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Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt), is a significant pest of horticultural crops in Australia. The sterile insect technique (SIT) is currently employed to eradicate outbreaks in fruit fly free regions and may also be used to suppress populations in endemic regions. For SIT to succeed, it is imperative that the released sterile males survive, disperse, attain sexual maturity, and are sexually competitive against their wild rivals. Q-fly SIT programmes have conventionally held adult flies for two to three days and have sometimes fed them only sugar before release, providing little time or nutrition for development prior to release. We investigated whether a 5-d pre-release holding period and provision of yeast hydrolysate (YH) together with sugar in the pre-release diet increase abundance of mature male Q-fly in the field. Indicating increased survivorship and/or maturation, the combination of YH feeding and 5-d pre-release holding period resulted in 6–8 times more recaptures of mature male flies in cuelure traps than was the case for flies released at 2 d with or without YH and for flies released at 5 d without YH. Flies held for 5 d and fed YH were relatively more abundant than flies from other treatments in traps close to the release point and were as abundant as other treatments in traps at the greatest assessed distances. These findings strongly support a recommendation that sterile Q-flies be provided a pre-release diet of YH and sugar and be held for 5-d post-eclosion before release.

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.

Queensland fruit fly is one of the most economically important horticultural pests in Australia. Sterile insect technique (SIT) is now being reconsidered and upscaled to combat this pest so reliable discrimination of released sterile Q-flies from wild flies in monitoring traps is important for effective SIT operations. Stable isotopes provide a permanent chemical marker to discriminate sterile and wild flies when dye marking is unclear. In this study, we compared the isotopic ratios of carbon and nitrogen between Q-flies reared on different larval diets and wild flies collected from diverse locations in Australia and New Caledonia. Finally, we conducted a release–recapture study to corroborate differences in stable isotope C and N ratios in laboratory-reared and wild Q-flies. The δ15N values obtained from wild and laboratory Q-flies showed high variability that is likely related to the food source of the larval and/or adult stage and do not offer an effective means to discriminate between sterile and wild Q-flies. The δ13C values of examined wild Q-flies ranged from − 27.46 to − 24.37‰ VPDB, whereas those from laboratory-reared, released and recaptured Q-flies ranged from − 25.73 to − 19.26‰ VPDB. Differences in δ13C values resulted in 100% correct classification of wild flies and 96.88% correct classification of released flies. Measurements of intrinsic δ13C values offer a precise tool to discriminate between sterile and wild Q-flies in SIT programs, regardless of the composition of the larval or adult pre-release diets.

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.

The sterile insect technique (SIT), a benign pest control strategy, is currently undergoing a substantial renewal for controlling the Queensland fruit fly (Q-fly), a major horticultural pest in Australia. Production and delivery of high-quality flies is fundamental to effective SIT. While efforts are commonly made to improve production at a factory level (both quantity and quality), the methods and conditions during post-production operations also need to be tuned to minimize reductions in fly quality. Hypoxia, irradiation and vibration are important stressors that are experienced by pupae during production, packing and transportation. However, little is known about the impacts of such stressors on Q-fly quality. We conducted two laboratory experiments to investigate the response of Q-fly to such post-production stressors, with the aim of then developing guidelines that minimize quality reductions for SIT programs. In Experiment 1, Q-fly pupae of different ages (3, 6 or 9-day old) were exposed to vibrations for 5, 30, 60 and 300 s. Flight ability tests revealed that 6- and 9-day-old pupae were the most sensitive to vibration, with significant reduction in emergence and percentage of fliers. In Experiment 2, the effect of hypoxia, irradiation and vibration, in isolation or in combination, was evaluated on 9-day-old pupae. Irradiation and vibration exhibited a multiplicative negative effect on flight ability parameters. It is recommended not to subject pupae to intense or extensive vibration during the late pupal stage and to avoid prolonged periods of hypoxia.

Pre‐release dietary treatment with methoprene, a juvenile hormone analogue, decreases the age at which male Queensland fruit flies mature and hence may decrease the post‐release delay until released sterile flies participate in sterile insect technique (SIT) programmes. However, if matings of young methoprene‐treated males are not effective at inducing sexual inhibition in their mates, then this treatment may not enhance SIT. The present study investigates efficacy of matings of methoprene‐treated males at inducing sexual inhibition in their mates. Methoprene incorporated into a diet of sugar and yeast hydrolysate (w/w 3:1) for 48 hr after emergence resulted in significantly increased male mating propensity when flies were

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.

Queensland fruit flies Bactrocera tryoni (Froggatt) have a long adult maturation phase which, together with high mortality rates, can substantially reduce number of released flies that survive to mature and contribute to sterile insect technique (SIT) programmes. This constraint on SIT can potentially be addressed by incorporating methoprene, a juvenile hormone analogue, into an adult diet of sugar and yeast hydrolysate for 2 days after emergence. Methoprene treatments have been found to accelerate sexual development of male Queensland fruit fly, resulting in increased mating propensity of 5–7-day-old males in no-choice laboratory trials. Before considering deployment of methoprene as a pre-release treatment in SIT, it is necessary to demonstrate mating competitiveness and compatibility of methoprene-treated flies under field-like conditions. In the present study, we assessed whether methoprene treatment increases ability of sterile males (5 and 7 days old) to compete with mature (wild or laboratory) males for matings with mature (wild or laboratory) females in field cages. We also investigated mating compatibility to test for sexual isolation between sterile flies and mature (wild or laboratory) fertile flies. In mating competitiveness tests, methoprene-treated males of either age outperformed mature wild or laboratory males for matings with mature wild or laboratory females, respectively. Untreated 5- and 7-day-old males were less competitive than mature wild or laboratory mature males and hence had lower relative sterility indexes. Methoprene-treated males mated earlier in the evening and continued mating for longer than untreated sterile males and mature wild or laboratory males. In mating compatibility trials, methoprene-treated males mated equally with methoprene-treated females and mature females, whereas methoprene-treated females tended to mate more often with mature males than with methoprene-treated males. However, untreated flies of both sexes exhibited substantial sexual isolation. Pairings that comprised methoprene-treated males and mature females had shorter mating latency and longer copulations than other pairings. Unlike males, methoprene-treated females did not exhibit changes in mating latency or duration. Overall, the present study supports the use of pre-release dietary methoprene treatment in Queensland fruit fly SIT.

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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.

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.

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.

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.

The dose–response effects of gamma radiation on the carob or date moth Ectomyelois ceratoniae (Zeller) (Lepidoptera: Pyralidae) were studied as a basic requirement for the initiation of an SIT/F1 sterility program. In this study, the effect of gamma radiation was determined on fecundity, egg hatch, longevity, and level of inherited sterility in the carob moth. Newly emerged virgin male and female carob moth adults were exposed to increasing irradiation doses (100, 150, 200, 250, 300, and 350 Gy) to determine the effective dose of gamma rays capable of sterilizing male and female adults of the carob moth. Parental fecundity and longevity was affected by increasing irradiation dose. The results indicated that a dose of 350 Gy could completely sterilize carob moth males. No larvae hatched from eggs deposited by 300 Gy irradiated females. The data indicate that 300 Gy was the fully sterilizing dose for female and sub-sterilizing dose for male carob moth. Radiation-induced deleterious inherited effects in offspring from irradiated males and females were expressed as reduction in fecundity, egg hatch, longevity, and decreased adult emergence over subsequent generations.

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.

We investigated the survival of Mediterranean fruit fly (Medfly) larval stages on yellow peppers stored for 21 d in cold environments (1.5, 4 and 7 °C), and the differences in larval mortality rates of Medflies exposed to 1.5 °C storage environments between yellow and red peppers. In addition, we investigated the effect of packing yellow peppers and exposing them to cold environments for 21 d on the quality of the fruits. The aim of the study was to investigate packing and shipping post-harvest protocols as a preliminary investigation to explore cold environments as a system to assure the export of quality peppers with low risks of surviving Medfly immature stages. This exploratory study showed that storing yellow peppers for 21 d in lined boxes (20-μm-thick macroperforated Xtend film) at 1.5 and 4 °C protected the quality of the fruit, and precluded the survival of Medfly eggs and larval stages (L1, L2 and L3). In contrast, peppers stored at 7 °C for 21 d did not completely kill immature stages of the Medfly. In addition, the study showed that Medfly larval stages exposed to 1.5 °C environments on yellow and red peppers have different tolerances to cold. Sensitivity to cold in red pepper was higher than in yellow for all larval stages. In addition, sensitivity to cold between larval stages varied between yellow and red peppers: for yellow, sensitivity preceded as follows L1 > L2 > L3, while in red peppers it was L2 > L1 > L3. Abilities to accumulate lipids and protein by feeding Medfly larvae suggest that palatability and nutrition may be partially responsible for the differences in cold tolerance.

Fruit flies in the genus Bactrocera are among the most destructive insect pests of fruits and vegetables throughout the world. A number of studies have identified volatiles from fruit flies, but few reports have demonstrated behavioral effects or sensitivities of fly antennae to these compounds. We applied a recently developed method of automated headspace analysis using SPME (Solid Phase Microextraction) fibers and GC–MS (gas chromatography mass spectrometry), termed SSGA, to reveal volatiles specific to each sex of B. zonata that are emitted in a diel periodicity. The volatiles released primarily at dusk were identified by GC–MS and chemical syntheses as several spiroacetals, pyrazines, and ethyl esters. Solvent extraction of male rectal glands or airborne collections from each sex, followed by GC–MS, showed that certain of the volatiles increase or decrease in quantity sex-specifically with age of the flies. Electroantennographic (EAG) analysis of dose-response indicates differences in sensitivities of male and female antenna to the various volatiles. Our study provides a comprehensive analysis of the volatile chemicals produced and released by B. zonata and their antennal responses. The possible pheromone and semiochemical roles of the various volatiles released by each sex and the difficulties of establishing behavioral functions are discussed.

a b s t r a c t The simulation of insect pest populations in agricultural and forest ecosystems is an important and useful tool for integrated pest management (IPM). Insect population models are mainly driven by envi- ronmental temperature data, which are usually collected from agrometeorological stations or derived from geographic statistical extrapolations. The present study describes the modeling of olive fly (Bac- trocera oleae) populations in the Eastern Mediterranean region using the MODIS (Moderate Resolution Image Spectro Radiometer) land surface temperature (LST) product from NASA TERRA satellite. These data, together with in situ temperature data, were used to estimate the tree-canopy temperatures at the pixel resolution (1 km). The estimated canopy temperature was used as input for the olive fly population model. Our main aim was to demonstrate the use of satellite-acquired information for modelling bio- logical and ecological phenomena. Eleven years (2001-2012) of olive fly population fluctuations were simulated for three different geographic locations, representing different geo-climatic conditions. The model successfully simulated the seasonal population fluctuations throughout the 11-year period and did a good job of connecting all of the life stages of the insect. To evaluate the validity of these findings, we compared them with adult olive-fly trapping data. We observed a high degree of correlation between the trapping data and our model's predictions. Here, we demonstrate that satellite thermal data can be used to predict insect pest population fluctuations for IPM purposes. The study also advances some new modelling concepts, such as the "window of opportunity" which links physiological development with chronological age.

As part of the development of a sterile insect technique (SIT) application for the Ethiopian fruit fly, Dacus ciliatus, we studied the mating behaviour of a laboratory-adapted strain (a 4-year-old colony kept for more than 40 generations) and a wild population. Effects of laboratory rearing and irradiation were assessed by carrying out mating compatibility and male mating competitiveness tests using a 1:1 ratio between irradiated (120 Gy) laboratory males and non-irradiated wild males. Mating behaviour was studied on host and non-host plants under field cage conditions. To assess the effect of mass rearing upon male performance, we repeated the mating competitiveness test using non-irradiated laboratory insects. The findings indicated a high degree of compatibility among the two populations and satisfactory competitiveness of the irradiated laboratory males (ca. 35%). The competitiveness of non-irradiated laboratory males was also ca. 35%, suggesting that no adverse effects resulted from their irradiation. Mating occurred only at twilight and mainly on the underside of leaves of non-host plants (lemon trees). Findings are discussed in view of their implications for a future application of SIT against this fruit fly pest.

The Ethiopian fruit fly, Dacus ciliatus (Loew) (Diptera: Tephritidae), is a significant pest of cucurbit crops in Asia and Africa and is currently controlled with insecticides. The sterilizing effect of gamma radiation on D. ciliatus adults was investigated to assess the suitability of sterile insect technique (SIT) for use as an alternative, non-chemical strategy for the control of this pest. Late pupae (48 h before emergence) were irradiated with 60, 80, 100, 120, and 140 Gy of gamma rays emitted by a 60Co source. Following emergence, the biological characteristics of the experimental cohorts (including all possible male-female combinations of irradiated and untreated flies) were recorded. No significant negative effects of irradiation on pupal eclosion or the ability of newly emerged flies to fly were observed. Samples of eggs at reproductive fly-ages (12-, 15-, and 17-day-old pairs) were collected and their hatch rates were assessed. At 60 Gy, females were completely sterilized, whereas complete sterilization of the males was observed only at 140 Gy (a small amount of fertility persisted even at 120 Gy). In addition to the above experiments, three fruit infestation trials were conducted with zucchini [Cucurbita pepo L. (Cucurbitaceae)] as the plant host and the pupae produced in those trials were collected and recorded. We observed significant (ca. 10%) infestation following treatment with up to 120 Gy and zero progeny only at 140 Gy, mirroring the egg-hatch results. Our findings support the feasibility of SIT for the control of D. ciliatus.

The olive fly Bactrocera oleae (Rossi) is the key pest for olive cultivation worldwide. Substantial effort has been invested in the development of the sterile insect technique (SIT) to control this pest. One of the limitations to develop SIT technology for olive fruit fly is the low ability of wild females to lay eggs in other medium than olive fruits, and their slow adaptation to oviposition in artificial substrates. In the present study, fruit grapes were used as an alternative egg collection medium to harvest eggs and young larvae from freshly colonized wild strains originating from France, Italy, Spain and Croatia. The larvae were allowed to develop into the fruits until the second instar, before they were extracted out and further reared on a standard artificial diet. Furthermore, F1 to F4 female flies were alternatively offered wax bottles to oviposit. Finally, the performance of hybrid strains created from crosses between wild and long colonised flies was assessed. The results showed that females of all 4 wild strains readily oviposited eggs in grapes and from the F2 generation onward, females from all strains were adapted to laying eggs in wax bottles. No difference was observed in eggs and pupae production among all strains tested. The findings are discussed for their implications on SIT application against olive fruit fly.

The present study investigated the patterns of microorganisms in an artificial larval diet during Dacus ciliatus (Diptera; Tephritidae) larval development. Microbial population contents in the diet of total heterotrophic bacteria, yeast and molds, coliform and lactobacilli, and their dynamics during development, were monitored. Initially, the microbial composition in diet trays failing to produce viable pupae and in trays successfully producing pupae and adult flies was characterized. The failing diet trays contained large populations of lactobacilli that increased during larval development, and low populations of coliforms. In contrast, the successful diet showed an increasing population of coliforms and a low, or undetected, population of lactobacilli. To study the hypothesis that lactobacilli affect D. ciliatus larval development, we conducted controlled inoculation experiments in which Lactobacillus plantarum was added into fresh diet at the time of egg seeding. L. plantarum inoculated trays showed no production of D. ciliatus. Control trays without lactobacilli inoculation showed variable results. One tray successfully produced viable pupae and adults, and showed a slight and slow increase in the indigenous populations of lactobacilli. The second tray, however, failed to produce pupae and showed a fast increase of the indigenous lactobacilli to very high levels. Monitored pH trends in L. plantarum-inoculated diet showed a sharp pH decrease during the first 4 days of larval development from 5 to less than 4 units, while successful diet, producing viable D. ciliatus pupae and adults, showed a moderate pH drop during most of the larval development period. The paper discusses the possible ecological interactions between D. ciliatus larvae, the microbial content of the diet and the physical properties of the diet. The discussion also points out at the usefulness of this approach in understanding and managing mass production parameters of tephritid fruit flies industrial diets used for Sterile Insect Technique.

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.

Research on the olive fruit fly Bactrocera oleae (Rossi) - rearing simplification, insect microflora and transgenic strain evaluation - yielded several findings: (1) incorporation of antibiotics in the adult diet is evidently not needed; (2) colonization appears to be easier when wild adults are collected from the field instead of using mature larvae emerging from field-collected infested olives; (3) a combination of standard solid starter with liquid (no cellulose powder) finisher impregnated in synthetic sponge larval diets was more promising compared with all-liquid diets; (4) molecular analysis revealed extensive differences in bacterial species associated with the fly between laboratory flies and strains from different olive varieties, as well as between strains originating from different seasons of the year; (5) when an enhanced green fluorescent protein transgenic strain was compared with the standard long mass-reared strain, it proved significantly inferior according to all quality control tests applied, i.e. egg production, egg hatch, larval-stage duration, pupal recovery, pupal weight, adult emergence and adult survival. The aforementioned findings are discussed in the context of mass rearing and quality requirements for more successful implementation of the sterile insect technique against this pest.

Management of the olive fly using environmentally friendly methods includes strategies based on lure and kill and/or mass-trapping. Despite a wealth of studies related to the efficacy of different lure and kill and mass-trapping systems in several olive producing areas, there are few known regarding the performance of such systems in low input olive farms of the desert-like areas of the Middle East. The present study reports on the control of the olive fly using both lure and kill and mass-trapping devices in low-input farms in the region of Tubas, Palestine, between 2010 and 2012. The effect of environmental factors, such as elevation, on general patterns of olive fly infestation and the lure and kill system's performance was also studied. Our final goal included the development of a general strategy to produce organic olive oil using alternative olive fly control methods. Initially we used a commercially available lure and kill device, the Eco-Trap®. The Eco-Trap performed relatively well during the three years of study, significantly reducing olive fly damage levels. Eco-Trap effectiveness was more evident at elevation above 400 m, where damage in the region of Tubas was more intense than at lower elevations. In a subsequent trial we tested a low-cost mass-trapping device. This device demonstrated similar levels of protection to the one seen with the Eco-Trap. Results showed that lure and kill and mass-trapping tactics can be effectively applied in the region of Tubas, and that accessible devices can be adopted to reduce control costs. The results also showed differential levels of damage throughout the region, which were related to elevation above sea level. The possibility of applying a regional management concept throughout the area by a centralized organization, such as a farmer's union based on mass-trapping systems seems feasible and is discussed.

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.

The effect of a sub-sterilizing gamma radiation dose on Dacus ciliatus adults was investigated to assess the suitability of the sterile insect technique (SIT) as an alternative method to control this pest. Late pupae (48 h prior to adult emergence) from a laboratory strain were irradiated with 120 Gy of gamma rays emitted by a 60Co source. Following adult emergence, the mortality of irradiated and non-irradiated cohorts was recorded. Over a period of 50 days after emergence, no significant negative effects of irradiation upon the longevity of male or female laboratory flies were observed. A laboratory competitiveness study (Fried test), using irradiated laboratory and wild males at a ratio of 3:1 was conducted to assess the ability of irradiated males to reduce the egg hatch rates of a wild population. The overall competitiveness was found to be ca. 0.32, suggesting a reduced, but satisfactory, quality of irradiated laboratory as compared with wild males. Based on the above findings, we calculated and proposed effective male release ratios for field application of SIT against D. ciliatus.

The present essay links historical socioeconomic processes with pest control activities and research and development (R&D) trends in plant protection. We selected the olive orchard agroecosystem, especially in Southern Europe, as a model system. We specifically followed the evolution of olive fly (Bactrocera oleae) control strategies and research activities and linked them with economic processes in the producing countries and with European policy directives. Our analysis includes the period following the Second World War and until recent times. Our main aim was to understand the socioeconomic forces that shape agroecosystem management, especially pest control. Although we only developed the case for the olive fly in Southern Europe, we believe that most human agricultural environments are subjected to similar economic, social, and environmental processes and forces. This historical account shows the complexity involved in the management of the agroecosystem and the effect of global and local factors on plant protection activities and developments, highlighting the need for a holistic approach when agricultural and research policies are formulated.

The microbiome of the olive fruit fly, Bactrocera oleae (Gmelin), a worldwide pest of olives (Olea europaea L.), has been examined for >100 yr as part of efforts to identify bacteria that are plant pathogens vectored by the fly or are beneficial endosymbionts essential for the fly's survival and thus targets for possible biological control. Because tephritid fruit flies feed on free-living bacteria in their environment, distinguishing between the transient, acquired bacteria of their diet and persistent, resident bacteria that are vertically transmitted endosymbionts is difficult. Several culture-dependent and -independent studies have identified a diversity of species in the olive fruit fly microbiome, but they have not distinguished the roles of the microbes. Candidatus Erwinia dacicola, has been proposed to be a coevolved endosymbiont of the olive fruit fly; however, this was based on limited samples from two Italian populations. Our study shows that C. Erwinia dacicola was present in all New ...

The dispersal of marked, irradiated olive fruit flies originating from a hybrid laboratory strain was studied in an olive groove located in the arid regions of southern Israel. Release–recapture experiments (eight in total) were conducted throughout a period of 5 months (July–December 2008). In each experiment, ca. 5000 flies were released. Recapture of flies was conducted using a grid of 30 yellow sticky traps set in expanding semicircles from the centre of release. Service of traps was conducted 3 and 15 days after the release. Fly quality and adult food type (only sucrose and protein + sucrose) before release was also investigated. Results point at an average dispersal distance of marked, irradiated olive flies of ca. 50 m. Pre-release adult diet did not affect dispersal ability. Fly recovery averaged ca. 3.5% during summer and ca. 1.5% during autumn. Most of the recovery concentrated during the first 3 days after releases, suggesting low survival of the released flies afterwards. As inferred from circular statistics, direction of dispersal was non-random with a significant directionality to the north-west. Results are discussed in view of environmental temperatures and wind direction.

The olive fly (OLF), Bactrocera oleae (Rossi), is an invasive tephritid fruit fly that causes extensive damage to olive crops around the world (especially in the Mediterranean basin and North America). Previous attempts to use the Sterile Insect Technique (SIT) for the OLF were not successful because of the inability to rear high quality OLF in the laboratory on an artificial diet. New improvements in rearing methods and additional understanding of the basic biology of the OLF have led to a renewal of interest in using SIT for OLF. This review discusses the history, difficulties, improvements and future directions of OLF mass-rearing. Issues include: the design of cages and oviposition substrates, cost and quality of artificial diets, maintenance of endosymbiotic microbiota, control of pathogenic microbes, collection of pupae, the fitness of adults, and the competitiveness of sterilized laboratory males released to the field.

The olive fruit fly [Bactrocera (Dacus) oleae (Rossi) (Diptera: Tephritidae)] adult diet since its development in the 1960’s regularly incorporates antibiotic. Considering recent findings on the importance and function of the indigenous microbial flora of insects, the effects of antibiotic removal were measured on the survival and egg laying of wild flies derived from McPhail trappings and from field infested olive fruits. In the first case wild flies fed no antibiotic laid significantly greater numbers of eggs for two generations (in 5 out of 10 and 2 out of 10 counting dates for G1 and G2 respectively), while there were no significant differences in survival in either test (P = 0.221 for P generation, P = 0.988 for G1 generation from McPhail traps, P = 0.056 for flies from infested fruits). Percent egg‐pupa recovery and adult emergence were not significantly affected by lack of antibiotic. An antibiotic-free strain has been maintained for eight generations, showing acceptable performance when compared to the long-reared standard ‘Laboratory’ strain. Overall results suggest the feasibility of an adult diet free of antibiotic without negative effects on colony survival and performance. J. Appl. Entomol.

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. Fil: Ordano, Mariano Andrés. Fundación Miguel Lillo; Argentina. Consejo Nacional de Investigaciones Científico y Técnicas. Centro Científico Tecnológico Tucumán. Unidad Ejecutora Lillo; Argentina Fil: Engelhard, Izhar. Institute of Plant Protection; Israel Fil: Rempoulakis, Polychronis. Institute of Plant Protection; Israel Fil: Nemny Lavy, Esther. Institute of Plant Protection; Israel Fil: Blum, Moshe. Bar-Ilan University; Israel Fil: Yasin, Sami. Nippon International Cooperation for Community Development; Palestina (ANP) Fil: Lensky, Itamar M.. Bar-Ilan University; Israel Fil: Papadopoulos, Nikos T.. University of Thessaly; Grecia Fil: Nestel, David. Institute of Plant Protection; Israel

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.

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.

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.

The microbiome of the olive fruit fly, Bactrocera oleae (Gmelin), a worldwide pest of olives (Olea europaea L.), has been examined for100 yr as part of efforts to identify bacteria that are plant pathogens vectored by the fly or are beneficial endosymbionts essential for the fly's survival and thus targets for possible biological control. Because tephritid fruit flies feed on free-living bacteria in their environment, distinguishing between the transient, acquired bacteria of their diet and persistent, resident bacteria that are vertically transmitted endosymbionts is difficult. Several culture-dependent and -independent studies have identified a diversity of species in the olive fruit fly microbiome, but they have not distinguished the roles of the microbes. Candidatus Erwinia dacicola, has been proposed to be a coevolved endosymbiont of the olive fruit fly; however, this was based on limited samples from two Italian populations. Our study shows that C. Erwinia dacicola was present in all New and Old World populations and in the majority of individuals of all life stages sampled in 2 yr. Olive fruit flies reared on olives in the laboratory had frequencies of C. Erwinia dacicola similar to that of wild populations; however, flies reared on artificial diets containing antibiotics in the laboratory rarely had the endosymbiont. The relative abundance of C. Erwinia dacicola varied across development stages, being most abundant in ovipositing females and larvae. This uniform presence of C. Erwini dacicola suggests that it is a persistent, resident endosymbiont of the olive fruit fly.