Sequestration of pyrrolizidine alkaloids by larvae ofTellervo zoilus (Lepidoptera: Ithomiinae) and their role in the chemical protection of adults against the spiderNephila maculata (Araneidae)

Summary Life stages of the primitive Australian ithomiine butterflyTellervo zoilus and its larval hostplant, the apocynaceous vineParsonsia straminea, were quantitatively assayed for pyrrolizidine alkaloids (PAs). PAs were found in all stages, mainly as N-oxides, being most concentrated in larvae and freshly-emerged adults. Although adults feed at various confirmed PA sources this probably does not compensate for losses, as wild-caught adults had considerably lower concentrations of PAs. The main alkaloid present in both freshly-emerged adults and in leaves of the host-plant was lycopsamine (1b), stored by butterflies in the N-oxide form. Its presence in higher proportion, in relation to intermedine (1a), in larvae, pupae and adults ofTellervo in relation to the host-plants suggests the inversion of intermedine to lycopsamine by the insects. No 14-member ring macrocyclic PAs were detected in either food-plant or butterflies. Several other PAs were found in wild-caught adults reflecting visits to other PA sources. PAs were also found in high concentrations in freshly-emerged individuals of the danaineEuploea core bred onParsonsia straminea. Wild-caughtDanaus affinis had high PA levels acquired from adult feeding. Freshly emergedEuploea raised onIschnocarpus frutescens andDanaus raised onIschnostemma carnosum (both PA-free) were preyed on by the orb weaving spiderNephila maculata, and showed no PAs. In all cases where PAs were present, most butterflies were liberated, usually cut out of the web unharmed, byNephila. The spider's response was not closely linked to PA concentration, however, and may also depend on hunger levels and previous experience with PA-containing butterflies. All control and other non-PA containing butterflies were consumed although rejection of some body parts of freshly-emergedDanaus affinis suggests that compounds other than PAs may be involved.     

Behavior and ecology of Old World Luperini beetles of the genusAulacophora (Coleoptera: Chrysomelidae)

Summary The attraction of Old World leaf beetles in the genusAulacophora to kairomones and parakairomones which are effective lures for New WorldDiabrotica andAcalymma were investigated. Beetles captured on sticky traps baited with single and multicomponent lures were no different from the control traps for two species ofAulacophora. Yellow colored traps and squash blossoms are attractive toAulacophora beetles which detect sub-microgram quantities of cucurbitacins on silica gel. Leaf feeding behavior and flight activity data are correlated with varietal preference of threeAulacophora species. The common response byDiabrotica andAulacophora to cucurbitacins reinforces the two groups' coevolutionary association with the Cucurbitaceae. The apparent lack of a common response toCucurbita blossom volatiles suggests recent evolutionary pathways are substantially different for these two groups of beetles.     

Antennal responses of four species of tree-killing bark beetles (Coleoptera: Scolytidae) to volatiles collected from beetles,and their host and nonhost conifers

Summary. Host selection in tree-killing bark beetles (Coleoptera: Scolytidae) is mediated by a complex of semiochemical cues. Using gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses, we conducted a comparative study of the electrophysiological responses of four species of tree-killing bark beetles, the Douglas-fir beetle, Dendroctonus pseudotsugae, Hopkins, the mountain pine beetle, D. ponderosae Hopkins, the spruce beetle, D. rufipennis Kirby, and the western balsam bark beetle, Dryocoetes confusus Swaine, to volatiles captured by aeration of 1) bole and foliage of four sympatric species of conifers, Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, lodgepole pine, Pinus contorta var. latifolia Engelm., interior spruce, Picea engelmannii x glauca, and interior fir, Abies lasiocarpa x bifolia, and 2) con- and heterospecific beetles at three stages of attack. We identified 13 monoterpenes in the conifers and nine compounds in the volatiles of beetles that elicited antennal responses. There was no qualitative difference in the terpene constitution of the four species of conifers and very little difference across beetle species in their antennal response to compounds from conifers or beetles. The lack of species-specific major or minor components in conifers suggests that beetles would need to detect differences in the ratios of different compounds in conifers to discriminate among them. Attraction to hosts and avoidance of nonhost conifers may be accentuated by perception of compounds emitted by con- and heterospecific beetles, respectively. The 22 compounds identified are candidate semiochemicals with potential behavioural roles in host location and discrimination.     

The effects of larval experience with a complex plant latex on subsequent feeding and oviposition by the cabbage looper moth: Trichoplusia ni (Lepidoptera: Noctuidae)

Summary. Leaf disc choice and oviposition bioassays were used to examine the effects of larval experience with a Hoodia gordonii latex on subsequent behaviors. The latex deterred feeding and oviposition by “naïve” cabbage looper (Trichoplusia ni, Noctuidae) larvae and moths with no previous exposure to the material. “Experienced” insects, reared on a diet with the H. gordonii latex (1000 ppm), exhibited lesser feeding deterrence relative to naïve insects. Experienced female moths actually preferred to lay eggs on treated rather than control leaves. There was no observed transfer of behavioral preferences from experienced parents to their offspring. Our results suggest that moths may be acquiring oviposition preferences from larval feeding experience as described by Hopkins’ host selection principal (HHSP) or through chemical legacy.     

Protein and amino acid composition of the tergal gland secretions ofBlatta orientalis andEurycotis floridana (Dictyoptera: Blattidae)

Summary Nymphs and adult females ofBlatta orientalis and nymphs ofEurycotis floridana produce a proteinaceous sticky secretion which accumulates on the last abdominal tergites. The proteic patterns do not differ between individuals of the same species. HPLC analyses show that all the common amino acids are found in both species, aspartic and glutamic acids representing 24 to 37% of the total amount of amino acids. InB. orientalis, glutamic acid is the more abundant amino acid whereas inE. floridana it is the aspartic acid. The secretion appears and accumulates rapidly on isolated insects. Behavioural assays revealed that these secretions have a defensive role.     

Sexually differentiated functions of female-produced pheromone of the black chafer <Emphasis Type="Italic">Holotrichia loochooana loochooana</Emphasis> (Sawada) (Coleoptera: Scarabaeidae)

Summary. Both male and female Holotrichia loochooana loochooana (Sawada) (Coleoptera: Scarabaeidae) were attracted with female-produced pheromone, anthranilic acid (2-aminobenzoic acid), in the field. Male chafers were observed to apparently directly locate cotton balls impregnated with 1 to 10 mg of pheromone. In contrast, females never directly oriented to the treated balls but landed 0.2-1.5 m away and exposed their abdominal glands in a calling posture, which occasionally resulted in aggregation of both females and males. This suggested the mating aggregation of this species could be primarily induced by pheromone released by females. A hypothesis for adaptability of female aggregation is proposed and discussed.     

Diet-induced chemical phytomimesis by twig-like caterpillars of Biston robustum Butler (Lepidoptera: Geometridae)

Summary. Polyphagous caterpillars of the giant geometer Biston robustum resemble the twigs of their respective food sources in color and shape. Common predatory ants, including Lasius and Formica, were often observed to freely prowl directly on caterpillars bodies, even after antennal contact. This suggests that the cuticular chemicals of the caterpillars resemble those of the twigs of the foodplants, so we analyzed both by GC and GC-MS. The chemical compositions differed among caterpillars fed on a cherry, Prunus yedoensis, a chinquapin Castanopsis cuspidata, and a camellia Camellia japonica. The cuticular chemicals of the caterpillars resembled those of their corresponding food sources. When the caterpillar diets were switched from the cherry to camellia or chinquapin at the 4^th instars, the caterpillars cuticular chemicals changed after molting to resemble those of their respective foods. Caterpillars also changed their cuticular chemicals when they perched on cherry twigs and fed on camellia or chinquapin leaves, but not when they perched on camellia or chinquapin twigs and fed on cherry leaves. The chemical similarities between the caterpillars and the twigs were due to the digestion of host leaves, which indicates that this is a diet-induced adaptation.     

Oviposition by butterflies on young leaves: Investigation of leaf volatiles

Summary Various butterflies select young foliage on which to lay their eggs; volatiles emitted by young and old leaves have been compared (by sorption enrichment, followed by GC-MS) to gauge possible qualitative and quantitative differences between the two age groups. The plants investigated are cabbage (Brassica oleracea), two milkweeds (Asclepias syriaca andA. curassavica), the bitter orange (Citrus aurantium) and the lime (C. aurantiifolia). The chemical compounds identified belong to three classes, isoprenoids, fatty acid derivatives and benzenoids. Quantitative differences were found between young and old leaves, of which a few may be characteristic of young leaves only. Thirty-four single trials withDanaus plexippus exposed to volatiles from young and old leaves are recorded.     

Aggregation pheromone ofCarpophilus freemani (Coleoptera: Nitidulidae): A blend of conjugated triene and tetraene hydrocarbons

Summary Males ofCarpophilus freemani Dobson (Coleoptera: Nitidulidae) produce an aggregation pheromone to which both sexes fly in a wind-tunnel bioassay. The major pheromone component (ca. 30 ng per male per day in volatile collections) was identified as (2E,4E,6E)-5-ethyl-3-methyl-2,4,6-nonatriene. A minor component, (2E,4E,6E,8E)-7-ethyl-3,5-dimethyl-2,4,6,8-undecatetraene, was 3–10% as abundant as the major triene and was 5–20% as active when compared at relative doses ranging from natural proportions to 1:1. These compounds act synergistically: a mixture of major and minor components in natural proportions attracted more than twice as many beetles as the major component alone, and the mixture fully accounted for the activity of male-derived volatile collections. Six other male-derived conjugated hydrocarbons, ranging from 2% down to 0.04% as abundant as the major component, were also identified. These are (in order of decreasing bioassay activity when compared on an equal-weight basis): (3E,5E,7E)-6-ethyl-4-methyl-3,5,7-decatriene, (2E,4E,6E)-5-ethyl-3-me-thyl-2,4,6-octatriene, (3E,5E,7E,9E)-8-ethyl-4,6-dimethyl-3, 5,7,9-dodecatetraene, (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6, 8-undecatetraene, (3E,5E,7E)-5-ethyl-7-methyl-3,5,7-undecatriene, and (2E,4E,6E)-3,5-dimethyl-2,4,6-nonatriene. All structure identifications were confirmed by synthesis. In the wind tunnel, the pheromone acted synergistically with host-type volatiles such as propyl acetate, valeric acid, and ethanol. This concept was verified by fields tests in California, in which there was dramatic synergism between the pheromone and fermenting host materials. Pheromone biosynthesis is discussed.     

The aggregation pheromone ofIps duplicatus and its role in competitive interactions withI. typographus (Coleoptera: Scolytidae)

Summary Ips duplicatus withI. typographus co-inhabiting Norway spruce (Picea abies (L.) Karst.) would benefit from a pheromone blend distinct from that of the larger competitorI. typographus. GC-MS analysis showed thatI. duplicatus males feeding in the host produced ipsdienol (Id),cis-verbenol (cV),trans-verbenol (tV), myrtenol (Mt), andE-myrcenol (EM) and traces of 2-methyl-3-buten-2-ol (MB).I. duplicatus produced Id in approximately racemic form (48.9-54.5% (+)-(S)-isomer). The amounts of Id and EM released over a 9 day period had a maximum of 250 and 5 ng/h/male, respectively, on day 2. Exposure ofI. duplicatus males to myrcene and -pinene resulted in the production of small amounts of Id, cV, tV, Mt, andtrans-pinocarveol, but not of EM. In laboratory bioassays with walking beetles, the pheromone component Id alone was weakly attractive while EM was inactive, but in binary combination with Id strongly synergized attraction. A combination of EM and Id at a release rate equivalent to 100–200 males was more attractive in the field than 70 unmated males in a spruce log. The addition of myrcene ( a suggested pheromone precursor of Id) to Id did not enhance trap caches, while addition of EM increased catches > 10-fold. Subtracting EM from a blend of Id, EM, cV and MB drastically reduced trap catches while subtraction of cV or MB or both had no significant effect. Addition of EM over a wide concentration range to the synthetic pheromone ofI. typographus did not reduce the attraction of females of this species in the laboratory. A two-species pheromone interaction field test releasingI. typographus pheromone components (MB + cV) at 10–1000 male equivalents (ME) andI. duplicatus pheromone (Id + EM) at 0, 10–1000 ME in all possible combinations showed both positive intraspecific dose-response effects and an interspecific inhibition. Higher release rates of EM appeared to inhibitI. typographus, especially males. In a tree colonization model, the response of the two competing species to their respective pheromones show a good separation during the mass-attack with a small initial cross-attraction. It remains to be shown whether either of the two pheromone systems have in fact evolved in the present sympatry, or if they are an incidental effect of ancestry of these phylogenetically distantIps.     

A distributed-delay model to predict ageing and survival rates of adults of Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae) in granaries filled with wheat

A time-varying distributed-delay model simulating effects of multifactors was developed. Prediction of the ageing rate and survival distribution of adults of the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae) in various environments found in wheat-filled granaries was conducted as an example to illustrate the application of this developed model. Published adult mortalities, determined at different temperatures, relative humidities, and food sources, were directly used to find the average ageing rate and family of cumulative function of adult mortality. The developed model could predict the adult survival rate at constant or transient temperatures with different relative humidities. This model could also simulate the effect of adult acclimation to their environment when they experience temperature and moisture fluctuations inside granaries. To validate the developed model, the simulation results were compared with available experimental data from the literature. There was no difference between predicted and measured mortalities in two granaries in which the mortalities were determined in a 4-month experiment.     

Fruit detection and discrimination by small fruit-eating bats (Phyllostomidae): echolocation call design and olfaction

We studied the role of echolocation and other sensory cues in two small frugivorous New World leaf-nosed bats (Phyllostomidae: Artibeus watsoni and Vampyressa pusilla) feeding on different types of fig fruit. To test which cues the bats need to find these fruit, we conducted behavioral experiments in a flight cage with ripe and similar-sized figs where we selectively excluded vision, olfaction, and echolocation cues from the bats. In another series of experiments, we tested the discrimination abilities of the bats and presented sets of fruits that differed in ripeness (ripe, unripe), size (small, large), and quality (intact(infested with caterpillars). We monitored the bats' foraging and echolocation behavior simultaneously. In flight, both bat species continuously emitted short (<2 ms), multi-harmonic, and steep frequency-modulated (FM) calls of high frequencies, large bandwidth, and very low amplitude. Foraging behavior of bats was composed of two distinct stages: search or orienting flight followed by approach behavior consisting of exploration flights, multiple approaches of a selected fruit, and final acquisition of ripe figs in flight or in a brief landing. Both bat species continuously emitted echolocation calls. Structure and pattern of signals changed predictably when the bats switched from search or orienting calls to approach calls. We did not record a terminal phase before final acquisition of a fruit, as it is typical for aerial insectivorous bats prior to capture. Both bat species selected ripe over unripe fruit and non-infested over infested fruit. Artibeus watsoni preferred larger over smaller fruit. We conclude from our experiments, that the bats used a combination of odor-guided detection together with echolocation for localization in order to find ripe fruit and to discriminate among them.     

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> L. (Asterales: Asteraceae), as feeding stimulants of <Emphasis Type="Italic">Ophraella communa</Emphasis> LeSage (Coleoptera: Chrysomelidae)

Summary. The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. -Amyrin acetate, -amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (-amyrin acetate or -amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.     

Sequestration of phorbolesters by the aposematically coloured bug Pachycoris klugii (Heteroptera: Scutelleridae) feeding on Jatropha curcas (Euphorbiaceae)

Summary. The phorbol ester DHPB has been detected in 5th instars and adults of Pachycoris klugii which feed on Jatropha curcas, a producer of phorbol esters with mollusc-, insect-, and vertebrate toxicity. DHPB from Pachycoris activates protein kinase C (PKC) which appears to be the main molecular target for phorbol esters. Phorbol esters of J. curcas exhibit a wide range of acute toxic effects in vertebrates and insects. It is therefore likely that the sequestration of DHPB, which would explain the aposematic colouration of the bugs, confers chemical protection to P. klugii against vertebrate predators. Received 26 April 2000; accepted 31 May 2000     

Variation of pyrrolizidine alkaloids in Ithomiinae: A comparative study between species feeding on Apocynaceae and Solanaceae

Summary The primitive, Apocynaceae-feeding Ithomiine,Tithorea harmonia, incorporates dehydropyrrolizidine alkaloids (PAs) from its larval foodplant (Prestonia acutifolia), rarely visiting PA sources pharmacophagously in the adult; females show higher concentrations of PAs than males, with similar variance. The close relativeAeria olena (feeding onP. coalita, without PAs) shows similar PA concentrations in both sexes and greater variation in males, like more advanced Solanaceae-feeding Ithomiine such asMechanitis polymnia, which likeA. olena obtain PAs by pharmacophagy in the adult (mainly males). This difference is due to the dynamics of PA incorporation in these species. Little variation in PA content was found among allopatric populations of the same species, but variation in available PA sources in different months was correlated with different average storage levels in the butterflies.     

Response of the predatorRhizophagus grandis to host plant derived chemicals inDendroctonus micans larval frass in wind tunnel experiments (Coleoptera: Rhizophagidae,Scolytidae)

Summary Adults of the predatory beetleRhizophagus grandis are strongly attracted to both adult and larval frass of its specific prey,Dendroctonus micans, in walking bioassays. Spruce bark and resin are relatively unattractive. Solvent extracts of larval frass that were attractive toR. grandis adults in a flight wind tunnel contained a mixture of monoterpenes of host plant origin. A synthetic mixture of these monoterpenes, (+)—-pinene, (–)—-pinene, -phellandrene,dl limonene and 3-carene, was responsible for 70–80% of the activity of the most attractive extract. We suggest that a blend of monoterpenes in frass acts as a kairomone forR. grandis but that attraction to monoterpenes is only the first of a sequence of behavioural responses that ensures successful host location and identification.     

Profiles of cuticular hydrocarbons mediate male mate choice and sexual isolation between hybridising <Emphasis Type="Italic">Chrysochus</Emphasis> (Coleoptera: Chrysomelidae)

Summary. Chemical signals frequently underlie sexual isolation between insect species. Our understanding of the evolutionary forces influencing these signaling systems is known for very few systems, challenging both our efforts to understand insect speciation, and our ability to predict long-term changes in the chemical communication systems of insects. Thus, we are in need of more systems in which both the chemical signals causing sexual isolation and the evolutionary forces driving sexual isolation are understood. Sexual isolation in the hybrid zone between Chrysochus cobaltinus and C. auratus has apparently increased in response to natural selection against hybridisation (i.e. reinforcement). Previous experiments suggested that this isolation was due, at least in part, to male preferences for conspecific females. Here, we confirm this role of male choice, and document that male mate choice in this system is influenced by cuticular hydrocarbon (CHC) profiles. Specifically, male C. cobaltinus responses to control cadavers and conspecific female cadavers painted with different cuticular hexane extracts, together with analyses of the composition of those extracts, revealed that male mate choice is governed by CHC profiles. Multivariate analyses of GC profiles demonstrated that those profiles are indeed both sex- and species-specific. Although GC-MS enabled identification and quantification of the specific cuticular hydrocarbons, we have not yet determined which individual compounds govern mate choice. Having established that CHCs influence sexual isolation in this system, we can now assess the evolutionary lability of these cues, which will inform both our understanding of speciation, and of the conditions under which the chemical signaling systems that influence mate choice in insects can evolve.     

Oviposition-stimulating activity of (E)-capsaicin identified in Capsicum annuum fruit and related compounds towards Helicoverpa assulta (Lepidoptera: Noctuidae)

Summary. The oviposition-stimulating activity of (E)-capsaicin identified in the fruits of red pepper, Capsicum annuum L. (Solanaceae), towards the oriental tobacco budworm, Helicoverpa assulta (Guenée), was examined in the laboratory by using choice and no-choice bioassays. Results were compared with those of the commercially available capsanthin and dihydrocapsaicin, another known constituents of C. annuum fruit, and (Z)-capsaicin. In no-choice oviposition bioassays at 10.5 μg/cm², (E)-capsaicin was the most active oviposition stimulant. Weak stimulatory activity was observed with capsanthin, (Z)-capsaicin and dihydrocapsaicin. In choice oviposition bioassays at 2.6 μg/cm², there were significant differences in oviposition-stimulating activity between (E)-capsaicin and each of the other three alkaloids. Structure-activity relationships indicate that the presence of double bond and its trans configuration appeared to play a crucial role in the oviposition-stimulating activity. On the basis of our results, (E)-capsaicin acts as a contact oviposition stimulant of female H. assulta.     

Chemical recognition of queen cells by honey bee workersApis mellifera (Hymenoptera: Apidae)

Summary Honey bee workers are able to nurse or to destroy and thus to recognize the capped queen cells containing a pupa. Fatty acid esters, especially methyl oleate, methyl palmitate and ethyl oleate were found in significant amounts on the queen pupal cuticle. Methyl oleate, the major component, along with smaller amounts of methyl linoleate and methyl linolenate, were involved in the recognition of queen cells by workers. In natural conditions of the colony, queen cells containing a paraffin pupal lure with methyl oleate were accepted 5.9 days by workers, releasing about 1.8 queen pupa equivalents during that period, when control cells (without ester) were kept only 2.1 days. Although these esters are non specific to honey bees, they are of great importance in social regulation of the honey bee colony.     

Chemical determinants of resistance in winter-dormant seedlings of European white birch (Betula pendula) to browsing by the mountain hare

Summary A sample of one-year-old seedlings of European white birch (Betula pendula) was analyzed to determine the content of sugars, phenolics and terpenoid compounds. Two vertical segments of each seedling were analyzed separately. The number of resin droplets, which correlates strongly and negatively with feeding by the mountain hare, was also counted on the bark of experimental seedlings. The variation in the palatability of birch seedlings to mountain hare was determined primarily by the most abundant terpenoid, papyriferic acid. On the other hand, the resistance to hare feeding at the seedling bases, which are frequently attacked by voles, appeared to be dependent on other components, apparently phenolic substances. Sugars did not affect the resistance of the tested seedlings. The variation among experimental seedlings was much greater for secondary substances, especially terpenoid compounds, than for sugars. It is suggested that this high variation in protective compounds may be an adaptive trait selected for by the feeding of generalist herbivores.