Aggressive chemical mimicry of moth pheromones by a bolas spider: how does this specialist predator attract more than one species of prey?

Summary. The bolas spider, Mastophora hutchinsoni, attracts Lacinipolia renigera and Tetanolita mynesalis males by mimicking the female moth sex pheromones. However, as the prey species use completely different pheromone blends we conducted experiments to determine how this is accomplished by the predator. The periodicity of L. renigera mate-seeking activities occurs early in the scotophase, whereas male T. mynesalis are active late at night, corresponding with periods when these moths are captured by the spider. The pheromone blend of early-flying L. renigera interferes with attraction of late-flying T. mynesalis to its pheromone in a dose-dependent manner, suggesting the spider must always produce a single sub-optimal “compromise” blend for both species or that it adjusts its allomonal blend to optimize capture of the respective prey species at different times during the night. We delayed (L. renigera) or advanced (T. mynesalis) the periodicity of male activity through photoperiodic manipulation and found that the bolas spider attracted both prey species outside their normal activity windows. These results support the idea that bolas spiders produce components of both species at all times rather than producing the pheromone of each prey species at different times of the night. However, using coupled gas chromatography-electroantennography, we also demonstrated that the spider decreases its emission of the L. renigera pheromone over the course of the night. This modification should reduce the behavioral antagonism of the L. renigera pheromone on T. mynesalis males and increase the predator's success of attracting T. mynesalis during this prey's normal activity window late at night. Received 13 October 2001; accepted 28 December 2001.     

Metabolic transformations of acquired lucibufagins by firefly “femmes fatales”

Summary. Female Photuris fireflies sequester defensive steroidal pyrones (lucibufagins) from male fireflies of the genus Photinus. Lucibufagin analyses of Photuris females and Photinus ignitus males show that the lucibufagin mixtures of predator and prey differ in their composition. Analyses of whole body extracts showed that P. ignitus males contain a mixture of eight non-glycosylated lucibufagins, composed mostly of compounds with two oxygenated positions in the steroidal A-ring (C-3, C-5). After feeding on P. ignitus males, Photuris females contain six major lucibufagins. Three of these compounds are not present in the prey, including the novel lucibufagin glycoside 5β,11α-dihydroxy-12-oxo-3β-O-β-D-xylopyranosylbufalin, and two other lucibufagins with a trioxygenated A-ring (C-2, C-3, C-5). These results indicate that Photuris females transform the sequestered lucibufagins both by glycosylation and oxidation, which could affect the systemic transportability of these compounds due to an increase in their polarity. Received 18 February 1999; accepted 19 April 1999.     

Lack of sequestration of host plant glucosinolates in <Emphasis Type="Italic">Pieris rapae</Emphasis> and <Emphasis Type="Italic">P. grarricae</Emphasis>

Summary. Sequestration of plant toxins in herbivores is often correlated with aposematic coloration and gregarious behaviour. Larvae of Pieris brassicae show these conspicuous morphological and behavioural characteristics and were thus suggested to sequester glucosinolates that are characteristic secondary metabolites of their host plants. P. rapaeare camouflaged and solitary, and are thus not expected to sequester. To test this hypothesis and to check the repeatabi-lity of a study that did report the presence of the glucosinolate sinigrin in P. brassicae, larvae were reared on three species of Brassicaceae (Sinapis alba, Brassica nigra and Barbarea stricta), and different leaf and insect samples were taken for glucosinolate analysis. The major host plant glucosinolates could only be found in traces or not at all in larval haemolymph, bled or starved larvae, faeces or pupae of both species or P. brassicae regurgitant. Haemolymph of both Pieris spp. was not rejected by the ant Myrmica rubra in dual-choice assays; the regurgitant of P. brassicae was rejected. This suggests the presence of compounds other than glucosinolates that might be sequestered in or produced by P. brassicae only. In faeces of both Pieris spp. a compound which yielded 4-hydroxybenzylcyanide (HBC) upon incubation with sulfatase was detected in high concentrations when larvae had been reared on S. alba. This compound may be derived from hydrolysis of sinalbin, the main glucosinolate of that plant. The unidentified HBC progenitor was apparently not sequestered in the two Pieris spp., and was not detected in faeces of larvae reared on B. nigra or B. stricta. Received 18 July 2002; accepted 11 September 2002.     

The roles of nicotine and natural enemies in determining larval feeding site distributions of Manduca sexta L. and Manduca quinquemaculata (Haworth) on tobacco

Summary. Field observations indicated that hornworms select feeding sites non-randomly on tobacco. We tested the hypotheses that differences in feeding site locations of larvae of Manduca sexta L. and Manduca quinquemaculata (Haworth) (Lepidoptera: Sphingidae) on tobacco could be explained by differential nicotine concentrations within plants and leaves, species-specific responses to nicotine, or pressure exerted by natural enemies. Results showed that third-instar larvae of M. sexta fed more proximally and centrally on the leaf, whereas M. quinquemaculata fed more distally. Within-plant selection of leaves did not differ; both species selected leaves in the middle region of the plant. Nicotine concentrations in a high nicotine genotype, NC95, varied within each leaf, increasing 2—3 fold from the basal to apical portion of the leaf, and within each plant, increasing 7—10 fold from the first fully expanded leaf to the twelfth (lowest) leaf. In laboratory bioassays, both Manduca species responded to nicotine as a feeding deterrent. Electrophysiological studies demonstrated that gustatory organs of both species responded to nicotine at concentrations found in tobacco leaves and that M. quinquemaculata generally showed a less vigorous response to nicotine than M. sexta. Field mortality of M. sexta due to parasitism by Cotesia congregata (Say) and to parasitism and predation combined differed among feeding sites; predation alone did not. Results suggest that although nicotine concentration and species specific responses to nicotine play a role in determining feeding site locations, pressure exerted by natural enemies, especially parasitism by C. congregata, is more important. Received 22 February 2000; accepted 20 July 2001.     

Two herbivore-deterrent iridoid glycosides reduce the in-vitro growth of a specialist but not of a generalist pathogenic fungus of Plantago lanceolata L

Summary. Many secondary plant compounds are involved in defense against both insect herbivores and pathogens. Two secondary plant compounds of Plantago lanceolata, the iridoid glycosides catalpol and its precursor aucubin, are well known for their deterrent effects on generalist and non-adapted specialist insect herbivores. We tested the effects of these compounds on the in-vitro growth of a specialist and generalist fungal pathogen of this host species. Two chemical forms of these iridoids were tested. The glycosides and their aglycones, the products of enzymatic conversion by specific $/Beta$-glucosidase enzymes. The glycosides enhanced growth of both the specialist fungus Diaporthe adunca and the generalist fungus Fusarium moniliforme var. subglutinans. The positive effect of these glycosides on the generalist fungus is in sharp contrast with the generally negative effects of these glysosides on generalist insect herbivores. The aglycones of aucubin and catalpol reduced the growth of the specialist fungus D. adunca, but, contrary to expectation, enhanced the growth of the generalist fungus F. moniliforme var. subglutinans. Effects of aucubin on D. adunca were stronger than effects of catalpol. This was true both for the growth stimulating effects of the glycosides and for the fungitoxic effects of the aglycones. We therefore expect that the effects of these iridoids in P. lanceolata on the specialist fungus will strongly depend on the ratio between catalpol and its precursor aucubin and the chemical form (glycoside or aglycone) in which these compounds are encountered by the fungus during growth. Our results suggest that iridoid glycosides in P. lanceolata can be used as defense against both herbivores and pathogens, but that their effects are highly specific with respect to the natural enemy species that is encountered. Received 11 April 2002; accepted 9 August 2002     

Interspecific transmission of Paederus endosymbionts: relationship to the genetic divergence among the bacteria associated with pederin biosynthesis

Summary. Endosymbiotic bacteria implicated in pederin production of Paederus (+)-females (Coleoptera: Staphylinidae) can be transmitted horizontally within and less frequently among the three species analyzed (P. melanurus, P. riparius, P. sabaeus). The 16S rDNA isolated from (+)-females reveals closely related bacterial sequences in the three species as well as in Paederus fuscipes and Paederidus ruficollis. This confirms the association of the undescribed endosymbiont and pederin biosynthesis in 5 of the 13 species that have been shown to contain the substance. In spite of the high sequence identities (> 99.5%), which suggest one species of endosymbiont, some of the heterospecific hosts were incompatible. This indicates adaptation and specific preferences of the endosymbiont for their natural host. Received 5 December 2001; accepted 11 March 2002.     

Pyrrolizidine alkaloid content in <Emphasis Type="Italic">Senecio</Emphasis>: ontogeny and developmental constraints

Summary. The ontogeny of pyrrolizidine alkaloid (PA) synthesis and constraints on defence level during the seedling stage were examined in the annual Senecio vulgaris and the monocarpic perennial Senecio jacobaea. In both species, PAs were actively synthesized from the onset of seedling growth so that juvenile stages did not go through an undefended stage. Roots are known to be the exclusive sites where PAs are produced. Root biomass was the single most important biomass parameter explaining variation in total PAs per seedling. All correlation coefficients between—relative growth rate and PA concentration were negative, but none was significant. However, a significant negative—correlation was found between shoot to root ratio and PA concentration in S. jacobaeaseedlings, suggesting a dilution effect of the PAs. Earlier studies have shown that the shoot to root ratio is positively correlated with relative growth—rate of established S. jacobaea plants. It is therefore suggested that young S. jacobaea plants with a high shoot to root ratio and hence a high growth capacity necessarily have lower PA defence levels than plants with a low shoot to root ratio. Received 10 July 2002; accepted 16 November 2002.     

Comparative sensitivity to and consumption of methyl eugenol in three <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis> (Diptera: Tephritidae) complex sibling species

Summary. Sensitivities to methyl eugenol of three sibling species in the Bactrocera dorsalis complex were compared. The degree of species sensitivity to methyl eugenol, i.e. B. dorsalis > B. papayae > B. carambolae (in decreasing order), was concomitant with the species age-related response to methyl eugenol as previously reported. The ability to consume methyl eugenol by the three sibling species showed similar trend - the average ME consumption per male was 0.70 ml for B. dorsalis, 0.58 ml B. papayae and 0.18 μl B. carambolae. Results obtained were discussed in relation to area-wide control of fruit fly. Received 21 April 2002; accepted 9 July 2002     

Uptake and metabolism of [<Superscript>14</Superscript>C]rinderine and [<Superscript>14</Superscript>C]retronecine in leaf-beetles of the genus <Emphasis Type="Italic"> Platyphora</Emphasis> and alkaloid accumulation in the exocrine defensive secretions

Summary. Sequestration and processing of pyrrolizidine alkaloids (PAs) by leaf beetles of the genus Platyphora were investigated. Tracer experiments with labeled alkaloids were performed with P. eucosma feeding on Koanophyllon panamense (Asteraceae, tribe Eupatorieae). P. eucosma catalyzes the same reactions previously demonstrated for P. boucardi specialized to Prestonia portobellensis (Apocynaceae): (i) epimerization of rinderine to intermedine; (ii) esterification of retronecine yielding insect-specific PAs; (iii) efficient transport of the PAs as free bases into the defensive secretions. P. bella feeding on Tournefortia cuspidata (Boraginaceae) shows the same sequestration behavior and ability to synthesize the specific retronecine esters. P. ligata, a species phylogenetically closely related to the PA adapted species and clustering in the same clade, but feeding on a host plant devoid of PAs, feeds easily on PA treated host-plant leaves, but does not sequester or metabolize PAs. P. kollari a species clustering outside the PA clade refused to feed on its food-plant leaves painted with PAs. The results are discussed in relation to host-plant selection of the PA adapted species and the role of PAs in chemical defense. Received 20 September 2002; accepted 18 November 2002.     

Lingual and biting responses to prey chemicals by ingestively naive scincid lizards: discrimination from control chemicals,time course,and effect of method of stimulus presentation

Summary. We tested responses to prey chemicals by lizard hatchlings of an oviparous species and neonates of a viviparous species, neither of which had never eaten. Both species responded more strongly to prey chemicals than to odorous and odorless control stimuli presented on cotton swabs. Although only a few species have been examined, all that have been tested have an innate capacity for prey chemical discrimination, suggesting that this innate response to prey chemicals is widespread among lizards that use the lingual-vomeronasal system to locate and identify prey. Innate prey chemical discrimination has the great advantage of permitting lizards lacking prior experience with food to respond appropriately to chemical cues associated with food. Both species discriminated prey chemicals from control substances at age three days, earlier than previously known. Our data hint that Mabuya macularia may be capable of discrimination on its day of birth, but further study is needed to determine the exact onset. A stronger tendency to attack swabs bearing prey chemicals by Scincella lateralis than by M. macularia may be explained by differences in defensiveness near an experimenter or by differences in the importance of visual prey cues for confirmation of chemical cues in the natural habitats of these species. In M. macularia responses to the control stimuli declined over days of testing, suggesting habituation, but responses to prey chemicals did not habituate by the third day of testing, which is interpreted as a possible adaptive response to permit location of food. In the standard method of stimulus presentation, a cotton swab bearing a chemical stimulus is placed anterior to a lizard's snout. We tested a new method in which the swab was placed in continuous contact with the lizard's anterior labial scales. The new method elicited significantly stronger responses from M. macularia. We discuss reasons for this finding and applications for the new method. Received 2 September 1999; accepted 15 December 1999     

Preferences of six leaf beetle species among qualitatively different leaf age classes of three Salicaceous host species

Summary. As Salicaceous plants produce new leaves for a prolonged period of time, they expose a wide range of differentially aged leaves to herbivores during the growing season. In this work, I show that young leaves of three Salicaceous species, Populus tremula L., Salix phylicifolia L. and S. pentandra L., contain more nitrogen than conspecific old leaves. In P. tremula and S. pentandra young leaves also contained more low-molecular weight secondary compounds, phenolic glucosides. Leaves of S. phylicifolia did not contain phenolic glucosides in detectable amounts. Furthermore, in P. tremula and S. pentandra young leaves contained less polymeric digestability-reducing phenolics, condensed tannins, than old leaves. In S. phylicifolia, higher concentrations of condensed tannins were found in young leaves. In laboratory feeding trials with six leaf beetle species, young leaves of the studied plants were invariably preferred in all tested herbivore × host species combinations. In particular, it is remarkable that three leaf beetle species with known different overall relationships to phenolic glucosides equally preferred more glucoside-containing young S. pentandra leaves over conspecific old ones. Four beetle species were found to prefer young leaves of S. phylicifolia despite the higher content of condensed tannins in young leaves. These results indicate that the general preference of leaf beetles for young leaves of Salicaceous plants probably does not primarily result from variable distribution of secondary compounds. Apparently, the preference for young leaves is fundamentally due to variation in leaf nutritive traits, such as nitrogen content. Received 9 February 2001.     

Antennal responses to floral scents in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae)

Summary. To better understand the biological role of floral scents for butterflies, electrophysiological responses to floral scents were investigated using combined gas chromatography and electroantennographic detection (GC-EAD). The antennal responses of three butterfly species, Aglais urticae L. (Nymphalidae), Inachis io L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae) to floral scent compounds from both natural and synthetic mixtures were examined. Floral scents were collected from the butterfly nectar plants Cirsium arvense (L.) (Asteraceae), and Buddleja davidii Franchet cv. (Loganicaeae) with dynamic head-space methods on Tenax-GR and eluted with pentane. These eluates, composed of natural floral scent blends, represent an array of compounds in their natural state. In the GC-EAD analyses eleven compounds were identified from C. arvense with the benzenoid compound phenylacetaldehyde in highest abundance. Seventeen compounds were identified from B. davidii with the irregular terpene oxoisophorone in highest abundance. Thirty-nine synthetic floral scent compounds were mixed in pentane, in equal amounts; about 35 ng were allowed to reach the antennae. The butterflies showed antennal responses to most of the floral scent compounds from both natural and synthetic blends except to the highly volatile monoterpene alkenes. Certain benzenoid compounds such as phenylacetaldehyde, monoterpenes such as linalool, and irregular terpenes such as oxoisophorone, were emitted in relatively large amounts from C. arvense and B. davidii, and elicited the strongest antennal responses. These compounds also elicited strong antennal responses when present in the synthetic scent blends. Thus, the butterflies seem to have many and /or sensitive antennal receptors for these compounds, which points to their biological importance. Moreover, these compounds are exclusively of floral scent origin. For B. davidii, which depends highly on butterflies for pollination, the exclusive floral scent compounds emitted in high abundance could be the result of an adaptive pressure to attract butterflies. Received 2 Septemter 2001; accepted 9 September 2002.     

Pheromone-based aggregation behaviour of the firebrat, <Emphasis Type="Italic">Thermobia domestica</Emphasis> (Packard) (Thysanura: Lepismatidae)

Summary. We tested the hypothesis that aggregation behaviour of the firebrat, Thermobia domestica (Packard) (Thysanura: Lepismatidae), an inhabitant of enclosed microhabitats, is mediated, at least in part, by a pheromone. Individual insects were released into the central chamber of a 3-chambered olfactometer and test stimuli were placed in lateral chambers. Paper discs previously exposed for 3 days to 10 female, male, or juvenile T. domestica were all preferred by female, male, or juvenile T. domestica over unexposed paper discs, indicating the presence of an aggregation/arrestment pheromone. In additional experiments, frass and scales from female T. domestica, tested singly and in combination, proved not to be the source of the pheromone. Physical contact was required for pheromone recognition, indicating that the pheromone arrests rather than attracts conspecifics. Arrestment by the long-tailed silverfish, Ctenolepisma longicaudata Escherich (Thysanura: Lepismatidae), but not by the common silverfish, Lepisma saccharina L. (Thysanura: Lepismatidae), to T. domestica exposed paper discs suggests closer phylogenetic relatedness between C. longicaudata and T. domestica, than between C. longicaudata and L. saccharina. Whether C. longicaudata or L. saccharina produce an aggregation signal, and whether T. domestica respond to this signal is unknown. Received 10 June 2002; accepted 30 September 2002.     

Foraging responses in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae) to floral scents

Summary. For butterflies to be efficient foragers, they need to be able to recognize rewarding flowers. Flower signals such as colours and scents assist this recognition process. For plant species to attract and keep butterflies as pollinators, species-specific floral signals are crucial. The aim of this study is to investigate foraging responses to floral scents in three temperate butterfly species, Inachis io L. (Nymphalidae), Aglais urticae L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae), in behavioural choice bioassays. The butterflies were allowed to choose bet-ween flower models varying in scent and colour (mauve or green). Flowers or vegetative parts from the plants Centaurea scabiosa L. (Asteraceae), Cirsium arvense (L.) (Asteraceae), Knautia arvensis (L.) (Dipsacaceae), Buddleja davidii Franchet (Loganicaeae), Origanum vulgareL. (Lamiaceae), Achillea millefolium L. (Asteraceae), and Philadelphus coronarius L. (Hydrangiaceae) were used as scent sources. All visits to the models — those that included probing and those that did not — were counted, as was the duration of these behaviours. Both flower-naive and flower-experienced (conditioned to sugar-water rewards, the colour mauve, and specific floral scents) butterflies were tested for their preference for floral versus vegetative scents, and to floral scent versus colour. The butterflies were also tested for their ability to switch floral scent preferences in response to rewards. Flower-naive butterflies demonstrated a preference for the floral scent of the butterfly-favourable plants C. arvense and K. arvensis over the floral scent of the non-favourable plants Achillea millefolium (Asteraceae), and Philadelphus coronarius cv. (Hydrangiaceae). Most of the butterflies that were conditioned to floral scents of either C. arvense, K. arvensis, or B. davidii readily switched theirfloral scent preferences to the one most recently associated with reward, thus demonstrating that floral scent constancy is a result from learning. These findings suggest that these butterflies use floral scent as an important cue signal to initially identify and subsequently recognize and distinguish among rewarding plants. Received 2 September 2001; accepted 9 September 2002.     

Predatory behavior and prey selection by army ants in a desert-grassland habitat

Summary Colonies of Neivamyrmex nigrescens conduct extensive nocturnal raids on other ants and termites in the desert-grassland of Arizona-New Mexico. We collected quantitative data on several aspects of raiding to pinpoint differences due to colony size and behavioral phase. In the nomadic phase, colonies began raiding at sunset and continued until dawn. Larger colonies covered more area, discovered more prey sites, and collected more booty than smaller colonies, but there were no systematic changes in raid intensity over the course of the nomadic phase. In the statary phase, raiding occurred less frequently and was less intense when it occurred; however, at the end of this phase, raiding was similar to nomadic phase raids in extent, duration, and booty captured. N. nigrescens preyed exclusively on termites and ants, and appeared to select certain species of Pheidole in preference to other ants. Pheidole was the most abundant genus, but was preyed upon twice as often as expected based on relative colony density. Pheidole attempted to avoid predation by fleeing or defending their nest, but rarely succeeded. Because they are about the same size as army ants and lack defensive chemicals, Pheidole made comparatively easy prey. N. nigrescens ignored or was repelled by other ants (Pogonomyrmex, Novomessor, Iridomyrmex, Myrmecocystus) during the early summer, when Pheidole was abundant; however, in late summer when Pheidole was less available, the army ants preyed upon Novomessor cockerelli. N. harrisi raided in close proximity to N. nigrescens, but preyed exclusively on Solenopsis xyloni. Selection of prey and partitioning of resources are now indicated in several army ant species; these processes have probably been important factors in the evolution of the ants' predatory behavior.     

Chemical camouflage by myrmecophilous beetles Zyras comes (Coleoptera: Staphylinidae) and Diaritiger fossulatus (Coleoptera: Pselaphidae) to be integrated into the nest of Lasius fuliginosus (Hymenoptera: Formicidae)

Summary. The myrmecophilous beetles, Zyras comes (Staphylinidae) and Diaritiger fossulatus (Pselaphidae) are guests of the black shining ant Lasius fuliginosus. Host worker ants never attacked these beetles, and often gave regurgitant to Z. comes following tactile communication with the beetle. By contrast, the workers from colonies without the myrmecophiles showed hostile responses towards Z. comes before tactile contact, but were not aware of D. fossulatus until contact. In L. fuliginosus, workers within a colony shared profiles, but the profiles differed among colonies. GC analyses showed that both Z. comes and D. fossulatus beetles had the same hydrocarbons as L. fuliginosus, and the profiles were more similar to those of the host colony workers than the foreign workers. Both Z. comes and D. fossulatus appear to imitate the hydrocarbon profile of their host workers, allowing integrating into the host nest. A Y-maze bioassay indicated that Z. comes can follow the trail pheromone of L. fuliginosus. This suggests that Z. comes may detect other chemical signals of L. fuliginosus to keep closer interactions with the workers. Received 22 June 2001; accepted 12 November 2001.     

Natterer’s bat (Myotis nattereri Kuhl, 1818) hawks for prey close to vegetation using echolocation signals of very broad bandwidth

We present a hitherto unknown prey perception strategy in bats: Myotis nattereri (Vespertilionidae, Chiroptera) is able to perceive prey by echolocation within a few centimeters of echo-cluttering vegetation, by using frequency-modulated search signals of very large bandwidth (up to 135 kHz). We describe the species’ search behavior and echolocation repertoire from the field and from experiments in a flight tent. In the field, bats varied signal parameters in relation to their distance from vegetation and usually flew close to vegetation. In the flight tent, M. nattereri detected and localized prey by echolocation alone as close as 5 cm from vegetation. Apparently, the bats were able to tolerate some overlap between prey and clutter echoes. Passive prey cues (vision, olfaction, prey-generated sounds) were not used in prey perception. The bats selected prey by size. The animals performed aerial catches and produced approach sequences typical for aerial hawking bats, but were able to do so within a few centimeters of the substrate. M. nattereri thus has access to silent, suspended prey very close to vegetation (e.g., spiders, and caterpillars on threads). Received: 29 September 1999 / Received in revised form: 12 February 2000 / Accepted: 12 February 2000     

Effects of hydrolysable tannins on a herbivorous insect: fate of individual tannins in insect digestive tract

Summary. We investigated the effects of four chemically characterised galloylglucoses (GGs, a subgroup of hydrolysable tannins) and their hydrolysis product, gallic acid (GA), on consumption and performance of larvae of the autumnal moth Epirrita autumnata. Larvae were fed with birch (Betula pubescens) leaves that had been painted individually with each of the compounds at two levels, 5 and 20 mg/g. In addition, we investigated the fates of the leaf-painted GGs and GA in the E. autumnata digestive tract by comparing phenolics in leaves consumed and in faeces. In general, GGs reduced leaf consumption by E. autumnata during the second and fourth instars, although there was high compound- and instar-specific variation. However, GGs did not affect the leaf consumption rates by the most voracious fifth instar larvae. This resulted in approximately the same loss of total biomass by the experimental tree, regardless of the nature and level of GGs enriched to its foliage. The characteristic fate of hydrolysable tannins, i.e. hydrolysis, was evidenced in the larval digestive tract for three of the four leaf-painted GGs. In addition to hydrolysis, the almost total absence of GGs in larval faeces was presumably related to the oxidation of GGs. The dose-dependent excretion percentage of ingested GA showed that it's faecal content should not be used, although it commonly is, to calculate the level of GG hydrolysis. Moreover, by comparing the non-uniform appearance of faecal tetragalloylglucoses, whether ingested as such or hydrolysed from pentagalloylglucose, we concluded that a major part of oxidation of GGs occurs before their hydrolysis in the digestive tract of E. autumnata. Criticism against the common use of tannic acid, a heterogeneous mixture of GA and GGs, in ecological studies is presented. Received 15 May 2002; accepted 16 July 2002     

Alkaloid tolerance in Manduca sexta and phylogenetically related sphingids (Lepidoptera: Sphingidae)

Summary. Nicotine tolerance is well known for Manduca sexta. It also occurs in several other sphingids of the subfamilies Macroglossinae and Sphinginae. Only members of the subfamily Smerinthinae appear to be more susceptible to nicotine intoxication. Phylogenetic trees have been reconstructed from mitochondrial 16S rDNA and nuclear DNA to map nicotine tolerance.?The nicotine binding site of both α-subunits of nicotinic acetylcholine receptors (nAChR) have been amplified and sequenced. No apparent amino acid substitution can be seen in the putative nicotine binding site of the α-subunits of nAChR from nicotine tolerant and nicotine sensitive sphingids. Thus, a simple target-site modification can be ruled out as a cause for nicotine tolerance. This finding agrees with feeding experiments: larvae of M. sexta and other sphingids of the Macroglossinae and Sphinginae not only tolerated nicotine, but also many other alkaloids that affect neuroreceptors other than acetylcholine receptors (nAChR, mAChR).?Only 10 to 20% of nicotine injected into larvae of nicotine-tolerant taxa could be recovered later as free nicotine, nicotine N-oxide or cotinine, i.e., 80 to 90% must have been converted to polar conjugates or degradation products which are not detectable with the methods applied. Usually more than 98% of the recoverable alkaloids were found in the faeces. Excretion reached a maximum 6 h after injection in tolerant taxa. Larvae of Manduca sexta, which were reared on a nicotine-rich diet, showed higher nicotine degradation and faster nicotine elimination than na?ve larvae. Application of the cytochrome P450 inhibitor SKF 525A (proadifen) reduced the formation of nicotine N-oxide and the rate of alkaloid degradation. Thus, an inducible detoxification mechanism, coupled with a rapid and inducible excretion, appear to be a strategy in Sphingidae that helps them to live on host plants rich in otherwise toxic secondary metabolites. Received 23 March 2001; accepted 4 August 2001.