Exploration for Asian longhorned beetle parasitoids in Korea using an improved sentinel log trap.

The Asian longhorned beetle (ALB), Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), is a destructive invasive woodboring insect pest, and efforts are being made to find parasitoids for ALB biological control. Through a four-year survey in Korea using a sentinel log trap associated with host chemical cues potentially important for host finding by parasitoids, two parasitoid species were discovered attacking ALB. One species is Spathius ibarakius Belokobylskij & Maetô, which is known to also parasitize citrus longhorned beetle, Anoplophora chinensis (Forster). The other parasitoid species, whose offspring were dead before imago, could not be morphologically identified at the adult stage. We attempted molecular and morphological identification of the larvae/pupae of the unidentified parasitoid; however, only superfamily-level identification was possible. The parasitism rate recovered in the logs was 0.3% by the unidentified parasitoid in Gapyeong-gun in 2019, while it reached 29.2% by S. ibarakius in Busan city in 2022. Future efforts for exploring ALB natural enemies in the pest's native range may focus on parasitoids with high parasitism rates.

Title: Exploration des parasitoïdes du longicorne asiatique en Corée à l'aide d'un piège à bûche sentinelle amélioré. Abstract: Le longicorne asiatique (LA), Anoplophora glabripennis (Motschulsky) (Coleoptera : Cerambycidae), est un insecte ravageur envahissant et destructeur du bois, et des efforts sont déployés pour trouver des parasitoïdes pour la lutte biologique contre lui. Au cours d'une étude de quatre ans en Corée utilisant un piège à bûche sentinelle associé à des signaux chimiques de l'hôte potentiellement importants pour la détection de l'hôte par les parasitoïdes, deux espèces de parasitoïdes ont été découvertes attaquant le longicorne. Une espèce est Spathius ibarakius Belokobylskij & Maetô, qui est connue pour parasiter également le longicorne des agrumes, Anoplophora chinensis (Forster). Les autres espèces de parasitoïdes, dont les descendants sont morts avant l'imago, n'ont pu être identifiées morphologiquement au stade adulte. Nous avons tenté une identification moléculaire et morphologique des larves/pupes du parasitoïde non identifié, mais seule une identification au niveau de la superfamille a été possible. Le taux de parasitisme observé dans les bûches était de 0,3 % par le parasitoïde non identifié à Gapyeong-gun en 2019, tandis qu'il atteignait 29,2 % par S. ibarakius dans la ville de Busan en 2022. Les efforts futurs pour explorer les ennemis naturels du capricorne dans l'aire de répartition naturelle du ravageur pourraient se concentrer sur les parasitoïdes à taux de parasitisme élevés.

Parasitoid-induced changes in metabolic rate and feeding activity of the emerald ash borer (Coleoptera: Buprestidae): implications for biological control.

Parasitoid-host interactions form the foundation of biological control strategies against many agriculture and forest insect pests. The emerald ash borer (EAB), Agrilus planipennis (Coleoptera: Buprestidae), is a serious invasive pest of ash (Fraxinus spp.) trees in North America. Tetrastichus planipennisi (Hymenoptera: Eulophidae) is a gregarious, koinobiont endoparasitoid, attacking late (3rd to 4th) instars of EAB larvae, which feed in the live phloem of ash trunks or branches, making serpentine-like galleries filled with larval frass. In the present study, we tested the hypothesis that T. planipennisi regulates the host metabolism and feeding activity to optimize its offspring development and fitness. We first compared the respiration rate of parasitized and unparasitized host larvae at different times after parasitism, and then measured feeding activity of both parasitized and unparasitized host larvae inside their feeding galleries. Although parasitized host larvae increased metabolic rate and feeding activity in the first few days of parasitism, T. planipennisi parasitism induced an overall reduction of the metabolic rate and decrease in feeding activity of parasitized host larvae over their development period. In addition, there was a negative relationship between feeding activity of parasitized hosts and brood sizes of the parasitoid progeny-i.e., the more parasitoid progeny a host larva received, the less feeding activity the host had. These findings suggest that T. planipennisi has limited ability to optimize its offspring development and fitness through regulations of the host metabolism and feeding activity and its parasitism reduces feeding damage of parasitized EAB larvae to infested ash trees.

Successful establishment, spread, and impact of the introduced parasitoid Spathius galinae (Hymenoptera: Braconidae) on emerald ash borer (Coleoptera: Buprestidae) populations in postinvasion forests in Michigan.

Spathius galinae is a larval parasitoid native to the Russian Far East that was approved for release in the United States in 2015 for biological control of the emerald ash borer (EAB), Agrilus planipennis, an invasive beetle from Asia responsible for widespread mortality of ash trees (Fraxinus spp.) in North America. From 2015 to 2017, 1,340-1,445 females of S. galinae along with males were released into each release plot, paired with a nonrelease control plot (1-12.5 km apart), at 6 postinvasion forested sites containing abundant pole-sized ash trees in Michigan. By 2018, S. galinae had spread to all but one control plot. Based on the first year that S. galinae was found in trees in each control plot and the distances of those trees to the parasitoid release point within each site, we estimated that S. galinae spread at 3.7 (±1.9) km per year after its initial releases in 2015. The proportion of sampled trees with S. galinae broods, brood densities within sampled trees, and parasitism of EAB larvae increased sharply in both control and release plots after the last field releases in 2017, with the highest parasitism rates (42.8-60.3%) in 2020. Life table analysis showed that S. galinae alone reduced EAB's net population growth rate by 35-55% across sites from 2018 to 2020. These results demonstrate that S. galinae has established an increasing population in Michigan and now plays a significant role in reducing EAB populations in the area.

Postrelease assessment of Oobius agrili (Hymenoptera: Encyrtidae) establishment and persistence in Michigan and the Northeastern United States.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is an invasive woodboring pest of ash trees (Fraxinus sp.) in North America. Among the Asiatic parasitoids being released for the management of EAB in North America, Oobius agrili Zhang and Huang (Hymenoptera: Encyrtidae) is the only EAB egg parasitoid. To date, more than 2.5 million O. agrili have been released in North America; however, few studies have examined its success as a biological control agent of EAB. We conducted studies to assess O. agrili establishment, persistence, spread, and EAB egg parasitism rates in Michigan at the earliest release sites (2007-2010), as well as at more recent release sites (2015-2016) in 3 Northeastern United States (Connecticut, Massachusetts, New York). In both regions, we documented successful O. agrili establishment at all but one release site. In Michigan, O. agrili has persisted at release sites for over a decade and spread to all control sites located 0.6-3.8 km from release sites. Overall, EAB egg parasitism ranged from 1.5% to 51.2% (mean of 21.4%) during 2016-2020 in Michigan and from 2.6% to 29.2% (mean of 16.1%) during 2018-2020 in the Northeastern states. Future research efforts should focus on factors affecting the spatiotemporal variation in EAB egg parasitism rates by O. agrili, as well as its potential range in North America.

Effects of emamectin benzoate trunk injections on protection of neighboring ash trees against emerald ash borer (Coleoptera: Buprestidae) and on established biological control agents.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire, is the most destructive invasive pest on ash (Fraxinus spp.) in the United States. We determined whether ash trees injected with emamectin benzoate (EB) could protect untreated neighboring ash trees. We also determined whether the selective treatment of ash trees with EB injections had adverse effects on the establishment of introduced larval parasitoids Tetrastichus planipennis Yang and Spathius galinae Belokobylskij & Strazenac. In experiment one, trees were treated with EB and then retreated 3 years later. Five years post initial treatment, we found that 90% of treated ash trees retained healthy crowns, significantly higher than those of untreated control ash trees (16%). For experiment two, trees only received one treatment of EB and after 2 years 100% of treated ash trees retained healthy crowns, significantly higher than those of untreated ash trees (50%). In both experiments, we found that distance from the central EB-treated tree was not a significant predictor for tree health or presence of EAB exit holes. Although distance from the EB-treated trees appeared to have a significant positive relationship with woodpecker feeding signs on neighboring trees, such relationships did not result in significant differences in the proportion of neighboring ash trees retaining healthy crowns between EB treatment and control plots. The introduced EAB parasitoids appeared to have established equally well between treatment and control plots. Findings are discussed in the context of integration of EB trunk injection with biological control for protection of North American ash against EAB.

Potential effects of RNA interference of Asian longhorned beetle on its parasitoid.

BACKGROUND: It is important to understand how non-target insects such as parasitoids may be impacted directly or indirectly by RNA interference with double-stranded RNA (dsRNA) that has emerged as a novel pest control tool. We examined the potential effects of a dsRNA targeting an inhibitor of apoptosis (IAP) of the Asian longhorned beetle Anoplophora glabripennis on its gregarious larval ectoparasitoid Ontsira mellipes, directly on adult wasp's survival via injection of 4 µg of dsIAP per wasp, and indirectly on the detectability and suitability of host larvae injected with 2, 4 or 8 µg of dsIAP per larva. RESULTS: Compared with no injection or injection with a control dsGFP targeting a region of gene coding for a green fluorescence protein (GFP), dsIAP did not affect adult wasp's survival. Ontsira mellipes locates hosts in the wood by sensing their movement. Host larvae did not completely cease movement after the injection of dsIAP and were still detected and parasitized. Clutch size was reduced and only 3.8% of the parasitoid offspring developed into adults on host larvae treated at the highest dose. However, clutch size was not affected and 25.5% of the parasitoid offspring developed into adults on host larvae treated at the lowest dose. The fitness of developed wasps (development time, sex ratio, body size, and fecundity) was not affected when compared to the control treatments. No dsIAP was detected in parasitoid larvae. CONCLUSION: The results show no direct effect of the dsRNA on its parasitoid, but the potential indirect effect of dsRNA-affected host on the parasitoid, which may be minimized through optimizing dsRNA dosage to promote compatible applications of both management options for this invasive forest pest. © 2022 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

An integrative phenology and climatic suitability model for emerald ash borer.

Introduction: Decision support models that predict both when and where to expect emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), are needed for the development and implementation of effective management strategies against this major invasive pest of ash (Fraxinus species) in North America and other regions such as Europe. We present a spatialized model of phenology and climatic suitability for EAB for use in the Degree-Days, Risk, and Phenological event mapping (DDRP) platform, which is an open-source decision support tool to help detect, monitor, and manage invasive threats. Methods: We evaluated the model using presence records from three geographic regions (China, North America, and Europe) and a phenological dataset consisting primarily of observations from the northeastern and midwestern United States. To demonstrate the model, we produced phenological event maps for a recent year and tested for trends in EAB's phenology and potential distribution over a recent 20-year period. Results: Overall, the model exhibited strong performance. Presence was correctly estimated for over 99% of presence records and predicted dates of adult phenological events corresponded closely with observed dates, with a mean absolute error of ca. 7 days and low estimates of bias. Climate stresses were insufficient to exclude EAB from areas with native Fraxinus species in North America and Europe; however, extreme weather events, climate warming, and an inability for EAB to complete its life cycle may reduce suitability for some areas. Significant trends toward earlier adult emergence over 20 years occurred in only some areas. Discussion: Near real-time model forecasts for the conterminous United States are available at two websites to provide end-users with decision-support for surveillance and management of this invasive pest. Forecasts of adult emergence and egg hatch are particularly relevant for surveillance and for managing existing populations with pesticide treatments and parasitoid introductions.

Rapid Spread of an Introduced Parasitoid for Biological Control of Emerald Ash Borer (Coleoptera: Buprestidae) in Maryland.

Emerald ash borer (Agrilus planipennis Fairmaire (Coleoptera: Buprestidae)), an invasive phloem-feeding beetle native to Asia, has devastated North American ash forests since its detection in Michigan, United States in 2002. As the emerald ash borer has continued to spread, the potential for successful long-term management hinges upon the release, establishment, and spread of introduced larval and egg parasitoids for biological control. Here, we focus on the establishment and evidence for spatial spread of introduced larval parasitoid, Spathius agrili Yang and Spathius galinae Belokobylskij & Strazanac (Hymenoptera: Braconidae) in the state of Maryland. To assess each species, we analyzed historical release and recovery data and resampled previous release sites and nonrelease sites for establishment. We found little evidence of establishment or spread for S. agrili, despite a comparatively large number of release locations, events, and individuals. By contrast, despite much lower propagule pressure and shorter history of releases, we detected multiple established populations of S. galinae at release sites and at sites up to 90 km from the nearest release point approximately 3 yr after its most current release. Our findings show that S. galinae has established and spread rapidly following field releases whereas its congener, S. agrili has not. Although it may still be too early to evaluate the level of population control and ash protection afforded by S. galinae, these findings indicate the need for continued investment in S. galinae for emerald ash borer classical biological control efforts.

Optimal Conditions for Diapause Survival of Aprostocetus fukutai, an Egg Parasitoid for Biological Control of Anoplophora chinensis.

Aprostocetus fukutai is a specialist egg parasitoid of the citrus longhorned beetle Anoplophora chinensis, a high-risk invasive pest of hardwood trees. The parasitoid overwinters as diapausing mature larvae within the host egg and emerges in early summer in synchrony with the egg-laying peak of A. chinensis. This study investigated the parasitoid's diapause survival in parasitized host eggs that either remained in potted trees under semi-natural conditions in southern France or were removed from the wood and held at four different humidities (44, 75, 85-93 and 100% RH) at 11 °C or four different temperature regimes (2, 5, 10 and 12.5 °C) at 100% RH in the laboratory. The temperature regimes reflect overwintering temperatures across the parasitoid's geographical distribution in its native range. Results show that the parasitoid resumed its development to the adult stage at normal rearing conditions (22 °C, 100% RH, 14L:10D) after 6- or 7-months cold chilling at both the semi-natural and laboratory conditions. It had a low survival rate (36.7%) on potted plants due to desiccation or tree wound defense response. No parasitoids survived at 44% RH, but survival rate increased with humidity, reaching the highest (93.7%) at 100% RH. Survival rate also increased from 21.0% at 2 °C to 82.8% at 12.5 °C. Post-diapause developmental time decreased with increased humidity or temperature. There was no difference in the lifetime fecundity of emerged females from 2 and 12.5 °C. These results suggest that 100% RH and 12.5 °C are the most suitable diapause conditions for laboratory rearing of this parasitoid.

Protective neighboring effect from ash trees treated with systemic insecticide against emerald ash borer.

BACKGROUND: The emerald ash borer (EAB) (Agrilus planipennis Fairmaire) (Coleoptera: Buprestidae) is now the most destructive invasive species in North America. While biocontrol using parasitoids shows promising results in natural forests, strategies are needed to protect high-value trees against invasive EAB populations. Emamectin benzoate is a commonly used systemic insecticide for the protection of valuable trees. Methods that optimize its use allow for reduced quantities of insecticide to be released in the environment and save time and money in efforts to protect ash trees from EAB. We hypothesize that a treated tree can also offer a protective neighboring effect to nearby untreated ash trees, allowing for an optimized spatial planning of insecticide applications. RESULTS: We sampled 896 untreated ash trees, in the vicinity of treated trees, in Maryland and Washington DC. We recorded signs of EAB infestation (canopy condition, exit holes, wood pecks, epicormic growth, and bark splits). Two subsequent yearly samplings were made of 198 and 216 trees, respectively. We also present a novel proximity index for this particular application. Results show consistent decrease in EAB infestation signs in untreated trees as proximity to treated trees increases. CONCLUSION: Results support that a neighboring effect occurs. However, proximity to treated trees must be high for a tree to be safely left untreated. This proximity seems rare in forests, but can happen in urban/planted landscapes. Future studies should test and validate these findings, and could lead to a more precise recommended safe index tailored across multiple ash species and geographic regions.

Effects of Low Temperature Exposure on Diapause, Development, and Reproductive Fitness of the Emerald Ash Borer (Coleoptera: Buprestidae): Implications for Voltinism and Laboratory Rearing.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), native to Northeast Asia, is the most destructive invasive ash (Fraxinus spp.) pest in the United States. In the present study, we evaluated the effect of exposure of diapausing mature fourth instars (J-shaped larvae, JL) of EAB to cool temperatures, either 1.7 or 12.8°C for 1-9 mo, on their post-chill development including adult emergence, longevity, and lifetime fecundity under standard rearing conditions (26 ± 0.5°C, 16:8 h L:D). In addition, we determined the effect of different stages of the larvae chilled at 12.8°C for 3 mo on the subsequent post-chill development to EAB adults. Findings from the study revealed that a period (≥2 mo) of chill at 12.8°C is required for the termination of the EAB diapause. However, chill treatment of the larvae at the near zero temperature (1.7°C) does not result in the post-diapause larval development to adults, regardless of the chill time (1-9 mo). In addition, our results showed that chill treatment of immature young larvae (L1-L4 prior to JL) results in little production of EAB adults, indicating that EAB diapause predominantly as JL. Findings of this study may be useful to laboratory rearing of EAB from eggs through continuous generations and help us understand the pest's voltinism resulting from the diapause and post-diapause development under different climatic conditions.

Optimizing Parasitoid and Host Densities for Efficient Rearing of Ontsira mellipes (Hymenoptera: Braconidae) on Asian Longhorned Beetle (Coleoptera: Cerambycidae).

Ontsira mellipes Ashmead is a gregarious larval ectoparasitoid of woodboring cerambycids. It is native to North America but can readily attack the exotic Asian longhorned beetle, Anoplophora glabripennis (Motschulsky). This study aimed to develop an efficient rearing system for this parasitoid, as a potential novel association biocontrol agent for the beetle, by investigating the effects of different densities of host (two, three, or four larvae) and parasitoid (one, two, three, four, five, six, seven, and eight female wasps) on Ontsira's parasitization efficiency and reproductive outcomes. Results showed that overall parasitism and total numbers of parasitized hosts or progeny produced increased with host and/or parasitoid densities, but the number of parasitized hosts or progeny produced per female parasitoid decreased with parasitoid density at each given host density. Nonlinear regression indicated a consistent pattern of mutual interference as parasitoid density increased. Additional experiments showed that superparasitism (indirect interference) did not occur probably because the parasitoid detects hosts through vibration cues from host feeding and attacked (thus paralyzed) hosts are no longer detectable. Thus, the interference probably results from direct or exploitative competition. Interestingly, female parasitoids responded to increased parasitoid density with a significant increase in clutch size. Overall, per capita parasitization efficiency or reproductive outcomes were optimized at a low parasitoid-host ratio but with large group size of hosts and parasitoids. Therefore, an optimal combination of exposing three or four parasitoids to four hosts is proposed for efficient mass-rearing of this parasitoid.

Discovery of Parasitoids of Anoplophora glabripennis (Coleoptera: Cerambycidae) and Their Seasonal Abundance in China Using Sentinel Host Eggs and Larvae.

Asian longhorned beetle, Anoplophora glabripennis (Motschulsky), is a high-risk invasive forest pest worldwide. We surveyed Asian longhorned beetle parasitoid guilds and their seasonal abundance using field-deployed sentinel host logs infested with Asian longhorned beetle eggs or newly hatched (early-instar) larvae in three different sites of China (Beijing City, Shanghai City, and Jilin Province) from 2015 to 2018. Our survey detected 12 species of hymenopteran parasitoids (four Pteromalidae, three Braconidae, two Eupelmidae, one Eurytomidae, one Ichneumonidae, and one Bethylidae) attacking sentinel Asian longhorned beetle larvae or eggs deployed in these sites. Total parasitism by all the parasitoid species varied with different sites and across different years of the survey (averaging 7-16% in Beijing, 4-11% in Shanghai, and 0-0.2% in Jilin Province). In addition, the seasonal pattern of parasitism also differed among different sites, with parasitism peaking in July in the northern site (Beijing, 19%) and June in the southern site (Shanghai, 16%). Among all the parasitoid species recovered, Oxysychus sp. (Hymenoptera: Pteromalidae) was the most abundant parasitoid species in both Beijing and Shanghai (with 42-66% relative abundance and an average of 6% Asian longhorned beetle parasitism). The second most abundant species was Bracon planitibiae Yang, Cao et Gould (Hymenoptera: Braconidae), which accounted for 35% of the species collected and caused an average of 5% Asian longhorned beetle parasitism. Relevance of our findings to Asian longhorned beetle biocontrol is discussed.

Effects of Extreme Low Winter Temperatures on the Overwintering Survival of the Introduced Larval Parasitoids Spathius galinae and Tetrastichus planipennisi: Implications for Biological Control of Emerald Ash Borer in North America.

Climate change has been linked to shifts in the distribution and phenology of species although little is known about the potential effects that extreme low winter temperatures may have on insect host-parasitoid interactions. In late January 2019, northern regions of the United States experienced a severe cold wave caused by a weakened jet stream, destabilizing the Arctic polar vortex. Approximately 3 mo later at six study sites in southern Michigan and three in southern Connecticut, we sampled the overwintering larvae of the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), and two larval parasitoids, Spathius galinae (Hymenoptera: Braconidae) and Tetrastichus planipennisi (Hymenoptera: Eulophidae), that are being introduced as emerald ash borer biocontrol agents in North America. At these nine study sites, emerald ash borer-infested ash trees and/or saplings were debarked and each overwintering emerald ash borer and parasitoid larva was then examined for cold-induced mortality, as indicated by a brown coloration, flaccid, and watery consistency. In early spring in Michigan, we found 4.5-26% of emerald ash borer larvae, 18-50% of S. galinae larvae, and 8-35% of T. planipennisi larvae were killed by cold. In Connecticut where temperatures were more moderate than in Michigan during the 2019 cold wave, <2% of the larval hosts and parasitoids died from cold injury. Our findings revealed that cold-induced mortality of overwintering larvae of emerald ash borer and its larval parasitoids varied by location and species, with higher mortality of parasitoid larvae in most Michigan sites compared to host larvae. The potential impacts of our findings on the management of emerald ash borer using biocontrol are discussed.

Gene content evolution in the arthropods.

BACKGROUND: Arthropods comprise the largest and most diverse phylum on Earth and play vital roles in nearly every ecosystem. Their diversity stems in part from variations on a conserved body plan, resulting from and recorded in adaptive changes in the genome. Dissection of the genomic record of sequence change enables broad questions regarding genome evolution to be addressed, even across hyper-diverse taxa within arthropods. RESULTS: Using 76 whole genome sequences representing 21 orders spanning more than 500 million years of arthropod evolution, we document changes in gene and protein domain content and provide temporal and phylogenetic context for interpreting these innovations. We identify many novel gene families that arose early in the evolution of arthropods and during the diversification of insects into modern orders. We reveal unexpected variation in patterns of DNA methylation across arthropods and examples of gene family and protein domain evolution coincident with the appearance of notable phenotypic and physiological adaptations such as flight, metamorphosis, sociality, and chemoperception. CONCLUSIONS: These analyses demonstrate how large-scale comparative genomics can provide broad new insights into the genotype to phenotype map and generate testable hypotheses about the evolution of animal diversity.

Cerambycid Communities and Their Associated Hymenopteran Parasitoids From Major Hardwood Trees in Delaware: Implications for Biocontrol of Invasive Longhorned Beetles.

Cerambycidae provide important ecological services in forests yet cause economic damage when they infest living trees. Parasitoids can regulate woodborer populations, providing considerable control of pest cerambycids. Identifying parasitoids of native cerambycids may be useful in managing cerambycid outbreaks and aid in new-association biocontrol of exotic invasive cerambycids. We investigated Cerambycidae and associated hymenopteran parasitoid communities infesting Acer rubrum, Pinus virginiana, and Carya tomentosa from a forest in Delaware from 2005 to 2012. Cerambycid abundance, diversity, and richness, as well as parasitoid abundance, were measured by collecting trees in different conditions: felled, girdled, and naturally infested. Effect of edge or interior red maple on cerambycid abundance, diversity, and richness was examined. Over 14,500 cerambycids of 56 species and 38 genera were collected during the 7-yr period. Eleven species represented 95% of all cerambycids collected. Treatment only affected red maple, showing increased cerambycid richness and diversity from naturally infested trees. Cerambycid richness and diversity were two times greater on hickory than other species when combining girdled and felled treatments. Over 19,000 parasitic Hymenoptera of 12 families emerged from woodborer-infested wood with >70% of individuals belonging to Braconidae. Thirteen known species, and two unknown species, of Braconidae were identified from a subsample of 495 specimens; Ontsira mellipes (Ashmead) (Hymenoptera: Braconidae) and Rhoptrocentrus piceus Marshall (Hymenoptera: Braconidae) were the most abundant. This study provides fundamental information on native parasitoids associated with Cerambycidae, including cerambycid larval host associations. Parasitoids identified herein should be investigated for potential adaptation to invasive Cerambycidae to benefit invasive woodborer management.

Establishment and Early Impact of Spathius galinae (Hymenoptera: Braconidae) on Emerald Ash Borer (Coleoptera: Buprestidae) in the Northeastern United States.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire, a buprestid beetle native to Asia, has become a serious pest of ash trees (Fraxinus spp.) in North America since the early 2000s. Due to the impracticality of applying insecticides in natural forests, biocontrol is the most viable method to manage EAB in natural ecosystems. Here, we report the first evidence for the establishment and impact of Spathius galinae Belokobylskij & Strazenac, a larval parasitoid first released in North America in 2016 and 2017 at six mixed-hardwood forest sites, in Connecticut, New York, and Massachusetts. We also report current levels of abundance and parasitism of another introduced larval EAB parasitoid, Tetrastichus planipennisi Yang (Hymenoptera: Eulophidae), released in 2015 and 2016 in these same sites. Spathius galinae was recovered at all release sites in 2018, and its density in sampled trees had increased 1.5- to 20-fold (relative to the first postrelease sample year), reaching a final density of 2.3-14.3 broods/m2 of phloem area and causing 13.1-49.2% marginal rate of parasitism at four of the six sites. In contrast, T. planipennisi was only recovered in 2018 at four of the six release sites, and both its density (0.1-2.3 broods/m2 of phloem area) and parasitism (0.1-5.6%) were lower than that of S. galinae throughout the study at the four sites where recoveries were made. Our data fill a critical gap in the development of a biocontrol-based EAB management plan to protect surviving ash trees capable of reaching maturity and producing replacement trees.

Influence of Density on Interspecific Competition Between Spathius galinae (Hymenoptera: Braconidae) and Tetrastichus planipennisi (Hymenoptera: Eulophidae), Larval Parasitoids of the Invasive Emerald Ash Borer (Coleoptera: Buprestidae).

The outcomes of interspecific interactions between parasitoids depend on a variety of factors. Understanding the influence of these factors is important for classical biological control, where the success of parasitoid releases partly depends on interactions with native and other introduced species. However, results from laboratory experiments may not always reflect those in the field, as densities may be artificially inflated. To mitigate this problem, we examined the effects of multiple densities on interspecific competition between two larval parasitoids of emerald ash borer (Agrilus planipennis Fairmaire): Spathius galinae Belokobylskij and Tetrastichus planipennisi Yang. Parasitoid species were housed individually or together at two different densities, and we measured the effects on percent parasitism and progeny production, before calculating the interaction strengths. We found no significant effects of parasitoid density on percent parasitism, but the effect of competition on parasitism generally was reduced at lower densities. However, there were significant differences in parasitism by species, with S. galinae parasitizing more larvae than T. planipennisi. There were also no significant effects of parasitoid density on the number of progeny produced by each species, though the effect of competition on progeny production was greater at higher densities. Similarly, though, there were significant differences between species in the number of progeny produced. Specifically, T. planipennisi consistently produced larger broods than S. galinae. Our findings complement existing research suggesting that competition between these two species in the field will likely be negligible.

Collecting Nontarget Wood-Boring Insects for Host-Specificity Testing of Natural Enemies of Cerambycids: A Case Study of Dastarcus helophoroides (Coleoptera: Bothrideridae), a Parasitoid of the Asian Longhorned Beetle (Coleoptera: Cerambycidae).

We describe approaches to addressing the perennial challenge of collecting a sufficient diversity of nontarget insects for host-specificity testing of candidate biocontrol agents of invasive wood-borers such as the Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae). Multifunnel and intercept traps, retrofitted to maintain live insects and baited with cerambycid-specific pheromone lures, were deployed in diverse forests in southeastern Massachusetts. We collected 1,288 adult beetles comprising 56 species, mostly from the subfamilies targeted by the lures (Cerambycinae and Lamiinae). The type of trap and tree species in which the trap was hung did not seem to affect the species caught. Methods used to induce egg laying and techniques to rear cerambycid larvae are described. Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae) is the most common Asian longhorned beetle parasitoid in China; therefore, we conducted tests to determine whether cerambycids native to North America would be at risk should this species be released. D. helophoroides attacked all six species tested: Monochamus scutellatus (Say) (Coleoptera: Cerambycidae: Lamiinae), Monochamus notatus (Drury) (Coleoptera: Cerambycidae: Lamiinae), Apriona rugicollis Chevrolat (Coleoptera: Cerambycidae: Lamiinae), Enaphalodes rufulus (Haldeman) (Coleoptera: Cerambycidae: Cerambycinae), Graphisurus fasciatus (DeGeer) (Coleoptera: Cerambycidae: Lamiinae), and Neoclytus acuminatus (F.) (Coleoptera: Cerambycidae: Cerambycinae). Parasitism of native cerambycids was not statistically different from parasitism of Asian longhorned beetle, except for N. acuminatus, which is a considerably smaller species than Asian longhorned beetle, and for M. notatus and M. scutellatus, which attack pine trees rather than hardwood trees like Asian longhorned beetle and the other native species tested. Our testing indicates that many native cerambycids would be vulnerable to D. helophoroides and we conclude that D. helophoroides should not be considered for release as a biocontrol agent in North America.

Identification of highly effective target genes for RNAi-mediated control of emerald ash borer, Agrilus planipennis.

Recent study has shown that RNA interference (RNAi) is efficient in emerald ash borer (EAB), Agrilus planipennis, and that ingestion of double-stranded RNA (dsRNA) targeting specific genes causes gene silencing and mortality in neonates. Here, we report on the identification of highly effective target genes for RNAi-mediated control of EAB. We screened 13 candidate genes in neonate larvae and selected the most effective target genes for further investigation, including their effect on EAB adults and on a non-target organism, Tribolium castaneum. The two most efficient target genes selected, hsp (heat shock 70-kDa protein cognate 3) and shi (shibire), caused up to 90% mortality of larvae and adults. In EAB eggs, larvae, and adults, the hsp is expressed at higher levels when compared to that of shi. Ingestion of dsHSP and dsSHI caused mortality in both neonate larvae and adults. Administration of a mixture of both dsRNAs worked better than either dsRNA by itself. In contrast, injection of EAB.dsHSP and EAB.dsSHI did not cause mortality in T. castaneum. Thus, the two genes identified cause high mortality in the EAB with no apparent phenotype effects in a non-target organism, the red flour beetle, and could be used in RNAi-mediated control of this invasive pest.