Combining Irradiation and Biological Control against Brown Marmorated Stink Bug: Are Sterile Eggs a Suitable Substrate for the Egg Parasitoid Trissolcus japonicus?

The brown marmorated stink bug (BMSB), Halyomorpha halys, is a phytophagous invasive pest native to south-eastern Asia, and it is now distributed worldwide. This species is considered to be one of the most damaging insect pests in North America and in Europe. In agriculture, the predominant approach to managing BMSB is based on the use of insecticides, specifically pyrethroids and neonicotinoids. Unfortunately, the biology of the species and its facility to develop mechanisms of resistance to available pesticides has induced farmers and scientists to develop different, least-toxic, and more effective strategies of control. In a territorial area-wide approach, the use of a classical biological control program in combination with other least-toxic strategies has been given prominent consideration. Following exploratory surveys in the native range, attention has focused on Trissolcus japonicus, a small scelionid egg parasitoid wasp that is able to oviposit and complete its larval development in a single egg of H. halys. A common method for detecting egg parasitoids in the native range involves the placement of so-called 'sentinel' egg masses of the pest in the environment for a short period, which are then returned to the laboratory to determine if any of them are parasitized. Outside of the area of origin, the use of fertile sentinel eggs of the alien species may lead to the further release of the pest species; an alternative is to use sterile sentinel eggs to record the presence of new indigenous egg parasitoids or to detect the dispersal of alien species (in this case, T. japonicus) released in a new environment to control the target insect pest species. This study evaluated the performance of three types of sterile sentinel eggs as a suitable substrate for the oviposition and larval development of the egg parasitoid T. japonicus in a context of combining classical biological control with a Sterile Insect Technique (SIT) approach.

Effects of Irradiation on Biology and Mating Behaviour of Wild Males of Brown Marmorated Stink Bug Using a 6 MV Medical Linear Accelerator.

The brown marmorated stink bug, Halyomorpha halys, is a pentatomid bug of Eastern Asian origin that became an economically relevant pest in the Eurasian and American continents. Management of this species is limited to use of chemical insecticides: an inefficient method due to the strong adaptability of the target pest. The sterile insect technique (SIT) is potentially a valid tactic in the search for nontoxic alternatives. In this work, we investigated the suitability of mass-trapped overwintering males, collected during the aggregation phase before the winter diapause, for their release as competitive sterile males in an SIT programme. Differently from previous studies, irradiation was applied with a linear accelerator device that produced high-energy photons. Following a similar scientific protocol with newly emerged irradiated males, the effects of X-ray irradiation on physiological parameters (longevity, fecundity and fertility) were assessed. In addition, behavioural bioassays were carried out in no-choice conditions to evaluate if irradiation interferes with mating processes. The results are very encouraging; the effects of the irradiation at 32 Gy did not differ from the controls in the longevity or fecundity of the exposed overwintering adults. The hatching rate of the eggs laid by the fertile females that had mated with the irradiated males was less than 5%. The results of behavioural bioassays showed that the irradiation did not cause a significant impact on the quality of the sterile males. More research is warranted to evaluate the mating competitiveness of sterile males in semi-field and field conditions.

Invasive potential of tropical fruit flies in temperate regions under climate change.

Tropical fruit flies are considered among the most economically important invasive species detected in temperate areas of the United States and the European Union. Detections often trigger quarantine and eradication programs that are conducted without a holistic understanding of the threat posed. Weather-driven physiologically-based demographic models are used to estimate the geographic range, relative abundance, and threat posed by four tropical tephritid fruit flies (Mediterranean fruit fly, melon fly, oriental fruit fly, and Mexican fruit fly) in North and Central America, and the European-Mediterranean region under extant and climate change weather (RCP8.5 and A1B scenarios). Most temperate areas under tropical fruit fly propagule pressure have not been suitable for establishment, but suitability is predicted to increase in some areas with climate change. To meet this ongoing challenge, investments are needed to collect sound biological data to develop mechanistic models to predict the geographic range and relative abundance of these and other invasive species, and to put eradication policies on a scientific basis.

Irradiation-induced sterility in an egg parasitoid and possible implications for the use of biological control in insect eradication.

Classical biological control is a pest control tool involving the release of imported natural enemies. The Sterile Insect Technique (SIT) comprises releasing sexually sterile insects of a pest into the wild population for suppression or eradication. Both these approaches are environmentally friendly and their combination can result in a synergistic impact on pest populations and improve eradication. However, stringent regulation surrounding the introduction of biological control agents limits their use in eradication owing to the perceived risk of effects on non-target organisms. We investigated the irradiation biology of the egg parasitoid Trissolcus basalis to ascertain whether sterile parasitoids could mitigate the risk of potential sustained non-target impacts. Mated female T. basalis were gamma-irradiated at doses between 120 and 150 Gy and exposed to egg masses of their host Nezara viridula throughout their lifespans. This resulted in host mortality, despite a substantial reduction in developing parasitoid offspring, which followed a negative dose-response. There was no emergence of parasitoid offspring at 140 Gy and above. Irradiation did not affect oviposition behaviour but caused an increase in longevity. Consequently, sterile parasitoids could possibly alleviate concerns regarding the irreversibility of biological control release, which promotes further investigation of their potential role in eradication.

Operational Parameters for the Aerial Release of Sterile Codling Moths Using an Uncrewed Aircraft System.

The codling moth is a serious pest of apples in most regions of the world where this fruit is produced. The sterile insect technique is one strategy used to control this pest and is employed as part of an area-wide integrated pest management program for the codling moth in British Columbia, Canada. Modified fixed wing aircraft are the most common method for the release of sterile insects in large area-wide pest management programs. However, aerial release with a full-size aircraft can be prohibitively expensive. We evaluated the use of small, uncrewed aircraft systems (UASs) for the release of sterile codling moths. Sterile codling moths released from greater altitudes were more broadly distributed and drifted more in strong winds, compared to those released from lower altitudes. Most of the released insects were recaptured in a 50 m wide swath under the release route. Recapture rates for aerially released insects were 40-70% higher compared to those released from the ground. UASs provide a promising alternative to ground release and conventional aircraft for the release of sterile codling moths.

Combined Effects of Mating Disruption, Insecticides, and the Sterile Insect Technique on Cydia pomonella in New Zealand.

Codling moth was introduced into New Zealand, and remains a critical pest for the apple industry. Apples exported to some markets require strict phytosanitary measures to eliminate the risk of larval infestation. Mating disruption and insecticide applications are the principal means of suppression in New Zealand. We tested the potential for the sterile insect technique (SIT) to supplement these measures to achieve local eradication or suppression of this pest. SIT was trialed in an isolated group of six integrated fruit production (IFP) orchards and one organic orchard (total 391 ha), using sterilized insects imported from Canada, with release by unmanned aerial vehicle and from the ground. Eradication was not achieved across the region, but a very high level of codling moth suppression was achieved at individual orchards after the introduction of sterile moths in combination with mating disruption and larvicides. After six years of releases, catches of wild codling moths at three IFP orchards (224 ha) were 90-99% lower than in 2013-2014, the year before releases began. Catches at three other IFP orchards (129 ha) decreased by 67-97% from the year before releases began (2015-2016), from lower initial levels. At a certified organic orchard with a higher initial population under only organic larvicides and mating disruption, by 2019-2020, there was an 81% reduction in wild moths capture from 2016-2017, the year before releases began.

Kairomone and Camera Trapping New Zealand Flower Thrips, Thrips obscuratus.

This project investigated how kairomone lures, camera traps, and counting software could together contribute to pest management. Images of cumulative daily catch of New Zealand Flower Thrips (NZFT) attracted to a ripe peach lactone (6-pentyl-2H-pyran-2-one; 6-PAP) were automatically loaded to the internet and compared with scanned bases checked weekly using in-house software and manual counting. Camera traps were able to provide thrips counts equivalent to delta traps, but daily and remotely. An 11-fold greater NZFT count occurred within 24 h in passive traps after polyethylene sachets loaded with 250 mg of 6-PAP were placed in trees. Intensive trapping, by placing 1, 2, 4, and 8 traps per tree (500 mg/trap), resulted in a maximum 32-fold increase in thrips per tree. While 6-PAP has proved to be a useful tool for monitoring NZFT numbers, our results suggest that it is not likely to be suitable for mass trapping. Future research should investigate NZFT behavior to better understand population movement on an area-wide basis. Camera traps can be a valuable tool for recording insect flight activity remotely, but the number of traps required for statistically reliable estimates may be prohibitive.

Egg Sterilisation of Irradiated Nezara viridula (Hemiptera: Pentatomidae).

Nezara viridula Linnaeus (Hemiptera: Pentatomidae) is a polyphagous pest of a wide range of economically important crops. Because the control of this species and other pentatomids relies primarily on insecticide application, investigation into the Sterile Insect Technique (SIT) is warranted. We aimed to investigate the irradiation biology of N. viridula for the potential application of SIT against this pest. Male and female N. viridula were gamma-irradiated at doses between 4 and 28 Gy and mated with both irradiated and nonirradiated conspecifics. Sterility of the resulting eggs followed a dose-response in each case. Irradiated males crossed with untreated females showed higher F1 egg sterility than crosses where the female was irradiated. The greatest F1 egg sterility was observed when both parents were irradiated. There was no obvious dose-response for the longevity of irradiated males, and for the fecundity of nonirradiated females mated with irradiated males. The fecundity of irradiated females appeared to decrease with irradiation dose. These results can be applied to a potential future application of SIT against N. viridula, but predominantly supports the ongoing development of SIT for Halyomorpha halys Stål (Hemiptera: Pentatomidae) and hemipteran pests in general.

Can Vibrational Playbacks Disrupt Mating or Influence Other Relevant Behaviours in Bactericera cockerelli (Triozidae: Hemiptera)?

Behaviours of insects can be manipulated by transmitting vibrational signals to host plants in order to develop pest management techniques. Bactericera cockerelli is an important pest and uses vibrations for mate-finding. In order to design a future control strategy for B. cockerelli, three different bioassays were performed to assess whether vibrational signals could affect relevant behaviours. Single males or pairs were treated with a female playback in test 1 and 2, respectively. In test 3, mixed sex groups received either different disturbance playbacks. The use of a female playback significantly reduced the mating success of males, since they were attracted towards the source of the stimulus. Moreover, test 2 revealed that B. cockerelli females are competitive, since they used their signals to cover the playback and to duet with males, while in test 3, the disturbance playback, consisting of broadband noises significantly reduced male signalling activity. However, none of the treatments of test 3 negatively affected the mating success of males, which tended to mount the other conspecifics present on the same leaf. The role of vibrations in sexual communication and their potential application as control technique for B. cockerelli are discussed as well.

Will Peri-Urban Cydia pomonella (Lepidoptera: Tortricidae) Challenge Local Eradication?

Codling moth, Cydia pomonella (Lepidoptera: Tortricidae), is a phytosanitary pest of New Zealand's export apples. The sterile insect technique supplements other controls in an eradication attempt at an isolated group of orchards in Hawke's Bay, New Zealand. There has been no attempt in New Zealand to characterize potential sources of uncontrolled peri-urban populations, which we predicted to be larger than in managed orchards. We installed 200 pheromone traps across Hastings city, which averaged 0.32 moths/trap/week. We also mapped host trees around the pilot eradication orchards and installed 28 traps in rural Ongaonga, which averaged 0.59 moths/trap/week. In Hastings, traps in host trees caught significantly more males than traps in non-host trees, and spatial interpolation showed evidence of spatial clustering. Traps in orchards operating the most stringent codling moth management averaged half the catch rate of Hastings peri-urban traps. Orchards with less rigorous moth control had a 5-fold higher trap catch rate. We conclude that peri-urban populations are significant and ubiquitous, and that special measures to reduce pest prevalence are needed to achieve area-wide suppression and reduce the risk of immigration into export orchards. Because the location of all host trees in Hastings is not known, it could be more cost-effectively assumed that hosts are ubiquitous across the city and the area treated accordingly.

Mazes to Study the Effects of Spatial Complexity, Predation and Population Density on Mate Finding.

The difficulty to locate mates and overcome predation can hamper species establishment and population maintenance. The effects of sparseness between individuals or the effect of predators on the probability of population growth can be difficult to measure experimentally. For testing hypotheses about population density and predation, we contend that habitat complexity can be simulated using insect mazes of varying mathematical difficulty. To demonstrate the concept, we investigated whether the use of 3D printed mazes of varying complexity could be used to increase spatial separation between sexes of Drosophila simulans, and whether the presence of a generalist predator hampered mate-finding. We then examined how increasing D. simulans population density might overcome the artificially created effects of increasing the distance between mates and having a predator present. As expected, there was an increase in time taken to find a mate and a lower incidence of mating as habitat complexity increased. Increasing the density of flies reduced the searching time and increased mating success, and overcame the effect of the predator in the maze. Printable 3D mazes offer the opportunity to quickly assess the effects of spatial separation on insect population growth in the laboratory, without the need for large enclosed spaces. Mazes could be scaled up for larger insects and can be used for other applications such as learning.

The Scent of Individual Foraging Bees.

Honey bees (Apis mellifera) forage by using their sense of smell and returning to floral odours that they have previously learned to associate with high-quality food rewards. Foraging bees communicate with other bees in the hive about food sources by exchanging chemical and locational information. It is well established that bees transfer non-volatile information regarding taste and quality of nectar via trophallaxis and communicate location information via directional dances. But to our knowledge, volatiles carried by returning forager bees on their bodies has not been explored as another source of chemical information. We investigated the cuticular-adsorbed odours of bees when foraging on three different crops and compared their odours with the crops' flower headspace. We found that cuticular extracts were in majority correlated with the flower headspace where bees were foraging, specific to the crop and field. Our results support the hypothesis that the scent of returning forager bees can be communicated to hivemates and is associated with information about current floral resources. Some of the floral volatiles that we identified in bee extracts had been previously found to be key compounds learned from the crop, thus supporting a mechanism for the selection of decisive compounds.

Electrophysiological and Behavioral Responses of Queensland Fruit Fly Females to Fruit Odors.

Females of the Queensland fruit fly (QFF), Bactrocera tryoni, are amongst the most damaging pests of horticulture in Australia and neighboring countries. Females can lay eggs into more than a hundred species of fruits and vegetables, resulting in large crop losses. Sexually mature males can be managed sustainably with traps baited with long-lasting synthetic lures, and sexually immature males and females can be attracted and killed by short-lived protein baits applied directly on surfaces, with a low success rate (< 20%). No long-lasting attractants for virgin or mated females exist. With the aim of developing a female attractant for surveillance, we collected and analyzed the odors of four ripe host fruits: orange, cherry guava, banana and feijoa. Virgin and mated female QFF were tested with gas-chromatography coupled with electro-antennographic detection to identify electrophysiologically (EAD)-active compounds. We detected 41 EAD-active compounds, with seven found common for more than one fruit. Overall, mated females responded more often and with higher intensity than virgin females. In particular, five compounds present either in cherry guava or feijoa triggered a significantly higher EAD response from mated females than from virgins. Twenty-six EAD-active compounds were selected and tested individually in a Y-tube olfactometer to measure attraction of both virgin and mated females. Behavioral responses differed significantly amongst the compounds, but not strongly between virgin and mated females. We did not find any correlation between electrophysiological and behavioral responses. Further field testing with behaviorally-active compounds is needed for the development of a new QFF female lure.

Odorant-Based Detection and Discrimination of Two Economic Pests in Export Apples.

Detection of pest infestations in fresh produce traded internationally could offer improved prospects for reducing the movement of unwanted pests. Because immature stages of some pests can be difficult to find visually, other cues such as herbivore-induced volatiles that can potentially be detected at the early stages of infestation are worth investigating. In this study, we artificially infested postharvested apples (Malus × domestica 'Royal Gala') with two economic apple pests, the specialist codling moth (CM, Cydia pomonella Linnaeus, Lepidoptera: Tortricidae) and the generalist Queensland fruit fly (QFF, Bactrocera tryoni, Froggatt, Diptera: Tephritidae) and collected volatile organic compounds (VOCs) over time (days 0, 6, and 14-15). In both infestation experiments, we found a strong and significant interaction between time and treatment. Apples infested with the QFF emitted lower total amounts of VOCs than uninfested apples, whereas apples infested with the CM released similar total amounts of VOCs. Apples infested with CM had increases in several hexyl and butyl esters, which were particularly noticeable after 15 d. In contrast, changes in ethyl esters were characteristics of QFF infestation and could be detected from day 6. Our multilevel and multivariate statistical analysis identified specific volatile biomarkers for each species at each sampling time that can be used to design a new tool for remote detection and surveillance of these invasive pests in harvested apples. Nevertheless, other information such as the cultivar as well as the storage condition needs to be taken into consideration to increase accuracy of future odorant-based sensors for pest identification.

Trapping Brown Marmorated Stink Bugs: "The Nazgȗl" Lure and Kill Nets.

Improvements to current brown marmorated stink bug (BMSB), Halyomorpha halys, surveillance and killing systems are needed to improve detection sensitivity and to reduce pesticide use. Detection of BMSB in New Zealand with traps is reliant on sticky panels with aggregation pheromone, which are low cost but inefficient compared with beating foliage. Trapping for BMSB adults and nymphs was conducted daily with lethal traps consisting of an aggregation pheromone-baited-coat hanger covered with dark-colored long-lasting insecticide-treated mesh, we termed "The Nazgȗl", based on its sinister appearance. A deep tray lined with white plastic was attached centrally at the base for collecting the dead BMSB. The lethal traps killed and caught up to 3.5-fold more nymphs and adult BMSB than identically-baited sticky panels in the 3 weeks of deployment, and provided a snapshot of phenology by instar. We expect that lure-and-kill stations could contribute to the suppression of a delimited population and could be included as part of a semiochemical-based eradication program. Attracting and killing females and nymphs, thus removing future offspring, could contribute to population suppression during an eradication.

Live Traps for Adult Brown Marmorated Stink Bugs.

Surveillance for detection of the brown marmorated stink bug, Halyomorpha halys, is reliant on sticky panels with aggregation pheromone, which are low cost, but very inefficient (est. 3%). Trapping for adults was conducted in Italy with novel live (or lethal) traps consisting of aggregation pheromone-baited cylinders with a wind vane, with the upwind end covered by mesh and the downwind end sealed by a removable entry-only mesh cone, admitting the attracted bugs. The novel traps caught up to 15-times more adult H. halys than identically-baited sticky panels in two weeks of daily checking (n = 6 replicates) (the new live traps were, in Run 1, 5-, 9-, 15-, 13-, 4-, 12-, 2-fold; and in Run 2, 7-, 1-, 3-, 7-, 6-, 6-, and 5-fold better than sticky traps, daily). The maximum catch of the new traps was 96 live adults in one trap in 24 h and the average improvement was ~7-fold compared with sticky panels. The rotating live traps, which exploit a mesh funnel facing the plume downwind that proved useful for collecting adults, could also be used to kill bugs. We expect that commercially-available traps could replace the crude prototypes we constructed quickly from local materials, at low cost, as long as the principles of a suitable plume structure were observed, as we discuss. The traps could be useful for the sterile insect technique, supporting rearing colonies, or to kill bugs.

The Competitive Mating of Irradiated Brown Marmorated Stink Bugs, Halyomorpha halys, for the Sterile Insect Technique.

The sterility of eggs and nymphs from gamma-irradiated male Halyomorpha halys was investigated to determine the potential for the sterile insect technique (SIT). Males irradiated at 0, 16, 24 and 32 Gy were placed with untreated virgin females, and egg sterility was determined, showing 54.3% at 16 Gy. The percentage of sterility from irradiation was 26 percent lower than previous results from the USA and the variance was very high. Competitive overflooding ratio trials between irradiated virgin males and fertile virgin males at a 5:1 ratio resulted in the expected egg sterility, indicating competitive performance by irradiated males. By July and August, older, irradiated overwintered males were significantly less competitive than similar, non-irradiated males. There is a need to revisit the irradiation delivery method to achieve proper precision around the paternal dose required for an expected >80% egg sterility and subsequent ~99% endpoint sterility estimated at adult emergence in the F1 phase. These results suggest that the mating competitiveness and competency of males after irradiation at 16 Gy is not limiting to the sterile insect technique for suppression. A wild harvest of overwintering males using the aggregation pheromone, followed by irradiation and male release, might replace rearing. Mass-collected, sterilized bugs could be transported from an area of high H. halys density and shipped for release to enable suppression or eradication elsewhere. This concept is under development but further work is needed now to understand the difference in results between the US and Italian irradiators and increase the reliability of dosimetry.

Peri-Urban Community Attitudes towards Codling Moth Trapping and Suppression Using the Sterile Insect Technique in New Zealand.

New, more socially-acceptable technologies are being developed to suppress horticultural pests, because suppression is technically difficult with current technologies, especially in urban areas. One technique involves the release of sterile insects to prevent offspring in the next generation. This technology involves aerial or ground release systems, but this could also create issues for the public. This study investigated community perceptions of a recently-introduced response to codling moth control in New Zealand-Sterile Insect Technique (SIT). Community attitudes to SIT were examined in Hastings, New Zealand, in April, 2018. Eighty-six detailed interviews were undertaken with a random sample of households. This community was very willing (98% agreement) to host a sex pheromone trap in their gardens, and condoned regular visits to monitor traps. Attitudes to SIT were very positive (98% in favor). Once explained, the concept of using unmanned aerial vehicles to deliver sterile insects was also acceptable (98%) to the community. Use of unmanned aerial vehicles to release sterile insects during a hypothetical incursion response of an exotic fruit fly was also supported at 98% by respondent householders. Investigation of community attitudes can be valuable to guide practitioners in determining suitable technologies before an area-wide programme is launched.

Multiple Mating in the Citrophilous Mealybug Pseudococcus calceolariae: Implications for Mating Disruption.

The citrophilous mealybug Pseudococcus calceolariae (Maskell) (Hemiptera, Pseudococcidae) is a primary pest of various crops, including grapevines. The use of insecticides against this species is difficult in most cases because its life cycle includes an extended duration of eggs, juveniles, and adults under the bark and on the roots. Pheromone-based control strategies can present new eco-friendly opportunities to manage this species, as in the case of Planococcus ficus (Signoret) and Planococcus citri (Risso). With this aim it is critical to understand behavioral aspects that may influence pheromone-based control strategies. Herein, the capability of males to fertilize multiple females was investigated, trying to understand whether this behavior could negatively impact the efficacy of mass trapping, mating disruption, or the lure and kill technique. Results showed that a P. calceolariae male can successfully mate and fertilize up to 13 females. The copulation time in subsequent mating events and the time between copulations did not change over time but the number of matings per day significantly decreased. In a further experiment, we investigated the mate location strategy of P. calceolariae males, testing the attractiveness of different loadings of sex pheromone on males in a flight tunnel. Males constantly exposed to 16 rubber septa loaded with the sex pheromone showed a significant decrease in female detection at 1 and 30 µg loadings (0.18 and 0.74 visits per female for each visit per septum, respectively), whereas in the control about 9.2-fold more of the released males successfully detected the female in the center of the array of 16 septa without pheromone. Male location of females in the control (45%) was significantly higher than in the arrays with surrounding pheromone (5% and 20% at 1 and 30 µg loadings, respectively). Mating only occurred in the control arrays (45%). This study represents a useful first step to developing pheromone-based strategies for the control of citrophilous mealybugs.

Effect of Lure Combination on Fruit Fly Surveillance Sensitivity.

Surveillance for invading insect pests is costly and the trapper usually finds the traps empty of the target pest. Since the successful establishment of new pests is an uncommon event, multiple lures placed into one trap might increase the efficiency of the surveillance system. We investigated the effect of the combination of the Tephritidae male lures - trimedlure, cuelure, raspberry ketone and methyl eugenol - on catch of Ceratitis capitata, Zeugodacus cucurbitae, Bactrocera tryoni, B. dorsalis, B. aquilonis and B. tenuifascia in Australia and the USA (not all species are present in each country). The increase in trap density required to offset any reduction in catch due to the presence of lures for other Tephritidae was estimated. The effect of increasing trap density to maintain surveillance sensitivity was modelled for a hypothetical population of B. tryoni males, where the effective sampling area of cuelure traps for this species has been estimated. The 3-way combination significantly reduced the catch of the methyl eugenol-responsive B. dorsalis. Unexpectedly, we found that trimedlure-baited traps that contained methyl eugenol had ×3.1 lower catch of C. capitata than in trimedlure-only-baited traps in Australia, but not in Hawaii where no difference in catch was observed, we cannot satisfactorily explain this result. Based on the data presented here and from previous research, combinations of some male lures for the early detection of tephritid flies appear compatible and where there is any reduction in surveillance sensitivity observed, this can be offset by increasing the density of traps in the area.