Comparing the biological effectiveness of low energy 100 kV and high energy 5 MeV X-rays against Oriental fruit fly (Diptera: Tephritidae).

Radioisotope irradiators (using cesium-137 or cobalt-60) are used as sources of ionizing radiation to control quarantine or phytosanitary insect pests in internationally traded fresh commodities and to sterilize insects used in sterile insect release programs. There are institutional initiatives to replace isotopic irradiators (producing γ-rays) with lower-energy X-ray machines due to concerns about radiological terrorism and increasingly stringent regulations on the movement of radioisotopes. Questions remain about whether the biological effects of low-energy X-rays are comparable to those of γ-rays since differences in energy levels and dose rates of X-rays may have different efficacies. We compared adult emergence, flight ability, and adult survival in the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritdae), after irradiation of third instar larvae with 100 kV or 5 MeV (5,000 kV) X-rays at 20 and 40 Gy in replicated studies. At 20 Gy, the adult emergence rate was significantly lower after irradiation with 100 kV compared to 5 MeV X-rays, suggesting higher efficacy at the lower energy level. In a follow-up study using 100 kV X-rays, applying 20 Gy using a slow dose rate (0.24 Gy min-1) resulted in significantly higher adult emergence than did a fast dose rate (3.3 Gy min-1), suggesting lower efficacy. Although our study suggests higher efficacy of low energy 100 kV X-rays, there is uncertainty in measuring the dose from an X-ray tube operating at 100 kV using an ionization chamber; we discuss how this uncertainty may change the interpretation of the results. Using a 100 kV X-ray irradiator to develop a phytosanitary treatment may underestimate the dose required for insect control using commercial high-energy γ-ray or X-ray systems.

Rearing and 60Co radiation do not affect attractiveness but alter the volatile profiles released by Anastrepha obliqua calling males.

Calling males of Anastrepha obliqua release volatile compounds to attract conspecific males to form leks and females to mate. Male volatiles from Mexican and Brazilian populations of A. obliqua have been previously identified. However, there are differences in the number and identity of volatile compounds between the populations. These differences in volatile profiles may be due to male origin (e.g. wild or mass-reared flies) or methodological issues (e.g. sampling techniques). In this study, we evaluated the attractiveness of wild, laboratory non-irradiated, and laboratory-irradiated flies under semi-field conditions. Male volatiles were collected using dynamic headspace sampling (DHS) and solid-phase microextraction (SPME) techniques, and identified using gas chromatography-coupled mass spectrometry. The results showed no difference in the attractiveness of wild, laboratory non-irradiated, and irradiated males to females. However, the number of captured females differed according to the origin; wild and non-irradiated females were captured more frequently than the irradiated flies. A total of 21 compounds were found using SPME, whereas only 12 were collected using DHS, although the relative amounts of these compounds were higher than those obtained using the former sampling technique. In addition, only laboratory non-irradiated males released α-pinene and menthol, which have not been previously reported in this fruit fly species. Additionally, we identified novel compounds in A. obliqua; however, certain compounds previously reported were not detected. This study suggests that despite the qualitative and quantitative variations in the volatile profiles of A. obliqua males, their attractiveness was unaffected.

Increasing radiation doses in Anastrepha obliqua (Diptera: Tephritidae) larvae improve parasitoid mass-rearing attributes.

Doses of 40, 80, 120, and 160 Gy were applied to 5-, 6-, 7-, and 8-day-old Anastrepha obliqua larvae, which were exposed to the Neotropical-native braconids Doryctobracon crawfordi and Utetes anastrephae and the Asian braconid Diachasmimorpha longicaudata. These tests were performed to know the effect of the increase in host radiation on the emergence of the aforementioned parasitoids and the related consequences of oviposition on the host. The study was based on the fact that higher radiation doses may cause a decrease in the host immune activity. There was a direct relationship between the increase in radiation dose and the parasitoid emergence. Both, the weight and the mortality of the host larvae were not affected by radiation. Although the larval weight of the larvae was lower and the mortality was higher in the younger larvae. Both, the number of scars and immature stages per host puparium originated from the younger larvae were lower than those from older larvae. Only U. anastrephae superparasitized more at lower radiation. Superparasitism by D. longicaudata was more frequent at 160 Gy. Qualitative measurements of melanin in the larvae parasitized showed that the levels were lower with increasing radiation. As radiation doses increased, the antagonistic response of the A. obliqua larva was reduced. Host larvae aged 5- and 6-day-old irradiated at 120-160 Gy significantly improve parasitoid emergence. This evidence is relevant for the mass production of the three tested parasitoid species.

60Co-γ Radiation Alters Developmental Stages of Zeugodacus cucurbitae (Diptera: Tephritidae) Through Apoptosis Pathways Gene Expression.

Radiation is considered as a promising insect pest control strategy for minimizing postharvest yield losses. Among various techniques, irradiation is a method of choice as it induces lethal biochemical or molecular changes that cause a downstream cascade of abrupt physiological abnormalities at the cellular level. In this study, we evaluated the effect of 60Co-γ radiation on various developmental stages of Zeugodacus cucurbitae Coquillett and subsequent carry-over effects on the progeny. For this purpose, we treated eggs with 30- and 50-Gy radiation doses of 60Co-γ. We found that radiation significantly affected cellular antioxidants, insect morphology, and gene expression profiles. Our results indicate that in response to various doses of irradiation reactive oxygen species, catalase, peroxidase, and superoxide dismutase activities were increased along with a significant increase in the malondialdehyde (MDA) content. We observed higher mortality rates during the pupal stage of the insects that hatched from irradiated eggs (50 Gy). Furthermore, the life span of the adults was reduced in response to 50 Gy radiation. The negative effects carried over to the next generation were marked by significantly lower fecundity in the F1 generation of the irradiation groups as compared to control. The radiation induced morphological abnormalities at the pupal, as well as the adult, stages. Furthermore, variations in the gene expression following irradiation are discussed. Taken together, our results signify the utility of 60Co-γ radiation for fruit fly postharvest management.

Mitochondrial superoxide dismutase overexpression and low oxygen conditioning hormesis improve the performance of irradiated sterile males.

The Sterile Insect Technique (SIT) is a successful autocidal control method that uses ionizing radiation to sterilize insects. However, irradiation in normal atmospheric conditions can be damaging for males, because irradiation generates substantial biological oxidative stress that, combined with domestication and mass-rearing conditions, may reduce sterile male sexual competitiveness and quality. In this study, biological oxidative stress and antioxidant capacity were experimentally manipulated in Anastrepha suspensa using a combination of low-oxygen conditions and transgenic overexpression of mitochondrial superoxide dismutase (SOD2) to evaluate their role in the sexual behavior and quality of irradiated males. Our results showed that SOD2 overexpression enhances irradiated insect quality and improves male competitiveness in leks. However, the improvements in mating performance were modest, as normoxia-irradiated SOD2 males exhibited only a 22% improvement in mating success compared to normoxia-irradiated wild type males. Additionally, SOD2 overexpression did not synergistically improve the mating success of males irradiated in either hypoxia or severe hypoxia. Short-term hypoxic and severe-hypoxic conditioning hormesis, per se, increased antioxidant capacity and enhanced sexual competitiveness of irradiated males relative to non-irradiated males in leks. Our study provides valuable new information that antioxidant enzymes, particularly SOD2, have potential to improve the quality and lekking performance of sterile males used in SIT programs.

Irradiation of early immature Anastrepha ludens stages for the rearing of Doryctobracon areolatus (Hymenoptera: Braconidae), a fruit fly parasitoid.

Doryctobracon areolatus is a native parasitoid of the Neotropical region that presents the highest percentages of natural parasitism of fruit flies of the genus Anastrepha. In the Moscafrut Program SADER-SENASICA, located in Metapa de Domínguez, Chiapas, Mexico, a laboratory colony of this species is maintained on Anastrepha ludens, the Mexican fruit fly, with the aim to scale the production of the parasitoid up to massive levels. In order to eliminate unwanted emergence of adult flies during the rearing process, this study evaluated the effect of irradiation (at doses of 20, 30, 40, and 50 Gy) applied to eggs, and first and second instar larvae of A. ludens; all irradiated stages were subsequently exposed as second instar larvae to adult females of D. areolatus. Irradiation did not affect the eclosion of A. ludens eggs but, at doses of 40 and 50 Gy, it did cause delayed larval development and pupation, as well as lower larval weight. Adult fly emergence was suppressed at all doses, except in eggs irradiated at 20 Gy. Doses of 20 and 30 Gy applied to the eggs and larvae did not affect the emergence, survival, fecundity or flight ability of the emerged parasitoids, but the second instar larvae were easily handled during the rearing process. Our results suggest that D. areolatus can be successfully produced in second instar larvae of A. ludens irradiated at 30 Gy.

Diet and irradiation effects on the bacterial community composition and structure in the gut of domesticated teneral and mature Queensland fruit fly, Bactrocera tryoni (Diptera: Tephritidae).

BACKGROUND: Mass-rearing, domestication and gamma irradiation of tephritid fruit flies used in sterile insect technique (SIT) programmes can negatively impact fly quality and performance. Symbiotic bacteria supplied as probiotics to mass-reared fruit flies may help to overcome some of these issues. However, the effects of tephritid ontogeny, sex, diet and irradiation on their microbiota are not well known. RESULTS: We have used next-generation sequencing to characterise the bacterial community composition and structure within Queensland fruit fly, Bactrocera tryoni (Froggatt), by generating 16S rRNA gene amplicon libraries derived from the guts of 58 individual teneral and mature, female and male, sterile and fertile adult flies reared on artificial larval diets in a laboratory or mass-rearing environment, and fed either a full adult diet (i.e. sugar and yeast hydrolysate) or a sugar only adult diet. Overall, the amplicon sequence read volume in tenerals was low and smaller than in mature adult flies. Operational taxonomic units (OTUs), belonging to the families Enterobacteriaceae (8 OTUs) and Acetobacteraceae (1 OTU) were most prevalent. Enterobacteriaceae dominated laboratory-reared tenerals from a colony fed a carrot-based larval diet, while Acetobacteraceae dominated mass-reared tenerals from a production facility colony fed a lucerne chaff based larval diet. As adult flies matured, Enterobacteriaceae became dominant irrespective of larval origin. The inclusion of yeast in the adult diet strengthened this shift away from Acetobacteraceae towards Enterobacteriaceae. Interestingly, irradiation increased 16S rRNA gene sequence read volume. CONCLUSIONS: Our findings suggest that bacterial populations in fruit flies experience significant bottlenecks during metamorphosis. Gut bacteria in teneral flies were less abundant and less diverse, and impacted by colony origin. In contrast, mature adult flies had selectively increased abundances for some gut bacteria, or acquired these bacteria from the adult diet and environment. Furthermore, irradiation augmented bacterial abundance in mature flies. This implies that either some gut bacteria were compensating for damage caused by irradiation or irradiated flies had lost their ability to regulate bacterial load. Our findings suggest that the adult stage prior to sexual maturity may be ideal to target for probiotic manipulation of fly microbiota to increase fly performance in SIT programmes.

Generic Irradiation and Hot Water Phytosanitary Treatments for Mango Fruits cv. 'Ataulfo' niño Infested by Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae).

The mango fruit cv. 'Ataulfo' niño is an underdeveloped fruit that has a split on the back and a pronounced peak, and among the current total supply of commercialized mangoes cv. 'Ataulfo', approximately 2% are classified as 'Ataulfo' niño, which are strongly infested by Anastrepha ludens (Loew) (Diptera: Tephritidae) and Anastrepha obliqua Mcquart. The objective of this study was to determine the tolerance to 150-Gy generic irradiation for fruit flies of the Anastrepha genus in comparison to the development of a hot water treatment (HWT) as phytosanitary treatments for mango fruits cv. 'Ataulfo' niño infested by A. ludens and A. obliqua. The results indicated that both treatments were effective; 150-Gy irradiation and HWT at 46.3-47°C for 51 min did not result in significant effects on the external and internal color, total sugar content, firmness, pH, or weight. The sensorial quality described by the appearance, flavor, color, and odor did not show any significant differences between treatments. For both A. ludens and A. obliqua, the third larval instar was the most thermotolerant. The efficacy test was conducted with an immersion time of 47 min. A. ludens did not survive, but for A. obliqua, three larvae out of a total of 6,890 did survive and pupate. Consequently, the confirmatory test consisted of submerging mangoes infested with third-instar A. ludens and A. obliqua in water at 46.3-47°C for 51 min. In total, 67,392 A. ludens and 22,086 A. obliqua larvae were treated, and no surviving larvae were observed.

Optimization of the sterilizing doses and overflooding ratios for the South American fruit fly.

The Sterile Insect Technique (SIT) is an autocidal control method that relies on inundative releases of sterilized mass-reared insects. This technology has been used in several area-wide programmes for the suppression/eradication of fruit fly populations. Choosing the optimum sterilizing dose and the sterile release density is an essential step of the SIT. Considering unsolved issues related to the application of this technique against Anastrepha fraterculus (Wiedemann), this study aimed to define accurately the central target dose for both sexes of this species and to verify the induction of sterility in fertile flies at different sterile:fertile ratios. The results from the regression analyses proved that the sterilization process for the A. fraterculus Brazilian-1 morphotype (the most common in southern Brazil and Argentina) could consist of irradiating pupae 72 h before adult emergence at 40 Gy, with no detrimental effects to standard quality control parameters. The ovarian development in irradiated females was characterized, demonstrating that doses equal to or higher than 25 Gy cause complete and irreversible ovarian atrophy. The laboratory and field cage tests showed that the sterility induction increased with the proportion of sterile flies, and a sterile:fertile ratio of 50:1 should be appropriate in SIT field trials. The sterile females apparently did not distract the sterile males, despite of the slightly higher reductions in pupal yield for all ratios in their absence. The data generated in this study have a great practical value and will help decision-makers in planning field trials to evaluate the efficacy of the SIT against A. fraterculus populations.

Feasibility of using the radiation-based sterile insect technique (SIT) to control the olive fruit fly, Bactrocera oleae Gmelin (Diptera: Tephritidae) in Iran.

Bactrocera oleae is one of the most hazardous pests threatening olive orchards in Iran. SIT is an environment-friendly system of pest control based on releasing sterile males able to compete with wild males to mate with wild females. To determine sterile doses of radiation, pupae were irradiated to the doses of 0-160 Gy. Doses of 90-100 Gy were found optimal providing the necessary sterilization without severely impairing the competitiveness of the irradiated males in mating.

Radiation of Bactrocera dorsalis (Diptera: Tephritidae) Eggs to Improve the Mass Rearing of Diachasmimorpha longicaudata (Hymenoptera: Braconidae).

This study explored the potential for Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) larvae hatched from irradiated eggs as hosts for Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). B. dorsalis eggs of three different ages (12-, 24-, and 36-h old) were analyzed for hatchability, pupation rate, pupal weight, emergence rate, and sex ratio after exposure to different doses of radiation (5 and 10 Gy) at different dose rates (1 and 6 Gy/min). For the eggs of different ages exposed to radiation, only the hatchability and pupal weight of 36-h-old eggs exposed to the dose rate of 1 Gy/min were not affected; therefore, 6 Gy/min was not suitable for irradiating eggs. The viability of the parents and progenies of D. longicaudata when the parents were reared from 36-h-old eggs irradiated at nine different doses (0, 5, 10, 15, 20, 25, 30, 35, and 40 Gy) under laboratory conditions were investigated. The emergence percentage, sex ratio, and longevity of parasitoids developed from irradiated eggs were similar to those reared from nonirradiated hosts. A significant increase in larva mortality was observed for the eggs irradiated at doses above 25 Gy, and no redundant adult flies emerged at doses above 15 Gy. Hence, for B. dorsalis eggs to be applied in the mass rearing of D. longicaudata, the age of 36 h and a dose of 20-25 Gy are the optimal parameters. The results reveal that hosts and parasitoids need not be separated, enabling a reduction in cost, labor, and time and resulting in an improved mass rearing procedure for D. longicaudata.

Packing and Postirradiation Handling of the Anastrepha ludens (Diptera: Tephritidae) Tapachula-7 Genetic Sexing Strain: Combined Effects of Hypoxia, Pupal Size, and Temperature on Adult Quality.

The production of genetic sexing strains (GSS) of tephritid flies for sterile insect technique (SIT) programs convey the need to determine new conditions for packing and shipment since these flies are more susceptible to stressors than standard bisexual strains. We studied the effect of hypoxia, pupae size, and temperature on the new GSS Tapachula-7 of Anastrepha ludens flies (Diptera: Tephritidae). In one experiment, we tested the interaction size hypoxia using three pupae sizes, 6 (11.6 ± 1.1 mg), 7 (15.3 ± 1.5 mg), and 8 (17.9 ± 1.3 mg) (95% of produced pupae exhibit these categories of size), and four hypoxia periods, 12, 24, 36, 48 h and a control. In a second experiment, we tested two periods of hypoxia (24 and 48 h) and four temperatures: 15, 20, 25, and 30°C and a control (without hypoxia at laboratory temperature). Our results showed that the emergence and percent of fliers from the pupae exposed to hypoxia were adversely affected; however, emergence was higher in pupae of size 7. Treatment for 12 and 24 h hypoxia led to a higher number of fliers. In the case of the interaction of hypoxia and temperature, it was observed that those flies that emerged from the pupae exposed to hypoxia at 15 and 20°C exhibited quality control parameters similar to those that were not exposed to hypoxia. We discuss our results on the basis of the metabolic response to these factors and its application in the SIT programs.

Sexual Competitiveness, Field Survival, and Dispersal of Anastrepha obliqua (Diptera: Tephritidae) Fruit Flies Irradiated at Different Doses.

The sterile insect technique (SIT) is used in area-wide pest management programs for establishing low pest prevalence and/or areas free of fruit flies (Diptera: Tephritidae). The aim of this technique is to induce high levels of sterility in the wild population, for this the released insects must have a high sexual competitiveness and field dispersal. However, radiation decreases these biological attributes that do not allow it to compete successfully with wild insects. In this study the sexual competitiveness, field survival and dispersal of Anastrepha obliqua (Macquart; Diptera: Tephritidae) irradiated at 0, 40, 50, 60, 70, and 80 Gy were evaluated in laboratory. A dose of 60 Gy produced 98% sterility, whereas doses of 70 and 80 Gy produced 99% sterility. Sexual competitiveness was assessed in field cages, comparing males irradiated at 0, 50, 60, 70, and 80 Gy against wild males for mating with wild fertile females. Males irradiated at 50 and 60 Gy achieved more matings than those irradiated at 70 and 80 Gy. Wild males were more competitive than mass-reared males, even when these were not irradiated (0 Gy). There was no effect of irradiation on mating latency, yet wild males showed significantly shorter mating latency than mass-reared males. Female remating did not differ among those that mated with wild males and those that mated with males irradiated with different doses. The relative sterility index (RSI) increased from 0.25 at 80 Gy to 0.37 at 60 Gy. The Fried competitiveness index was 0.69 for males irradiated at 70 Gy and 0.57 for those irradiated at 80 Gy, which indicates that a 10 Gy reduction in the irradiation dose produces greater induction of sterility in the wild population. There were no significant differences in field survival and dispersal between flies irradiated at 70 or 80 Gy. Reducing the irradiation dose to 60 or 70 Gy could improve the performance of sterile males and the effectiveness of the SIT. Our results also distinguish between the effects of irradiation and mass-rearing on the performance of sterile males.

Low-Dose Irradiation With Modified Atmosphere Packaging for Mango Against the Oriental Fruit Fly (Diptera: Tephritidae).

Irradiation is used to disinfest the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) and other pests on mango fruits before export from Thailand to foreign markets. Modified atmosphere packaging (MAP) used during export of mangoes creates a low-oxygen environment that may reduce the efficacy of quarantine irradiation treatment against B. dorsalis. 'Nam Dok Mai' mangoes infested with third-instar larvae of B. dorsalis, wrapped with three different kinds of MAP bags (CF1, FF5, and H34M) or no MAP, were treated with gamma radiation at 0 (control), 30, 60, 90, 120, and 150 Gy. The average O2 and CO2 concentrations in MAP bags with mangos were 7.2 and 8.7% in the H34M bag, 8.6 and 21.2% in the CF1 bag, and 9.6 and 26.7% in the FF5 bag, respectively. The use of MAP on infested mangoes significantly increased mortality of B. dorsalis under irradiation treatment. The estimated lethal doses to cause 99% mortality (LD99) for no MAP and MAP (CF1, FF5, and H34M bags) treatments were 58.1, 41.6, 43.8, and 47.4 Gy, respectively. Therefore, MAP acted as an additional stressor rather than providing radioprotection in irradiated B. dorsalis. Large-scale confirmatory testing of 35,000 B. dorsalis larvae treated at a radiation dose of 150 Gy in mangoes with H34M MAP bags produced no survivors to the adult stage. Commercial use of MAP producing the O2 levels that we observed for mangos in this study will not reduce the efficacy of the approved 150 Gy quarantine irradiation treatment for B. dorsalis.

Blue light-induced immunosuppression in Bactrocera dorsalis adults, as a carryover effect of larval exposure.

Detrimental effects of ultraviolet (UV) light on living organisms are well understood, little is known about the effects of blue light irradiation. Although a recent study revealed that blue light caused more harmful effects on insects than UV light and blue light irradiation killed insect pests of various orders including Diptera, the effects of blue light on physiology of insects are still largely unknown. Here we studied the effects of blue light irradiation on cuticular melanin in larval and the immune response in adult stage of Bactrocera dorsalis. We also evaluated the effects of blue light exposure in larval stage on various age and mass at metamorphosis and the mediatory role of cuticular melanin in carryover effects of larval stressors across metamorphosis. We found that larvae exposed to blue light decreased melanin contents in their exoskeleton with smaller mass and delayed metamorphosis than insects reared without blue light exposure. Across metamorphosis, lower melanotic encapsulation response and higher susceptibility to Beauveria bassiana was detected in adults that had been exposed to blue light at their larval stage, thereby constituting the first evidence that blue light impaired adult immune function in B. dorsalis as a carryover effect of larval exposure.

Irradiation of Anastrepha fraterculus (Diptera: Tephritidae) Eggs to Inhibit Fly Emergence in the Mass-Rearing of Diachasmimorpha longicaudata (Hymenoptera: Braconidae).

As the incidence of Anastrepha fraterculus (Wiedemann) has increased in Southern Brazil in the past 3 yr, an initiative to release sterile flies and parasitoids has started. In order to make feasible the mass-rearing of the parasitoid Diachasmimorpha longicaudata (Ashmed), this study investigated the suitability of A. fraterculus larvae derived from irradiated eggs as host for D. longicaudata Two different ages of A. fraterculus eggs (24 and 48 h old) were analyzed for hatchability after the exposure to a range of radiation doses. The hatchability of 48-h-old eggs was not affected by radiation, and no fly emerged at doses higher than 27.5 Gy. The larvae derived from irradiated eggs proved to be suitable hosts for the parasitoid development, with observed parasitism rates higher than 70% and sex ratio values above 0.6. The parasitism capability and longevity of D. longicaudata reared on larvae derived from irradiated eggs were also assessed. During the 10 d of parasitism evaluated, D. longicaudata from the treatments were able to parasitize nonirradiated larvae similarly as the parasitoids from controls and the laboratory colony. The longevity of D. longicaudata from the treatments was not affected either, with survival rates higher than 80% after 20 d of evaluation. The age of 48 h and a dose of 30 Gy could be considered the best age and dose for A. fraterculus eggs to be used in the mass-rearing of D. longicaudata The results of this study will decrease the costs of mass-rearing D. longicaudata on A. fraterculus.

LARVAL X-RAY IRRADIATION INFLUENCES PROTEIN EXPRESSION IN PUPAE OF THE ORIENTAL FRUIT FLY, BACTROCERA DORSALIS.

The sterile insect technique (SIT) was developed to eradicate the new world screwworm from the southern United States and Mexico, and became a component of many area-wide integrated pest management programs, particularly useful in managing tephritid fruit flies. SIT is based on the idea of rearing and sterilizing male pests, originally by ionizing radiation, and then releasing into field, where they compete for and mate with wild females. Mating with sterile males leads to reduced fecundity to lower pest populations. There are concerns with the use and distribution of radioisotopes for SIT programs, which have led to developing X-ray irradiation protocols to sterilize insects. We considered the possibility that X-ray irradiation exerts sublethal impacts aside form sterilizing insects. Such effects may not be directly observable, which led us to the hypothesis that X-ray irradiation in one life stage creates alterations in biological fitness and protein expression in the subsequent stage. We tested our hypothesis by irradiating larvae of Bactrocera dorsalis. There are two major points. One, exposing larvae to X-ray treatments led to reduced adult emergence, fecundity, fertility, and flight capacity from the corresponding pupae and emerged adults. Two, the X-ray treatments led to substantial expression changes in 27 pupal proteins. We assorted the 67 spots representing these proteins into three groups, metabolism, development, and structure. Our interpretation is these X-ray induced changes in biological performance and protein expression indicate their adult counterparts may be disabled in their abilities to successfully compete for and mate wild females in native habitats.

Laboratory longevity and competitiveness of Dacus ciliatus Loew (Diptera: Tephritidae) following sub-sterilizing gamma irradiation.

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.

Male irradiation affects female remating behavior in Anastrepha serpentina (Diptera: Tephritidae).

Female remating in target pest species can affect the efficacy of control methods such as the Sterile Insect Technique (SIT) but very little is known about the postcopulatory mating behavior of these pests. In this study, we investigated the remating behavior of female Anastrepha serpentina (Diptera: Tephritidae), an oligophagous pest of Sapotaceae. First, we tested how long the sexual refractory period of females lasted after an initial mating. Second, we tested the effect of male and female sterility, female ovipositing opportunities and male density on female propensity to remate. Lastly, we tested if the amount of sperm stored by females was correlated to the likelihood of females to remate. We found that receptivity of mass-reared A. serpentina females had a bimodal response, with up to 16% of mass-reared A. serpentina females remating five days after the initial copulation, decreasing to 2% at 10 and 15 days and increasing to 13% after 20 days. Compared to fertile males, sterile males were less likely to mate and less likely to inhibit females from remating. Copula duration of sterile males was shorter compared to fertile males. Remating females were less likely to mate with a sterile male as a second mate. Sterile females were less likely to mate or remate compared to fertile females. Opportunity to oviposit and male density had no effect on female remating probability. Sperm numbers were not correlated with female likelihood to remate. Information on the post-copulatory behavior of mass-reared A. serpentina will aid fruit fly managers in improving the quality of sterile males. We discuss our results in terms of the differences this species presents in female remating behavior compared to other tephritids.

Gamma Irradiation as a Phytosanitary Treatment of Bactrocera tau (Diptera: Tephritidae) in Pumpkin Fruits.

The fruit fly Bactrocera tau (Walker) is an important quarantine pest that damages fruits and vegetables throughout Asian regions. Host commodities shipped from infested areas should undergo phytosanitary measures to reduce the risk of shipping viable flies. The dose-response tests with 1-d-old eggs and 3-, 5-, 7-, 8-d-old larvae were initiated to determine the most resistant stages in fruits, and the minimum dose for 99.9968% prevention of adult eclosion at 95% confidence level was validated in the confirmatory tests. The results showed that 1) the pupariation rate was not affected by gamma radiation except for eggs and first instars, while the percent of eclosion was reduced significantly in all instars at all radiation dose; 2) the tolerance to radiation increased with increasing age and developmental stage; 3) the estimated dose to 99.9968% preventing adult eclosion from late third instars was 70.9 Gy (95% CL: 65.6-78.2, probit model) and 71.8 Gy (95% CL: 63.0-87.3, logit model); and iv) in total, 107,135 late third instars cage infested in pumpkin fruits were irradiated at the target dose of 70 Gy (62.5-85.0, Gy measured), which resulted in no adult emergence in the two confirmatory tests. Therefore, a minimum dose of 85 and 72 Gy, which could prevent adult emergence at the efficacy of 99.9972 and 99.9938% at the 95% confidence level, respectively, can be recommended as a minimum dose for phytosanitary treatment of B. tau in any host fruits and vegetables under ambient atmospheres.