Interactions Between syn- and anti-2,3-Hexanediol Lures on Trap Catches of Woodboring Beetles and Associates in Southeastern United States.

In 2016, we conducted three experiments to clarify the effects of 2,3-hexanediols isomers on trap catches of Neoclytus acuminatus (F.) (Coleoptera: Cerambycidae). We also noted the effects of the isomers on trap catches of other cerambycids and associated species of predators and competitors. Catches of N. acuminatus in traps baited with ethanol + syn-2,3-hexanediol + racemic 3-hydroxyoctan-2-one were reduced with the addition of anti-2,3-hexanediol, an attractant for Curius dentatus Newman (Coleoptera: Cerambycidae). A fourth experiment conducted in 2017 verified that racemic 3-hydroxyoctan-2-one increases catches of N. acuminatus in traps baited with ethanol + syn-2,3-hexanediol. The addition of anti-2,3-hexanediol increased catches of Knulliana cincta (Drury) (Coleoptera: Cerambycidae) in traps baited with ethanol + racemic 3-hydroxyoctan-2-one, whereas attraction of Neoclytus scutellaris (Olivier) to traps baited with ethanol + racemic 3-hydroxyhexan-2-one was reduced by syn-2,3-hexanediol. Trap catches of the beetle predators Chariessa pilosa (Forster), Enoclerus ichneumonus (F.), and Madoniella dislocata (Say) (Coleoptera: Cleridae) were affected by 2,3-hexanediol isomers, whereas other common predators were unaffected by the isomers. Attraction of the bostrichid Xylobiops basilaris (Say) (Coleoptera: Bostrichidae) was increased by the 2,3-hexanediols; the relative effect of the two isomeric blends was dependent on trap co-baits of 3-hydroxy-2-ketones. The two enantiomeric blends of 2,3-hexanediol had minimal effects on catches of most species of ambrosia beetles, whereas the 3-hydroxy-2-ketones affected trap catches of some species.

Interactions between Ethanol, syn-2,3-Hexanediol, 3-Hydroxyhexan-2-one, and 3-Hydroxyoctan-2-one Lures on Trap Catches of Hardwood Longhorn Beetles in Southeastern United States.

The effectiveness of a four-component "super lure" consisting of ethanol (E) and the cerambycid pheromones syn-2,3-hexanediol (D6), racemic 3-hydroxyhexan-2-one (K6), and racemic 3-hydroxyoctan-2-one (K8) on trap catches of Cerambycidae (Coleoptera) was determined in southeast United States with seven trapping experiments in 2011-2013. We captured 74 species of longhorn beetles in our three-year study. Ethanol significantly increased the mean catches of seven species and increased the number of cerambycid species detected. Traps with the "super lure" were effective for 8 of 13 species of Cerambycidae previously shown to be attracted to binary combinations of ethanol plus one of the three pheromones. However, the "super lure" was less effective for the remaining five species with catch reductions of 40-90% compared with combinations of ethanol and one or two of the pheromones. For example, K6 + K8 lures reduced catches of Anelaphus villosus (F.) in traps with E + D6 by 90%. Similarly, catches of Anelaphus pumilus (Newman) in traps with E + K6 + D6 were reduced by 50% with the addition of K8. Catches of Knulliana cincta (Drury) in traps with K6 + K8 lures were interrupted by D6, an effect negated by the addition of ethanol. Given the interruptive effects on trap catches of some species when lures are combined in a single trap, developing optimal lure blends to maximize detection efficacy will be a challenge for managers of detection programs for non-native invasive species of longhorn beetles.

Unsaturated Cuticular Hydrocarbons Enhance Responses to Sex Pheromone in Spruce Budworm, Choristoneura fumiferana.

The primary sex pheromone components of the female spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), are (E)- and (Z)-11-tetradecenal, produced in 95:5 ratio. However, male flight responses to calling females in a wind tunnel were faster and maintained longer than responses to any synthetic aldehyde blend. Analyses of cuticular extracts from spruce budworm adults revealed series of n-alkanes and n-monoalkenes with predominantly odd numbers of carbon atoms from C23- C29 in both sexes. (Z,Z,Z)-3,6,9-tricosatriene and (Z,Z,Z)-3,6,9-pentacosatriene were identified only in cuticular extracts from females. Pheromonally naïve males showed wing fanning and circling responses to forewing scales from females but not to scales from males. Males also exhibited the same strong responses to scales excised from pharate females, indicating that the pheromone components are produced by females prior to emergence. (Z)-11-hexadecenal and (Z)-5-tricosene enhanced male responses to the primary sex pheromone aldehydes in wind tunnel bioassays, including higher proportions of in-flight and copulatory responses by males and increased time on the source. Addition of (Z,Z,Z)-3,6,9-tricosatriene to the 95/5 blend of (E)- and (Z)-11-tetradecenal released close-range copulatory responses including abdomen curling on treated septa. We propose that the sex pheromone blend of C. fumiferana is composed of the 95/5 blend of (E)- and (Z)-11-tetradecenal as primary components, with (Z)-11-hexadecenal, (Z)-5-tricosene and (Z,Z,Z)-3,6,9-tricosatriene fulfilling secondary roles in orientation and close-range courtship.

Responses of Cerambycidae and Other Insects to Traps Baited With Ethanol, 2,3-Hexanediol, and 3,2-Hydroxyketone Lures in North-Central Georgia.

In north-central Georgia, 13 species of woodboring beetles (Coleoptera: Cerambycidae: Cerambycinae) were attracted to multiple-funnel traps baited with ethanol and one of the following pheromones: (1) racemic 3-hydroxyhexan-2-one; (2) racemic 3-hydroxyoctan-2-one; and (3) syn-2,3-hexanediol. The following species were attracted to traps baited with ethanol and 3-hydroxyhexan-2-one: Anelaphus pumilus (Newman), Eburia quadrigeminata (Say), Euderces pini (Olivier), Knulliana cincta (Drury), Neoclytus mucronatus (F.), Neoclytus scutellaris (Olivier), and Xylotrechus colonus (F.). Clytus marginicollis Castelnau & Gory, and Anelaphus parallelus (Newman) were attracted to traps baited with ethanol and 3-hydroxyoctan-2-one, whereas traps baited with ethanol and syn-2,3-hexanediol were attractive to Anelaphus villosus (F.), A. parallelus, Neoclytus acuminatus (F.), Neoclytus jouteli jouteli Davis, and Megacyllene caryae (Gahan). Ethanol enhanced catches of seven cerambycid species in traps baited with syn-2,3-hexanediol and 3,2-hydroxyketones. Catches of bark and ambrosia beetles (Curculionidae: Scolytinae) in ethanol-baited traps were largely unaffected by the addition of syn-2,3-hexanediol and 3,2-hydroxyketone lures, except for two species. The mean catches of Hypothenemus rotundicollis Wood & Bright and Dryoxylon onoharaensum (Murayama) in ethanol-baited traps increased and decreased, respectively, with the addition of racemic 3-hydroxyoctan-2-one. Traps baited with ethanol and syn-2,3-hexanediol were attractive to Xylobiops basilaris (Say) (Bostrichidae) and Chariessa pilosa (Forster) (Cleridae), whereas Temnoscheila virescens (F.) (Trogossitidae) were attracted to traps baited with ethanol and 3-hydroxyhexan-2-one. The assassin bug, Apiomerus crassipes (F.) (Hemiptera: Reduviidae), was attracted to traps baited with ethanol and 3,2-hydroxyketones.

A Biologically Active Analog of the Sex Pheromone of the Emerald Ash Borer, Agrilus planipennis.

The emerald ash borer, Agrilus planipennis (Coleoptera: Buprestidae) (EAB), is an invasive species causing unprecedented levels of mortality to ash trees in its introduced range. The female-produced sex pheromone of EAB has been shown to contain the macrocyclic lactone (3Z)-dodecen-12-olide. This compound and its geometrical isomer, (3E)-dodecen-12-olide, have been demonstrated previously to be EAG active and, in combination with a host-derived green leaf volatile, (3Z)-hexenol, to be attractive to male EAB in green prism traps deployed in the ash tree canopy. In the current study, we show that the saturated analog, dodecan-12-olide, is similarly active, eliciting an antennal response and significant attraction of EAB in both olfactometer and trapping bioassays in green traps with (3Z)-hexenol. Conformational modeling of the three lactones reveals that their energies and shapes are very similar, suggesting they might share a common receptor in EAB antennae. These findings provide new insight into the pheromone ecology of this species, highlighting the apparent plasticity in response of adults to the pheromone and its analog. Both of the unsaturated isomers are costly to synthesize, involving multistep, low-yielding processes. The saturated analog can be made cheaply, in high yield, and on large scale via Mitsunobu esterification of a saturated ω-hydroxy acid or more simply by Baeyer-Villiger oxidation of commercially available cyclododecanone. The analog can thus provide an inexpensive option as a lure for detection surveys as well as for possible mitigation purposes, such as mating disruption.

Identification of sex pheromone components of blueberry spanworm Itame argillacearia (Lepidoptera: Geometridae).

Blueberry spanworm, Itame argillacearia (Packard), is an important defoliator of lowbush (syn. 'wild') blueberry, Vaccinium angustifolium Aiton, in north-eastern North America. The goal of the present study was to identify the female I. argillacearia sex pheromone, which could be used in traps for monitoring or mass-trapping this pest. Gas chromatography/mass spectrometry (GC/MS) and electroantennogram (EAG) recordings of sex pheromone gland extracts, in combination with chemical synthesis, a Y-tube olfactometer study and field experiments confirmed (2R,3S)-2-ethyl-3-((Z,Z)-tridecadi-2,5-enyl) oxirane (hereafter (Z,Z)-(3R,4S)-3,4-epoxy-6,9-heptadecadiene) and (Z,Z,Z)-3,6,9-heptadecatriene as female-produced sex pheromone components. (Z,Z)-(3R,4S)-3,4-Epoxy-6,9-heptadecadiene elicited a response from male I. argillacearia antennae during EAG recording, and in the Y-tube olfactometer tests males did not discriminate between a live female and (Z,Z)-(3R,4S)-3,4-epoxy-6,9-heptadecadiene. Field-trapping experiments showed that a blend of (Z,Z)-(3R,4S)-3,4-epoxy-6,9-heptadecadiene and (Z,Z,Z)-3,6,9-heptadecatriene was more attractive to male moths than (Z,Z)-(3R,4S)-3,4-epoxy-6,9-heptadecadiene alone.

Stereoselective biosynthesis of chloroarylpropane diols by the basidiomycete Bjerkandera adusta.

Previously we have shown that 1-arylpropane-1,2-diols are catabolic products of L-phenylalanine during idiophasic metabolism of B. adusta that are stereoselectively biosynthesized from a C(7)-unit (ring+benzylic carbon) and a C(2)-unit as predominantly erythro 1R, 2S enantiomers.In order to probe the mechanism of 1-arylpropane-1,2-diol formation, the products of the incubation of isotopically labelled aromatic aldehydes as substrates with Bjerkandera adusta (DAOM 215869) have been characterized. The aromatic aldehydes were benzaldehyde (ring D(5)) and 4-methoxy- and 4-hydroxybenzaldehydes (ring 13C(6)). These aldehydes were all stereoselectively incorporated into the corresponding 1-arylpropane-1,2-diols, including the chloro analogues, as well as into the corresponding alpha-ketols (phenyl acetyl carbinols (PAC's) and 2-hydroxy propiophenones (2-HPP's)) the presumed precursors of the diols. Benzoic acid (ring D(5)) was likewise incorporated into the diols, chlorodiols and alpha-ketols. These results lead us to conclude that the aromatic aldehydes benzaldehyde, 4-hydroxybenzaldehyde and 4-methoxybenzaldehyde are likely C(7)-unit precursors in the carboligation reaction(s) that leads to 1-arylpropane-1,2-diol biosynthesis. The metabolic role of the diols remains to be elucidated but they may be important intermediates in CAM (chlorinated anisyl metabolite) aldehyde-alcohol cycling and also act as substrates for the chlorination/hydroxylation enzymes yet to be identified in white rot fungi.

Chlorometabolite production by the ecologically important white rot fungus Bjerkandera adusta.

Two strains of the basidiomycete, Bjerkandera adusta (DAOM 215869 and BOS55) produce in static liquid culture, phenyl, veratryl, anisyl and chloroanisyl metabolites (CAM's) (alcohols, acids and aldehydes) as well as a series of compounds not previously known to be produced by Bjerkandera species: 1-phenyl, 1-anisyl, 1-(3-chloro-4-methoxy) and 1-(3,5-dichloro-4-methoxy) propan-1,2-diols, predominantly as erythro diastereomers with IR, 2S absolute configurations. 1-Anisyl-propan-1,2-diol and 1-(3,5-dichloro-4-methoxy)-propan-1,2-diol are new metabolites for which the names Bjerkanderol A and B, respectively, are proposed. Experiments with static liquid cultures supplied with 13C6- and 13C9-L-phenylalanine showed that all identified aromatic compounds (with the exception of phenol) can be derived from L-phenylalanine. For the aryl propane diols, the 13C label appeared only in the phenyl ring and the benzylic carbon, suggesting a stereoselective re-synthesis from a C7 and a C2-unit, likely aromatic aldehyde and decarboxylated pyruvate, respectively. Other compounds newly discovered to be derived from phenylalanine by this white rot fungus include phenylacetaldehyde and phenylpyruvic, phenylacetic, phenyllactic, mandelic and phenyl glyoxylic (benzoyl formic) acids. For both strains, cultures supplied with Na37Cl showed incorporation of 37Cl in all identified chlorometabolites. Veratryl alcohol and the CAM alcohols, which occur in both strains and can be derived from L-phenylalanine (all 13C-labelled), have reported important physiological functions in this white rot fungus. Possible mechanisms for their formation through the newly discovered compounds are discussed.

Identification of sex pheromone component of spruce budmothZeiraphera canadensis.

The analyses of virgin female sex pheromone gland extracts by gas chromatography (GC), GC-electroantennographic detection (GC-EAD) and GC-mass spectrometry (GC-MS) followed by field-trapping experiments, have identified (E)-9-tetradecenyl acetate (E9-14:Ac) as the primary sex pheromone component of the spruce budmoth,Zeiraphera canadensis. Dosages of 1.0-100.0 [Symbol: see text]g ofE9-14:Ac impregnated in rubber septa provide effective trap baits.

Reduction in diet breadth results in sequestration of plant chemicals and increases efficacy of chemical defense in a generalist grasshopper.

The lubber grasshopper,Romalea guttata, is a generalist feeding on a broad diet of many herbaceous plant species and has a metathoracic defensive secretion normally containing phenolics and quinones synthesized by the insect. When insects were reared on a restricted diet of wild onion, they sequestered sulfur volatiles from the plant into their defensive secretions. These compounds were not detected by gas chromatography-mass spectroscopy in secretions of insects on an artificial diet or a natural, generalist diet of 26 plants that included wild onion as a component, nor were they present in secretions from field-collected insects. Defensive secretions of insects reared on wild onion were significantly more deterrent, by as much as an order of magnitude, to two species of ant predators than secretions from insects on either of the other two diets, despite a reduction in the concentration of autogenous defensive chemicals in secretions of insects on the onion diet. Sequestration of plant chemicals that increased defensive efficacy occurred when diet breadth was reduced. We suggest that this occurs because under conditions of specialization, plant secondary metabolites are more likely to be ingested and bioaccumulated in sufficient concentrations to have biological activity against predators. What we define as casual bioaccumulation of bioactive plant chemicals following dietary specialization may lead to evolution of sequestered defense syndromes in insects, and this process may not necessarily require specific adaptation to or coevolution with a toxic host plant.

Sex pheromone of purplestriped shootworm,Zeiraphera unfortunana powell.

The analyses of virgin female sex pheromone gland extracts and gland volatiles by GC, GC-EAD and GC-MS, followed by field trapping experiments, have identified (E)-9-dodecenyl acetate (E9-12∶Ac) as the primary sex pheromone component of the purplestriped shootworm,Zeiraphera unfortunana. Dosages of 1.0-10.0 µg ofE9-12∶Ac impregnated in rubber septa provide an effective trap bait and can be used for monitoring purposes.

Effects of diet breadth on autogenous chemical defense of a generalist grasshopper.

The lubber grasshopper,Romalea guttata, produces a metathoracic defensive secretion containing primarily phenolics and quinones. This insect feeds on a wide range of plant species. Insects reared on an artificial diet and a diet of onion,Allium canadense, had secretions that contained fewer compounds, lower concentrations of compounds, and markedly altered relative composition of components compared to insects reared on a varied diet of 26 plant species that included onion. The study demonstrates that diet breadth has a major effect on the quality and quantity of the autogenous defensive secretion of this generalist herbivore. The results are compared to diet effects known in chemically defended specialists. Two possible mechanisms explaining the effects of diet breadth are proposed: one involves changes in precursor availability with changing diet breadth; the other suggests that physiological stress due to diet restriction changes allocation of resources to chemical defense.

Chemical identification and behavioral characterization of male wing pheromone ofEphestia elutella (Pyralidae).

Behavioral and chemical evidence is presented for the identity of the male wing pheromone ofEphestia elutella (Hübner) and the role of this pheromone in courtship success is evaluated. Males with the forewing removed experienced a mating success rate less than half of that of either intact males or males that had only the wing gland area remaining of their forewings. GC-MS analysis and microchemical reactions indicated the presence of (E)-phytol and a series of saturated γ-lactones in a methylene chloride extract of the wings. Using an assay of female courtship behavioral response, (E)-phytol was found to evoke an intermediate level of response in females when presented alone, while the complete array of insect-derived γ-lactones produced no significant response. The combination of either γ-decalactone or γ-undecalactone with (E)-phytol in a 1∶2 ratio, however, elicited a female response equivalent to that produced by the unfractionated wing extract.

Spruce budworm (Choristoneura fumiferana) pheromone chemistry and behavioral responses to pheromone components and analogs.

This paper reviews the sex pheromone chemistry and pheromone-mediated behavior of the spruce budworm and related coniferophagous (Choristoneura) budworms. InC. fumiferana, temporal changes in pheromone-gland monounsaturated fatty acids (pheromone precursors) enable the prediction of the primary sex pheromone components. This technique may also be applicable for predicting additional pheromone components. Tetradecanal (14∶ Ald), previously shown to enhance close-range precopulatory behavior, lowers the threshold of response by males for upwind flight to a pheromone-component source. Spruce budworm males maintain upwind flight to 95∶5 (E/Z)-1,12-pentadecadiene (diolefin analog) after initiating upwind flight to a primary-component pheromone source (95∶5E/Z11-14∶Ald). This is the first demonstration of apparently normal male flight responses to a pheromone analog.

Idiosyncratic variation in chemical defenses among individual generalist grasshoppers.

The defensive secretion of the lubber grasshopper,Romalea microptera, shows extreme chemical variation among individual adults of the same sex within a single wild population. Certain phenolic compounds were absent in some individuals and present in others. Concentrations of compounds, when present, varied over two to three orders of magnitude. Chemical variation attributable to individuals accounted for 60-88% of the total quantitative variation and was evenly contributed by all individuals in both sexes. Cluster and regression analyses showed no discernible predictable patterns in the defensive secretion variation. The specificity of chemical cues used by predators may explain why these defenses are so idiosyncratic.

Identification of sex pheromone components of jack pine budworm,Choristoneura pinus pinus freeman.

Chemical identification and field-trapping experiments have shown that a blend of 85∶15 (E,Z)-11-tetradecenyl acetates and 85∶15 (E,Z)-11-tetradecen-1-ols (in a 9∶1 ratio) are female sex pheromone components for jack pine budworm,Choristoneura pinus pinus. This blend of chemicals, formulated in PVC (0.1 %, w/w) sources is as effective a trap bait as virgin females. Preliminary wind tunnel observations have indicated that this blend, effective as a trap bait, is not equivalent to females.

Double-bond location in monounsaturated fatty acids by dimethyl disulfide derivatization and mass spectrometry: Application to analysis of fatty acids in pheromone glands of four lepidoptera.

A rapid analytical procedure for the determination of the position of double bonds in mixtures of monounsaturated fatty acid methyl esters has been developed. The method is based on direct capillary GC-MS-EI analysis of dimethyl disulfide adducts. The procedure was applied to mixtures of monounsaturated fatty esters from pheromone gland extracts of three tortricids from theChoristoneura genus,C. fumiferana, C. occidentalis, andC. pinus pinus, and one noctuid from the Plusiinae subfamily,Plusia chalcites. A correlation was found between the known major pheromone components in the four species with the corresponding fatty acids. Some of the additional fatty acids may be precursors to as yet unidentified minor pheromone components, present in extremely small quantities, in these species.

Polychlorinated biphenyls, organochlorine pesticides and chlorobenzenes content of livers from Atlantic cod (Gadus morhua) caught off Halifax Nova Scotia.

The mean levels ± s.d., (ranges) of organochlorines, pesticides, chlorobenzenes and polychlorinated biphenyl were determined in the livers of 100 Atlantic cod (Gadus morhua) caught off the east coast of Canada in 1980 and were as follows: PCB 1.71±0.90 (0.24-4.33); p,p'-DDE 0.28±0.15 (0.01-0.84); p,p'-DDD 0.10±0.09 (trace-0.84); p,p'-DDT 0.15±0.09 (ND-0.58); α-HCH 0.06±0.02 (trace-0.10); α-HCH (ND-trace); heptachlor 0.02±0.01 (ND-0.04); heptachlor epoxide 0.15±0.37 (ND-2.16); dieldrin 0.06±0.11 (ND-0.71); hexachlorobenzene 0.02±0.01 (trace-0.05) mg kg(-1) (wet wt.). The levels of these compounds in 1980 have not changed over the previous eight years with the exception of PCB and the DDT group where there appears to be a general decline between 1972 and 1975 with no significant change thereafter.

Behavioral effects of secondary components of sex pheromone of western spruce budworm (Choristoneura occidentalis) free.

In a laboratory wind tunnel, upwind flight and close-range orientation to a pheromone source by maleC. occidentalis were facilitated by the addition of the secondary componentsE/Z11-tetradecen-1-ol acetate (89∶11) andE/Z11-tetradecen-1-ol (85∶15) to low source concentrations of the primary components,E/Z11-tetradecenal (92∶8). Male responses to the blends tested never equalled their responses to virgin females. The primary components alone, when released at a rate similar to that of a "calling" female, never elicited male upwind flight or source location. However, the addition of the secondary components enhanced these behavioral sequences.

Sex pheromone chemistry of the western spruce budwormChoristoneura occidentalis Free.

SCOT capillary Chromatographic and SCOT capillary chromatographic-mass spectrometic analyses of gland washes and effluvia of virgin femaleChoristoneura occidentalis Free, have been conducted with both a diapausing and nondiapausing strain of this insect. The following compounds were identified in gland washes and effluvia in both strains:E andZ11-14∶Ald,E andZ11-14∶Ac,E andZ11-14∶OH and 14∶Ald, 14∶Ac, and 14∶OH. The average aldehyde: acetate: alcohol ratio found by analysis of single glands by virgin females (nondiapausing strain) was 1∶7∶0.73. Analysis of virgin female effluvia gave this ratio as 10∶3∶8 (diapausing strain: %Z=8, 11, 15, respectively) and 10∶3∶6 (nondiapausing strain: %Z=8, 11, 12, respectively). The saturated components were generally 1-2% of theE isomer in each case. Comparisons of EAG responses of bothC. occidentalis andC. fumiferana toE11-14∶Ald,E11-14∶Ac andE11-14∶OH were made. Correlations with both laboratory and field data previously published were also made betweenC. fumiferana andC. occidentalis.