Field Test of the Propheromones of the Whitemarked Tussock Moth (WMTM) Orgyia leucostigma (J.E. Smith) (Lepidoptera: Erebidae).

The whitemarked tussock moth (WMTM), Orgyia leucostigma (J.E. Smith) (Lepidoptera: Erebidae), is an economic pest in Nova Scotia, Canada, where it undergoes periodic outbreaks defoliating several tree species of economic value, including balsam fir, Abies balsamea (L.) Miller (Pinaceae). Herein is described a releasing device for the WMTM sex pheromone (Z,Z)-6, 9-heneicosadien-11-one based on a rubber septum, which converts pheromone precursors, such as acetals, namely (Z,Z)-11,11-dimethoxy-6,9-heneicosadiene and (Z,Z)-6,9-heneicosadien-11-one ethylene ketal, to the pheromone itself by the action of acetic acid and releases it over time. The pheromone is unstable in nature and, consequently, lures made with this compound will only attract WMTM for a day or two. The two pheromone precursors, however, are more stable, and are converted slowly into the pheromone by acetic acid impregnated in the releasing device, or by acidic conditions in the environment. The two pheromone precursors were synthesized in 2019 using a modified, previously published approach. Field trapping studies conducted from 2019-22 showed that traps baited with rubber septa loaded with either (Z,Z)-11,11-dimethoxy-6,9-heneicosadiene by itself or (Z,Z)-6,9-heneicosadien-11-one ethylene ketal plus acetic acid consistently caught significantly more WMTM than traps baited with blank septa in most experiments.

Courtship behavior and identification of a sex pheromone in Ibalia leucospoides (Hymenoptera: Ibaliidae), a larval parasitoid of Sirex noctilio (Hymenoptera: Siricidae).

BACKGROUND: Ibalia leucospoides (Hymenoptera: Ibaliidae) is a larval parasitoid that has been widely introduced as a biological control agent for the invasive woodwasp,Sirex noctilio (Hymenoptera: Siricidae) in the Southern Hemisphere. In this study, the courtship behavior and identificaion of sex pheromones are described for I. leucospoides under laboratory conditions. METHODS: For courtship behavior, both sexes were observed in a wire mesh observation cylinder (75 cm length ×10 cm diameter) for 15 minutes. The female body washes were analyzed using Gas Chromatography- Electroantennographic Detection (GC-EAD). Then the EAD-active compounds were tentatively identified using GC-Mass Spectrometry (GC-MS) and examined in olfactometer assays. RESULTS: The courtship behavior included rhythmic lateral movements, mounting, head-nodding cycles in males, and wing-fanning in females. GC-EAD analysis of female body washes with male antennae revealed seven compounds which elicited antennal responses, four of which are straight-chain alkanes (C23, C25, C26, and C27). The identities of these alkanes were confirmed by matching the retention times, mass spectra, and male antennal activity to those of commercially obtained chemicals. In olfactometer assays, a blend of the four straight-chain alkanes was attractive to I. leucospoides males, and there was no response to blends that lacked any of these four compounds. Female body wash was no more attractive than the four-component blend. The ratios of EAD-active components differ between hydrocarbon profiles from males and females. CONCLUSION: This study is the first investigation of cuticular hydrocarbons in the family Ibaliidae. It provides evidence that the ubiquitous alkanes (C23, C25, C26, and C27) in sex-specific ratios attract I. leucospoides males.

Evaluation of Trapping Schemes to Detect Emerald Ash Borer (Coleoptera: Buprestidae).

Management responses to invasive forest insects are facilitated by the use of detection traps ideally baited with species-specific semiochemicals. Emerald ash borer, Agrilus planipennis Fairmaire, is currently invading North American forests, and since its detection in 2002, development of monitoring tools has been a primary research objective. We compared six trapping schemes for A. planipennis over 2 yr at sites in four U.S. states and one Canadian province that represented a range of background A. planipennis densities, canopy coverage, and ash basal area. We also developed a region-wide phenology model. Across all sites and both years, the 10th, 50th, and 90th percentile of adult flight occurred at 428, 587, and 837 accumulated degree-days, respectively, using a base temperature threshold of 10°C and a start date of 1 January. Most trapping schemes captured comparable numbers of beetles with the exception of purple prism traps (USDA APHIS PPQ), which captured significantly fewer adults. Trapping schemes varied in their trap catch across the gradient of ash basal area, although when considering trap catch as a binary response variable, trapping schemes were more likely to detect A. planipennis in areas with a higher ash component. Results could assist managers in optimizing trap selection, placement, and timing of deployment given local weather conditions, forest composition, and A. planipennis density.

Effects of Topical Tebufenozide Application to Choristoneura fumiferana Pupae (Lepidoptera: Tortricidae).

Choristoneura fumiferana (Clemens) (Lepidoptera: Tortricidae) is a defoliating pest in Canada and the northeastern United States. Given its important ecological and economic effects in affected regions, several direct management techniques have been developed, including the application of the insect growth regulator tebufenozide (Mimic™, RH-5992) to feeding larval stages. While the effectiveness of tebufenozide, in this capacity, is understood, management programs of other lepidopteran pests have demonstrated the effectiveness of tebufenozide application when utilized against other life stages. Here, we investigated the toxicity of topically-applied tebufenozide to C. fumiferana pupae to determine if such a strategy could be feasible. We observed significant dose-dependent decreases in the likelihood of adult emergence, increases in the likelihood of pupal death or adult deformity at eclosion, and significant decreases in mean adult longevity. Estimated LD 50 (lethal dose) values for adult male and female C. fumiferana treated as pupae ≤ 4 days after pupation were approximately 1-3 and 2-3.5% ACI (active commercial ingredient) respectively. Estimated L-SD (lethal-sublethal) 50 doses for adult male and female C. fumiferana treated as pupae ≤4 days after pupation were <1, and <2% ACI, respectively. Mating success was also significantly lower in mating pairs containing adults treated as pupae. Although, the amounts required to cause appreciable pupal mortality were much higher than those currently applied operationally in the C. fumiferana system, our study illustrates the potential of tebufenozide to utilized against additional developmental stages in other lepidopteran pests.

Semiochemical and Communication Ecology of the Emerald Ash Borer, Agrilus planipennis (Coleoptera: Buprestidae).

Knowledge of buprestid chemical ecology is sparse but the appearance of the invasive pest Agrilus planipennis Fairmaire in North America has provided the impetus to study in detail the semiochemistry and ecology of this important buprestid. The macrocyclic lactone (3Z)-12-dodecenolide [(3Z)-lactone] is identified as a key antennally-active compound that is produced by females and attracts males. Though a weak trap attractant alone, when combined with the host kairomone (3Z)-hexenol and the important visual cue of a green canopy trap, significant increases in male trap capture occur, thus defining (3Z)-lactone as both a sex pheromone of A. planipennis as well as the first and only known buprestid pheromone. The non-natural stereoisomer (3E)-12-dodecenolide and the saturated analog, 12-dodecanolide also exhibit mimetic activities towards male A. planipennis, suggesting a notable plasticity in this pheromonal structural motif. Efficient synthetic routes to these compounds have been developed. A series of fluoro-12-dodecanolides has also been synthesized containing CF2 groups as a strategy to bias the conformational space accessed by these macrolides and to assess if the analogs may act as mimetics for 12-dodecanolide pheromones associated in A. planipennis. These compounds also afford a unique opportunity to study the binding affinities of lactone surrogates with A. planipennis chemosensory proteins and olfactory receptors. Some progress has also been made in identifying the genes involved in the reception, processing and degradation of volatiles in this invasive insect. It is now evident that the behavior and ecology of A. planipennis involves a complex pattern of sensory modalities, including visual, tactile, olfactory and potentially acoustic components. Earlier reviews focused on studies of attractive host volatiles in development of a trapping system for early detection and visual and contact phenomena in A. planipennis mate finding. This review will update the semiochemistry and chemical ecology of A. planipennis and discuss studies on chemistry and behavior that have identified female-produced pheromone components and host kairomones.

First Report of Mating Disruption With an Aggregation Pheromone: A Case Study With Tetropium fuscum (Coleoptera: Cerambycidae).

Tetropium fuscum (F.), native to Europe and established in Nova Scotia, Canada, since at least 1990, is considered a low-to-moderate threat to spruce (Picea spp.) forests in North America and regulated as a quarantine pest by the Canadian Food Inspection Agency. We tested broadcast applications of the aggregation pheromone racemic (5E)-6,10-dimethyl-5,9-undecadien-2-ol (fuscumol), formulated at 10% concentration in Hercon Bio-Flakes (Hercon International, Emigsville, PA), for efficacy in disrupting T. fuscum mating and suppressing populations. Two applications of 2.5-2.75 kg Bio-Flakes (250-275 g a.i.) per ha per season significantly reduced trap catches and mating success (2009, 2010, 2012): about 30% of females trapped in treated plots had mated compared with 60% of females trapped in untreated plots. Similar reductions in mating success were observed in 2011 with one or two 4.5 kg/ha applications of Bio-Flakes. Mean densities of T. fuscum colonizing sentinel bait logs or girdled trees were 36% lower in pheromone-treated plots than in untreated plots, but the difference was not statistically significant. Lack of population suppression may have been because mated females immigrated into treated plots or because populations were so high that despite a 50% reduction in mating success, absolute numbers of mated females were sufficient to infest our bait logs or trees. This is the first demonstration of insect mating disruption via broadcast application of an aggregation pheromone. Pheromone-mediated mating disruption has potential to slow the spread of invasive cerambycids by targeting low-density outlier populations near or beyond the leading edge of an infestation.

Interaction of Visual and Chemical CUES in Promoting Attraction of Agrilus planipennis.

Female emerald ash borers, Agrilus planipennis (Coleoptera: Buprestidae), emit a macrocyclic lactone, (Z)-3-dodecen-12-olide, that increases field trap captures on large-panel prism traps when co-emitted with the green leaf volatile (Z)-3-hexenol. We assessed attraction to these compounds by using visual decoy-baited branch traps, which attract males by mimicking a living female resting upon a leaf. Pairs of branch traps, with and without visual decoy beetles, were placed on green ash, Fraxinus pennsylvanica, trees, which were assigned different odor treatments: 1) no odor, 2) (Z)-3-hexenol alone, and 3) (Z)-3-hexenol-plus-lactone. Male captures were positively affected by the presence of decoys and the emission of either (Z)-3-hexenol or (Z)-3-hexenol plus lactone. The decoy-baited traps with the combination of (Z)-3-hexenol plus lactone caught more males than any other treatment. Greater male captures were associated with continuing captures later in the season, suggesting that decoy and odor attractants remain attractive throughout the flight period. Female captures were not affected by the visual decoys, but odors did influence captures, with the (Z)-3-hexenol plus lactone treatment catching the greatest number of females. The rare female trap captures were negatively correlated with the more common male captures on the odorless and (Z)-3-hexenol-baited traps, but were not correlated with male captures when the lactone was added. Thus, in the absence of the lactone, the visual signal of other conspecifics can inhibit female attraction. However, the pheromone attracts both sexes independently of the visual signal on the trap.

Labeling Feral Spruce Budworm (Lepidoptera: Tortricidae) Populations With Rubidium.

Rubidium (Rb) is a trace element that occurs naturally in low concentrations and is easily absorbed by plants, making it a useful tool for labeling insect defoliators, such as spruce budworm (Choristoneura fumiferana Clemens). Balsam fir trees (Abies balsamea (L.) Miller) injected with either 8 or 16 g per tree of rubidium chloride (RbCl) showed quick uptake and distribution throughout the crown, with no negative effects on tree shoot growth or spruce budworm survival and development. Adult spruce budworm that fed as larvae on trees injected with RbCl were clearly labeled, with significantly higher Rb concentrations than the background levels found in adults that fed as larvae on control trees. Rb concentrations in feral spruce budworm adults for both the 8 g (9 µg/g) and 16 g (25 µg/g) per tree treatments were at least five times lower than those in laboratory-reared adults on 1,000 µg/g RbCl diet (125 µg/g); survival, development, pupal weight, sex ratio, and mating status of spruce budworm were not adversely affected by Rb treatment. Egg masses laid by feral females that fed as larvae on Rb-labeled trees were also labeled with Rb. Injecting trees with RbCl is a viable technique for labeling feral spruce budworm populations to help distinguish local populations from immigrants to better evaluate the success of early intervention strategies such as mating disruption.

Spruce Budworm (Lepidoptera: Tortricidae) Oral Secretions II: Chemistry.

As sessile organisms, plants have evolved different methods to defend against attacks and have adapted their defense measures to discriminate between mechanical damage and herbivory by insects. One of the ways that plant defenses are triggered is via elicitors from insect oral secretions (OS). In this study, we investigated the ability of second-instar (L2) spruce budworm [SBW; Choristoneura fumiferana (Clemens)] to alter the volatile organic compounds (VOCs) of four conifer species [Abies balsamea (L.) Mill., Picea mariana (Miller) B.S.P., Picea glauca (Moench) Voss, Picea rubens (Sargent)] and found that the emission profiles from all host trees were drastically changed after herbivory. We then investigated whether some of the main elicitors (fatty acid conjugates [FACs], ß-glucosidase, and glucose oxidase) studied were present in SBW OS. FACs (glutamine and glutamic acid) based on linolenic, linoleic, oleic, and stearic acids were all observed in varying relative quantities. Hydroxylated FACs, such as volicitin, were not observed. Enzyme activity for ß-glucosidase was also measured and found present in SBW OS, whereas glucose oxidase activity was not found in the SBW labial glands. These results demonstrate that SBW L2 larvae have the ability to induce VOC emissions upon herbivory and that SBW OS contain potential elicitors to induce these defensive responses. These data will be useful to further evaluate whether these elicitors can separately induce the production of specific VOCs and to investigate whether and how these emissions benefit the plant.

Spruce Budworm (Lepidoptera: Tortricidae) Oral Secretions I: Biology and Function.

The potential roles of the oral secretions (OS) of spruce budworm (SBW; Choristoneura fumiferana Clemens) larvae and factors that may affect the volume of OS disgorged were investigated in the laboratory. Experiments revealed that diet-fed SBW larvae readily disgorge OS when induced ("milked"), with minimal overall cost to their development and eventual pupal weight. Exposure of conspecific larvae to OS throughout larval development negatively affected survival and male pupal weight; however, male development time was faster when exposed to OS. Female pupal weight and development time were not affected. Preliminary experiments suggested that OS had a repellent effect on a co-occurring herbivore, the false hemlock looper, Nepytia canosaria (Walker). OS produced by larvae that fed on three host tree species and on artificial diet significantly increased the grooming time of ants (Camponotus sp.), indicating that SBW OS have an anti-predator function. The volume of OS is significantly greater in L6 than in L4 or L5, with the volume produced by L6 depending on weight and age as well as feeding history at time of milking. These findings indicate that SBW OS function as both an intra- and interspecific epideictic pheromone and as an anti-predator defensive mechanism, while incurring minimal metabolic costs.

Effects of Pheromone Release Rate and Trap Placement on Trapping of Agrilus planipennis (Coleoptera: Buprestidae) in Canada.

The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is a devastating insect pest in its introduced range. A trapping system that increases trap catches or detection rates in low-density populations would be beneficial for survey programs. Five trapping experiments were conducted to investigate factors influencing capture rates of male beetles on dark green traps baited with the A. planipennis pheromone, (3Z)-dodecen-12-olide ((3Z)-lactone), plus the green leaf volatile, (3Z)-hexenol. Low doses (0.001-1.0 mg) of (3Z)-lactone + (3Z)-hexenol did not consistently increase captures of male A. planipennis. In other experiments, mean captures of males were significantly higher on traps baited with a moderate dose (3.0 mg/septum) of (3Z)-lactone + (3Z)-hexenol, compared with lower doses (0.001, 0.1, and 1.0 mg) or (3Z)-hexenol alone. Next, we demonstrated that addition of (3Z)-lactone to traps baited with (3Z)-hexenol resulted in significantly greater increases in male captures when pairs of traps were placed on the same tree, than when traps were placed on adjacent trees. Moreover, significantly more A. planipennis were captured on pheromone-baited traps placed in the southern versus northern aspect of the crown. These results highlight the importance of experimental set-up for elucidating lure treatment effects and also suggests the (3Z)-lactone may be more active at close range. Our findings increase our understanding of the pheromone ecology of this species and lend support toward the use of dark green traps baited with 3.0 mg (3Z)-lactone + (3Z)-hexenol deployed in the south aspect of the canopy for detection programs for this insect.

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.

A simulation approach to assessing sampling strategies for insect pests: an example with the balsam gall midge.

Estimation of pest density is a basic requirement for integrated pest management in agriculture and forestry, and efficiency in density estimation is a common goal. Sequential sampling techniques promise efficient sampling, but their application can involve cumbersome mathematics and/or intensive warm-up sampling when pests have complex within- or between-site distributions. We provide tools for assessing the efficiency of sequential sampling and of alternative, simpler sampling plans, using computer simulation with "pre-sampling" data. We illustrate our approach using data for balsam gall midge (Paradiplosis tumifex) attack in Christmas tree farms. Paradiplosis tumifex proved recalcitrant to sequential sampling techniques. Midge distributions could not be fit by a common negative binomial distribution across sites. Local parameterization, using warm-up samples to estimate the clumping parameter k for each site, performed poorly: k estimates were unreliable even for samples of n ∼ 100 trees. These methods were further confounded by significant within-site spatial autocorrelation. Much simpler sampling schemes, involving random or belt-transect sampling to preset sample sizes, were effective and efficient for P. tumifex. Sampling via belt transects (through the longest dimension of a stand) was the most efficient, with sample means converging on true mean density for sample sizes of n ∼ 25-40 trees. Pre-sampling and simulation techniques provide a simple method for assessing sampling strategies for estimating insect infestation. We suspect that many pests will resemble P. tumifex in challenging the assumptions of sequential sampling methods. Our software will allow practitioners to optimize sampling strategies before they are brought to real-world applications, while potentially avoiding the need for the cumbersome calculations required for sequential sampling methods.

Steps in the biosynthesis of fuscumol in the longhorn beetles Tetropium fuscum (F.) and Tetropium cinnamopterum Kirby.

Fuscumol [(2S,5E)-6,10-dimethyl-5,9-undecadien-2-ol] was recently identified as the male-produced aggregation pheromone of the brown spruce longhorn beetle, Tetropium fuscum (F.), and the eastern larch borer, Tetropium cinnamopterum Kirby. Several other species use this homoterpenoid alcohol motif, its ketone, or its acetate as part of their pheromone system. Investigation of the biosynthesis of this compound in these two Tetropium species demonstrated that geranylacetone [(5E)-6,10-dimethyl-5,9-undecadien-2-one] and farnesol [(2E,6E)-3,7,11-trimethyl-2,6,10-dodecatrien-1-ol] are both intermediates in this process. This was accomplished by applying deuterium-labeled geranylacetone and deuterium-labeled farnesol in separate experiments to the abdominal sterna of live T. fuscum and T. cinnamopterum and analyzing the deuterium labeling in the fuscumol and geranylacetone emitted by the insects with solid-phase microextraction (SPME) and GC/MS analysis. Deuterium labeling studies also showed that nerolidol[(3S,6E)-3-hydroxy-3,7,11-trimethyl-1,6,10-dodecatriene] and 2,3-epoxyfarnesol are not intermediates in fuscumol or geranylacetone synthesis in T. fuscum or T. cinnamopterum. Tissue-specific expression of T. fuscum farnesyl diphosphate synthase (TfFPPS), an enzyme expected to provide a key fuscumol precursor, was measured. TfFPPS transcripts were relatively abundant in male midguts, but were also present at significant levels in other tissues.

Attraction of Agrilus planipennis (Coleoptera: Buprestidae) to a volatile pheromone: effects of release rate, host volatile, and trap placement.

Attraction of emerald ash borer, Agrilus planipennis Fairmaire, to a volatile pheromone was demonstrated in three field experiments using baited green sticky traps. A dose-response curve was generated for male A. planipennis to increasing release rates of (3Z)-dodecen-12-olide ((3Z)-lactone) in combination with the green leaf volatile, (3Z)-hexenol. Only the lowest release rate (<2.50 µg/d) of (3Z)-lactone significantly increased captures of male A. planipennis, as compared with traps baited with (3Z)-hexenol alone. Effect of trap height, (3Z)-lactone, and (3Z)-hexenol and their interactions on the trap capture of A. planipennis was determined in a factorial experiment. Number of males per trap was significantly and positively affected by (3Z)-lactone, (3Z)-hexenol, and trap height whereas number of females per trap was affected by trap height only; none of the interactions were significant. As predicted, the greatest mean catch of males was in traps baited with (3Z)-lactone and (3Z)-hexenol placed high in the canopy. Electroantennogram tests on the bark volatile, 7-epi-sesquithujene, demonstrated the ability of male and female A. planipennis antennae to detect and respond to this compound, particularly the (+)-7-epi-sesquithujene isomer. Results from an olfactometer bioassay and field testing did not demonstrate attraction of either males or females to (+)-7-epi-sesquithujene. These data increase our understanding of the pheromone ecology of the invasive emerald ash borer, provide further confirmation of the behavioral activity of the female-produced lactone pheromone, and should increase the ability to detect A. planipennis infestations where they are present.

New Coleoptera records from New Brunswick, Canada: Cerambycidae.

Five species of Cerambycidae, Acmaeops discoideus (Haldeman), Anelaphus villosus (Fabricius), Phymatodes species (CNC sp. n. #1), Sarosesthes fulminans (Fabricius), and Urgleptus signatus (LeConte) are newly recorded for New Brunswick, Canada. All but Acmaeops villosus are new to the Maritime provinces. Phymatodes testaceus (Linnaeus) is removed from the faunal list of the province as a result of mislabeled specimens, records of Phymatodes maculicollis LeConte are presented confirming the presence of this species in New Brunswick, and the first recent records ofNeospondylis upiformis (Mannerheim) are presented. Additional records are given for the recently recorded Phymatodes aereus (Newman), indicating a wider distribution in the province. Collection data, habitat data, and distribution maps are presented for each species.

Evidence for limited spatial spread in an exotic longhorn beetle, Tetropium fuscum (Coleoptera: Cerambycidae).

The longhorn beetle Tetropium fuscum F. (Coleoptera: Cerambycidae) has become established in Nova Scotia, Canada, where it coexists with Tetropium cinnamopterum Kirby. The two Tetropium species share a similar ecological niche and use the same volatile cues for mate attraction. Exotic T. fuscum was introduced near Halifax, Nova Scotia, in approximately 1990, but the rate of its spread 20 yr later has not been documented. We report a large-scale, 3-yr study that investigates the distribution of T. fuscum relative to its site of introduction. Traps baited with male-produced pheromone and host volatiles were used to estimate the relative abundance of the two Tetropium species. Adult T. fuscum emerged 1-2 wk earlier than T. cinnamopterum each year between 2008 and 2010. The spatial distribution of T. fuscum was characterized by a sharp decline in abundance in relation to its point of introduction, up to a threshold distance of approximately 80 km beyond which T. fuscum is rare in comparison with native T. cinnamopterum. The restricted range of T. fuscum 20 yr after its introduction may be attributed to limited dispersal of adults or reproductive failures of low-density populations. The distribution of T. fuscum seemed stable between 2008 and 2010. In 1 of 3 yr, the abundance of T. cinnamopterum increased with the distance to the site of introduction of T. fuscum, which suggests competitive interactions between the two Tetropium species.

Contact sex pheromones identified for two species of longhorned beetles (Coleoptera: Cerambycidae) Tetropium fuscum and T. cinnamopterum in the subfamily Spondylidinae.

Male Tetropium fuscum (F.) and T. cinnamopterum Kirby mated with live and dead (freeze-killed) conspecific females upon antennal contact, but did not respond to dead females after cuticular waxes were removed by hexane rinsing. Significantly fewer males of each species attempted to copulate with live or dead heterospecific females than with conspecifics, indicating that mate recognition was mediated by species-specific contact sex pheromones in the female's cuticular hydrocarbons. GC/MS analysis of T. fuscum elytra identified n-alkanes and mono-methyl branched alkanes of which 11-methylheptacosane and 3- and 5-methyltricosanes were dominant in females. Full male responses, including copulatory behavior, were restored with application of enantiomerically pure synthetic (S)-11-methyl-heptacosane at 40 µg/female (one female equivalent) but not with racemic or (R)-11-methyl-heptacosane. The cuticular hydrocarbons on T. cinnamopterum elytra included 11-methyl-heptacosane as well as n-alkanes, methyl-branched alkanes, mono-alkenes, and (Z, Z)-6, 9-alkadienes. (Z)-9-pentacosene, (Z)-9-heptacosene, and 11-methyl-heptacosane were female dominant, but only (Z)-9-pentacosene elicited precopulatory behaviors in conspecific males at levels similar to those behaviors elicited by unrinsed females, but elicited copulation in fewer than half of males. At female equivalent dosages (10 µg), neither (Z)-9-heptacosene nor (S)-11-methyl- heptacosane elicited responses in males that were significantly different from those responses to a rinsed female but when applied together, the proportion of males responding was significantly increased. 11-methyl-heptacosene is thus a contact pheromone component common to both species, which may explain the heterospecific mating attempts by some males.

Evidence for a volatile pheromone in Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) that increases attraction to a host foliar volatile.

Analysis by gas chromatography/mass spectrometry (GC/MS) of volatiles from virgin female emerald ash borer, Agrilus planipennis Fairmaire confirmed the emission of (3Z)-lactone [(3Z)-dodecen-12-olide] but not its geometric isomer, (3E)-lactone [(3E)-dodecen-12-olide]. Gas chromatographic/electroantennographic (GC/EAD) analysis of synthetic (3Z)-lactone, which contained 10% (3E)-lactone, showed a strong response of male and female antennae to both isomers. EAG analysis with 0.01- to 100-µg dosages showed a positive dose response, with females giving significantly higher responses than males. In field experiments with sticky purple prism traps, neither lactone isomer affected catches when combined with ash foliar or cortical volatiles (green leaf volatiles or Phoebe oil, respectively). However, on green prism traps, the (3Z)-lactone significantly increased capture of male A. planipennis when traps were deployed in the canopy. Captures of males on traps with both (3E)-lactone and (3Z)-hexenol or with (3Z)-lactone and (3Z)-hexenol were increased by 45-100%, respectively, compared with traps baited with just (3Z)-hexenol. In olfactometer bioassays, males were significantly attracted to (3E)-lactone, but not the (3Z)-lactone or a 60:40 (3E):(3Z) blend. The combination of either (3E)- or (3Z)-lactone with Phoebe oil was not significantly attractive to males. Males were highly attracted to (3Z)-hexenol and the (3Z)-lactone + (3Z)-hexenol combination, providing support for the field trapping results. These data are the first to demonstrate increased attraction with a combination of a pheromone and a green leaf volatile in a Buprestid species.

Effect of chirality, release rate, and host volatiles on response of Tetropium fuscum (F.), Tetropium cinnamopterum Kirby, and Tetropium castaneum (L.) to the aggregation pheromone, fuscumol.

The male-produced aggregation pheromones of Tetropium fuscum (F.) and T. cinnamopterum Kirby were identified as (2S,5E)-6,10-dimethyl-5,9-undecadienol by chemical analysis, synthesis, electronantennography, and field trapping; the compound is here renamed "fuscumol". The effect of fuscumol chirality, alone or with host volatiles, and fuscumol release rate on Tetropium spp. was tested in field-trapping experiments in Nova Scotia and Poland. Both (S)-fuscumol and racemic fuscumol synergized trap catches of male and female T. fuscum, T. cinnamopterum, and T. castaneum (L.) when combined with a blend of host monoterpenes and ethanol. Without added host volatiles, fuscumol was either unattractive (in Nova Scotia) or only slightly so (in Poland). (R)-Fuscumol, alone or in combination with host volatiles, did not elicit increases in trap capture of any Tetropium species, relative to the controls. Fuscumol synergized attraction of both sexes to host volatiles, thus indicating it acts as an aggregation pheromone. Sex ratio was often female-biased in traps baited with fuscumol plus host volatiles, and was either unbiased or male-biased in traps with host volatiles alone. In traps with host volatiles and racemic fuscumol, mean catches of Tetropium species were unaffected by fuscumol release rates ranging from 1 to 32 mg/d. The attraction of three different Tetropium species to the combination of (S)-fuscumol and host volatiles suggests that cross-attraction may occur where these species are sympatric, and that reproductive isolation possibly occurs via differences in close-range cues. These results have practical applications for survey and monitoring of T. fuscum, a European species established in Nova Scotia since at least 1980, and for early detection of T. castaneum, a European species not presently established in North America.