Biosecurity and Management Strategies for Economically Important Exotic Tephritid Fruit Fly Species in Australia.

Exotic tephritid incursions are of high concern to Australia's biosecurity and its horticultural industries. It is vital that Australia remains ready to respond to incursions as they arise, as an incursion of tephritid fruit fly species will result in significant economic losses. In this review, we compared Australian incursion management strategies for fruit flies with global management strategies and identified possible areas where improvements could be made in an Australian context. Overall, Australia has a good understanding of the main tephritid threats, of which Bactrocera species from across the Torres Strait (northern Australia) are of most concern. Effective tools for tephritid detection and early warning surveillance at points of entry are in place at ports and in horticultural areas Australia-wide and provide the basis for initiating biosecurity responses in the event of an incursion. Area-wide control measures used in successful eradication attempts globally are available for use in Australia. However, a specific tephritid emergency response plan identifying suitable response measures and control options for species of concern is not yet available. We have identified that Australia has the policies and management tools available to respond to an exotic tephritid incursion, but the speed at which this could be accomplished would be greatly improved by the development of species-specific emergency response plans.

Raspberry ketone feeding makes Queensland fruit fly, Bactrocera tryoni (Froggatt), more vulnerable to desiccation but not starvation.

BACKGROUND: Queensland fruit fly (Qfly) males exhibit accelerated sexual maturation when their diet is supplemented with raspberry ketone (RK) for 48 h following emergence, which is beneficial for sterile insect technique operation. The present study tests whether RK supplementation makes Qfly more vulnerable to starvation or desiccation. RESULTS: Flies were fed for 48 h with a yeast hydrolysate and sugar diet (1:3) that contained 0% RK (control), 1.25% RK (low dose) or 5% RK (high dose) to test subsequent vulnerability to starvation and desiccation. RK feeding decreased body weight and water content in males and increased lipid levels in both sexes before exposure to any stress treatment. Under nutritional stress, flies fed the low RK dose, but not the high RK dose, had higher survival than controls. Under desiccation stress, flies fed both the low and high RK doses had lower survival than the controls. Body weight, water content and lipid reserves at death were all affected by RK dose when under nutritional stress, but not when under desiccation stress. In the absence of stress, body weight at death was higher than controls in flies provided with the high RK dose and lipids were lower than controls in flies provided with the low RK dose. CONCLUSION: Feeding with RK makes Qflies more vulnerable to desiccation but not starvation. In most conditions, it is expected that the disadvantage of increased desiccation vulnerability would be outweighed by the benefits of accelerated sexual maturation in RK-fed young adult Qflies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Methoprene treatment increases activity, starvation and desiccation risk of Queensland fruit fly.

Juvenile hormone is an important regulator of sexual development in insects, and application of methoprene, a juvenile hormone analogue, together with access to a protein-rich diet, has been found to accelerate sexual maturation of several tephritid fruit fly species including Queensland fruit fly Bactrocera tryoni ('Q-fly'). Such accelerated development is a potentially valuable means to increase participation of released males in sterile insect technique programs. However, there is a risk that benefits of accelerated maturation might be countered by increased vulnerability to starvation and desiccation. The present study investigates this possibility. After emergence, flies were treated with three levels of methoprene (0, 0.05%, and 0.5%) incorporated into a diet of sugar and yeast hydrolysate for two days after emergence. Survival of groups and individual flies was assessed under conditions of food stress, food and water stress, and ad libitum access to diet, and survival of individual flies was also assessed under desiccation stress. Most flies provided ad libitum access to diet were still alive at 7 days, whereas all stressed flies died within 4 days. Desiccation stressed flies had the shortest survival followed by food and water stress, and then food stress. Methoprene supplements increased susceptibility of flies to each stress. Flies subjected to food and water stress had the least lipid reserves at death, whereas flies subjected to desiccation stress retained the least water reserves. To investigate mechanisms that might underlie reduced survival under stress; we also quantified activity level of flies that were subjected to food and water stress and desiccation stress. Activity level was greater for flies provided methoprene, but did not vary with stress type or sex, suggesting that increased vulnerability of flies to stress is related to elevated metabolism associated with elevated activity. Deleterious effects of methoprene supplements on stress tolerance indicate a need for careful consideration of the conditions that will be encountered by flies in the field before deploying methoprene as a pre-release treatment in Q-fly sterile insect technique programs.

Pre-release dietary supplements of methoprene and raspberry ketone increase field abundance of sterile Queensland fruit flies (Diptera: Tephritidae).

The sterile insect technique (SIT) is a sustainable pest management tool based on the release of millions of sterile insects that suppress reproduction in targeted populations. Success of SIT depends on survival, maturation, dispersal, and mating of released sterile insects. Laboratory and field cage studies have demonstrated that dietary supplements of methoprene and raspberry ketone (RK) promote sexual maturation of adult Queensland fruit fly, Bactrocera tryoni (Froggatt), and may hence shorten the delay between release and maturity in the field. We investigated the effects of methoprene and RK dietary supplements on field abundance of sexually mature sterile Q-flies relative to untreated flies fed only sugar and yeast hydrolysate before release at 2 d of age. Compared with untreated flies, more methoprene- and RK-treated flies were recaptured in cuelure traps to which only sexually mature males are attracted. At distances of 100 and 200 m from the release point, recapture rates were higher for methoprene- and RK-treated flies than for untreated flies, but at 300 m recapture rates were low and were similar for treated and untreated flies. Rainfall, relative humidity, wind speed, and wind direction did not affect recapture rates, but temperature was positively correlated with recapture rates for all treatments. There was a strong correlation between the number of sterile and wild flies caught in traps, indicating co-location in the field. Dietary supplements of methoprene and RK can substantially increase abundance of sexually mature sterile male Q-flies in the field following release as 2-d-old immature adults.

Canopy distribution and microclimate preferences of sterile and wild Queensland fruit flies.

Insects tend to live within well-defined habitats, and at smaller scales can have distinct microhabitat preferences. These preferences are important, but often overlooked, in applications of the sterile insect technique. Different microhabitat preferences of sterile and wild insects may reflect differences in environmental tolerance and may lead to spatial separation in the field, both of which may reduce the control program efficiency. In this study, we compared the diurnal microhabitat distributions of mass-reared (fertile and sterile) and wild Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Flies were individually tagged and released into field cages containing citrus trees. We recorded their locations in the canopies (height from ground, distance from canopy center), behavior (resting, grooming, walking, feeding), and the abiotic conditions on occupied leaves (temperature, humidity, light intensity) throughout the day. Flies from all groups moved lower in the canopy when temperature and light intensity were high, and humidity was low; lower canopy regions provided shelter from these conditions. Fertile and sterile mass-reared flies of both sexes were generally lower in the canopies than wild flies. Flies generally fed from the top sides of leaves that were lower in the canopy, suggesting food sources in these locations. Our observations suggest that mass-reared and wild B. tryoni occupy different locations in tree canopies, which could indicate different tolerances to environmental extremes and may result in spatial separation of sterile and wild flies when assessed at a landscape scale.

Effect of Chilling on Quality Control Parameters of Sterile Queensland Fruit Fly (Diptera: Tephritidae).

Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt), presents a major threat to Australian fruit production and trade. The sterile insect technique (SIT) is increasingly employed to manage Q-fly. Quality of sterile males released in SIT programs, and hence program efficacy, can be affected by pre- and post-production processes, such as mass rearing, packing, irradiation, transportation, and release. Given long distances from rear-out facilities to release sites, adult flies are usually chilled to reduce metabolism and stress during transportation. To guide SIT procedures, it is important to understand the impact of such practices on performance of sterile Q-fly. The present study assesses the effect of chilling temperature and exposure period on quality parameters of sterile Q-fly. We considered the effects of two temperature regimes (4 and 6°C) and six exposure periods (0, 1, 2, 4, 6, and 12 h) on chill-coma recovery time, flight ability, survival under nutritional stress, and longevity of both males and females. Flies chilled at 4°C took longer to recover than that those chilled at 6°C. Flight ability, survival under nutritional stress, and longevity all decreased as chilling period increased but did not differ between the two tested temperatures. We recommend that periods of chilling during transportation from rear-out facilities to release sites be minimized in order to retain quality of sterile Q-fly and that increased release rates be considered when longer chilling periods are required.

Spatio-temporal distribution of sexual calling behaviour in domesticated, sterile and wild Queensland fruit fly males under field cage conditions.

BACKGROUND: The sterile insect technique (SIT) is used in Australia to contain and eradicate outbreaks of Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) in fruit-fly-free regions, and to manage populations in some endemic regions. To assess the likely impact of SIT on wild B. tryoni populations it is important to assess the colocation and synchrony of male calling between sterile and wild flies. We observed the location and timing of calling behaviours of marked mass-reared (fertile and sterile) and wild B. tryoni males in walk-in field cages. RESULTS: We found that wild males called further from the canopy centre than mass-reared (fertile or sterile) males. Mass-reared (fertile or sterile) males called earlier in the evening than wild males and, consequently, mass-reared males called when temperature and light intensity were higher than when wild males called. CONCLUSION: Male calling is a prerequisite to mating among dacine fruit flies. Therefore, our observations of spatio-temporal divergence in male calling behaviour may lead to assortative mating between mass-reared and wild B. tryoni in SIT applications. The importance of these spatio-temporal differences warrants further inquiry, with particular focus on how environmental conditions modify calling behaviour and avenues to ameliorate differences between sterile and wild flies. © 2021 Society of Chemical Industry.

Electrophysiological Responses to Cuelure of Raspberry Ketone-Fed Queensland Fruit Flies.

The sterile insect technique (SIT) and male annihilation technique (MAT) are important tools for the control of Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major insect pest of horticultural crops in Australia. In MAT, mature Q-fly males are attracted to a toxic bait using Cuelure, a synthetic analog of raspberry ketone (RK). Substantial improvements in control could be achieved by simultaneous use of SIT and MAT, but this requires suppression of the Cuelure response in released sterile flies. Recent studies report that prerelease feeding with RK during the first 48 h after emergence can reduce the response of mature Q-fly males to Cuelure, but the mechanism underpinning this is unknown. Here, to test whether reduced sensory sensitivity to Cuelure is involved, we evaluated the effects of RK supplements, adult diet (yeast-supplemented diet throughout adult stage vs yeast-supplemented diet only for 48 h), and age on electroantennogram (EAG) and electropalpogram (EPG) responses of Q-flies to Cuelure stimuli. EAG responses did not vary with RK supplements, sex, or age of Q-flies fed yeast-supplemented diet throughout the adult stage, but the responses of Q-flies fed other diet regime decreased with age. EPG responses of both sexes of Q-flies were affected by RK supplements, age, and their interaction, but without patterns that might indicate reduced maxillary palp response of RK supplemented flies to Cuelure. Our findings do not support the hypothesis that reduced Cuelure response of male Q-flies fed RK supplements is explained by reduced electrophysiological response in antennae or maxillary palps.

Caffeine as a promotor of sexual development in sterile Queensland fruit fly males.

Sterile insect technique (SIT) is an environmentally benign pest management technique that involves releasing millions of sterile insects to suppress reproduction of pest populations. Many fruit flies, including Queensland fruit fly (Bactrocera tryoni Froggatt, 'Q-fly'), have long adult maturation periods such that pre-maturation mortality can greatly reduce abundance of sexually active sterile males and impede SIT efficacy. Q-fly is the most difficult and costly challenge to market access for Australia's horticulture industries, and has been targeted for intensive use of SIT program. We here demonstrate potential of pre-release caffeine supplements as a novel means to accelerate sexual maturation in male Q-fly. In mating trials, analytical caffeine was very effective at accelerating sexual maturation, while no positive effects of caffeine-containing instant coffee or guarana supplements were detected. In parallel, development of testes and ejaculatory apodemes was accelerated in males provided analytical caffeine but not instant coffee or guarana. High doses of guarana and instant coffee reduced longevity while even the highest doses of analytical caffeine did not affect longevity. Pre-release caffeine supplements promote sexual maturation in Q-flies, and similar benefits are expected in other fruit flies having long adult maturation periods.

Accelerated Sexual Maturation in Methoprene-Treated Sterile and Fertile Male Queensland Fruit Flies (Diptera: Tephritidae), and Mosquito Larvicide as an Economical and Effective Source of Methoprene.

Queensland fruit flies Bactrocera tryoni ('Q-fly') have long adult prereproductive development periods, which can present challenges for sterile insect technique (SIT) programs. Holding the sterile flies in release facilities is expensive for control programs. Alternatively, releases of sexually immature males can lead to substantial mortality of sterile males before they mature. Recent studies have reported effectiveness of dietary supplementation with a mosquito larvicide (NOMOZ) that contains S-methoprene, a juvenile hormone analogue, for accelerating sexual development of fertile Q-fly males. However, it is not known whether effects on sterile flies are comparable to effects on fertile flies, or whether effects of methoprene-containing larvicide are comparable to effects of analytical standard methoprene such has been used in most studies. Here we address both knowledge gaps, investigating the effects of analytical standard methoprene and NOMOZ mixed with food and provided for 48 h following emergence on sexual development and longevity of fertile and sterile Q-flies. Compared with controls, fertile and sterile male Q-flies that were provided diets supplemented with methoprene from either source exhibited substantially accelerated sexual development by 2-3 d and longer mating duration. Unlike males, females did not respond to methoprene treatment. Although fertile and sterile flies were generally similar in sexual development and response to methoprene treatment, sterile flies of both sexes tended to have shorter copula duration than fertile flies. Neither methoprene supplements nor sterilization affected longevity of flies. The present study confirms effectiveness of dietary methoprene supplements in accelerating sexual development of both fertile and sterile male (but not female) Q-flies, and also confirms that low-cost mosquito larvicides that contain methoprene can achieve effects similar to those for high-cost analytical grade methoprene as prerelease supplements for Q-fly SIT.

Effect of natural predators on Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) control by sterile insect technique (SIT).

BACKGROUND: Queensland fruit fly (Q-fly) is a destructive insect pest that infests a wide variety of agricultural plants in Australia. The sterile insect technique (SIT) is used to manage Q-flies, but the effectiveness of SIT has not been tested in the presence of natural predators. The objective of this study was to investigate the effect of natural predators and SIT on the survival and reproduction of laboratory reared Q-flies under semi-natural conditions. We altered the presence of predators and irradiated Q-fly males, and measured survival, number of eggs laid and egg-hatching rate. RESULTS: The presence of natural predators significantly affected the survival of Q-flies and appeared to decrease the number of eggs laid. Interestingly, we found that both sterile and fertile males were more prone to predation than females, but we found no difference among males. The presence of sterile males significantly reduced Q-fly fertility, but the interaction of natural predators and sterile males did not significantly reduce the number of fertile eggs. CONCLUSION: Our findings highlight the important role of natural predators in controlling Q-flies together with SIT and provide a solid foundation for similar large-scale field trials using wild counterparts. © 2019 Society of Chemical Industry.

One for all: Mating compatibility among various populations of olive fruit fly (Diptera: Tephritidae) for application of the sterile insect technique.

The olive fruit fly, Bactrocera oleae (Rossi), is the most important insect pest for the cultivation of olives worldwide. Considerable research efforts have been invested in the past decades to develop eradication or suppression tactics for use within an area-wide integrated pest management (AW-IPM) approach that includes a sterile insect technique (SIT) component. One of the major obstacles encountered in the development of SIT for olive fruit fly was the inferior quality of the mass-reared flies, expressed among others evident primarily by sterile males having a different timing of peak mating and a lower mating propensity in comparison with their wild counterparts. In this study we assessed the mating behaviour and mating compatibility of olive fruit flies originating from four countries of the Mediterranean region (Croatia, France, Italy, Spain) in walk-in field cages and post zygotic compatibility (expressed as % egg hatch) under laboratory conditions. Furthermore, we tested the hypothesis whether a hybrid strain (Greece (domesticated)/Israel (wild)) adapted to laboratory rearing conditions showed any mating barriers with all the four "wild" populations. Finally, we examined the effect of colonization on the mating compatibility of the four newly established populations over three consecutive generations. The results showed no pre-zygotic (mating barriers) or post-zygotic isolations (measured by egg hatch%) among the olive fruit fly populations from the four countries tested. Also, there was no evidence of mating barriers between the hybrid strain and the wild populations of the Mediterranean region.

Correction: Suppression of cuelure attraction in male Queensland fruit flies provided raspberry ketone supplements as immature adults.

[This corrects the article DOI: 10.1371/journal.pone.0184086.].

Suppression of cuelure attraction in male Queensland fruit flies provided raspberry ketone supplements as immature adults.

Tephritid fruit flies are amongst the most damaging insect pests of horticulture globally. Some of the key fruit fly species are managed using the sterile insect technique (SIT), whereby millions of sterile males are released to suppress reproduction of pest populations. Male annihilation technique (MAT), whereby sex specific lures are used to attract and kill males, is often used to reduce wild male numbers before SIT programs commence, providing released sterile males an increased numerical advantage. Overall program efficacy might be improved if MAT could be deployed simultaneously with SIT, continuously depleting fertile males from pest populations and replacing them with sterile males. However, such 'male replacement' requires a means of suppressing attraction of released sterile males to lures used in MAT. Previous studies have found that exposure of some fruit flies to lure compounds as mature adults can suppress subsequent response to those lures, raising the possibility of pre-release treatments. However, this approach requires holding flies until after maturation for treatment and then release. The present study takes a novel approach of exposing immature adult male Queensland fruit flies (Bactrocera tryoni, or 'Qfly') to raspberry ketone (RK) mixed in food, forcing these flies to ingest RK at ages far younger than they would naturally. After feeding on RK-supplemented food for two days after emergence, male Qflies exhibited a reduction in attraction to cuelure traps that lasted more than 20 days. This approach to RK exposure is compatible with current practises, in which Qflies are released as immature adults, and also yields advantages of accelerated reproductive development and increased mating propensity at young ages.

Evaluation of Quality Production Parameters and Mating Behavior of Novel Genetic Sexing Strains of the Mediterranean Fruit Fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae).

The Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is one of the most important pest of fruits and vegetables in tropical and subtropical countries. The sterile insect technique (SIT) as a component of area-wide integrated pest management (AW-IPM) approaches is being used for the successful management of this pest. VIENNA 8 is a genetic sexing strain (GSS) that has a white pupae (wp) and temperature sensitive lethal (tsl) mutation, the latter killing all female embryos when eggs are exposed to high temperatures (34°C). The use of this GSS permits production and the release of only males which has increased the cost effectiveness of the SIT several fold for this pest. An efficient method of identification of recaptured sterile males can further increase the cost effectiveness of the SIT for this pest. Therefore, VIENNA 8-Sergeant2 (Sr2) strain and the transgenic strain VIENNA 8-1260 having visible markers were constructed. All three strains were evaluated for egg production, egg hatch, and egg sterility parameters under semi mass-rearing conditions and mating competitiveness in field cages. VIENNA 8-1260 females produced significantly fewer eggs as compared with the two other strains, which produced similar numbers of eggs. However, egg hatch of all strains was similar. Egg hatch of eggs produced by untreated females that had mated with adult males that had been irradiated with 100 Gy as pupae 2 days before emergence, was different for the three strains, i.e., egg hatch of 0.63%, 0.77%, 0.89% for VIENNA 8, VIENNA 8-1260, and VIENNA 8-Sr2, respectively. Differences in male mating competitiveness of the three strains against wild-type males were gradually reduced with successive generations under semi mass-rearing conditions. However, VIENNA 8 males adapted faster to laboratory conditions as compared with VIENNA 8-Sr2 and VIENNA 8-1260 males with respect to mating competitiveness. VIENNA 8 males of the F10 generation were equally competitive with wild-type males, whereas the mating competitiveness of VIENNA 8-Sr2 and VIENNA 8-1260 males was similar but lower as compared with wild-type males. Males from all three strains copulated earlier than wild-type males. Results are discussed in relation with the potential benefits of incorporating novel strains for more effective SIT application.

Olive Fruit Fly (Bactrocera oleae) Population Dynamics in the Eastern Mediterranean: Influence of Exogenous Uncertainty on a Monophagous Frugivorous Insect.

Despite of the economic importance of the olive fly (Bactrocera oleae) and the large amount of biological and ecological studies on the insect, the factors driving its population dynamics (i.e., population persistence and regulation) had not been analytically investigated until the present study. Specifically, our study investigated the autoregressive process of the olive fly populations, and the joint role of intrinsic and extrinsic factors molding the population dynamics of the insect. Accounting for endogenous dynamics and the influences of exogenous factors such as olive grove temperature, the North Atlantic Oscillation and the presence of potential host fruit, we modeled olive fly populations in five locations in the Eastern Mediterranean region. Our models indicate that the rate of population change is mainly shaped by first and higher order non-monotonic, endogenous dynamics (i.e., density-dependent population feedback). The olive grove temperature was the main exogenous driver, while the North Atlantic Oscillation and fruit availability acted as significant exogenous factors in one of the five populations. Seasonal influences were also relevant for three of the populations. In spite of exogenous effects, the rate of population change was fairly stable along time. We propose that a special reproductive mechanism, such as reproductive quiescence, allows populations of monophagous fruit flies such as the olive fly to remain stable. Further, we discuss how weather factors could impinge constraints on the population dynamics at the local level. Particularly, local temperature dynamics could provide forecasting cues for management guidelines. Jointly, our results advocate for establishing monitoring programs and for a major focus of research on the relationship between life history traits and populations dynamics.

Conserved metallomics in two insect families evolving separately for a hundred million years.

Μetal cofactors are required for enzymatic catalysis and structural stability of many proteins. Physiological metal requirements underpin the evolution of cellular and systemic regulatory mechanisms for metal uptake, storage and excretion. Considering the role of metal biology in animal evolution, this paper asks whether metal content is conserved between different fruit flies. A similar metal homeostasis was previously observed in Drosophilidae flies cultivated on the same larval medium. Each species accumulated in the order of 200 µg iron and zinc and approximately ten-fold less manganese and copper per gram dry weight of the adult insect. In this paper, data on the metal content in fourteen species of Tephritidae, which are major agricultural pests worldwide, are presented. These fruit flies can be polyphagous (e.g., Ceratitis capitata) or strictly monophagous (e.g., Bactrocera oleae) or oligophagous (e.g., Anastrepha grandis) and were maintained in the laboratory on five distinct diets based on olive oil, carrot, wheat bran, zucchini and molasses, respectively. The data indicate that overall metal content and distribution between the Tephritidae and Drosophilidae species was similar. Reduced metal concentration was observed in B. oleae. Feeding the polyphagous C. capitata with the diet of B. oleae resulted in a significant quantitative reduction of all metals. Thus, dietary components affect metal content in some Tephritidae. Nevertheless, although the evidence suggests some fruit fly species evolved preferences in the use or storage of particular metals, no metal concentration varied in order of magnitude between these two families of Diptera that evolved independently for over 100 million years.

Genetic elimination of field-cage populations of Mediterranean fruit flies.

The Mediterranean fruit fly (medfly, Ceratitis capitata Wiedemann) is a pest of over 300 fruits, vegetables and nuts. The sterile insect technique (SIT) is a control measure used to reduce the reproductive potential of populations through the mass release of sterilized male insects that mate with wild females. However, SIT flies can display poor field performance, due to the effects of mass-rearing and of the irradiation process used for sterilization. The development of female-lethal RIDL (release of insects carrying a dominant lethal) strains for medfly can overcome many of the problems of SIT associated with irradiation. Here, we present life-history characterizations for two medfly RIDL strains, OX3864A and OX3647Q. Our results show (i) full functionality of RIDL, (ii) equivalency of RIDL and wild-type strains for life-history characteristics, and (iii) a high level of sexual competitiveness against both wild-type and wild-derived males. We also present the first proof-of-principle experiment on the use of RIDL to eliminate medfly populations. Weekly releases of OX3864A males into stable populations of wild-type medfly caused a successive decline in numbers, leading to eradication. The results show that genetic control can provide an effective alternative to SIT for the control of pest insects.

Sex allocation in a polyembryonic parasitoid with female soldiers: an evolutionary simulation and an experimental test.

Parasitoid wasps are convenient subjects for testing sex allocation theory. However, their intricate life histories are often insufficiently captured in simple analytical models. In the polyembryonic wasp Copidosoma koehleri, a clone of genetically identical offspring develops from each egg. Male clones contain fewer individuals than female clones. Some female larvae develop into soldiers that kill within-host competitors, while males do not form soldiers. These features complicate the prediction of Copidosoma's sex allocation. We developed an individual-based simulation model, where numerous random starting strategies compete and recombine until a single stable sex allocation evolves. Life-history parameter values (e.g., fecundity, clone-sizes, larval survival) are estimated from experimental data. The model predicts a male-biased sex allocation, which becomes more extreme as the probability of superparasitism (hosts parasitized more than once) increases. To test this prediction, we reared adult parasitoids at either low or high density, mated them, and presented them with unlimited hosts. As predicted, wasps produced more sons than daughters in all treatments. Males reared at high density (a potential cue for superparasitism) produced a higher male bias in their offspring than low-density males. Unexpectedly, female density did not affect offspring sex ratios. We discuss possible mechanisms for paternal control over offspring sex.

Control of the olive fruit fly using genetics-enhanced sterile insect technique.

BACKGROUND: The olive fruit fly, Bactrocera oleae, is the major arthropod pest of commercial olive production, causing extensive damage to olive crops worldwide. Current control techniques rely on spraying of chemical insecticides. The sterile insect technique (SIT) presents an alternative, environmentally friendly and species-specific method of population control. Although SIT has been very successful against other tephritid pests, previous SIT trials on olive fly have produced disappointing results. Key problems included altered diurnal mating rhythms of the laboratory-reared insects, resulting in asynchronous mating activity between the wild and released sterile populations, and low competitiveness of the radiation-sterilised mass-reared flies. Consequently, the production of competitive, male-only release cohorts is considered an essential prerequisite for successful olive fly SIT. RESULTS: We developed a set of conditional female-lethal strains of olive fly (named Release of Insects carrying a Dominant Lethal; RIDL®), providing highly penetrant female-specific lethality, dominant fluorescent marking, and genetic sterility. We found that males of the lead strain, OX3097D-Bol, 1) are strongly sexually competitive with wild olive flies, 2) display synchronous mating activity with wild females, and 3) induce appropriate refractoriness to wild female re-mating. Furthermore, we showed, through a large proof-of-principle experiment, that weekly releases of OX3097D-Bol males into stable populations of caged wild-type olive fly could cause rapid population collapse and eventual eradication. CONCLUSIONS: The observed mating characteristics strongly suggest that an approach based on the release of OX3097D-Bol males will overcome the key difficulties encountered in previous olive fly SIT attempts. Although field confirmation is required, the proof-of-principle suppression and elimination of caged wild-type olive fly populations through OX3097D-Bol male releases provides evidence for the female-specific RIDL approach as a viable method of olive fly control. We conclude that the promising characteristics of OX3097D-Bol may finally enable effective SIT-based control of the olive fly.