Volatiles Production and Attractiveness to the Mexican Fruit Fly of Enterobacter agglomerans Isolated from Apple Maggot and Mexican Fruit Flies

We investigated two strains of uricase (+) Enterobacter agglomerans, one isolated from the apple maggot fly (AMF) and one from the Mexican fruit fly (MFF), for 1) attractiveness to MFF, and 2) production of attractive chemicals. Regarding chemicals demonstrated attractive to the MFF, the MFF bacterial strain produced more 2,5-dimethylpyrazine, 2-phenylethanol, and indole than the AMF strain, whereas the AMF, but not the MFF strain, produced 3-hydroxybutanone. Cell types that predominated in plated subcultures varied from batch to batch resulting in variation in volatiles production, especially by the AMF strain where indole was sometimes a major component of the odor and at other times not detectable. Despite the greater production of attractive chemicals by the MFF strain, the AMF strain was consistently more attractive and the MFF strain was not different from uninoculated control plates. Statistical analyses indicated negative correlations of attractiveness with production of indole, 2,5-dimethylpyrazine, and 2-phenylethanol, and positive correlation with 3-hydroxybutanone. Results support previous findings with the Mexican fruit fly that showed combinations of attractive chemicals sometimes are not attractive.     

Chemicals Attractive to Mexican Fruit Fly from Klebsiella pneumoniae and Citrobacter freundii. Cultures Sampled by Solid-Phase Microextraction

Headspace above tryptic soy broth culture filtrates of Klebsiella pneumoniae contained greater amounts of ammonia, methylamine, 3-methylbutanamine, 1-pyrroline, 2,3,4,5-tetrahydropyridine, and two pyrazines than were found above tryptic soy broth. It also contained chemicals not found above tryptic soy broth, including dimethyldisulfide and several alcohols and ketones. Headspace above tryptic soy broth culture filtrates of Citrobacter freundii contained greater amounts of ammonia, 1-pyrroline, and several pyrazines than were found above tryptic soy broth. It also contained chemicals not found above tryptic soy broth including dimethyldisulfide, some of the same alcohols as above K. pneumoniae filtrates, a different ketone, and phenol. Additional chemicals were detected above filtrates that were saturated with sodium chloride or had their pH adjusted up or down. In laboratory bioassays with protein-starved, sugar-fed Mexican fruit flies, chemicals that did not contain protonizable nitrogen were not attractive. All chemicals containing protonizable nitrogen, except 2-methylpyrazine, were attractive. Synthetic mixtures of ammonia, trimethylamine, 1-pyrroline, 3-methylbutanamine, pyrazine, 2,3,4,5-tetrahydropyridine, 2,5-dimethylpyrazine, and trimethylpyrazine in concentrations similar to those in filtrates of the two bacteria were 73–87% as attractive as bacterial filtrates.     

Volatiles from Duck Feces Attractive to Mexican Fruit Fly

Aqueous extracts of three types of bird feces were attractive to laboratory-colony Mexican fruit flies in laboratory bioassays. Extracts of black-bellied whistling duck feces were chosen for further analysis. Duck feces extract was attractive to both sugar-fed and sugar-starved flies but was more attractive to sugar-fed flies. Protein deprivation of flies had no effect on their response. Duck feces incubated in water for 24 hr were more attractive than those incubated for longer periods. Duck feces extract was attractive at pH 5–9, but was most attractive at pH 9, indicating that more than one class of chemicals contributed to attractiveness of the feces. Major components of headspace of duck feces extracts collected by solid-phase microextraction and identified by GC-MS, GC-FID, and GC-FTD included ethanol, propanol, phenol, ammonia, low-molecular weight amines, and pyrazines. In general, chemicals containing nitrogen proved most attractive to sugar-fed flies. However, dimethylamine and 1-pyrroline were nearly equally attractive to sugar-fed and sugar-deprived flies. Two chemicals without nitrogen were attractive to sugar-deprived flies; none were attractive to sugar-fed flies. A synthetic mixture was prepared containing ammonia, methylamine, dimethylamine, trimethylamine, 1-pyrroline, phenol, and 2-ethylhexanol in amounts found in odor of duck feces extract. The synthetic mixture was 96% as attractive as duck feces extract to sugar-fed flies and 80% as attractive to sugar-deprived flies.     

Attractiveness to mexican fruit flies of combinations of acetic acid with ammonium/amino attractants with emphasis on effects of hunger

Ammonium acetate was more attractive than other ammonium salts to Mexican fruit flies (Anastrepha ludens) in an orchard test. We hypothesized that acetic acid enhanced the attractiveness of ammonia in the orchard test and that acetic acid may similarly enhance attractiveness of AMPu, an attractant consisting of a mixture of ammonium bicarbonate or ammonium carbonate, methylamine HCl, and putrescine. In laboratory experiments, acetic acid was attractive to flies deprived of either yeast hydrolysate or both sugar and yeast hydrolysate but not to flies fed both sugar and yeast hydrolysate. AMPu/acetic acid combinations were more attractive than AMPu alone to flies deprived of both sugar and yeast hydrolysate but not to flies fed sugar, regardless of yeast hydrolysate deprivation status. Acetic acid is the first attractant found that has become more attractive with both sugar and protein deprivation in studies withA. ludens. It is also the first that has enhanced the attractiveness of another attractant type. In orchard tests, yellow sticky panels baited with either AMPu or 17 mg of acetic acid were at least six times more attractive than unbaited panels. However, panels baited with both acetic acid (17–68 mg) and AMPu were less attractive than AMPu alone. These results differed from the laboratory data in which combinations were never less attractive than AMPu alone.     

Grapefruit Oil Enhances Attraction of Mexican Fruit Flies to a Synthetic Food-Odor Lure

We investigated the attractiveness of grapefruit oil to the Mexican fruit fly. Only high concentrations were attractive in laboratory wind-tunnel bioassays. Attraction of flies to grapefruit oil was not enhanced if they had previous experience with grapefruit. In citrus orchard experiments, undiluted grapefruit oil attracted Mexican fruit flies and enhanced attraction to traps baited with a synthetic food-odor lure emitting ammonia and other nitrogenous chemicals. This is the first demonstration of host fruit odor increasing attraction to another type of attractive blend in Mexican fruit fly. These results indicate differences in the way the flies respond to undiluted grapefruit oil compared with previously tested fruit odors.     

Identification of odors from overripe mango that attract vinegar flies,Drosophila melanogaster

Bioassays with a variety of overripe fruits, including mango, plum, pear, and grape, and their extracts showed that odors from overripe mango were most attractive to adult vinegar flies, Drosophila melanogaster. Combined gas chromatography–electroantennographic detection (GC-EAD) analyses of solid-phase microextraction (SPME) and Tenax extracts of overripe mango odors showed that several volatile compounds, including ethanol, acetic acid, amyl acetate, 2-phenylethanol, and phenylethyl acetate elicited significant EAG responses from antennae of female flies. Most of the volatile compounds in the extracts were identified by mass spectral and retention index comparisons with synthetic standards. In cage bioassays, lures with a blend of ethanol, acetic acid, and 2-phenylethanol in a ratio of 1:22:5 attracted six times more flies than any single EAG-active compound. This blend also attracted four times more flies than traps baited with overripe mango or unripe mango. However, in field trials, the blend was not as attractive as suggested by the laboratory bioassay.     

Attractants fromStaphylococcus aureus cultures for Mexican fruit fly,Anastrepha ludens

Volatile chemicals from tryptic soy broth cultures ofStaphylococcus aureus that attract sugar-fed, protein-hungry adult Mexican fruit flies were identified. Chemicals identified from the headspace above the filtrate of the bacterial cultures were ammonia, trimethylamine, isoamylamine, 2-methylbutylamine, 2,5-dimethylpyrazine, and acetic acid. Each chemical attracted flies. A mixture of the chemicals in the same concentrations as were found in the bacterial filtrate was 89% as effective in attracting flies as the bacterial filtrate in laboratory bioassays. Additional chemicals were identified from various concentrated or pH altered preparations made from the filtrate. Many of these chemicals also attracted flies. One of these chemicals, dimethylamine, was the most effective chemical identified. The use of solid-phase microextraction for volatile collection and of thick-film (5-µm) capillary GC columns was essential to the success of this work.     

Attraction of Female Oriental Fruit Fly, Bactrocera dorsalis, to Volatile Semiochemicals from Leaves and Extracts of a Nonhost Plant, Panax (Polyscias guilfoylei) in Laboratory and Olfactometer Assays

Fresh whole leaves and solvent–water leaf extracts of the hedgerow plant panax, Polyscias guilfoylei (Bull), were tested for their attractiveness to male and female Oriental fruit flies, Bactrocera dorsalis, in laboratory flight tunnel and cage olfactometer bioassays. Fresh mature whole panax leaves were found to be attractive to mated female oriental fruit flies in the flight tunnel. Response of males and virgin females was low and in most instances not significantly different from controls. Attraction of mated female flies to the layers resulting from a methylene chloride–water partition or a hexane–water partition of freshly ground leaves using small McPhail traps was greatest in the methylene chloride fraction. When methylene chloride and water layers were tested competitively in a multiple-choice rotating olfactometer, the methylene chloride fraction was more attractive. Tests involving the methylene chloride–water interface (an emulsion of the two partitioned layers) with and without a standard attractant NuLure, showed the emulsion layer to be significantly more attractive than the other fractions or NuLure. In outdoor cage olfactometer assays of methylene chloride and water fractions, activity was greatest in the methylene chloride fraction. The results suggests that volatile semiochemicals from this nonhost plant are attractive to mated female Oriental fruit flies. The results are discussed in relation to the chemical ecology of B. dorsalis and the potential use of this nonhost plant for detection and control of female Oriental fruit flies in the field.     

A novel attractant for Mexican fruit fly,Anastrepha ludens,from fermented host fruit

Chemicals from fermented chapote fruit were identified and evaluated as attractants for hungry adult Mexican fruit flies in laboratory and greenhouse bioassays. Twenty-eight chemicals identified from an attractive gas-chromatography fraction were as attractive as a chapote volatiles extract (CV) when mixed in the same amounts found in CV. Sixteen of the chemicals were slightly attractive to flies when tested individually. A mixture containing 15 of the chemicals by design and the 16th as an impurity, in arbitrary concentrations, was at least as attractive as the original CV. In a series of experiments, the number of chemicals was reduced to three by elimination of unnecessary components. The three-component mixture retained the attractiveness of the 15-component mixture. The three chemicals were 1,8-cineole, ethyl hexanoate, and hexanol (CEH). Attractiveness of the three-chemical mixture was equal to the sum of the attractiveness of the three individual components, suggesting that each chemical binds to a different receptor type that independently elicits partial attraction behavior. Optimal ratios were 1011 of the three chemicals, respectively. Optimal test quantities ranged between 0.4–4g of 1,8-cineole and 40–400 ng each of ethyl hexanoate and hexanol applied to filter paper in the laboratory bioassays. A neat 1011 mixture of the chemicals was 1.8 times more attractive than aqueous solutions ofTorula dried yeast and borax to starved 2-day-old flies when the lures were tested in competing McPhail traps in a large greenhouse cage.Diptera: Tephritidae.     

Attraction of both sexes of Mexican fruit fly,Anastrepha ludens,to a mixture of ammonia,methylamine, and putrescine

Eight chemicals were evaluated in laboratory experiments as attractants for sugar-fed adult Mexican fruit flies. Ammonium bicarbonate, methylamine HCl, ethanolamine, pyrrolidine, putrescine, and monomethyl succinate were slightly attractive when tested singly. A mixture containing all eight chemicals was much more attractive than any of the individual chemicals. Through a series of experiments, a mixture of three of the chemicals was found that was at least as attractive as the original eight-component mixture. The final mixture consisted of ammonium bicarbonate, methylamine HCl, and putrescine in a 10101 ratio. Ratios were less important than actual concentrations of individual components over the range of component concentrations tested. The three-component mixture was equally attractive to male and female flies over at least a 1000-fold range of concentrations and was slightly more attractive thanTorula yeast over the upper 10-fold range in competing McPhail traps in a greenhouse flight chamber.     

Partial characterization and HPLC isolation of bacteria-produced attractants for the Mexican fruit fly,Anastrepha ludens

Methods were developed to collect and isolate volatile chemicals produced by aStaphylococcus bacterium in tryptic soy culture that are attractive to protein-hungry adult Mexican fruit flies. Centrifugation of bacteria culture yielded a slightly attractive pellet containing most of the bacteria cells and a highly attractive supernatant. Supernatant filtered to remove the remaining bacteria was as attractive as the unfiltered supernatant. Filtrate at pH 7 and above was much more attractive than filtrate at pH 5 and below. Most of the attractiveness was retained on strong cation exchange media under acidic conditions and eluted with base. Attractive principles could not be trapped on adsorbents such as Porapak Q or extracted with organic solvents from aqueous preparations, but they were easily collected by headspace sweeping with steam. The attractive components were efficiently concentrated by rotary evaporation of steam distillate at pH 5, but at higher pH much of the attractiveness distilled. A reverse-phase HPLC method using a negative counter-ion was developed to separate and collect attractive components of concentrated steam distillate. Attractive fractions collected using this method were concentrated and injected onto silica HPLC. Activity eluted from silica in two distinct bands. Results suggest that the most attractive components of the bacterial odor are highly polar, low-molecular-weight amines.Diptera: Tephritidae.     

Mexican Fruit Fly Attractants: Effects of 1-Pyrroline and Other Amines on Attractiveness of a Mixture of Ammonia, Methylamine, and Putrescine

Several amines were tested alone and in combination with AMPu, an attractant mixture containing ammonium bicarbonate or ammonium carbonate, methylamine hydrochloride, and putrescine, for attractiveness to Mexican fruit flies (Anastrepha ludens Loew). In laboratory bioassay, 1-pyrroline, 3-pyrroline, 2-(methylamino)ethanol, spermidine, spermine, and indole-3-acetic acid were significantly more attractive than solvent controls. In orchard tests, traps baited with combinations of AMPu with dimethylamine hydrochloride, ethylamine, 2,5-dimethylpyrazine, or pyrrolidine captured fewer flies than traps baited with AMPu alone. Traps containing AMPu plus additional ammonium bicarbonate were much less attractive than AMPu alone. Combinations of AMPu with 1-pyrroline were about 50% more attractive than AMPu alone to both males and females. Combinations of AMPu with 3-pyrroline were not significantly more attractive than AMPu alone.     

Decreased Attraction of Anastrepha ludens to Combinations of Two Types of Synthetic Lures in a Citrus Orchard

Combinations of the previously developed attractants CEHO from host fruit aroma and AMPu from volatile metabolites of amino acids were evaluated for attractiveness to gamma-irradiated Mexican fruit flies, Anastrepha ludens, in a citrus orchard. In one experiment, McPhail traps with polyvinyl chloride (PVC) lures loaded with CEHO (10:1:1:50 mixture of 1,8-cineole, ethyl hexanoate, hexanol, and ethyl octanoate) were more attractive than blank traps. McPhail traps with AMPu (10:10:1 mixture of ammonium bicarbonate, methylamine HCl, and putrescine) were more attractive than blank and CEHO traps. Traps containing both AMPu and CEHO lures were less attractive than traps containing AMPu alone. In another experiment, sticky yellow panels and spheres were used to compare attractiveness of the same two attractants in different formulations. Sticky traps baited with membrane formulations of CEHO (10:1:1:10 mixture of the chemicals listed above) were not significantly attractive. Sticky traps with polypropylene tubes containing an agar formulation of AMPu (6:10:1 mixture of ammonium carbonate, methylamine HCl, and putrescine) were more attractive than blank and CEHO baited traps. As in the first experiment, traps with both AMPu and CEHO lures were less attractive than traps with AMPu alone. Results indicate that attractive host-odor volatiles and attractive amino-acid metabolites interact negatively with regard to attractiveness to the Mexican fruit fly.     

Field Attraction of Mediterranean Fruit Fly, Ceratitis capitata (Wiedemann) to Synthetic Stereoselective Enantiomers of the Ceralure B1 Isomer

Stereoselectively synthesized enantiomers of ethyl cis-5-iodotrans-2-methylcyclohexane-1-carboxylate (ceralure B1), a potent lure for male Mediterranean fruit flies, were tested in the laboratory and the field against laboratory reared sterile flies. The (–)-ceralure B1 enantiomer was significantly more attractive than the (+)-ceralure B1 antipode. Dose–response studies of the above compounds demonstrated that (–)-ceralure B1 and to a lesser extent, racemic ceralure B1, captured consistently more male flies than trimedlure, the current male attractant used in detection programs. Longevity tests demonstrated that, over a three-week period, both (–)-ceralure B1 and racemic ceralure B1 caught significantly more flies than trimedlure. The synthesis of specific enantiomers of ceralure B1 shows great promise as a male attractant that could replace trimedlure for detection and delimitation in action programs aimed at this exotic pest.     

Methyl Salicylate,a Soybean Aphid-Induced Plant Volatile Attractive to the Predator <Emphasis Type="Italic">Coccinella septempunctata</Emphasis>

Induced volatiles provide a signal to foraging predatory insects about the location of their prey. In Iowa, early in the growing season of soybean, Glycine max, many predacious seven-spotted lady beetles, Coccinella septempunctata, were observed on plants with heavy infestations of soybean aphid, Aphis glycines. We studied whether the attraction of this beetle is caused by the release of specific volatile compounds of soybean plants infested by aphids. Volatile compounds emitted by soybean plants infested by aphids were compared with those of undamaged, uninfested, and artificially damaged plants. Gas chromatography–mass spectrometry analyses revealed consistent differences in the profiles of volatile compounds between aphid-infested soybean plants and undamaged ones. Significantly more methyl salicylate was released from infested plants at both the V1 and V2 plant growth stages. However, release patterns of two other induced plant volatiles, (d)-limonene and (E,E)-α-farnesene, differed between the two plant growth stages. Gas chromatographic–electroantennographic detection of volatile extracts from infested soybean plants showed that methyl salicylate elicited significant electrophysiological responses in C. septempunctata. In field tests, traps baited with methyl salicylate were highly attractive to adult C. septempunctata, whereas 2-phenylethanol was most attractive to the lacewing Chrysoperla carnea and syrphid flies. Another common lady beetle, the multicolored Asian lady beetle, Harmonia axyridis, showed no preference for the compounds. These results indicate that C. septempunctata may use methyl salicylate as the olfactory cue for prey location. We also tested the attractiveness of some selected soybean volatiles to alate soybean aphids in the field, and results showed that traps baited with benzaldehyde caught significantly higher numbers of aphids.     

A four-component attractant for the mexican fruit fly,Anastrepha ludens (Diptera: Tephritidae), from host fruit

Sixteen chemicals found in fermented chapote fruit odor were evaluated as attractants for hungry adult Mexican fruit flies. Ethyl octanoate, ethyl benzoate, terpinyl acetate, ethyl salicylate, and (–)--copaene proved slightly attractive. Several of the chemicals also were tested for their ability to increase the attractiveness of the previously developed chapote-derived attractant (CEH) consisting of 1,8-cineole, ethyl hexanoate, and hexanol. Combinations containing CEH with ethyl octanoate, ethyl benzoate, 4-terpineol, (–)--cubebene, or-terpineol were significantly more attractive than CEH alone. The two most attractive four-component combinations were ethyl octanoate with CEH (CEHO) and ethyl benzoate with CEH. No combinations containing greater numbers of chemicals were significantly more attractive than CEHO. Therefore, CEHO was selected for further study in this paper. Of CEHO component ratios that were tested, the most attractive was 1011100 for the chemicals 1,8-cineole, ethyl hexanoate, hexanol, and ethyl octanoate, respectively. Formulations of CEHO into rubber septa and polyvinyl chloride (PVC) were aged 0–15 days and tested againstTorula yeast in competing McPhail traps in a flight chamber. Summed over all lure ages, rubber septa and PVC dispensers, respectively, were 1.2 and 1.5 times more attractive thanTorula yeast. PVC dispensers aged 10–15 days were approximately 2.1 times more attractive thanTorula yeast.     

Field Attraction of Hoplia communis to 2-Phenylethanol, A Major Volatile Component from Host Flowers, Rosa spp.

The attractiveness of volatile compounds from the floral scent of Rosa, one of the most preferred plants for adult Hoplia communis, was evaluated under field conditions. The beetles were attracted to most compounds tested, but 2-phenylethanol exhibited the highest capture rate. Catches increased with increasing emissions of between 9.1 and 287.2 mg/day. Catches in white traps were significantly larger (17.4-fold) than those in green traps when both were baited with anethole, an already known attractant; however, the trap color was not significant when a more attractive lure, 2-phenylethanol, was used. The use of a single funnel trap baited with 2 g of 2-phenylethanol at a heavily infested nursery exhibited promising results for mass trapping. Approximately 90,000 beetles of both sexes, which nearly corresponds to the estimated maximum population per 1000 m², were captured within six days.     

(7R,8S)-cis-7,8-epoxy-2-methyloctadec-17-ene: a novel trace component from the sex pheromone gland of gypsy moth, Lymantria dispar

Considering the vast Eurasian distribution of gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae), the many subspecies, and their presence in different lymantriid communities, we tested the hypothesis that L. dispar populations in eastern Asia employ one or more pheromone components in addition to the previously known single component pheromone (7R,8S)-cis-7,8-epoxy-2-methyloctadecane [= (+)-disparlure]. Coupled gas chromatographic–electroantennographic detection (GC–EAD) analyses of pheromone gland extracts of female L. dispar sensu lato (including both AGM and NAGM) on four GC columns (DB-5, DB-23, DB-210, and SP-1000) revealed a new trace component that eluted just before (DB-5; DB-210) or after (DB-23, SP-1000) disparlure, and elicited strong antennal responses. Isolation of this compound by high-performance liquid chromatography and hydrogenation produced disparlure, suggesting that the new component had the molecular skeleton of disparlure, with one or more double bonds. Of all possible monounsaturated cis-7,8-epoxy-2-methyloctadecenes, only cis-7,8-epoxy-2-methyloctadec-17-ene co-chro- matographed with the insect-produced compound on all GC columns and elicited comparable antennal responses. In field experiments in Honshu (Japan) with enantioselectively synthesized compounds, (7R,8S)-cis-7,8-epoxy-2- methyloctadec-17-ene (7R8S-epo-2me-17-ene-18Hy) was weakly attractive to male L. dispar, but was less effective as a trap bait than (+)-disparlure, and failed to enhance attractiveness of (+)-disparlure when tested in blends. The antipode, (7S,8R)-cis-7,8-epoxy-2-methyloctadec-17-ene, was not attractive, and when added to (+)-disparlure and/or 7R8S-epo-2me-17-ene-18Hy reduced attractiveness. Thus, the biological role of 7R8S-epo-2me-17-ene-18Hy remains unclear. It may enhance pheromone attractiveness or specificity in other L. dispar populations within their vast Eurasian distribution.     

Sex Pheromone of the Citrus Flower Moth <Emphasis Type="Italic">Prays nephelomima</Emphasis>: Pheromone Identification,Field Trapping Trials,and Phenology

Analysis of sex pheromone gland extract of the citrus flower moth, Prays nephelomima (Lepidoptera: Yponomeutidae) by coupled gas chromatography–electroantennogram detection, revealed one electrophysiologically active compound. Structural analysis using gas chromatography, gas chromatography–mass spectrometry, and dimethyldisulfide derivatization identified this as the monounsaturated aldehyde (Z)-7-tetradecenal. Field trials in commercial citrus orchards on the North Island of New Zealand showed that (Z)-7-tetradecenal was highly attractive to male P. nephelomima. Phenology data, collected over 19 months in three commercial orchards, from traps baited with the sex pheromone at a lure loading of 300 μg on a red rubber septum, indicated that male moths may be present throughout the year, with numbers peaking in late summer and autumn.     

Reproductive Character Displacement in Lymantria monacha from Northern Japan?

Our objective was to test the hypothesis that the pheromone blend and/or diel periodicity of pheromonal communication differ in populations of the nun moth, Lymantria monacha(Lepidoptera: Lymantriidae), from eastern Asia (northern Honshu, Japan) and Central Europe (Bohemia, Czech Republic). Coupled gas chromatographic–electroantennographic detection (GC-EAD) analyses of pheromone gland extract of female L. monachafrom Japan confirmed the presence of compounds previously identified in pheromone extracts of L. monachafrom Bohemia, as follows: (Z)-7-octadecene, 2-methyl-(Z)-7-octadecene (2me-Z7–18Hy), cis-7,8-epoxy-octadecane (monachalure), and cis-7,8-epoxy-2-methyloctadecane (disparlure). Field experiments in Honshu suggested that (+)-monachalure is the major pheromone component of L. monacha. 2me-Z7–18Hy significantly enhanced attractiveness of (+)-monachalure. Addition of (+)-disparlure to (+)-monachalure plus 2me-Z7–18Hy in Honshu and Bohemia increased attractiveness of lures by 1.2 and 20 times, respectively, indicating that (+)-disparlure is of least and most significance in the respective L. monachapopulations. Moreover, capture of male L. monachain pheromone-baited traps between 18:00 and 24:00 hr in Bohemia and 2:00 and 5:00 hr in Honshu revealed a markedly different diel periodicity of pheromonal communication. Pheromonal communication late at night and use of (+)-monachalure, rather than (+)-disparlure, as the major pheromone component by L. monachain Honshu may have resulted from interspecific competition with coseasonal L. fumida, which uses the early night for pheromonal communication and (+)-disparlure as major pheromone component. Whether communication channel divergence of L. monachain Honshu indeed constitutes a case of reproductive character displacement is difficult to prove. The evolution of such divergence in sympatricpopulations of L. fumidaand L. monachawould have to be demonstrated.