Deterrent and insecticidal effects of extracts of pithraj,Aphanamixis polystachya (Meliaceae), againstTribolium castaneum in storage

Laboratory experiments were carried out to investigate the efficacy of seed extracts of pithraj,Aphanamixis polystachya (Wall & Parker), a locally grown plant in Bangladesh, against the red flour beetle,Tribolium castaneum (Herbst.). Results of three different bioassays showed that crude extracts of pithraj seeds have strong repellent effects and moderate feeding deterrent and insecticidal (direct-contact) effects on adultTribolium castaneum.     

Olfactory behavior of red flour beetleTribolium castaneum (Herbst) (Coleoptera,Tenebrionidae) towards natural fatty acid esters

The olfactory responses of red flour beetles,Tribolium castaneum (Herbst), to methyl and ethyl ester of C₁₄–C₁₈ fatty acid prepared from tallow were studied. Methyl octadecanoate and ethyl octadecanoate discouraged aggregation of beetle adults. Ethyl 9.12-octadecadienoate acted as a repellent. Methyl pentadecanoate and ethyl tetradecanoate induced copulation at the level of 5 × 10² and 2 × 10³ g, respectively. The maximum response was observed at the level of 5 × 10² g of the same compounds, and these compounds appear to stimulate males only.     

Sexually dimorphic setiferous sex patch in the male red flour beetle,Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae): Site of aggregation pheromone production

Evidence for the existence of a male-produced aggregation pheromone secreted from the prothoracic femoral setiferous sex patch ofTribolium castaneum is reported. Both sexes were attracted toca. 60 ng of crude secretion. Males and females perceive the pheromone on the day of emergence while perception differs between the sexes: male response reaches a maximum on day 1 posteclosion, when tested at <1, 1, and 30 days; females show a maximum response at 30 days posteclosion. Behavioral responses to pheromone odors and a complex Chromatographic profile are reported.Supported in part by a Graduate Student Alumni Research Award from the Graduate School, The Ohio State University; the National Science Foundation; and the College of Agriculture and Life Sciences, University of Wisconsin, Madison, Wisconsin.Dedicated to Dr. Alexander Sokoloff for his monumental contributions to the studies onTribolium and upon the occasion of his 60th birthday.     

Behavioral responses to food volatiles by two species of stored-product coleoptera,Sitophilus oryzae (curculionidae) andTribolium castaneum (tenebrionidae)

Laboratory experiments were conducted to study the behavioral activity of grain-derived volatiles as attractants and pheromone synergists forSitophilus oryzae, an internal-feeding pest of sound grain, andTribolium castaneum, an external-feeding pest of damaged grains and flour. Behavioral studies with two-choice pitfall bioassays determined that the fresh grain volatiles valeraldehyde, maltol, and vanillin were attractive toS. oryzae at various doses, butT. castaneum were not attracted to any dose of any of these three compounds. When oils from pressed grains were bioassayed, sesame oil was significantly repellent and oat and wheat germ oils were attractive toS. oryzae. However, rice, soybean, oat, wheat germ, and corn oils were all attractive toT. castaneum. A commercial food product composed primarily of soybean oil and wheat germ was highly attractive toT. castaneum, but elicited no response fromS. oryzae. A combination of the three grain volatiles valeraldehyde, maltol, and vanillin with the synthetic pheromone sitophinone was more attractive toS. oryzae than either the pheromone alone or the tripartite grain volatile mix. Similarly, a combination of the commercial food product with the pheromone 4,8-dimethyldecanal was more attractive toT. castaneum than either food alone or pheromone alone. Behavioral responses to grain volatiles may reflect the ecological niche of the granivore:S. oryzae colonizes sound grain and is attracted to volatiles characteristic of fresh grain, whileT. castaneum utilizes damaged or deteriorated grains and responds best to oils characteristic of damaged or fungus-infested grain. Synergism of food odors and pheromones suggests that more effective traps can be devised for management of these pest insects.     

Olfactory responses ofIps plastographus maritimus lanier (coleoptera: Scolytidae) to insect and host-associated volatiles in the laboratory

Attraction of both sexes ofIps plastographus maritimus Lanier to bark-phloem-xylem discs of Monterey pine,Pinus radiata D. Don, was demonstrated in the laboratory. Increasing concentrations of male and female volatiles trapped separately and released in a one-to-one ratio decreased attraction for both sexes combined. Attraction of both sexes to volatiles derived from males and females tunneling together in a one-to-one ratio increased with increasing concentration of extract. Attraction of males and females to male-infested discs and to trapped male volatiles increased with increasing dose of males or male extract. Attraction of males and females to female-infested discs and to trapped female volatiles was also demonstrated. The presence of females in male galleries reduced the attractiveness of infested disks to both sexes combined. Increasing numbers of females, tunneling separately from males in the same disc, reduced attraction of males, but not females. When a constant attractive dose of male volatiles was released with increasing doses of female volatiles, there was no difference in response of either sex when female volatiles were present compared with the response to male volatiles alone. When a constant attractive dose of male volatiles was released with increasing concentrations of volatiles derived from males and females tunneling together in a one-to-one ratio, attraction ofI. p. maritimus decreased. Response of females was frequently higher than that of males to the same attractant source. Hence, both sexes produce an attractant, and both sexes tunneling together in the same gallery reduce attraction of males and females to an attractive dose of male attractant.     

A new bioassay for testing plant extracts and pure compounds using red flour beetleTribolium castaneum Herbst

We designed a new bioassay to test plant extract activity against stored product pests. Plant compounds were added to feed disks composed of wheat flour and yeast and fed to the red flour bettle (Tribolium castaneum). By measuring insect mass, disk mass, and insect mortality over time it was possible to calculate a phagodepression index, an antifeedant index, the amount of treatment chemical ingested by the beetles, the mortality rate, and the efficiency of conversion of ingested food. The assay was performed for 60 hr to allow for possible habituation effects and to discriminate between phagodeterrency and physiological stress caused by treatments. - and -Pinene, eugenol, kaurenic acid, sparteine, essential oils ofMinthostachis mollis andMelaleuca quinquenervia, and extracts ofSapindus saponaria were tested. Using this assay we detected the presence of both phagodepressant and phagostimulant compounds inS. saponaria extracts, and we quantified the pronounced effects of sparteine onT. castaneum.     

Microbe inhibition by Tribolium flour beetles varies with beetle species,strain, sex,and microbe group

Tribolium flour beetles produce defensive compounds, including quinones, putatively aimed at deterring predators and inhibiting microbes. Here we examine how effective the defensive secretions of Tribolium confusum and T. castaneum are at inhibiting growth of various microbes and how this varies with species, geographic strain, and sex of the beetles. We explore differences at both the kingdom and species level of common flour microbes in their susceptibility to defensive compounds. Beetle species and strains vary in their ability to inhibit microbial growth. In addition, microbes vary in their sensitivity to the beetles' defense compounds. The capability to suppress microbial growth is likely under stabilizing selection with optimum quinone production varying among populations and may be dependent on several environmental factors including temperature, humidity, and predators.     

Olfactory responses ofOrius insidiosus (Hemiptera: Anthocoridae) to volatiles of corn silks

A synomone present in hexane extracts of corn silks was found to attractOrius insidiosus (Say). The attraction was a diurnal, innate behavior, independant of sex. A portion of the prey-searching behavior ofO. insidiosus apparently relies on olfactory perception when corn is the prey's host.     

Attraction of adult sweet potato weevils,Cylas formicarius elegantulus (Summers), (Coleoptera: Curculionidae), to sweet potato leaf and root volatiles

A dual-choice olfactometer was developed to study the responses of sweet potato weevils,Cylas formicarius elegantulus (Summers), to volatiles from the sweet potato,Ipomoea batatas (L.) Lam. Both males and females were attracted by volatiles from sweet potato leaves and a methylene chloride leaf extract. Females, but not males, responded to volatiles from storage roots and a methylene chloride root extract. Leaves and storage roots from four sweet potato cultivars (Centennial, Jewel, Resisto, and Regal) were attractive to female weevils; however, the attractant response varied with cultivar. GC profiles from leaf and root extracts, and GC-MS analysis of leaf extract, for Jewel cultivar enabled the volatile peaks to be identified as sesquiterpenes.     

Olfactory responses of a specialist and a generalist grasshopper to volatiles ofArtemisia ludoviciana nutt. (Asteraceae)

Hypochlora alba is a specialist grasshopper that lives and feeds almost exclusively onArtemisia ludoviciana, a plant that produces large amounts of allelochemics including a variety of monoterpenes. This plant is not a host for generalist grasshoppers such asMelanoplus sanguinipes. The role of olfaction in the grasshopper-plant relationship was investigated by comparing electroantennograms (EAGs) of males and females of both species generated by solvent-extracted volatiles from plant leaves and by major individual components. Volatiles ofA. ludoviciana were identified by gas chromatography-mass spectrometry. The major components were 1,8-cineole, camphor, camphene, and borneol, while minor identified compounds were - and -pinene, -thujene, myrcene,p-cymene, Artemisia ketone, -thujone, and bornyl acetate. The EAGs (mV) ofH. alba males to a range of concentrations of individual volatiles or the total plant extract were nearly double those of conspecific females or both sexes ofM. sanguinipes. However, both sexes ofM. sanguinipes were more sensitive than either sex ofH, alba to geraniol, a monoterpene that commonly occurs in many plant species but is absent or is present in only trace amounts inA. ludoviciana. The increased sensitivity ofH. alba males to the odor components of their host plant appears to be related to the greater number of certain olfactory chemoreceptors on male versus female antennae. The significance of this phenomenon is discussed.     

Sites of production and occurrence of volatiles in Douglas-fir beetle,Dendroctonus pseudotsugae hopkins

Volatiles were found to be distributed throughout adult Douglasfir beetles,Dendroctonus pseudotsugae Hopkins, suggesting that the hindgut may not be the sole site of production. The volatile content of individual beetles increased dramatically with feeding; qualitative changes reflected the different bark habitats of newly emerged and established beetles. All detected volatiles were also found in beetle frass. Myrcene was the predominant monoterpene of emergent beetles and -pinene of fed beetles. Linalool and bornyl acetate occurred in significant amounts after feeding, and diacetone alcohol and sulcatol were also detected for the first time. The presence of sulcatol explains the reported cross-attraction withGnathotrichus spp.cis-Verbenol was also found to occur in addition totrans-verbenol. The volatile content of male beetles was similar to that of females but amounts of individual compounds were less. Acetic acid was found in both emerged and fed beetles and in lesser amounts in frass, suggesting that microbial metabolism could be a potential source of volatile production.     

Behavioral and electrophysiological responses of the banana weevilCosmopolites sordidus to host plant volatiles

Male and femaleCosmopolites sordidus were attracted to freshly cut banana rhizome and pseudostem in a still-air olfactometer. Females responded similarly to odors from a comparatively resistant and from a susceptible cultivar of banana, when presented as either freshly cut tissue or as Porapak-trapped volatiles. Females were also attracted to rotting banana pseudostem and to volatiles collected from it. Males and females gave similar responses to host tissue in both the behavioral bioassay and to collected volatiles in EAG recordings. Weevils did not respond, either behaviorally or electrophysiologically, to a synthetic mixture of mono- and sesqiterpenes, which made up over 9% of the volatiles collected from pseudostem.     

Olfactory responses of Ips duplicatus from inner Mongolia, China to nonhost leaf and bark volatiles

Leaf and bark volatiles from nonhost angiosperm trees were tested on Ips duplicatus by gas chromatographic–electroantennographic detection (GC-EAD) and by pheromone-baited traps in Sweden and Inner Mongolia, China, respectively. GC-EAD analysis of the headspace volatiles from fresh bark chips of Betula pubescens revealed trans-conophthorin, two green leaf volatiles (GLVs): 1-hexanol and (Z)-3-hexen-1-ol, and two C₈ alcohols: 3-octanol and 1-octen-3-ol, that consistently elicited antennal responses by I. duplicatus. The identification of these EAD-active compounds was confirmed in further GC-EAD recordings with synthetic mixtures. Antennal responses were also found to synthetic (E)-2-hexen-1-ol and linalool, which have been identifed from the leaves of nonhost birch and aspen species. No antennal responses of I. duplicatus were found to hexanal, (E)-2-hexenal, and (Z)-3-hexyl acetates. In field trapping experiments, blends of EAD-active green leaf alcohols or C₈ alcohols, or trans-conophthorin alone resulted in significant reductions (27–60%) in the number of I. duplicatus captured compared with pheromone-baited traps. The unsuitable host compound, verbenone (Vn), also significantly reduced trap catches by up to 60% in both experiments. The strongest disruptive effect resulted from the addition of the combination of green leaf alcohols, C₈ alcohols, and verbenone to the pheromone trap, which caused an 84% reduction in trap catch. The blend of two green leaf aldehydes plus the acetate increased the trap catches in 1998 and had no negative or positive effects in 1999. Our results suggest that these nonhost volatiles (NHVs) are important olfactory signals used by I. duplicatus in host selection. They may have great significance in developing semiochemical-based management programs for I. duplicatus by reducing or stopping attacks on suitable hosts.     

Chemosensory responses of cowpea weevil,Callosobruchus chinensis to an aquatic weed,water hyacinth,Eichhornia crassipes (Mart) Solms

The behavioral responses of cowpea weevil,Callosobruchus chinensis to the leaf extracts of the aquatic weed, water hyacinth,Eichhornia crassipes (Mart) Solms. was evaluated by three different methods. The petroleum ether-soluble fraction ofE. crassipes induced high weevil attraction in all methods employed. The olfactometer assay provided a reading on weevil chemosensory responses, whereas the free-choice tests measured response and preference of weevils to the untreated or extract-treated food grain of their choice over a longer period. The extracts were also presented under no choice test conditions to assay them for the presence of oviposition stimulants. In all these methods the extracts of water hyacinth evoked a quick directional response. Another interesting feature is that the extract, besides producing attraction, also caused mortality of the insects.     

4,8-Dimethyldecanal,the Aggregation Pheromone of <Emphasis Type="Italic">Tribolium castaneum</Emphasis>, is Biosynthesized Through the Fatty Acid Pathway

4,8-Dimethyldecanal (4,8-DMD) is the aggregation pheromone produced by male red flour beetles (RFB), Tribolium castaneum. To elucidate the biosynthetic origin of 4,8-DMD, the following studies were performed: (1)effects of juvenile hormone (JH) III, and pathway inhibitors mevastatin, an inhibitor of the mevalonate pathway, and 2-octynoic acid, an inhibitor of the fatty acid pathway, were tested to determine whether 4,8-DMD is derived from the fatty acid pathway or the mevalonate pathway; (2) incorporation of ¹³C-labeled acetate, propionate, and mevalonolactone into 4,8-DMD was measured to directly determine the biosynthetic origin of 4,8-DMD; and (3) incorporation of deuterium-labeled precursors, including 2-methylbutanoate (C5D), 4-methylhexanoate (C7D), 2,6-dimethyloctanoate (C10D), and 4,8-dimethyldecanoate (C12D) was tested to determine whether 4,8-DMD is biosynthesized in the sequence AcPrAcPrAc (Ac; acetate, Pr; propionate). JH III was topically applied to males at various doses. Inhibitors and isotopically labeled substrates were administered orally by feeding the beetles flour treated with the substrates of interest, after which volatiles were collected from both sexes of RFBs. The amount of 4,8-DMD produced was significantly reduced with increasing doses of JH III. Also, 2-octynoic acid inhibited the production of 4,8-DMD, but mevastatin did not. Exposure of RFBs to [1-¹³C]acetate and [1-¹³C]propionate, but not [2-¹³C]mevalonolactone, resulted in incorporation of the labeled compounds into 4,8-DMD. RFBs fed flour treated with deuterium-labeled C5D, C10D, and C12D, but not C7D, incorporated these compounds into 4,8-DMD. The findings that the production of 4,8-DMD was inhibited by 2-octynoic acid but unaffected by mevastatin, combined with the high incorporation of [1-¹³C]acetate and [1-¹³C]propionate into 4,8-DMD and the incorporation of deuterated precursors, unambiguously demonstrated that 4,8-DMD is of fatty acid rather than terpene biosynthetic origin, and that the biosynthesis of 4,8-DMD proceeds in the sequence Ac-Pr-Ac-Pr-Ac.     

Olfactory receptor cell responses ofIps grandicollis (eichhoff) (Coleoptera: Scolytidae) to intra- and interspecific behavioral chemicals

Electrophysiological recordings from antennal olfactory receptor cells were obtained fromIps grandicollis. Recordings were made from olfactory receptor cells from nine regions of the antennae in response to stimulation with the semiochemicals-pinene, frontalin,endo-brevicomin, verbenone,trans-verbenol,cis-verbenol, ipsdienol, and ipsenol. In many cases, up to two cells were recorded concurrently from the same location. When compared to males, females had a greater percentage of cells responsive to the primary pheromones ofDendroctonus frontalis, frontalin andtrans-verbenol, and ofIps spp., ipsdienol and ipsenol. Among females, more cells responded totrans-verbenol and theIps-produced volatiles than to host or otherD. frontalis-produced compounds. Olfactory cells of males responded mostly tocis-verbenol, followed by-pinene, verbenone,trans-verbenol, andendo-brevicomin. Of those cells responsive primarily to one compound, the greatest percentage were responsive totrans-verbenol in females and to verbenone in males. The response of the antennal olfactory receptor cells to semiochemicals used by male and femaleI. grandicollis is consistent with the presence of these compounds during the host colonization period for each sex. Our results, which show a lack of specificity in most pheromone and host odor receptor cells, is in contrast with previously published accounts of olfactory receptor cell specificity in otherIps species.     

Electroantennogram responses ofTrirhabda bacharides (weber) (Coleoptera: Chrysomelidae) to plant volatiles

Electroantennograms (EAGs) were recorded fromTrirhabda bacharides Le Conte (Coleoptera: Chrysomelidae) females to 28 odorants and volatiles emanating from leaves of 26 plant species including three known host plants,Baccharis halimifolia L.,B. neglecta Britt, andB. salicina T. & G. (Compositae: Asterae). Antennal receptors were highly responsive to components of the green leaf volatile complex, especially 6-carbon saturated and monounsaturated alcohols andtrans-2-hexenal. EAGs elicited by heptanal were greater than those elicited by any other saturated aldehyde. Oxygenated monoterpenes were more active than monoterpene hydrocarbons. -Bisabolol was the most active sesquiterpene. In general, EAGs to volatiles emanating fromBaccharis and other composite species were greater than those elicited by species from other families. Furthermore, when one considers only plants occurring in the environs ofT bacharides hostB. neglecta, EAGs elicited by its host plant were at least two times greater than those elicited by other coinhabiting plants.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Olfactory sensitivity of two sympatric species of rice leaf folders (Lepidoptera: Pyralidae) to plant volatiles

Electroantennograms (EAGs) were recorded from male and female moths of two sympatric leaf folder species,Cnaphalocrocis medinalis (Gue-née) andMarasmia patnalis Bradley (Lepidoptera: Pyralidae) to 91 volatile chemicals of plant origin. Responses of both leaf folder species were similar to all compounds except to three monoterpenes--myrcene, menthone, and isomenthone- and two sesquiterpenes-cis-nerolidol and isophytol. Response ofM. patnalis, an oligophagous leaf folder, to these compounds was higher compared with that of polyphagousC. medinalis. EAG responses of males to saturated and unsaturated aliphatic aldehydes were significantly higher than those of conspecific females in both species. A higher response ofC. medinalis males also was observed for 1-nonanol, 3-nonen-2-one, andtrans, trans-2,6-dimethyl-2,4,6-octatriene. In contrast, females of both species responded more to monoterpenes, borneol, isoborneol, and fenchyl alchohol. Response ofC. medinalis female was higher for terpinen-4-ol, carveol, dihydrocarveol, (–)-myrtenal, and perillaldehyde. In both species and sexes, high EAG responses were recorded for compounds of the green leaf odor complex. EAG responses to nonanal and hexanal were maximum among the aliphatic aldehydes while 1-hexanol elicited the highest response among the alcohols tested. EAG responses to terpene compounds-citronellal,-terpineol, and (–)-myrtenal-were equal to the response to 1-hexanol. While all compounds tested elicited a negative potential, thymol and carvacrol elicited a positive EAG potential. The EAG data are discussed with regard     

Electroantennogram responses of the cabbage seed weevil,Ceutorhynchus assimilis,to oilseed rape,Brassica napus ssp.Oleifera,volatiles

Electroantennograms (EAGs) were recorded from male and female cabbage seed weevils (Ceutorhynchus assimilis Payk.) in response to volatiles isolated and identified from the odor of oilseed rape (Brassica napus ssp.oleifera DC. cv. Ariana). Relatively large EAGs were obtained on stimulation with volatiles produced by the oilseed rape crop at the time when seed weevils were actively searching for host plants. Artificial rape odor without certain key volatile compounds was in most cases significantly less stimulatory than odor containing these volatiles. There were significant differences in the EAG response of the sexes ofC. assimilis to the green leaf volatiles of oilseed rape and several terpenes present in rape flower odor. The importance of the qualitative and quantitative composition of host-plant odor in host location byC. assimilis is discussed.     

Antennal olfactory responses of black turpentine beetle,Dendroctonus terebrans (Olivier), to bark beetle pheromones and host terpenes

Electroantennograms (EAGs) were recorded from male and female black turpentine beetles,Dendroctonus terebrans (Olivier), exposed to bark beetle pheromones and host terpenes. The dose-response curves indicated similarities in the receptor mechanisms for both sexes for each compound. Antennal sensitivity was greatest toendo-brevicomin, which correlates with the importance of the compound in the behavior of the beetles. At above-threshold concentrations, EAGs were greatest toendo-brevicomin and frontalin, suggesting a large population of antennal receptors for these compounds. A large population of receptors would be expected for compounds that play such a significant role in this beetle's behavior. Beetles were also shown to have receptors that respond to theIps pheromones, ipsenol, and ipsdienol.Research done in part while in the Department of Entomology, Texas Agricultural Experiment Station (TAES), Texas A&M University, and supported in part through McIntire-Stennis project 1525, USDA-CR Grant 85-CRCR-1-1856, and NATO Collaborative Research Grant 86-0710. All programs of the Agricultural Experiment Station are available without regard to race, ethnic origin, sex, or age. Virginia Polytechnic Institute and State University does not discriminate against employees, students, or applicants on the basis of race, sex, handicap, age, veteran status, national origin, religion or political affiliation.