Diel flight activity in response to attractants in the predatory beetle, Cathartus quadricollis (Coleoptera: Silvanidae) in macadamia nut in Hawaii.

The diel flight activity in Cathartus quadricollis (Guerin-Meneville) (Coleoptera: Silvanidae), a predator of two important pests in Hawaii, coffee berry borer, Hypothenemus hampei (Ferrari) and tropical nut borer, Hypothenemus obscurus (F.) (Coleoptera: Curculionidae: Scolytinae) was studied in a macadamia nut orchard using yellow sticky traps baited with pheromone and fungal volatile attractants. The study was conducted at different months throughout the year and at different times during the lunar cycle (new moon and full moon). Flight activity peaked in the late hours of the photophase into the early hours of the scotophase, between 1830 and 2000 h; flight activity also occurred but to a lesser extent in the early morning hours between 0700 and 1030 h. Numbers of captured C. quadricollis during periods of flight activity were negatively correlated with wind speed. The implications of these findings for the development of optimal pest management strategies including biological control are discussed.

Physical, Chemical, and Sensory Properties of a Turmeric-Fortified Pineapple Juice Beverage.

Beverage mixtures based on pineapple (Ananas comosus) and turmeric (Curcuma longa) juice as a ready-to-drink product were developed, and their physicochemical, nutritional, and sensory properties were evaluated. Four different concentrations of turmeric juice (5%, 10%, 15%, and 20% (v/v)) were added to pineapple juice to make turmeric-fortified pineapple (TFP) juice samples. Pineapple juice without turmeric was the control. The L*, a*, b*, titratable acidity (TA), total antioxidant capacity, and %DPPH scavenging values, as well as the concentrations of the phenolic compounds curcumin and demethoxycurcumin, were significantly increased with increasing turmeric concentration. Thirty volatile compounds were detected in the mixed juice samples with turmeric. Most of the turmeric-specific compounds, including monoterpenes, sesquiterpenes and turmerones, were detected in the TFP juice samples. While the antioxidant activity of the juice samples increased with increasing turmeric concentration, the pineapple juice fortified with 10% turmeric (10%T) had the best overall quality as determined by panelists. Greater concentrations of turmeric were associated with decreased palatability due to reduced mouthfeel and sweetness and increased aftertaste and sourness. These results suggest that the 10%T juice could be developed into a commercial functional beverage with increased overall flavor and nutritional quality.

Evaluation of bioactive low-density polyethylene (LDPE) nanocomposite films in combined treatment with irradiation on strawberry shelf-life extension.

A low-density polyethylene (LDPE) film reinforced with cellulose nanocrystals (CNCs) with an encapsulated bioactive formulation (cinnamon essential oil + silver nanoparticles) was developed for preservation of fresh strawberries. Antimicrobial activity of the active LDPE films was tested against Escherichia coli O157:H7, Salmonella typhimurium, Aspergillus niger, and Penicillium chrysogenum by agar volatilization assay. The optimal condition of the films showed ≥75% inhibitory capacity against the tested microbes. Strawberries were stored with different types of films: Group 1 (control): (LDPE + CNCs + Glycerol), Group 2: (LDPE + CNCs + Glycerol + AGPPH silver nanoparticles), Group 3: (LDPE + CNCs + Glycerol + cinnamon), Group 4: (LDPE + CNCs + Glycerol + active formulation), and Group 5: (LDPE + CNCs + Glycerol + active formulation + 0.5 kGy γ-radiation) at 4°C for 12 days. Weight loss (WL) (%), decay (%), firmness (N), color, and total phenolics and anthocyanin content of the strawberries were measured. Results showed that the most effective LDPE-nanocomposite film for reducing the microbial growth was LDPE + CNCs + Glycerol + active formulation film (Group 4). When combined with γ-irradiation (0.5 kGy), the LDPE + CNCs + Glycerol + active formulation (Group 5) significantly reduced both decay and WL by 94%, as compared to the control samples after 12 days of storage. Total phenols (from 952 to 1711 mg/kg) and anthocyanin content (from 185 to 287 mg/kg) increased with storage time under the different treatments. The mechanical properties, water vapor permeability (WVP), and surface color of the films were also tested. Though the WVP of the films were not influenced by the types of antimicrobial agents, they did significantly (p ≤ 0.05) change color and mechanical properties of the films. Therefore, combined treatment of active film and γ-irradiation has potential as an alternative method for extending the shelf-life of storage strawberries while maintaining fruit quality. PRACTICAL APPLICATION: Bioactive Low-density polyethylene (LDPE) nanocomposite film was developed in the study by incorporating active formulation (essential oil and silver nanoparticle) to extend the shelf life of stored strawberries. The bioactive LDPE-based nanocomposite film along with γ-irradiation could be used to preserve fruits for long-term storage by controlling the growth of foodborne pathogenic bacteria and spoilage fungi.

Susceptibility of 'Malama' Avocados to Bactrocera dorsalis (Diptera: Tephritidae) in Hawai'i and a Proposed Systems Approach for Market Access.

Avocados are generally poor hosts or nonhosts for tephritid fruit flies, which can influence the type of mitigation measures required by export protocols for trade. We conducted experiments to determine the susceptibility of the avocado, Persea americana, cv. 'Malama' to infestation by Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in Hawai'i using laboratory and field no-choice cage tests, together with field sampling. In no-choice cage tests, infestation rates increased with decreasing fruit firmness over time after harvest. Although risk of infestation was negatively correlated with fruit firmness, about 50% of mature hard fruit exposed immediately after harvest became infested. In field cage tests, mature hard fruit on the tree at the 25% purple color stage were not infested, whereas fruit at the 50, 75, and 100% purple color stages were infested at relatively low rates. Field sampling of fallen 'Malama' fruit over two seasons showed no natural infestation by B. dorsalis and trap captures in avocado orchards using McPhail traps containing protein bait lures averaged 0.13 ± 0.03 (mean ± SE) flies per trap per week, underscoring the low prevalence of fruit flies in avocado orchards and the poor host quality of this cultivar. A systems approach based on poor host status, low prevalence, and a limited harvest period (October to March) and distribution area (northern tier states) is proposed to export 'Malama' avocados from Hawai'i to the continental United States.

Poor Host Status of Australian Finger Lime, Citrus australasica, to Ceratitis capitata, Zeugodacus cucurbitae, and Bactrocera dorsalis (Diptera: Tephritidae) in Hawai'i.

We examined the host status of the Australian finger lime, Citrus australiasica F. Muell. (Rutaceae), to Hawai'i's tephritid fruit fly pests using laboratory and field studies. In high-density (500 flies, 1:1 males and females) no-choice cage exposures (25 × 25 × 25-cm cage size), both undamaged and punctured finger limes were infested by Ceratitis capitata and Zeugodacuscucurbitae at a low rate compared to papaya controls, whereas Bactrocera dorsalis did not infest undamaged fruit, suggesting finger lime is a nonhost. In low-density (50 females) no-choice cage exposures, C. capitata and Z. cucurbitae readily oviposited in undamaged fruit but individuals rarely developed to the pupal or adult stage. For C. capitata, 274 finger limes exposed to 2000 gravid females, which laid an estimated 14,384 eggs, produced two pupae and no adults. For Z. cucurbitae, 299 fruit exposed to 2000 gravid females, which laid an estimated 4484 eggs, produced four pupae and one adult. Field sampling of undamaged fruit from the tree and off the ground from commercial farms produced five C. capitata pupa and one adult from 1119 fruit, for an infestation rate of 0.05 flies per kilogram of fruit; field collections found no natural Z. cucurbitae or B. dorsalis infestation, but the number of fruit available was too low to demonstrate nonhost status with a high degree of confidence.

Silver nanoparticles-essential oils combined treatments to enhance the antibacterial and antifungal properties against foodborne pathogens and spoilage microorganisms.

Plant-derived essential oils (EOs) and commercial silver nanoparticles (AgNPs) were tested to evaluate their antibacterial and antifungal efficiency against two pathogenic bacteria (Escherichia coli O157:H7 and Salmonella Typhimurium) and three spoilage fungi (Aspergillus niger, Penicillium chrysogenum, and Mucor circinelloides). A broth microdilution assay was used to determine the minimal inhibitory concentration (MIC) of EOs and AgNPs. In the MIC assay, the cinnamon EO, Mediterranean formulation, citrus EO and spherical-shaped silver nanoparticles (AgNPs) (AGC 1, AGC 0.5, AGPP and AGPPH) showed moderate to high antibacterial and antifungal properties, with MIC ranging from 7.8 to 62.5 ppm for AgNPs and 312.5-1250 ppm for EOs against the tested bacteria and fungi. The possible interaction between the EOs and the AgNPs was determined using a checkerboard method by evaluating fractional inhibitory concentration (FIC) values. The combination of two or more EOs and AgNPs (Active combination 1: AGPPH+cinnamon EO, Active combination 2: AGC 0.5+Mediterranean formulation+citrus EO, Active combination 3: AGPP+cinnamon EO+Asian formulation+lavang EO) showed synergistic effects (FIC <1.0) against all tested bacteria and fungi. A modified Gompertz model was used to evaluate growth parameters including maximum colony diameter (A), maximum growth rate (Vm), and lag phase (λ), under the three active combinations suggested by the checkerboard method using a vapor assay. The three active combinations 1, 2 and 3 reduced the growth rate and maximum colony diameter of E. coli, S. Typhimurium, A. niger, P. chrysogenum, and M. circinelloides, and extended their lag phase from 1 to 5 days. In in situ tests with inoculated rice, the three active combinations showed a significant reduction of all tested bacteria and fungi at 27 °C for 28 days.

Mixture design methodology and predictive modeling for developing active formulations using essential oils and citrus extract against foodborne pathogens and spoilage microorganisms in rice.

The antibacterial and antifungal effects of six plant-derived essential oils (EOs) and two types of citrus extracts (CEs) were studied against two pathogenic bacteria (Salmonella Typhimurium and Escherichia coli O157:H7) and three fungi (Aspergillus niger, Penicillium chrysogenum, and Mucor circinelloides). A broth microdilution assay and checkerboard method were used to measure the minimal inhibitory concentration (MIC) of each extract and the possible interactions between them. The MIC assay showed that cinnamon EO, Mediterranean EO, Southern formulation, citrus EO, organic citrus extract (OCE), and natural citrus extract (NCE) had the highest antimicrobial and antifungal activity. The checkerboard method showed that the Mediterranean EO+OCE combination acted in synergy against all tested pathogens. A centroid mixture design was used to develop active formulations by predicting optimal concentrations of EO/CEs for increased antibacterial/antifungal activity. A mixture of four formulations (625 ppm OCE, 313 ppm Mediterranean EO, 625 ppm citrus EO, and 313 ppm cinnamon EO) named as active formulation 1, and the mixture from five formulations (625 ppm NCE, 625 ppm Asian formulation, 313 ppm Southern formulation, 625 ppm cinnamon EO, and 313 ppm savory thyme EO) named as active formulation 2, were formulated and tested because of their high microbicidal effectiveness. In situ tests with rice showed a significant reduction (P ≤ 0.05) of all tested pathogenic bacteria and fungi from the vapor of active formulations 1 and 2 after 28 days of storage. PRACTICAL APPLICATION: Active formulations (essential oils and citrus extracts) developed in the study are highly effective against foodborne pathogens. Active formulations in this study could be used as natural preservatives in the food industry for controlling foodborne diseases and spoilage organisms in stored foods.

Host Suitability Index for Polyphagous Tephritid Fruit Flies.

Tephritid fruit flies are major economic pests for fruit production and are an impediment to international trade. Different host fruits are known to vary in their suitability for fruit flies to complete their life cycle. Currently, international regulatory standards that define the likely legal host status for tephritid fruit flies categorize fruits as a natural host, a conditional host, or a nonhost. For those fruits that are natural or conditional hosts, infestation rate can vary as a spectrum ranging from highly attractive fruits supporting large numbers of fruit flies to very poor hosts supporting low numbers. Here, we propose a Host Suitability Index (HSI), which divides the host status of natural and conditional hosts into five categories based on the log infestation rate (number of flies per kilogram of fruit) ranging from very poor (<0.1), poor (0.1-1.0), moderately good (1.0-10.0), good (10-100), and very good (>100). Infestation rates may be determined by field sampling or cage infestation studies. We illustrate the concept of this index using 21 papers that examine the host status of fruits in five species of polyphagous fruit flies in the Pacific region: Bactrocera tryoni Froggatt, Bactrocera dorsalis (Hendel), Bactrocera latifrons (Hendel), Zeugodacus cucurbitae (Coquillett), and Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). This general-purpose index may be useful in developing systems approaches that rely on poor host status, for determining surveillance and detection protocols for potential incursions, and to guide the appropriate regulatory response during fruit fly outbreaks.

Coffee berry borer (Hypothenemus hampei) (Coleoptera: Curculionidae) development across an elevational gradient on Hawai'i Island: Applying laboratory degree-day predictions to natural field populations.

Coffee berry borer (CBB, Hypothenemus hampei) (Coleoptera: Curculionidae: Scolytinae) is the most destructive pest of coffee worldwide. Information on CBB development times can be used to predict the initiation of new infestation cycles early in the coffee-growing season and thus inform the timing of insecticide applications. While laboratory estimates of CBB development under constant conditions exist, they have not been applied under the heterogeneous environmental conditions that characterize many coffee-growing regions. We measured CBB development times and abundance in commercial coffee farms across an elevational gradient on Hawai'i Island and applied thermal accumulation models from previous laboratory studies to test their fit to field data. Artificial lures were used to infest coffee berries at five farms ranging in elevation from 279-792 m, and weather variables were monitored at macro (farm-level) and micro (branch-level) scales. CBB development was followed in the field from the time of initial berry infestation by the founding female through the development of F1 mature adults. Mean development time from egg to adult across all sites was 38.5 ± 3.46 days, while the mean time required for the completion of a full life cycle (from time of infestation to presence of mature F1 females) was 50.9 ± 3.35 days. Development time increased with increasing elevation and decreasing temperature. Using macro-scale temperature data and two different estimates for the lower temperature threshold (14.9°C and 13.9°C), we estimated a mean requirement of 332 ± 14 degree-days and 386 ± 16 degree-days, respectively, from the time of berry infestation to the initiation of a new reproductive cycle in mature coffee berries. Similar estimates were obtained using micro-scale temperature data, indicating that macro-scale temperature monitoring is sufficient for life-cycle prediction. We also present a model relating elevation to number of CBB generations per month. Our findings suggest that CBB development times from laboratory studies are generally applicable to field conditions on Hawai'i Island and can be used as a decision support tool to improve IPM strategies for this worldwide pest of coffee.

ß-cyclocitral synergizes the response of adult Drosophila suzukii (Diptera: Drosophilidae) to fruit juices and isoamyl acetate in a sex-dependent manner.

Semiochemicals play a pivotal role in the location, evaluation, and utilization of hosts by herbivorous insects. Mixtures of host plant-derived compounds are often required to elicit appropriate levels of response to olfactory stimuli. In multiple-choice bioassays, we characterized the response of adult Drosophila suzukii to foliage- and fruit-based synthetic compounds tested alone and in association with grape and tart cherry juices, and assessed whether synergistic interactions among olfactory stimuli are involved in the olfactory-driven behavior of D. suzukii. Our results established (1) significant attraction of females (but not males) to ß-cyclocitral and isoamyl acetate when tested singly, (2) the presence of a synergistic interaction between ß-cyclocitral and cherry juice only for females, and (3) the presence of a synergistic interaction between ß-cyclocitral and isoamyl acetate but only in the case of males. Our findings increase our understanding of male and female D. suzukii olfactory responses to synthetic compounds and fruit juices as sources of attractants. Combinations of foliage- and fruit-based compounds may be needed to increase SWD attraction.

Short-Term Physiological Response of a Native Hawaiian Plant, Hibiscus arnottianus, to Injury by the Exotic Leafhopper, Sophonia orientalis (Hemiptera: Cicadellidae).

Sophonia orientalis (Matsumura), also known as the two-spotted leafhopper, is a widespread exotic pest of many economically important crop plants and ornamental plants in Hawaii. Sophonia orientalis is highly polyphagous and is a major threat to some of the native endemic plants. Despite the successful establishment in Hawaii, interactions of S. orientalis with its host plants remain poorly understood. Previous studies primarily focused on distribution, parasitism, and oviposition of S. orientalis in Hawaii, whereas plant physiological responses to the leafhopper's injury, and, specifically, gas exchange rates in plants, have not yet been described. In this study, we examined a short-term physiological response of a native Hawaiian plant, Hibiscus arnottianus (A. Gray), to injury by S. orientalis. We also explored whether Camellia sinensis (L.) Kuntze, a native host plant of S. orientalis in Asia, exhibits a similar response. We found that H. arnottianus plants demonstrated a rapid (2-d) physiological response to injury accompanied by 40% reduction in rate of photosynthesis and 42% reduction in rate of transpiration, whereas C. sinensis did not exhibit any reduction in gas exchange rates. We did not record any changes in plant chlorophyll levels after plant injury in either species. Our results suggest that H. arnottianus is responding to the leafhopper feeding with a generalized wound response predicted for novel plant-insect herbivore associations. We discuss potential future directions for studies which might focus on host plant responses to S. orientalis in its native versus introduced range.

Comparative Cold Tolerance in Ceratitis capitata and Zeugodacus cucurbitae (Diptera: Tephritidae).

Cold tolerance studies were conducted with the egg and larval stages of Mediterranean fruit fly, Ceratitis capitata (Wiedemann) and melon fly, Zeugodacus (Bactrocera) cucurbitae (Coquillett) in Navel oranges to determine whether quarantine cold treatments approved for C. capitata might also be effective against Z. cucurbitae. Navel orange is a good host for C. capitata and a poor host for Z. cucurbitae, and therefore, artificial infestation of fruit was used to facilitate comparisons. Laboratory-reared eggs and larvae were inserted in the center of Navel oranges, placed in cold storage chambers at 1.5-2.0°C for 1, 2, 4, 6, 8, 10, 12, or 14 d, then removed, and evaluated for egg hatch or larval survival. Time-response data were analyzed using linear mixed-effects models, probit analysis, and visual inspection of survivorship graphs. C. capitata eggs were significantly more cold tolerant than Z. cucurbitae eggs, and Z. cucurbitae larvae were generally more cold tolerant than C. capitata larvae. C. capitata eggs and Z. cucurbitae second instar larvae were the most cold-tolerant life stages, and they were not significantly different from each other. Results suggest that cold treatment at ≤1.5°C for a minimum of 14 d would be sufficient to achieve disinfestation of C. capitata and Z. cucurbitae. The inherent cold tolerance in Z. cucurbitae is equal to or higher than that of C. capitata, and therefore, cold treatment protocols developed for C. capitata may not always be effective against Z. cucurbitae.

Irradiation for Quarantine Control of Coffee Berry Borer, Hypothenemus hampei (Coleoptera: Curculionidae: Scolytinae) in Coffee and a Proposed Generic Dose for Snout Beetles (Coleoptera: Curculionoidea).

Coffee berry borer (CBB), Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), is the most serious insect pest of coffee worldwide. Green coffee used in blending and roasting is traded between countries and may be subjected to fumigation for disinfestation of CBB. For example, green coffee shipped to Hawaii from the U.S. mainland must be treated with methyl bromide. Irradiation is an alternative disinfestation treatment option. Dose-response tests were conducted with adult beetles to identify a sterilizing dose, followed by large-scale confirmatory tests with adults infesting coffee berries at 100 Gy (measured doses 84-102 Gy). In total, 6,598 adult CBBs naturally infesting dried coffee berries were irradiated at 100 Gy and produced no viable offspring, whereas 1,033 unirradiated controls produced 327 eggs, 411 larvae, and 58 pupae at 3 wk post treatment. This is the first study to develop a postharvest irradiation treatment for a scolytine bark beetle and supports other studies suggesting 150 Gy is sufficient to prevent reproduction in snout beetles in the superfamily Curculionoidea.

Postharvest Irradiation Treatment for Quarantine Control of Western Flower Thrips (Thysanoptera: Thripidae).

The western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), is an important pest of fresh horticultural produce and as such is considered a biosecurity risk in many countries from which it is absent. Information is needed on the radiation tolerance of important surface pests of quarantine importance such as F. occidentalis so that phytosanitary irradiation treatments for exported fresh commodities can be lowered to below the 400 Gy generic treatment currently approved for most insects in the United States and Australia. Lowering the dose will help minimize any product quality problems, reduce costs, and shorten treatment time. In large-scale confirmatory trials conducted in two independent laboratories in Hawaii and Australia, a dose of 250 Gy (measured doses 222-279 Gy) applied to adult F. occidentalis on green beans resulted in no reproduction in 5,050 treated individuals. At 250 Gy, the effective dose is significantly below the 400 Gy generic dose, demonstrating that irradiation at this lowered level is an effective method for the disinfestation of F. occidentalis from fresh horticultural produce.

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.

Effect of Low-Oxygen Conditions Created by Modified Atmosphere Packaging on Radiation Tolerance in Drosophila suzukii (Diptera: Drosophilidae) in Sweet Cherries.

Modified atmosphere packaging (MAP) creates a low-oxygen (O2) environment that can increase the shelf life of fresh produce by decreasing respiration and the growth of pathogens. Low oxygen may also increase insect tolerance to irradiation (IR), and the use of MAP with products treated by IR to control quarantine pests before export may inadvertently compromise treatment efficacy. Spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae) is a quarantine pest of stone and small fruits and a potential target for postharvest IR treatment. The effect of low oxygen generated by MAP at ambient temperatures on the radiation tolerance of D. suzukii infesting sweet cherries was examined. Early pupal stage D. suzukii were inserted into ripe sweet cherries and treated by 1) MAP + IR, 2) IR alone, 3) MAP alone, or 4) no MAP and no IR and held for adult emergence. Three types of commercially available MAP products were tested that produced different oxygen concentrations between 3 and 15%, and a sublethal radiation dose (60 Gy) was used to allow comparisons between the treatments. Xtend PP61 bags (3.2-4.8% O2), Xtend PP71 bags (5.4-8.6% O2), and Xtend PP53 bags (13.6-15.4% O2) did not enhance survivorship to the adult stage in D. suzukii pupae irradiated at 60 Gy in sweet cherries. MAP use should not compromise phytosanitary IR treatment against D. suzukii in exported sweet cherries or other fruit.

Postharvest Irradiation Treatment for Quarantine Control of the Invasive Lobesia botrana (Lepidoptera: Tortricidae).

Irradiation is a postharvest treatment option for exported berries and berry-like fruits to prevent movement of the quarantine pest European grape vine moth, Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae). The effects of irradiation on egg, larval, and pupal development in L. botrana were examined. Eggs, neonates, third and fifth instars, and early- and late-stage pupae were irradiated at target doses of 50, 100, 150, or 200 Gy or left untreated as controls in replicated factorial experiments, and survival to the adult stage was recorded. Tolerance to radiation generally increased with increasing age and developmental stage. A dose of 150 Gy prevented adult emergence in eggs and larvae. Pupae were more radiotolerant than larvae, and late-stage pupae were more tolerant than early-stage pupae. In large-scale validation tests, 150 Gy applied to fifth instars in diet prevented adult emergence, but some survival occurred in fifth instars irradiated in table grapes; however, 250 Gy prevented fifth instar survival in grapes. For most commodities, the fifth instar is the most radiotolerant life stage likely to occur with the commodity; a minimum radiation dose of 250 Gy will prevent adult emergence from this stage. For traded commodities such as table grapes that may contain L. botrana pupae, 325 Gy applied to mature female pupae sterilized emerging adults and may provide quarantine security. Radiotolerance in L. botrana is comparable to other tortricids, and the data reported here support a generic dose of 250 Gy for eggs and larvae of this group.

Phytosanitary Treatments Against Bactrocera dorsalis (Diptera: Tephritidae): Current Situation and Future Prospects.

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is arguably the most important tephritid attacking fruits after Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). In 2003 it was found in Africa and quickly spread to most of the sub-Saharan part of the continent, destroying fruits and creating regulatory barriers to their export. The insect is causing new nutritional and economic losses across Africa, as well as the losses it has caused for decades in infested areas of Asia, New Guinea, and Hawaii. This new panorama represents a challenge for fruit exportation from Africa. Phytosanitary treatments are required to export quarantined commodities out of infested areas to areas where the pest does not exist and could become established. This paper describes current phytosanitary treatments against B. dorsalis and their use throughout the world, the development of new treatments based on existing research, and recommendations for further research to provide phytosanitary solutions to the problem.

Effect of Irradiation on Queen Survivorship and Reproduction in the Invasive Fire Ant Solenopsis invicta (Hymenoptera: Formicidae) and a Proposed Phytosanitary Irradiation Treatment for Ants.

We studied radiation tolerance in queens of the red imported fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) to identify a dose that prevents reproduction. Virgin or fertile queens were collected from Santa Fe and Formosa provinces in Argentina and reared in the laboratory in microcolonies. Virgin queens were irradiated at 0 (control), 70, 90, 120, or 150 Gy, and fertile queens were irradiated at 0, 60, 125, and 190 Gy, and then followed for 11 wk in the microcolonies to evaluate survival and reproduction. Virgin queens lay trophic eggs that do not hatch, whereas fertile queens lay eggs that hatch and develop into brood. In general, queen oviposition and survival decreased with increasing irradiation dose. For virgin queens, no eggs were laid by irradiated queens after the third week, whereas the control queens continued laying eggs throughout the 11-wk experiment. For fertile queens, only one larva and no pupae was observed in the 60 Gy treatment and no larvae or pupae were observed in the 125 and 190 Gy treatments, whereas a total of 431 larvae and 83 pupae were produced by untreated control queens during 11 wks. Survivorship of virgin and fertile queens was similarly reduced by irradiation treatment. These results with S. invicta are consistent with previous findings for three other invasive ants, Wasmannia auropunctata (Roger), Pheidole megacephala (F.), and Linephithema humile (Mayr), that are hitchhiker pests on fresh horticultural commodities. A radiation dose of 150 Gy is proposed as a phytosanitary treatment to prevent reproduction in ants.

Molecular Markers Detect Cryptic Predation on Coffee Berry Borer (Coleoptera: Curculionidae) by Silvanid and Laemophloeid Flat Bark Beetles (Coleoptera: Silvanidae, Laemophloeidae) in Coffee Beans.

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is a serious pest of coffee worldwide. It was first detected in Hawai'i in 2010. Two predatory beetles, Cathartus quadricollis (Coleoptera: Silvanidae) and Leptophloeus sp. (Coleoptera: Laemophloeidae), have been observed in H. hampei-infested coffee. Under laboratory conditions, colony-reared C. quadricollis and Leptophloeus sp. prey upon all life stages of H. hampei. However, the H. hampei life cycle occurs almost exclusively within a coffee bean obscured from direct observation. Thus, it is unknown if C. quadricollis and Leptophloeus sp. consume H. hampei as prey in the wild. To demonstrate predation of H. hampei by C. quadricollis and Leptophloeus sp., a molecular assay was developed utilizing species-specific primers targeting short regions of the mitochondrial COI gene to determine species presence. Using these primers, wild C. quadricollis and Leptophloeus sp. were collected and screened for the presence of H. hampei DNA using PCR. Analysis of collections from five coffee farms revealed predation of C. quadricollis and Leptophloeus sp. on H. hampei. Further laboratory testing showed that H. hampei DNA could be detected in predators for as long as 48 h after feeding, indicating the farm-caught predators had preyed on H. hampei within 2 d of sampling. This study demonstrates the utility of molecular markers for the study of the ecology of predators and prey with cryptic behavior, and suggests C. quadricollis and Leptophloeus sp. might be useful biocontrol agents against H. hampei.