm or other ejaculatory materials, and to the availability and reproductive status of females. Here, we investigated the reproductive strategy and sperm management of male and virgin female Aedes albopictus, a mosquito vector of chikungunya and dengue viruses. The dynamics of semen transfer to the female bursa inseminalis and spermathecae were observed. Double-mating experiments were conducted to study the effect of time lapsed or an oviposition event between two copulations on the likelihood of a female double-insemination and the use of sperm for egg fertilization; untreated fertile males and radio-sterilised males were used for this purpose. Multiple inseminations and therefore the possibility of sperm competition were limited to matings closely spaced in time. When two males consecutively mated the same female within a 40 min interval, in ca. 15% of the cases did both males sire progeny. When the intervals between the copulations were longer, all progeny over several gonotrophic cycles were offspring of the first male. The mating behavior of males was examined during a rapid sequence of copulations. Male Ae. albopictus were parceling sperm allocation over several matings; however they would also attempt to copulate with females irrespective of the available sperm supply or accessory gland secretion material. During each mating, they transferred large quantities of sperm that was not stored for egg fertilization, and they attempted to copulate with mated females with a low probability of transferring their genes to the next generation. The outcomes of this study provided in addition some essential insights with respect to the sterile insect technique (SIT) as a vector control method.

Effects of irradiation, presence of females, and sugar supply on the longevity of sterile males Aedes albopictus (Skuse) under semi-field conditions on Reunion Island.

BACKGROUND The development of the sterile insect technique (SIT) for reducing populations of Aedes albopictus (Skuse), (the vector of Chikungunya and Dengue fever), was studied in Reunion Island. For some mosquito species the sterilization process and mating activity may alter male survival. Most previous studies were carried out in the laboratory and may inadequately reflect the field situation. We conducted a semi-field experiment to evaluate the impact of sugar supply and mating activity under natural climatic conditions on wild and sterile male Ae. albopictus longevity, using large cages set up in an open clearing between trees and shrubs in Reunion Island. RESULTS Wild males had a mean longevity of 15.5 days in the absence of females and with an immediate sugar supply; longevity in sterile males was similar. The presence of females greatly reduced both wild and especially sterile male lifespan; however, an immediate sugar supply could counteract this effect and allow sterile males to live an average of 11.6 days. CONCLUSION The outcomes indicate that sugar feeding could compensate for sterilization-induced damage, and that mating activity is not deleterious for well-fed males. This study stresses the critical importance of providing suitable sugar sources prior to release during SIT programmes.

X-Ray-Induced Sterility in Aedes albopictus (Diptera: Culicidae) and Male Longevity Following Irradiation

ABSTRACT The mosquito Aedes albopictus (Skuse, 1895) is a potent vector of several arboviral diseases, most notably chikungunya and dengue fever. In the context of the sterile insect technique (SIT), the sterilization of the male mosquitoes before their release can be achieved by gamma-ray irradiation. As gamma-ray irradiators are becoming increasingly problematic to purchase and transport, the suitability of an X-ray irradiator as an alternative for the sterilization of Ae. albopictus males was studied. The sterilization of up to 200,000 pupae at one time can be achieved with relative ease, and the sterility results obtained were comparable with those achieved by gamma irradiation, where 99% sterility is induced with a dose of 40 Gy. A significant reduction of longevity was observed in the latter stages of the males’ life after irradiation treatments, especially at doses >40 Gy, which is consistent with the negative effects on longevity induced by similar radiation doses using gamma rays. Females irradiated at 40 Gy were not only 100% sterile, but also failed to oviposit entirely, i.e., all of the females laid 0 eggs. Overall, it was found that the X-ray irradiator is generally suitable for the sterilization process for sterile insect technique programs, as it showed a high processing capacity, practicality, high effectiveness, and reproducibility.

Laboratory selection for an accelerated mosquito sexual development rate

BackgroundSeparating males and females at the early adult stage did not ensure the virginity of females of Anopheles arabiensis (Dongola laboratory strain), whereas two years earlier this method had been successful. In most mosquito species, newly emerged males and females are not able to mate successfully. For anopheline species, a period of 24 h post-emergence is generally required for the completion of sexual maturation, which in males includes a 180° rotation of the genitalia. In this study, the possibility of an unusually shortened sexual maturity period in the laboratory-reared colony was investigated.MethodsThe effect of two different sex-separation methods on the virginity of females was tested: females separated as pupae or less than 16 h post-emergence were mated with males subjected to various doses of radiation. T-tests were performed to compare the two sex-separation methods. The rate of genitalia rotation was compared for laboratory-reared and wild males collected as pupae in Dongola, Sudan, and analysed by Z-tests. Spermatheca dissections were performed on females mated with laboratory-reared males to determine their insemination status.ResultsWhen the sex-separation was performed when adults were less than 16 h post-emergence, expected sterility was never reached for females mated with radio-sterilized males. Expected sterility was accomplished only when sexes were separated at the pupal stage. Observation of genitalia rotation showed that some males from the laboratory strain Dongola were able to successfully mate only 11 h after emergence and 42% of the males had already completed rotation. A small proportion of the same age females were inseminated. Wild males showed a much slower genitalia rotation rate. At 17 h post-emergence, 96% of the laboratory-reared males had completed genitalia rotation whereas none of the wild males had.ConclusionThis colony has been cultured in the laboratory for over one hundred generations, and now has accelerated sexual maturation when compared with the wild strain. This outcome demonstrates the kinds of selection that can be expected during insect colonization and maintenance, particularly when generations are non-overlapping and similar-age males must compete for mates.

Current status and future challenges for controlling malaria with the sterile insect technique: Technical and social perspectives

The intolerable burden of malaria, when faced with high levels of drug resistance, increasing insecticide resistance and meagre resources at the national level, remains a great public health challenge to governments and the research/control community. Efficient control methods against the vectors of malaria are desperately needed. Control strategies for malaria that integrate the transfer of sterile sperm by released males to wild virgin females with other control tactics are currently being developed, and optimised mass-rearing, irradiation and release techniques are being validated in several field sites. However, the success of this strategy as part of wider pest control or health management programmes strongly depends on gaining public understanding and acceptance. Here we attempt to review what progress has been made and the remaining challenges surrounding the use of the sterile insect technique against malaria from technical and social perspectives.

Genetic sex separation of the malaria vector, Anopheles arabiensis, by exposing eggs to dieldrin

BackgroundThe sterile insect technique (SIT) has been used with success for suppressing or eliminating important insect pests of agricultural or veterinary importance. In order to develop SIT for mosquitoes, female elimination prior to release is essential as they are the disease-transmitting sex. A genetic sexing strain (GSS) of Anopheles arabiensis was created based on resistance to dieldrin, and methods of sex separation at the egg stage were developed. The use of this strain for SIT will require sexually sterile males: useful radiation doses for this purpose were determined for pupae and adults.MethodsFor the creation of the sexing strain, dieldrin-resistant males were irradiated with 40 Gy using a 60Co source and were subsequently crossed to homozygous susceptible virgin females. Individual families were screened for semi-sterility and for male resistance to dieldrin. For sex separation, eggs of a resulting GSS, ANO IPCL1, were exposed to varying concentrations of dieldrin for different durations. Percent hatch, larval survival, and male and female emergence were recorded. Radiation induced sterility was determined following adult and pupa exposure to gamma rays at 0–105 Gy. Mortality induced by dieldrin treatment, and levels of sterility post radiation were investigated.ResultsANO IPCL1 contains a complex chromosome aberration that pseudo-links the male-determining Y chromosome and dieldrin resistance, conferring high natural semi-sterility. Exposure of eggs to 2, 3, and 4 ppm dieldrin solutions resulted in complete female elimination without a significant decrease of male emergence compared to the controls. A dose of 75 Gy reduced the fertility to 3.8 and 6.9% when males were irradiated as pupae or adults respectively, but the proportions of progeny of these males reaching adulthood were 0.6 and 1.5% respectivelyConclusionThe GSS ANO IPCL1 was shown to be a suitable strain for further testing for SIT though high semi-sterility is a disadvantage for mass rearing.

Ovipositional behavior in the context of mass rearing of Anopheles arabiensis.

Abstract Large-scale production of mosquitoes is a key factor for a successful sterile insect technique program. A manageable mass-production cage must contain appropriate features for adult resting, mating, feeding, and ovipositional activities. In order to maximize egg collections, tests were conducted to determine the physical characteristics of ovipositional sites for caged Anopheles arabiensis. Effects of texture, shade, height, and shape of the ovipositional container on female behavior were investigated. Results indicate a strong preference for oviposition on humid substrates over free-standing water. The shade and texture of the cups walls also influenced site choice, with black rough inner vertical walls of the cup resulting in the largest number of eggs. Ovipositional sites with square shape were preferred rather than circular cups, and in the square cups, >60% of the eggs were laid in the corners. Height also affected oviposition, as An. arabiensis significantly favored the lowest sites even though some oviposition occurred at higher sites. Based on this study and from available literature, we determined the characteristics of an artificial ovipositional site that includes all these characteristics, which will yield large numbers of eggs required for mass production of this species.

Comparisons of Life-History Characteristics of a Genetic Sexing Strain with Laboratory Strains of Anopheles arabiensis (Diptera: Culicidae) from Northern Sudan

ABSTRACT A genetic sex separation strain (GSS) has been created for Anopheles arabiensis (Patton) (Diptera: Culicidae), one of the major African malaria vectors, for use in controlling wild populations of this species via the sterile insect technique (SIT). This GSS strain, “ANO IPCL1,” allows sex separation by a translocation linking a dieldrin resistance allele and the Y chromosome. Differences between ANO IPCL1 relative to wild strains might reflect its field performance and therefore are of concern. Of more immediate interest is how differences might affect production during mass rearing. Life-history parameters were measured for the ANO IPCL1 strain and the two wild strains from which it originated. Although developmental rate differences were found among them, none were large. However, a major observed variation was the very low intrinsic fertility of ANO IPCL1 because of the translocation itself. This resulted in a much lower rate of increase: ANO IPCL1 was able to double its population size, in 7.8 ± 0.4 d, whereas Dongola and Sennar strains could do so in 4.9 ± 0.5 and 5.6 ± 0.4 d. The presence of the Y-autosome translocation mainly affected the natural fertility of the males, and this will require amplification steps during mass rearing.

Advancing vector biology research: a community survey for future directions, research applications and infrastructure requirements

Vector-borne pathogens impact public health, animal production, and animal welfare. Research on arthropod vectors such as mosquitoes, ticks, sandflies, and midges which transmit pathogens to humans and economically important animals is crucial for development of new control measures that target transmission by the vector. While insecticides are an important part of this arsenal, appearance of resistance mechanisms is increasingly common. Novel tools for genetic manipulation of vectors, use of Wolbachia endosymbiotic bacteria, and other biological control mechanisms to prevent pathogen transmission have led to promising new intervention strategies, adding to strong interest in vector biology and genetics as well as vector–pathogen interactions. Vector research is therefore at a crucial juncture, and strategic decisions on future research directions and research infrastructure investment should be informed by the research community. A survey initiated by the European Horizon 2020 INFRAVEC-2 consortium set out to canvass priorities in the vector biology research community and to determine key activities that are needed for researchers to efficiently study vectors, vector-pathogen interactions, as well as access the structures and services that allow such activities to be carried out. We summarize the most important findings of the survey which in particular reflect the priorities of researchers in European countries, and which will be of use to stakeholders that include researchers, government, and research organizations. Graphical Abstract