Identification of a potential third component of the male-produced pheromone of Anoplophora glabripennis and its effect on behavior.

The Asian longhorned beetle, Anoplophora glabripennis, is considered to be one of the most serious invasive pests of deciduous trees in North America. An efficient monitoring trap is needed to detect and delimit new introductions and assess population densities of established infestations. Previous studies on A. glabripennis have shown that males produce a two-component aggregation pheromone that consists of a 1:1 blend of 4-(n-heptyloxy)butan-1-ol and 4-(n-heptyloxy)butanal. Moderate attraction in field trapping studies suggested that there may be additional chemical cues missing. Volatiles from male A. glabripennis were examined to identify other potential pheromone components. Gas chromatographic / electroantennographic (GC/EAD) analyses of male aerations detected a consistent EAD-active response to a previously unidentified compound. This compound was identified as (3E,6E)-α-farnesene. Both male and female beetles were antennally responsive to this sesquiterpene, and both sexes were attracted to it in olfactometer bioassays at different doses. When (3E,6E)-α-farnesene was combined with 4-(n-heptyloxy)butan-1-ol and 4-(n-heptyloxy)butanal, attraction of both sexes increased compared to assays using 4-(n-heptyloxy)butan-1-ol and 4-(n-heptyloxy)butanal alone.

A comparison of electrophysiologically determined spectral responses in six subspecies of Lymantria.

The spectral sensitivity of the compound eye in three gypsy moth species from six different geographical regions (Lymantria dispar asiatica Vnukovskij [Asian gypsy moth], Lymantria dispar japonica Motschulsky [Japanese gypsy moth], and Lymantria dispar dispar L. [North American gypsy moth]) was tested electrophysiologically in the wavelength region 300-700 nm. For all moths examined, a maximum response occurred in the 480-520-nm range (blue-green region) with a shoulder peak occurring at 460 nm. A smaller, secondary peak was observed for both sexes at the 340-380-nm range, which is in the region considered behaviorally maximal in night-flying insects. No peaks in sensitivity were observed between 520 and 700 nm (red region) for any of the moths tested. Based on our retinal recording data, a short wavelength blocking filter with a transition wavelength near 500 nm should reduce gypsy moth attraction to artificial lighting sources. This would help reduce the number of Lymantria-infested ships traveling to and from foreign ports.

Improving detection tools for emerald ash borer (Coleoptera: Buprestidae): comparison of multifunnel traps, prism traps, and lure types at varying population densities.

The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is a serious invasive pest of North American ash (Fraxinus spp.) that has caused devastating mortality since it was first identified in North America in 2002. In 2012, we conducted field trapping assays that tested the efficacy of purple prism and fluon-coated green multifunnel (Lindgren funnel) traps. Traps were baited with combinations of several lures that were previously shown to be attractive to A. planipennis: manuka oil--a sesquiterpene-rich oil, (3Z)-hexenol--a green leaf volatile, or (3Z)-dodecen-12-olide [= (3Z)-lactone], a sex pheromone. Eighty-nine blocks (trap lines) were tested throughout nine states along the outer edges of the currently known A. planipennis infestation in North America. Trap catch was highest on fluon-coated green multifunnel traps, and trap detections at sites with low A. planipennis population density ranged from 72 to 76% for all trap and lure types tested. (3Z)-hexenol and (3Z)-lactone baited traps functioned as well as (3Z)-hexenol and manuka oil-baited traps. Independent of the lure used, detection rates on green fluon-coated multifunnel traps were comparable with glued purple prism traps in areas with low A. planipennis population densities.

Improving detection tools for the emerald ash borer (Coleoptera: Buprestidae): comparison of prism and multifunnel traps at varying population densities.

The current emerald ash borer survey trap used in the United States is a prism trap constructed from a stock purple corrugated plastic. In recent years, several colors (particularly shades of green and purple) have been shown to be more attractive to the emerald ash borer than this stock color. Our goal was to determine if plastics produced with these colors and incorporated into prism traps can improve and serve as a new alternative to plastics already in use for the emerald ash borer survey. The plastics were tested in moderate to heavily infested areas in Michigan in two initial studies to test their effectiveness at catching the emerald ash borer. Because results from studies performed in heavily infested sites may not always correspond with what is found along the edges of the infestation, we compared trap catch and detection rates (recording at least one catch on a trap over the course of the entire trapping season) of several trap types and colors at sites outside the core of the currently known emerald ash borer infestation in a nine-state detection tool comparison study. Two of the new plastics, a (Sabic) purple and a medium-dark (Sabic) green were incorporated into prism traps and tested alongside a standard purple prism trap and a green multifunnel trap. In areas with lower emerald ash borer density, the new purple (Sabic) corrugated plastic caught more beetles than the current purple prism trap, as well as more than the medium-dark green (Sabic) prism and green multifunnel traps. Sabic purple traps in the detection tools comparison study recorded a detection rate of 86% compared with 73, 66, and 58% for the standard purple, Sabic green, and green multifunnel traps, respectively. These detection rates were reduced to 80, 63, 55, and 46%, respectively, at low emerald ash borer density sites.

Influence of trap color and host volatiles on capture of the emerald ash borer (Coleoptera: Buprestidae).

Field trapping assays were conducted in 2009 and 2010 throughout western Michigan, to evaluate lures for adult emerald ash borer, A. planipennis Fairmaire (Coleoptera: Buprestidae). Several ash tree volatiles were tested on purple prism traps in 2009, and a dark green prism trap in 2010. In 2009, six bark oil distillate lure treatments were tested against manuka oil lures (used in 2008 by USDA APHIS PPQ emerald ash borer cooperative program). Purple traps baited with 80/20 (manuka/phoebe oil) significantly increased beetle catch compared with traps baited with manuka oil alone. In 2010 we monitored emerald ash borer attraction to dark green traps baited with six lure combinations of 80/20 (manuka/phoebe), manuka oil, and (3Z)-hexenol. Traps baited with manuka oil and (3Z)-hexenol caught significantly more male and total count insects than traps baited with manuka oil alone. Traps baited with manuka oil and (3Z)-hexenol did not catch more beetles when compared with traps baited with (3Z)-hexenol alone. When compared with unbaited green traps our results show that (3Z)-hexenol improved male catch significantly in only one of three field experiments using dark green traps. Dark green traps caught a high number of A. planipennis when unbaited while (3Z)-hexenol was seen to have a minimal (nonsignificant) trap catch effect at several different release rates. We hypothesize that the previously reported kairomonal attractancy of (3Z)-hexenol (for males) on light green traps is not as obvious here because of improved male attractancy to the darker green trap.

Electrophysiologically Determined Spectral Responses in Lobesia botrana (Lepidoptera: Tortricidae).

Electrophysiological methods were used to test the visual sensitivity of European grapevine moth, Lobesia botrana (Lepidoptera: Tortricidae) to wavelengths ranging from 300 to 700 nm. For male and females tested, a main, peak response occurred in the 460-540 nm range (blue-green wavelengths) with females having a generally lower response to wavelengths in that range. A second smaller peak was observed for both sexes at the 340-420 nm range. A general linear model indicated that males, virgin females, and mated females did not react differently to changes in wavelength. No moths showed any obvious sensitivity to wavelengths between 580 and 700 nm. Based on our retinal recording data we suggest that UV light traps (≤480 nm) could be utilized alongside pheromone traps when monitoring L. botrana in high risk areas.

Absolute configuration of 7-epi-sesquithujene.

7-epi-sesquithujene (1) is a bicyclic sesquiterpene isolated from phoebe oil, an essential oil of the Brazilian walnut tree, Phoebe porosa. It is also produced by stressed ash trees and has been shown to elicit strong electrophysiological responses on emerald ash borer, Agrilus planipennis, antennae. In the course of the development of a synthetic 7-epi-sesquithujene lure for field testing against the emerald ash borer, we found that the absolute configuration of this compound had not been determined. We isolated >95% pure 7-epi-sesquithujene from phoebe oil via successive fractionation and conventional and argentation (HPLC) chromatographies. The specific optical rotation of this compound matched that of a synthetic product of known configuration. We also synthesized two other stereoisomers of sesquithujene and developed a chiral GC method to separate all four. Based on the specific rotation, stereoselective syntheses, and chiral GC analyses, 7-epi-sesquithujene present in phoebe oil and white ash was found to have the 2S,6S,7R absolute configuration.

Chemical ecology of the emerald ash borer Agrilus planipennis.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) is a serious invasive pest that has caused devastating mortality of ash trees (Fraxinus sp., Oleaceae) since it was first identified in North America in 2002. Shortly after its discovery, surveys were conducted, based on the visual inspection of trees. The shortcomings of visual surveys have led to a critical research need to find an efficient survey method for detecting A. planipennis infestations. Here, we present a review of research that has led to the development of effective trapping methods for A. planipennis. Studies on the insect's biology and behavior have led to the identification of several potential attractants as well as the design of a visually attractive trap. The ongoing challenge in developing an optimally efficient trapping methodology for A. planipennis will involve finding the best combination of variables, such as trap shape, trap color (or other visual properties), trap placement, lure components, as well as the ratios and release rates of those components.

Optimization of trap color for emerald ash borer (Coleoptera: Buprestidae).

Field assays were performed to determine the optimal color for Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) traps. Previous studies have found that more A. planipennis are caught on purple or green traps than traps of other colors. In three studies, we evaluated various shades of purple, wavelengths of green (500-570 nm), and greens of different reflectance (from 9 to 66%). In all tests, traps of corrugated plastic in standard, commercially available purple (currently used to survey A. planipennis) and a customized green color were used as bases for comparison. Among purple traps, a paint color previously shown to be generally attractive to buprestids caught significantly more A. planipennis adults than traps coated with paints containing more blue or red, or traps constructed of the standard purple plastic. Among traps with maximum reflectance at varying green wavelengths, those ranging in wavelength from 525 to 540 nm caught significantly more adult A. planipennis than traps of other wavelengths. In the 530-540 nm range of the electromagnetic spectrum, there was no significant difference among traps in the 23-66% reflectance range, but traps painted with a peak reflectance of 49% caught more beetles than purple or the custom green plastic traps. Male to female ratio was highest on green traps.

Monoalkenes as contact sex pheromone components of the woodwasp Sirex noctilio.

A pheromone on the cuticle of females of the woodwasp Sirex noctilio, a recently introduced pest of pines in North America, induces conspecific males to attempt copulation. Dead females washed with hexane did not elicit copulation attempts from males, whereas reapplication of a female hexane body wash onto the cuticle of dead females elicited copulation attempts by 65% of males tested. Analysis of the hexane extract revealed saturated and unsaturated hydrocarbons as major components of the female cuticle. Behavior-guided fractionation of the female body wash led to the identification of three components, (Z)-7-heptacosene, (Z)-7-nonacosene, and (Z)-9-nonacosene, of the sex pheromone of S. noctilio that elicited copulatory responses from males.

Laboratory and field response of the emerald ash borer (Coleoptera: Buprestidae), to selected regions of the electromagnetic spectrum.

Retinal sensitivity of Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) was examined with an aim to improve trap efficacy for the beetle. Electroretinogram (ERG) recordings from dark-adapted compound eyes of male and female were measured at different wavelengths across the spectrum ranging from 300 to 700 nm. The spectral sensitivity curves revealed peaks in the UV (340 nm), the violet/purple (420-430 nm), blue (460 nm), and green (540-560 nm) regions of the spectrum. Females were sensitive to red regions of the spectrum (640-670 nm), whereas males were not. A spectrophotometer was used to measure the wavelength and reflectance for ash foliage, purple corrugated plastic traps, as well as the elytra and abdomen of adult A. planipennis. Traps were painted using colors based on ERG and spectrophotometer measurements and compared with purple corrugated plastic traps currently used by the USDA-APHIS-PPQ-EAB National Survey. In a field assay conducted along the edges of several A. planipennis-infested ash stands, there were no significant differences in trap catch among green, red, or purple treatments. Dark blue traps caught significantly fewer A. planipennis than red, light green, or dark purple traps. In a second assay where purple and green treatments were placed in the mid canopy of ash trees (approximately 13 m in height), trap catch was significantly higher on green treatments. We hypothesize that when placed in the mid-canopy, green traps constitute a foliage-type stimulus that elicits food-seeking and/or host seeking behavior by A. planipennis.

Isolation and identification of a male-produced aggregation-sex pheromone for the velvet longhorned beetle, Trichoferus campestris.

The velvet longhorned beetle, Trichoferus campestris (Faldermann) ("VLB"; Coleoptera: Cerambycidae), is native to eastern Asia where it infests and damages a wide range of deciduous and coniferous tree species, including orchard and timber species. Immature stages of VLB are transported to new countries via international commerce, and populations have established outside the native range of the species. Here, we show that identification of pheromones of invasive pest species can be expedited by knowledge of the semiochemistry of related taxa. Histological sectioning revealed subcuticular, male-specific prothoracic glands connected to pits in the cuticle, which, in related species, are diagnostic for production of male-produced aggregation-sex pheromones, usually characterized by 2,3-alkanediol/hydroxyketone structural motifs. However, in preliminary field bioassays, beetles were not attracted by any known cerambycid pheromones. Subsequently, we identified a novel variant of the hydroxyketone motif ("trichoferone") from headspace volatiles of males. In field bioassays, synthetic trichoferone was more attractive to both sexes of VLB than previously developed high-release-rate ethanol lures, and attraction was strongly female biased. This study demonstrated the utility of the prothoracic gland trait for predicting pheromone use in cerambycid species in the subfamily Cerambycinae, and that identification of pheromones of novel species can be expedited by knowledge of pheromones of related species. Trichoferone should prove to be a valuable tool for detection of VLB in regions where the beetle is or may become established.

Development of a host-based semiochemical lure for trapping emerald ash borer Agrilus planipennis (Coleoptera: Buprestidae).

Bark volatiles from green ash Fraxinus pennsylvanica were tested for electrophysiological activity by Agrilus planipennis using gas chromatographic-electroantennographic detection (GC-EAD) and for behavioral activity using baited purple traps in Michigan. GC-EAD analysis of the headspace volatiles of bark tissue samples from 0- and 24-h-old fully girdled (stressed) ash trees showed that the latter had elevated sesquiterpene levels. Six of the elevated compounds consistently elicited antennal responses by both male and female A. planipennis. Five of the antennally active compounds were identified as alpha-cubebene, alpha-copaene, 7-epi-sesquithujene, trans-beta-caryophyllene, and alpha-humulene (alpha-caryophyllene). The sixth EAD-active compound remains unidentified. We monitored capture of adult A. planipennis on traps baited with several combinations of ash tree volatiles. Treatments included two natural oil distillates (Manuka and Phoebe oil) that were found to contain, respectively, high concentrations of four and five of the six antennally active ash bark volatiles. A four-component leaf lure developed by the USDA Forest Service and Canadian Forest Service was also tested. In three separate field studies, Manuka oil-baited traps caught significantly more adult beetles than unbaited traps. Lures designed to release 5, 50, and 500 mg of Manuka oil per day all caught more insects than unbaited traps. In a field test comparing and combining Phoebe oil with Manuka oil, Phoebe oil-baited traps caught significantly more beetles than either Manuka oil-baited traps or unbaited traps. We hypothesize that the improved attractancy of Phoebe oil to A. planipennis over Manuka oil is caused by the presence of the antennally active sesquiterpene, 7-epi-sesquithujene.

Partial life tables from three generations of Enaphalodes rufulus (Coleoptera: Cerambycidae).

We used life table analyses to investigate age specific mortality and to better understand the population dynamics of the red oak borer, Enaphalodes rufulus (Haldeman) (Coleoptera: Cerambycidae). We continually sampled populations within 177 trees at primarily two sites in the Ozark National Forest in Arkansas throughout three (2-yr) generations. The first cohort (adults emerged in 2003) was sampled during a severe population outbreak, whereas the second and third (2005 and 2007) were sampled during the population crash that followed. Generation mortality was 94% in 2003 and 99% in both 2005 and 2007. Estimates of apparent mortality indicated that the E. rufulus population crash likely occurred during or before the first overwintering period (2003-2004) of the generation that emerged as adults in 2005. We found limited evidence for density dependent mortality, which suggest that intraspecific competition after the first active feeding period was apparently not an important mortality factor during E. rufulus development. Life tables revealed that E. rufulus larvae generally experienced the greatest apparent mortality during the second summer of active feeding (80-94%) when larvae were feeding in, and moving between phloem and sapwood. The least apparent mortality was incurred during the following spring and early summer (26-67%) when late stage larvae and pupae were deepest and most protected within sapwood or heartwood tunnels. We found very little evidence for mortality from associated species. Scarring of vascular tissue in response to E. rufulus feeding occurred during early life stages and may be an important tree resistance mechanism and E. rufulus mortality factor.