Mating Disruption as a Suppression Tactic in Programs Targeting Regulated Lepidopteran Pests in US.

Mating disruption, the broadcast application of sex-attractant pheromone to reduce the ability of insects to locate mates, has proven to be an effective method for suppressing populations of numerous moth pests. Since the conception of mating disruption, the species-specificity and low toxicity of pheromone applications has led to their consideration for use in area-wide programs to manage invasive moths. Case histories are presented for four such programs where the tactic was used in the United States: Pectinophora gossypiella (pink bollworm), Lymantria dispar (gypsy moth), Epiphyas postvittana (light brown apple moth), and Lobesia botrana (European grapevine moth). Use of mating disruption against P. gossypiella and L. botrana was restricted primarily to agricultural areas and relied in part (P. gossypiella) or wholly (L. botrana) on hand-applied dispensers. In those programs, mating disruption was integrated with other suppression tactics and considered an important component of overall efforts that are leading toward eradication of the invasive pests from North America. By contrast, L. dispar and E. postvittana are polyphagous pests, where pheromone formulations have been applied aerially as stand-alone treatments across broad areas, including residential neighborhoods. For L. dispar, mating disruption has been a key component in the program to slow the spread of the infestation of this pest, and the applications generally have been well tolerated by the public. For E. postvittana, public outcry halted the use of aerially applied mating disruption after an initial series of treatments, effectively thwarting an attempt to eradicate this pest from California. Reasons for the discrepancies between these two programs are not entirely clear.

Genetic structure, admixture and invasion success in a Holarctic defoliator, the gypsy moth (Lymantria dispar, Lepidoptera: Erebidae).

Characterizing the current population structure of potentially invasive species provides a critical context for identifying source populations and for understanding why invasions are successful. Non-native populations inevitably lose genetic diversity during initial colonization events, but subsequent admixture among independently introduced lineages may increase both genetic variation and adaptive potential. Here we characterize the population structure of the gypsy moth (Lymantria dispar Linnaeus), one of the world's most destructive forest pests. Native to Eurasia and recently introduced to North America, the current distribution of gypsy moth includes forests throughout the temperate region of the northern hemisphere. Analyses of microsatellite loci and mitochondrial DNA sequences for 1738 individuals identified four genetic clusters within L. dispar. Three of these clusters correspond to the three named subspecies; North American populations represent a distinct fourth cluster, presumably a consequence of the population bottleneck and allele frequency change that accompanied introduction. We find no evidence that admixture has been an important catalyst of the successful invasion and range expansion in North America. However, we do find evidence of ongoing hybridization between subspecies and increased genetic variation in gypsy moth populations from Eastern Asia, populations that now pose a threat of further human-mediated introductions. Finally, we show that current patterns of variation can be explained in terms of climate and habitat changes during the Pleistocene, a time when temperate forests expanded and contracted. Deeply diverged matrilines in Europe imply that gypsy moths have been there for a long time and are not recent arrivals from Asia.

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.

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.

Efficacy of multifunnel traps for capturing emerald ash borer (Coleoptera: Buprestidae): effect of color, glue, and other trap coatings.

Tens of thousands of adhesive-coated purple prism traps are deployed annually in the United States to survey for the invasive emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae). A reusable, more user-friendly trap is desired by program managers, surveyors, and researchers. Field assays were conducted in southeastern Michigan to ascertain the feasibility of using nonsticky traps as survey and detection tools for emerald ash borer. Three nonsticky trap designs, including multifunnel (Lindgren), modified intercept panel, and drainpipe (all painted purple) were compared with the standard purple prism trap; no statistical differences in capture of emerald ash borer adults were detected between the multifunnel design and the prism. In subsequent color comparison assays, both green- and purple-painted multifunnel traps (and later, plastic versions of these colors) performed as well or better than the prism traps. Multifunnel traps coated with spray-on adhesive caught more beetles than untreated traps. The increased catch, however, occurred in the traps' collection cups and not on the trap surface. In a separate assay, there was no significant difference detected between glue-coated traps and Rain-X (normally a glass treatment)-coated traps, but both caught significantly more A. planipennis adults than untreated traps.

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.

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.

Sex pheromone of browntail moth, Euproctis chrysorrhea (L.): synthesis and field deployment.

The browntail moth, Euproctis chrysorrhea (L.), is native to Eurasia, where periodic outbreaks result in defoliation of forest, shade, and ornamental trees. In addition to the damage caused by defoliation, human contact with larval urticating hairs often results in severe dermatitis. Hence, tools for monitoring and controlling the moth populations are desirable. The female-produced sex pheromone of the browntail moth was identified previously, but the synthesis had not been published. This paper reports the synthesis of the pheromone of the browntail moth, (7Z,13Z,16Z,19Z)-docosatetraenyl isobutyrate, using in a key step a Wittig olefination of (6Z)-13-(tetrahydo-2H-pyran-2-yloxy)tridecenal. Field trapping studies were conducted with rubber septa and string formulations of the pheromone and included dose-response, pheromone purity, and dispenser-aging trials. It was found that traps baited with 250 microg of pheromone of 91-94% isomeric purity (main impurity presumably being the 13E isomer) on rubber septa are suitable for monitoring moth populations during the entire flight season.

Influence of trap placement and design on capture of the emerald ash borer (Coleoptera: Buprestidae).

The key to an effective pest management program for the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera Buprestidae), is a survey program equipped with tools for detecting and delimiting populations. We studied the effects of trap design, color, and placement on the efficacy of sticky traps for capturing the emerald ash borer. There were significant differences in trap catch along a transect gradient from wooded to open field conditions, with most beetles being caught along the edge, or in open fields, 15-25 m outside an ash (Fraxinus spp. L.) (Oleaceae) woodlot. Greater emerald ash borer catch occurred on purple traps than on red or white traps. Traps placed in the mid-canopy of ash trees (13 m) caught significantly more beetles than those placed at ground level. We also describe a new trap design, a three-sided prism trap, which is relatively easy to assemble and deploy.

Assessing Flight Potential of the Invasive Asian Longhorned Beetle (Coleoptera: Cerambycidae) With Computerized Flight Mills.

The Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae: Lamiinae), is an invasive woodborer that poses a serious threat to urban and natural landscapes. In North America, this beetle is a quarantine pest, and populations are subject to eradication efforts that consist of the identification, removal, and destruction of infested host material, and removal or prophylactic treatment of high-risk host plant species. To enhance Asian longhorned beetle eradication protocols in landscapes with extensive host availability, we assessed the dispersal potential of male and female adults of varying age, mating, and nutritional status using computerized flight mills. In total, 162 individuals were tethered to computerized flight mills for a 24-h trial period to collect information on total distance flown, flight times and velocities, and number and duration of flight bouts. Adult Asian longhorned beetles (in all treatments) flew an average of 2,272 m within a 24-h period, but are capable of flying up to 13,667 m (8.5 miles). Nutrition and age had the greatest impacts on flight, with Asian longhorned beetle adults >5 d of age that had fed having greater overall flight performance than any other group. However, mating status, sex, and body size (pre-flight weight and elytron length) had a minimal effect on flight performance. This information will be useful for refining quarantine zones surrounding areas of infestation, and for providing greater specificity as to the risk the Asian longhorned beetle poses within invaded regions.

Genome of the Asian longhorned beetle (Anoplophora glabripennis), a globally significant invasive species, reveals key functional and evolutionary innovations at the beetle-plant interface.

BACKGROUND: Relatively little is known about the genomic basis and evolution of wood-feeding in beetles. We undertook genome sequencing and annotation, gene expression assays, studies of plant cell wall degrading enzymes, and other functional and comparative studies of the Asian longhorned beetle, Anoplophora glabripennis, a globally significant invasive species capable of inflicting severe feeding damage on many important tree species. Complementary studies of genes encoding enzymes involved in digestion of woody plant tissues or detoxification of plant allelochemicals were undertaken with the genomes of 14 additional insects, including the newly sequenced emerald ash borer and bull-headed dung beetle. RESULTS: The Asian longhorned beetle genome encodes a uniquely diverse arsenal of enzymes that can degrade the main polysaccharide networks in plant cell walls, detoxify plant allelochemicals, and otherwise facilitate feeding on woody plants. It has the metabolic plasticity needed to feed on diverse plant species, contributing to its highly invasive nature. Large expansions of chemosensory genes involved in the reception of pheromones and plant kairomones are consistent with the complexity of chemical cues it uses to find host plants and mates. CONCLUSIONS: Amplification and functional divergence of genes associated with specialized feeding on plants, including genes originally obtained via horizontal gene transfer from fungi and bacteria, contributed to the addition, expansion, and enhancement of the metabolic repertoire of the Asian longhorned beetle, certain other phytophagous beetles, and to a lesser degree, other phytophagous insects. Our results thus begin to establish a genomic basis for the evolutionary success of beetles on plants.

Aerial application of pheromones for mating disruption of an invasive moth as a potential eradication tool.

Biological invasions can cause major ecological and economic impacts. During the early stages of invasions, eradication is desirable but tactics are lacking that are both effective and have minimal non-target effects. Mating disruption, which may meet these criteria, was initially chosen to respond to the incursion of light brown apple moth, Epiphyas postvittana (LBAM; Lepidoptera: Tortricidae), in California. The large size and limited accessibility of the infested area favored aerial application. Moth sex pheromone formulations for potential use in California or elsewhere were tested in a pine forest in New Zealand where LBAM is abundant. Formulations were applied by helicopter at a target rate of 40 g pheromone per ha. Trap catch before and after application was used to assess the efficacy and longevity of formulations, in comparison with plots treated with ground-applied pheromone dispensers and untreated control plots. Traps placed at different heights showed LBAM was abundant in the upper canopy of tall trees, which complicates control attempts. A wax formulation and polyethylene dispensers were most effective and provided trap shut-down near ground level for 10 weeks. Only the wax formulation was effective in the upper canopy. As the pheromone blend contained a behavioral antagonist for LBAM, 'false trail following' could be ruled out as a mechanism explaining trap shutdown. Therefore, 'sensory impairment' and 'masking of females' are the main modes of operation. Mating disruption enhances Allee effects which contribute to negative growth of small populations and, therefore, it is highly suitable for area-wide control and eradication of biological invaders.

Improved synthesis of (3E,6Z,9Z)-1,3,6,9-nonadecatetraene, attraction inhibitor of bruce spanworm, Operophtera bruceata, to pheromone traps for monitoring winter moth, Operophtera brumata.

The winter moth, Operophtera brumata (Lepidoptera: Geometridae), is an early-season defoliator that attacks a wide variety of hardwoods and, in some cases, conifers. The insect is native to Europe but has become established in at least three areas of North America including southeastern New England. The female-produced sex attractant pheromone of the winter moth was identified as (3Z,6Z,9Z)-1,3,6,9-nonadecatetraene (1), which also attracts a native congener, the Bruce spanworm, Operophtera bruceata . Dissection, or (for certainty) DNA molecular testing, is required to differentiate between males of the two species. Thus, a trapping method that is selective for winter moth would be desirable. A geometric isomer of the pheromone, (3E,6Z,9Z)-1,3,6,9-nonadecatetraene (2), can reportedly inhibit attraction of Bruce spanworm to traps without affecting winter moth catch, but use of the pheromone and inhibitor together has not been optimized, nor has the synthesis of the inhibitor. This paper presents two new syntheses of the inhibitor (3E,6Z,9Z)-1,3,6,9-nonadecatetraene based on the intermediate (3Z,6Z)-3,6-hexadecadien-1-ol (4), which has also been utilized in the synthesis of the pheromone. The syntheses combine traditional acetylenic chemistry and Wittig olefination reactions. In one approach, 2 was synthesized in 80% purity (20% being pheromone 1), and in the second, tetraene 2 of 96% purity (and free of 1) was produced in 25% overall yield from dienol 4. The last method benefitted from a refined TEMPO-mediated PhI(OAc)(2) oxidation of 4 and a two-carbon homologation of the corresponding aldehyde 7.