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.
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.
Review of the male reproductive biology of the disease vector Aedes mosquito. Which parameters are of greatest importance to improve genetic control programmes?
•Aedes mosquito can be vector of ...serious pathogens to humans.•We review the literature on the reproductive biology of male Aedes mosquito.•Which parameters are of greatest importance to improve genetic control programmes?
Among Aedes mosquitoes are species responsible for transmission of serious pathogens to humans. To cope with the current threats to long-term effectiveness of the traditional vector control methods, non-conventional control strategies are being developed. These include autocidal control such as the release of sterile males (sterile insect technique) and the release of Wolbachia-infected males to induce sexual sterility (incompatible insect technique) and pathogen-refractory strain replacement variations using Wolbachia. Sterile male types of techniques particularly depend on released males’ ability to successfully mate with wild females. For that reason, a good understanding of male mating biology, including a thorough understanding of the reproductive system and mating capacity, increases the likelihood of success of such genetic vector control programmes. Here we review the literature concerning the reproduction of Aedes mosquitoes with an emphasis on the male biology. We consider sexual maturation, mate finding, insemination, male reproductive capacity, and the occurrence of multiple matings. We also discuss which parameters are of greatest importance for the successful implementation of autocidal control methods and propose questions for future research.
The sterile insect technique (SIT) can be an efficient solution for reducing or eliminating certain insect pest populations. It is widely used in agriculture against fruit flies, including the ...Mediterranean fruit fly (medfly), Ceratitis capitata. The re-mating tendency of medfly females and the fact that the released sterile males may have some residual fertility could be a challenge for the successful implementation of the SIT. Obtaining the right balance between sterility level and sterile male quality (competitiveness, longevity, etc) is the key to a cost-efficient program. Since field experimental approaches can be impacted by many environmental variables, it is difficult to get a clear understanding on how specific parameters, alone or in combination, may affect the SIT efficiency. The use of models not only helps to gather knowledge, but it allows the simulation of a wide range of scenarios and can be easily adapted to local populations and sterile male production. In this study, we consider single- and double-mated females. We first show that SIT can be successful only if the residual fertility is less than a threshold value that depends on the basic offspring number of the targeted pest population, the re-mating rates, and the parameters of double-mated females. Then, we show how the sterile male release rate is affected by the parameters of double-mated females and the male residual fertility. Different scenarios are explored with continuous and periodic sterile male releases, with and without ginger aromatherapy, which is known to enhance sterile male competitiveness, and also taking into account some biological parameters related to females that have been mated twice, either first by a wild (sterile) male and then a sterile (wild) male, or by two wild males only. Parameter values were chosen for peach as host fruit to reflect what could be expected in the Corsican context, where SIT against the medfly is under consideration. Our results suggest that ginger aromatherapy can be a decisive factor determining the success of SIT against medfly. We also emphasize the importance of estimating the duration of the refractory period between matings depending on whether a wild female has mated with a wild or sterile male. Further, we show the importance of parameters, like the (hatched) eggs deposit rate and the death-rate related to all fertile double-mated females. In general, re-mating is considered to be detrimental to SIT programs. However, our results show that, depending on the parameter values of double-mated females, re-mating may also be beneficial for SIT. Our model can be easily adapted to different contexts and species, for a broader understanding of release strategies and management options.
The 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.
For 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.
ANO 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% respectively
The 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.
Male Anopheles mosquitoes that swarm rely in part on features of the environment including visual stimuli to locate swarms. Swarming is believed to be the primary behaviour during which mating occurs ...in the field, but is not a common behaviour in the laboratory. Features that stimulate male Anopheles gambiae G3 strain swarming were created in novel large indoor cages.
The following visual features were tested in all combinations to determine which were important for swarm formation. Large cages and fading ceiling lights at dusk alone did not stimulate swarming while a dark foreground and contrasting illuminated background with a contrasting landmark stimulated and localized swarm formation during artificial twilight. Given the need to test transgenic strains in as natural a setting as possible, in this study it was investigated whether induced swarm behaviour and cage size would affect relative mating performance of wild-type and transgenic β2Ppo1 and β2Ppo2 A. gambiae sexually sterile males.
Even using a mosquito colony that has been in laboratory culture for 39 years, swarming behaviour was induced by this novel arrangement. The presence of swarming stimuli was associated with an increase in insemination frequency from 74.3 to 97.7% in large cages. Transgenic males showed a lower competitiveness in large cages compared to small cages regardless of the presence of swarming stimuli.
The results of the present study are discussed in view of the progressive evaluation of genetically modified A. gambiae strains and the potential applications of reproducing swarms in controlled conditions to dissect the mating behaviour of this species and the mechanisms controlling it.
Separating 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.
The 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.
When 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.
This 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.
Review of the technical advances and remaining challenges for genetic control of malaria vectors, and the importance of and different approaches to social science studies for such programmes.
•We ...present the history of malaria vector management leading to the SIT.•Technical advances and remaining challenges for genetic control are reviewed.•Importance and different approaches to related social science studies are discussed.
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.
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► We evaluated the longevity of males Aedes albopictus under semi-field conditions. ► The impact of radio-sterilization, sugar supply, and mating activity was tested. ► Sterilization ...and mating activity reduced male longevity. ► Immediate sugar feeding could compensate for these impairments. ► Providing sugar sources prior to release during SIT programmes appeared critical.
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.
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.
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.
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are invasive mosquito species that impose a substantial risk to human health. To control the abundance and spread of these arboviral pathogen vectors, the sterile insect technique (SIT) is ...emerging as a powerful complement to most commonly-used approaches, in part, because this technique is ecologically benign, specific, and non-persistent in the environment if releases are stopped. Because SIT and other similar vector control strategies are becoming of increasing interest to many countries, we offer here a pragmatic and accessible 'roadmap' for the pre-pilot and pilot phases to guide any interested party. This will support stakeholders, non-specialist scientists, implementers, and decision-makers. Applying these concepts will ensure, given adequate resources, a sound basis for local field trialing and for developing experience with the technique in readiness for potential operational deployment. This synthesis is based on the available literature, in addition to the experience and current knowledge of the expert contributing authors in this field. We describe a typical path to successful pilot testing, with the four concurrent development streams of Laboratory, Field, Stakeholder Relations, and the Business and Compliance Case. We provide a graphic framework with criteria that must be met in order to proceed.
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