Role of 3-Carene in Host Location and Colonization by Dendroctonus pseudotsugae (Coleoptera: Curculionidae).

The Douglas-fir beetle (Dendroctonus pseudotsugae Hopkins) occasionally colonizes western larch [Larix occidentalis Nutt. (Pinales: Pinaceae)] growing in close proximity to its primary host, Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco (Pinales: Pinaceae)], but brood have never been found to survive in live western larch. Western larch produces the monoterpene 3-carene in higher concentrations than Douglas-fir. In this study, the toxicity and repellency of 3-carene to Douglas-fir beetle was evaluated in a series of laboratory and field tests. In a laboratory bioassay, 3-carene was one of the most toxic monoterpenes to Douglas-fir beetles among those tested. In a field trial, addition of 3-carene to multiple-funnel traps baited with frontalin (the primary component of Douglas-fir beetle aggregation pheromone) or frontalin and α-pinene significantly reduced the number of Douglas-fir beetles collected. In another field study, live western larch, felled western larch, live Douglas-fir, felled Douglas-fir, and live Douglas-fir surrounded by 3-carene releasers were baited with Douglas-fir beetle aggregation pheromones. There were significantly fewer Douglas-fir beetle entrance holes and egg galleries excavated on both live western larch and live Douglas-fir surrounded by 3-carene compared with live Douglas-fir. Most egg galleries excavated in live western larch were heavily impregnated with resin and no eggs hatched. There were no significant differences in egg galleries excavated or eggs hatched between felled western larch and felled Douglas-fir. Collectively, these data support the hypothesis that 3-carene slows the colonization process in live western larch allowing more time for host trees to respond to a colonization attempt and a higher likelihood of successfully resisting infestation.

Wild Bee Response to Application of the Douglas-fir Beetle Anti-Aggregation Pheromone, 3-Methylcyclohex-2-En-1-One.

Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (DFB), is the most damaging insect pest of Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, in western North America. Individual high-value trees and stands can be protected during DFB outbreaks using the beetle's anti-aggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH), which is available in several commercial formulations. However, other methylcyclohexanones similar in molecular structure to MCH have been shown to repel bees from agricultural areas. In forest systems, wild bees (Hymenoptera: Apiformes) provide essential pollination services to many forb and shrub species. Therefore, it is important to determine whether resident wild bee communities are affected in forests treated with MCH. To address this, the bee community was sampled within experimental sites located in Idaho and Montana, USA in 2016. At both sites, there was no significant difference in overall bee abundance, species richness, or diversity between MCH-treated plots and untreated control plots. Overall, these results indicate that treatment of Douglas-fir with MCH does not negatively impact the resident wild bee community.

Changes in the Summer Wild Bee Community Following a Bark Beetle Outbreak in a Douglas-fir Forest.

The status of wild bees has received increased interest following recent estimates of large-scale declines in their abundances across the United States. However, basic information is limited regarding the factors affecting wild bee communities in temperate coniferous forest ecosystems. To assess the early responses of bees to bark beetle disturbance, we sampled the bee community of a Douglas-fir, Pseudotsuga menziesii (Mirb.), forest in western Idaho, United States during a Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (Coleoptera: Curculionidae), outbreak beginning in summer 2016. We resampled the area in summer 2018 following reductions in forest canopy cover resulting from mortality of dominant and codominant Douglas-fir. Overall, results from rarefaction analyses indicated significant increases in bee diversity (Shannon's H) in 2018 compared to 2016. Results from ANOVA also showed significant increases in bee abundance and diversity in 2018 compared to 2016. Poisson regression analyses revealed percent tree mortality from Douglas-fir beetle was positively correlated with increases in total bee abundance and species richness, where community response variables displayed a cubic trend with percent tree mortality. Percent reduction in canopy cover from 2016 to 2018 was also correlated with bee species richness and diversity. These findings suggest that wild bee communities may benefit from changes in forest structure following bark beetle outbreaks.

Temporal Asynchrony of Adult Emergence Between Leucopis argenticollis and Leucopis piniperda (Diptera: Chamaemyiidae), Predators of the Hemlock Woolly Adelgid (Hemiptera: Adelgidae), with Implications for Biological Control.

Two species of silver fly, Leucopis argenticollis (Zetterstedt) and Leucopis piniperda (Malloch) (Diptera: Chamaemyiidae), from the Pacific Northwest region of North America have been identified as potential biological control agents of hemlock woolly adelgid (Hemiptera: Adelgidae: Adelges tsugae Annand) in eastern North America. The two predators are collectively synchronized with A. tsugae development. To determine whether adult emergence of the two species of silver fly are also synchronized with one another, we collected adult Leucopis which emerged from A. tsugae-infested western hemlock [Pinaceae: Tsuga heterophylla (Raf.) Sarg.] from four sites in the Pacific Northwest over a 29-d period. Specimens were collected twice daily in the laboratory and identified to species using DNA barcoding. The study found that more adult Leucopis were collected in the evening than the morning. Additionally, the daily emergences of adults over the 29-d sampling period exhibited sinusoidal-like fluctuations of peak abundance of each species, lending evidence to a pattern of temporal partitioning. This pattern could have logistical implications for their use as biological control agents in eastern North America, namely the need to release both species for maximum efficacy in decreasing A. tsugae populations.

A Biodegradable Formulation of MCH (3-Methylcyclohex-2-en-1-one) for Protecting Pseudotsuga menziesii from Dendroctonus pseudotsugae (Coleoptera: Curculionidae) Colonization.

Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, trees and stands can be protected from Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (DFB)-caused mortality by application of synthetic formulations of the beetle's antiaggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH). A biodegradable formulation of MCH, SPLAT MCH, was developed and evaluated for protecting individual Douglas-fir trees and small stands from colonization and mortality by DFB. In an individual-tree experiment in Idaho, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of treated trees colonized and killed by DFB compared to untreated controls. SPLAT MCH was as effective as MCH bubble capsules for protecting individual trees. Both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB within 0.04-ha circular plots surrounding each treated tree compared to untreated controls. In 0.41 ha stands in New Mexico, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB compared to untreated controls, again with no differences observed between MCH treatments. In a similar stand level trial in Idaho, neither MCH treatment significantly reduced the proportion of trees colonized by DFB, and only MCH bubble capsules significantly reduced levels of tree mortality compared to untreated controls, but no significant difference was observed between SPLAT MCH and MCH bubble capsules. Overall, the results indicate that SPLAT MCH is as effective as MCH bubble capsules for protecting individual trees and small stands of Douglas-fir from DFB-caused mortality.

Coexistence of three specialist predators of the hemlock woolly adelgid in the Pacific Northwest USA.

The hemlock woolly adelgid (Hemiptera: Adelgidae: Adelges tsugae Annand) is an invasive insect, introduced from Japan to eastern North America, where it causes decline and death of hemlock trees. There is a closely related lineage of A. tsugae native to western North America. To inform classical biological control of A. tsugae in the eastern USA, the density and phenology of three native western adelgid specialist predators, Leucopis argenticollis (Zetterstedt), Le. piniperda (Malloch) (Diptera: Chamaemyiidae), and Laricobius nigrinus Fender (Coleoptera: Derodontidae), were quantified in the Pacific Northwest. Infested branches were collected from western hemlock (Pinaceae: Tsuga heterophylla (Raf.) Sarg.) at four sites around the Puget Sound, Washington and three sites in Oregon. Immature Leucopis were identified to species using DNA barcodes. Leucopis argenticollis was roughly twice as abundant as Le. piniperda. Laricobius nigrinus larvae were more abundant than the two species of Leucopis during the egg stage of the first adelgid generation, but Leucopis were present as feeding larvae during the second adelgid generation when La. nigrinus was aestivating in the soil, resulting in Leucopis being more abundant than La. nigrinus across the entire sampling period. Adelges tsugae and La. nigrinus densities were not correlated, while A. tsugae and Leucopis spp. densities were positively correlated. Leucopis spp. and La. nigrinus densities were negatively correlated. These results support the complementary use of La. nigrinus and the two Leucopis species for biological control of A. tsugae in the eastern USA, and point to the need for further investigation of spatial and temporal niche partitioning among the three predator species.

Evaluating High Release Rate MCH (3-Methylcyclohex-2-en-1-one) Treatments for Reducing Dendroctonus pseudotsugae (Coleoptera: Curculionidae) Infestations.

Current recommendations for applying the antiaggregation pheromone 3-methylcyclohex-2-en-1-one (MCH) to protect live trees from Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins, infestation are to space individual passive releasers (MCH bubble capsules) on a 12- by 12-m grid throughout areas to be protected. Previous field studies and a theoretical study using a puff dispersion model to predict pheromone concentrations have shown that releasers emitting higher rates of MCH spaced farther apart may be as effective as the established standard treatment. During 2012 and 2013, we tested higher release rates of MCH at correspondingly wider spacings to keep the total amount of MCH released per unit area equal in all treatments. In 2012 near Challis, ID, treatments included the established standard release rate and spacing, four and six times the standard release rate at correspondingly wider spacings, and an untreated control. In 2013 near Ketchum, ID, treatments included the established standard release rate and spacing, five and seven times the standard release rate at correspondingly wider spacings, and an untreated control. Results from both years indicated that all MCH treatments were equally effective in reducing Douglas-fir beetle infestation. Using higher release rate formulations at wider spacings will reduce labor costs of installing MCH treatments, and, in cases where it is necessary, retrieving the releasers as well. In addition to reducing labor costs, the revised treatment protocol may increase the feasibility of treating areas that currently may not be possible due to treatment costs.

Predicting Dendroctonus pseudotsugae (Coleoptera: Curculionidae) antiaggregation pheromone concentrations using an instantaneous puff dispersion model.

An instantaneous puff dispersion model was used to assess concentration fields of the Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins, antiaggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH), within a 1-ha circular plot. Several combinations of MCH release rate and releaser spacing were modeled to theoretically analyze optimal deployment strategies. The combinations of MCH release rate and releaser spacing used in the modeling exercise were based on results of previous field studies of treatment efficacy. Analyses of model results suggest that a release rate up to six times the initial standard, at a correspondingly wider spacing to keep the total amount of pheromone dispersed per unit area constant, may be effective at preventing Douglas-fir beetle infestation. The model outputs also provide a visual representation of pheromone dispersion patterns that can occur after deployment of release devices in the field. These results will help researchers and practitioners design more effective deployment strategies.

Prey suitability and phenology of Leucopis spp. (Diptera: Chamaemyiidae) associated with hemlock woolly adelgid (Hemiptera: Adelgidae) in the Pacific Northwest.

Leucopis spp. (Diptera: Chamaemyiidae) from the Pacific Northwest previously were identified as potential biological control agents for the hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), in the eastern United States. We collected Leucopis spp. larvae from A. tsugae infested western hemlocks in Oregon and Washington and reared them on an unidentified Pineus spp., Pineus strobi (Hartig), Adelges cooleyi (Gillette), Adelges piceae (Ratzeburg), and A. tsugae in three no-choice tests. Leucopis spp. survival on A. tsugae was significantly higher than on A. piceae during the 2010 progrediens generation test and significantly higher than on P. strobi and A. cooleyi during the 2010 sistens generation test. However, across all three tests, some larvae completed development to adult on all four of the alternative adelgid species. Larvae that survived to the adult stage were identified as Leucopis argenticollis Zetterstedt and Leucopis piniperda Malloch. These results suggest that populations of L. argenticollis and L. piniperda in the Pacific Northwest may not be specific to A. tsugae. We also studied the phenology of Leucopis spp. on fourteen A. tsugae infested western hemlock trees in Oregon and Washington over a period of 14 mo. Leucopis spp. larvae were collected year-round, but highest densities coincided with the presence of progrediens and sistens eggs and adults of A. tsugae. There was a positive correlation between Leucopis spp. and A. tsugae abundance.

Orientation behavior of the predator Laricobius nigrinus (Coleoptera: Derodontidae) to hemlock woolly adelgid and host tree odors in a multi-chambered olfactometer.

We studied the adult ambulatory response of the predator, Laricobius nigrinus Fender (Coleoptera: Derodontidae), to odors from its prey, Adelges tsugae Annand, the hemlock woolly adelgid, and foliage of hemlock woolly adelgid, host hemlocks (Tsuga spp.), and other conifers. Both the predator and hemlock woolly adelgid are apparently native to western North America, but the predator is being released in the eastern United States, which has different hemlock species, for biological control of a lineage of hemlock woolly adelgid inadvertently introduced from Japan. L. nigrinus responded to odors from hemlock woolly adelgid host trees, but not to odors from hemlock woolly adelgid. L. nigrinus collected from hemlock woolly adelgid-infested western hemlock were more strongly attracted to odors from western hemlock [Tsuga heterophylla (Rafinesque) Sargent] than eastern hemlock [Tsuga canadensis (L.) Carrière] in most trials. Odors from western white pine (Pinus monticola Douglas ex D. Don) and white spruce [Picea glauca (Moench) Voss] were as attractive as western hemlock odors whereas odors from Douglas-fir [Pseudotsuga menziesii variety menziesii (Mirbel)] and ponderosa pine (Pinus ponderosa Douglas ex Lawson) were avoided. L. nigrinus reared on hemlock woolly adelgid-infested eastern hemlock in the laboratory were lethargic and were not attracted to either eastern or western hemlock odors. Predators collected in the field and tested monthly from December to March responded similarly each month, except February, when they flew rather than walked in the olfactometer, suggesting a period of dispersal or mate finding at that time of year. The implications of these results for programs to release L. nigrinus in the eastern United States for control of hemlock woolly adelgid are discussed.

Elution rate and spacing of antiaggregation pheromone dispensers for protecting live trees from Dendroctonus pseudotsugae (Coleoptera: Scolytidae).

The antiaggregation pheromone 3-methylcyclohex-2-en-1-one (MCH) is highly effective in preventing the infestation of high-risk trees by Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins. A large portion of the cost of an MCH treatment is related to the time applicators spend walking through an area dispersing the formulated pheromone. Application of fewer MCH dispensers eluting at a higher rate than those currently registered for operational use could potentially reduce treatment costs. Two higher elution rates, 6 and 18 mg/d per dispenser, were compared with the current standard of 2 mg/d per dispenser and an untreated control on 1-ha circular plots. Dispensers were spaced 5,15, and 44 m apart around the plot perimeters eluting 2, 6, and 18 mg/d, respectively. The nominal dose of MCH was 144 mg/ha/d on all plots. Percentages of Douglas-fir trees > or = 20 cm diameter at breast height mass attacked by Douglas-fir beetle were significantly lower on plots treated with dispensers eluting 2 and 6 mg/d and spaced 5 and 15 m apart, respectively, compared with the untreated control. Infestation rate on plots treated with dispensers eluting 18 mg/d and spaced 44 m apart was not significantly different from the control. Douglas-fir beetle abundance and host tree availability were similar on all plots. These results indicate that MCH dispensers eluting 6 mg/d (three times the current standard rate) and spaced 15 m apart (three times existing standard distance) can effectively prevent Douglas-fir beetle infestations.