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.

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.

Influence of modified atmosphere packaging on radiation tolerance in the phytosanitary pest melon fly (Diptera: Tephritidae).

Modified atmosphere packaging (MAP) produces a low-oxygen (O2) environment that can increase produce shelf life by decreasing product respiration and growth of pathogens. However, low O2 is known to increase insect tolerance to irradiation, and the use of MAP with products treated by irradiation before export to control quarantine pests may inadvertently compromise treatment efficacy. Melon fly, Bactrocera cucurbitae Coquillet (Diptera: Tephritidae), is an important economic and quarantine pest of tropical fruits and vegetables, and one of the most radiation-tolerant tephritid fruit flies known. The effect of low O2 generated by MAP on the radiation tolerance of B. cucurbitae was examined. Third-instar larval B. cucurbitae were inoculated into ripe papayas and treated by 1) MAP + irradiation, 2) irradiation alone, 3) MAP alone, or (4) no MAP and no irradiation, and held for adult emergence. Three types of commercially available MAP products were tested that produced O2 concentrations between 1 and 15%, and a sublethal radiation dose (50 Gy) was used to allow comparisons between treatments. Ziploc storage bags (1-4% O2) increased survivorship to adult from 14 to 25%, whereas Xtend PP61 bags (3-8% O2) and Xtend PP53 bags (11-15% O2) did not enhance survivorship to the adult stage in B. cucurbitae irradiated at 50 Gy. Radiation doses approved by the United States Department of Agriculture and the International Plant Protection Commission for B. cucurbitae and Ceratitis capitata (Wiedemann) (Mediterranean fruit fly) are 150 and 100 Gy, respectively. In large-scale tests, 9,000 B. cucurbitae and 3,800 C. capitata larvae infesting papayas in Ziploc bags were irradiated at 150 and 100 Gy, respectively, with no survivors to the adult stage. MAP can increase insect survivorship during irradiation treatment at certain doses and O2 concentrations, but should not compromise the efficacy of the 150-Gy generic radiation treatment for tephritid fruit flies or the 100-Gy radiation treatment for C. capitata.

Exposure of insect cells to ionising radiation in vivo induces persistent phosphorylation of a H2AX homologue (H2AvB).

The response of eukaryotic cells to ionising radiation (IR)-induced double-strand DNA breaks is highly conserved and involves a DNA repair mechanism characterised by the early phosphorylation of histone protein H2AX (producing the active form γH2AX). Although the expression of an induced γH2AX variant has been detected in Drosophila melanogaster, the expression and radiation response of a γH2AX homologue has not been reported in economically important fruit flies. We use Bactrocera tryoni (Diptera: Tephritidae, Queensland fruit fly or 'Q-fly') to investigate this response with a view to developing molecular assays to detect/quantify exposure of fruit flies to IR and consequent DNA damage. Deep sequencing confirmed the presence of a H2AX homologue that we have termed H2AvB (i.e. variant Bactrocera) and has an identical sequence to a histone reported from the human disease vector Glossina morsitans. A linear dose-response of γH2AvB (0-400 Gy IR) was observed in whole Q-fly pupal lysates 24h post-IR and was detected at doses as low as 20 Gy. γH2AvB signal peaked at ~20min after IR exposure and at 24h post-IR the signal remained elevated but declined significantly by 5 days. Persistent and dose-dependent γH2AvB signal could be detected and quantified either by western blot or by laser scanning cytometry up to 17 days post-IR exposure in histone extracts or isolated nuclei from adult Q-flies (irradiated as pupae). We conclude that IR exposure in Q-fly leads to persistent γH2AvB signals (over a period of days) that can easily be detected by western blot or quantitative immunofluorescence techniques. These approaches have potential as the basis for assays for detection and quantification of prior IR exposure in pest fruit flies.

Assessment effect of gamma radiation on the flight ability of the peach fruit fly, Bactrocera zonata (Saunders).

The sterile insect technique is one of the most methods of fruit flies control. Flight ability of the Peach Fruit Fly (PFF), Bactrocera zonata was conducted under laboratory conditions to evaluate the effect of gamma radiation on flight ability of PFF, B. zonata. Pupae of PFF, B. zonata, were irradiated in an air atmosphere at 24, 48 and 72 h before adult emergence with three doses of Cobalt 60 (10, 30 and 50 Gray) and tested against 6, 12 and 20 cm tube heights. Flight Ability Percentage (FAP) of PFF was carried out for newly emerged flies and six-days-old of adult flies. FAP of newly emerged-and six- days-old of adult flies was inversely proportional to the tube heights, doses of gamma rays and with progress the age of flies. The FAP value was significantly higher at 6 cm tube height, followed by 12 cm then 20 cm tube heights for all tested levels of gamma rays, respectively.

Generic phytosanitary radiation treatment for tephritid fruit flies provides quarantine security for Bactrocera latifrons (Diptera: Tephritidae).

Bactrocera latifrons (Hendel) (Diptera: Tephritidae) is a quarantine pest of several solanaceous crops and tropical fruits that are treated using irradiation before export from Hawaii to the U.S. mainland. A dose of 150 Gy is approved as a generic irradiation treatment for tephritid fruit flies, but no confirmation of efficacy has been reported for B. latifrons. Dose response of B. latifrons was used to determine the most tolerant life stage and identify a dose that prevents adult emergence. Data indicated doses (plus 95% confidence limits) required to prevent adult emergence of 13.4 (10.0-29.6), 17.5 (14.4-24.8), and 88.1 (68.0-133.8) Gy for eggs, first instars and third instars, respectively. In large-scale confirmatory tests of the most radiotolerant life stage, a radiation dose of 150 Gy applied to B. latifrons late third instars in bell peppers (Capsicum annuum L.) resulted in no survival to the adult stage of 157,112 individuals, a treatment efficacy consistent with Probit 9-level mortality. The relative radiotolerance of melon fly Bactrocera cucurbitae Coquillet, and B. latifrons also was tested using a diagnostic radiation dose of 30 Gy. In diet, a mean of 6.9% of irradiated B. cucurbitae third instars developed to the adult stage, whereas no B. latifrons third instars developed to adults. In papaya, Carica papaya L., fruit, a mean of 3.3% of irradiated B. cucurbitae third instars developed to the adult stage, whereas 0.5% B. latifrons third instars developed to adults. This report supports the use of a generic radiation dose of 150 Gy in quarantine scenarios to control tephritid fruit flies on fresh commodities.

Ionizing radiation as a phytosanitary treatment against fruit flies (Diptera: Tephritidae): efficacy in naturally versus artificially infested fruit.

Some phytosanitary irradiation treatment research against tephritid fruit flies (Diptera: Tephritidae) has used artificially infested fruit with the unstated and untested assumption that the method adequately simulated a natural situation. We compare grapefruit, Citrus paradisi Macfayden, naturally infested by Mexican fruit fly, Anastrepha ludens (Loew), via oviposition until larvae reached the late third instar versus insertion of diet-reared third instars into holes made in grapefruits 24 h before irradiation; the latter technique has been used in other studies. Both infestation techniques resulted in statistically indistinguishable results, indicating that insertion of diet-reared third instar Mexican fruit fly into holes bored into grapefruit and subsequently sealed 24 h before irradiation would adequately represent natural infestation and could be used to develop a radiation phytosanitary treatment of the insect in grapefruit when prevention of adult emergence is used as the measure of efficacy. Nevertheless, it may not be advisable to extend this conclusion to other fruit fly/fruit combinations without doing appropriate comparison studies. Dissection of puparia from nonirradiated control insects that failed to emerge as adults showed a relatively even distribution of mortality among the developmental stages within the puparium. In contrast, dissection of puparia from irradiated third instars that did not emerge as adults revealed a sharp attenuation in development from cryptocephalic to phanerocephalic pupae demonstrating this transition to be the developmental step most affected by radiation.

Generic radiation quarantine treatments: the next steps.

In 2006, U.S. Department of Agriculture-Animal and Plant Health Inspection Service published a landmark rule providing generic radiation quarantine treatments. The rule approved radiation doses of 150 Gy for any tephritid fruit fly and 400 Gy for all other insects except the pupa and adult stages of Lepidoptera. The generic radiation treatments apply to all fresh horticultural commodities. Therefore, if a pest risk assessment demonstrates that no pupae or adult Lepidoptera are associated with a commodity, export approval can be forthcoming with no further research. Generic treatments are the culmination of decades of research but not an end point. Future research on quarantine and phytosanitary uses of radiation should focus on 1) development of specific doses for quarantine Lepidoptera not covered by the generic treatments, 2) reduction of dose levels for specific pests and commodities to shorten treatment time and minimize any deleterious effects of radiation treatment on commodity quality, 3) development of generic doses below 400 Gy for important groups of quarantine arthropods other than fruit flies, and 4) development of information on commodity tolerance and development of value-added irradiated fresh products that use generic radiation treatments. Generic treatments will facilitate safe trade between countries that have approved phytosanitary uses of radiation for fresh agricultural commodities.

Generic ionizing radiation quarantine treatments against fruit flies (Diptera: Tephritidae) proposed.

Tephritid fruit flies comprise the most important group of quarantined pests of fresh produce. Most quarantine treatments of fresh agricultural commodities are directed against these pests, and considerable effort in detection, trapping, and population control is expended worldwide to prevent these pests from invading new territories. Ionizing radiation has been studied for 70 yr for its possible use as a quarantine treatment against fruit flies, but has only been applied commercially on a limited basis since 1995. The treatment has great potential and will probably be used extensively in the future as it is tolerated by more species of fruits than any other major treatment. The U.S. Department Agriculture, Animal and Plant Health Inspection Service only recently proposed allowing irradiation for fresh agricultural imports from other countries, and other countries are studying proposals to do likewise. In 1991, the International Consultative Group on Food Irradiation recommended a generic dose against all tephritid fruit flies of 150 Gy. This article examines the literature dealing with irradiation quarantine treatments against fruit flies and recommends minimum absorbed doses of 70 Gy for Anastrepha spp., 101 Gy for Bactrocera jarvisi and B. tryoni, and 150 Gy for all Tephritidae except when fruits have been stored in hypoxic atmospheres.