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.     

Biochemical transformation of birch leaf phenolics in larvae of six species of sawflies

Summary. We investigated the biochemical transformation of individual phenolic compounds of mountain birch leaves in larvae of six birch-feeding sawfly species: Amauronematus amplus, Pristiphora alpestris, Nematus brevivalvis, Priophorus pallipes, Arge sp. and Nematus viridis by comparing the phenolic residues in larval faeces to those of their leaf diet. Partial hydrolysis of hydrolysable tannins, isomerisation of chlorogenic acid and glycosylation of flavonoid aglycones were observed in all studied species. Moreover, we found considerable among-species variation in the composition of phenolic compounds in larval faeces. In addition to foliar phenolics, seventeen non-foliar phenolic metabolites, including eight phenolic acids and nine flavonoid glycosides were detected from the faeces. Of the non-foliar phenolic acids, four were egested species-specifically and only two by all six sawfly species. We also detected differences in the ratios of chlorogenic acid isomers in the faeces of different species, which can indicate different physiological conditions in the guts of studied larvae. In addition to the qualitative differences, quantitative differences were detected in the egestion of chlorogenic acids, possible o-quinone precursors in the larvae. Detected differences, either qualitative or quantitative, could not be explained by seasonal changes in the content of compounds in the leaf diet.     

Cuticular volatiles,attractivity of worker larvae and invasion of brood cells by Varroa mites. A comparison of Africanized and European honey bees

Summary. Africanized honey bees (AHBs) of Brazil and Mexico have proven to be tolerant to Varroa destructor mites. In contrast, European honey bees (EHBs: Apis mellifera carnica) at the same tropical study site are highly intolerant to these ectoparasites. A lower attractiveness of Varroa-tolerant AHB larvae has been hypothesised to be an important trait in reducing the susceptibitlity of AHBs to these mites. Thus, selection for EHB brood that is less attractive to mites is thought to be one possibility for limiting mite population growth and thus increase the tolerance of EHBs to the mite.?In Ribeir?o Preto, Brazil, European A. m. carnica bees and AHBs were tested with respect to their rate of brood infestation and brood attractiveness to Varroa mites. For the comparison of brood infestation rates, we introduced combs with pieces of EHB and AHB brood into honey bee colonies (18 repetitions). The relative infestation rate of EHB brood was significantly higher compared to AHB brood.?The preference behaviour of single Varroa mites was tested in a laboratory bioassay where either living host stages were offered or host extracts were presented on dummies. By these tests we could confirm the preference of Varroa females for certain developmental host stages and for their corresponding extracts. In contrast to the within-colony results, Varroa mites in the laboratory bioassay showed a slight preference for AHB compared to EHB larvae.?The gas chromatographic analysis revealed differences in the chemical spectrum of extracts obtained from different larvae. In accord with the results of the bioassays, we could detect stage-specific odour differences in larval cuticular compounds, including methyl esters and hydrocarbons that have been described as kairomones. None of these substances, however, revealed significant race-specific differences. Therefore, the quantity and composition of certain cuticular compounds seem to be responsible only for the recognition of a suitable host stage by Varroa females. The different infestation rates in the colonies, however, seem to be caused neither by race-specific differences in attractiveness of bee larvae nor by an extended attractive period of EHB larvae: both AHB and EHB larvae become attractive approximately 21 h before capping of the brood cell, and thus have the same window of time when they can be parasitised.?Therefore differential Varroa-infestation rates are not related to larval attraction but probably are determined by other race-specific and colony-related factors. Received 11 June 2001; accepted 19 November 2001.     

Effects of three years’ increase in density of the geometrid Epirrita autumnata on the change in metabolome of mountain birch trees (Betula pubescens ssp. czerepanovii)

The principal objective of the study was to characterize effects of increasing density of the autumnal moth, Epirrita autumnata (Lepidoptera, Geometridae), for 3 successive years on the change in metabolome of mountain birch trees (Betula pubescens ssp. czerepanovii). During the three study years (2000–2002), the larval density varied from low to very high (outbreak) density. Samples of leaves were collected from the same 12 trees at the same phenological stage of the trees each year. The leaves were collected from undamaged trees in the first year, from trees slightly damaged by larvae in the second year, and from trees heavily damaged by larvae in the third year. Metabolome analysis showed that the increase in density of E. autumnata larvae and degree of damage of birch trees caused multiple biochemical changes in the leaves, including increased concentrations of phenolic compounds (proanthocyanidins and hydrolysable tannins) and reduced concentrations of nutritive metabolites (monosaccharides, amino acids and some organic acids). These changes reduced the quality of leaves as food for larvae and, probably, were associated with induced chemical resistance of the birches to herbivorous insects. Additionally, the concentration of α-tocopherol was significantly higher in both slightly and heavily damaged trees. Mechanisms of changes in the metabolism of phenolic compounds and carbohydrates, and the role of α-tocopherol in their regulation are discussed.     

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.     

Seaweed secondary metabolites as antifoulants: effects of Dictyota spp. diterpenes on survivorship, settlement, and development of marine invertebrate larvae

Summary. A recent investigation showed that the brown seaweed Dictyota menstrualis was unfouled relative to co-occurring seaweeds, and that larvae of fouling invertebrates avoided settling on D. menstrualis due to chemicals on its surface. The secondary metabolites dictyol E and pachydictyol A are among the compounds found on this alga's surface. In the present study, we tested the effects of specific diterpenes from Dictyota on the survivorship, growth, and development of invertebrate larvae and developing juveniles that could foul seaweeds. Exposure to dictyol E, dictyol B acetate, pachydictyol A, and dictyodial from Dictyota menstrualis and D. ciliolata caused significant larval mortality, abnormal development, and reduce growth rates for three species of co-occurring invertebrates when their larvae were forced into contact with these metabolites. Larvae were damaged at metabolite concentrations as low as 5% of maximum possible surface concentrations of these compounds for the populations of Dictyota we studied. The negative effects of these secondary metabolites on potential foulers, in conjunction with data demonstrating larval avoidance of dictyol-covered surfaces, suggest that these compounds could function as chemical defenses against fouling, and could select for larvae that avoid hosts producing these metabolites. Received 25 May 1998; accepted 22 June 1998.     

Temporal and spatial variation in mountain birch foliar enzyme activities during the larval period of <Emphasis Type="Italic">Epirrita autumnata</Emphasis>

Summary. Mountain birches are deciduous trees consisting of several clonal and partly autonomous ramets. Autumnal moth, Epirrita autumnata, is an extremely harmful pest of mountain birches that during outbreak years may cause the death of trees over large areas. During the larval development of E. autumnata, leaf characters and nutritional suitability vary considerably both temporally and spatially among and within mountain birches. Compared to variation between trees, however, the extent of within-tree variation is known for only a limited number of potentially defensive traits. Plant oxidases, polyphenoloxidases (PPOs) and peroxidases (PODs) may play an important part in dictating the suitability of a tree for plant-eating animals, including E. autumnata. In this study, we observed changes in the activities of oxidative and antioxidative enzymes along leaf development during the larval period of E. autumnata. We also estimated the relative amounts of spatial variation among and within trees. Our results show that POD activities were remarkably high during the rapid elongation growth phase of leaves but decreased with leaf maturation. Thus PODs are suggested to take part in leaf elongation growth. Time and within tree variation accounted for the most of variation in POD activities whereas there was no among tree variation. In contrast, the activities of antioxidative CATs, which act as antagonists to PODs, were low in young leaves and increased towards leaf maturation, reflecting an increase in the oxidative status of source leaves. Within tree variation accounted for the most of the variation in CAT activities. The absolute PPO activities decreased along leaf growth due to the dilution effect whereas the specific activity of PPOs, which has been shown to be defensive against E. autumnata larvae, did not vary temporally that might reflect the importance of these enzymes in the defence of birches. Acidic PPOs showed marked within- and among-tree variation, which may impact herbivores performance on certain genotypes and increase larval mobility within the tree canopy.     

Aristolochic acid content of South-East Asian troidine swallowtails (Lepidoptera: Papilionidae) and of Aristolochia plant species (Aristolochiaceae)

Summary. Analysis of South-East Asian troidine swallowtails revealed high variability in the content of aristolochic acids among individuals. The presence or absence of these compounds depends on the Aristolochia species available as food plant for the larvae. Only one plant species (Aristolochia philippinensis) contained a high concentration of aristolochic acids, while other species from various localities contained none or only marginal amounts. Whether aristolochic acids have a distinct function in chemical defense of these swallowtails is still an open question. Received 11 December 2000; accepted 4 August 2001.     

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.     

Ingestion of bivalve larvae by<Emphasis Type="Italic"> Mytilus edulis</Emphasis>: experimental and field demonstrations of larviphagy in farmed blue mussels

Ingestion of bivalve larvae by Mytilus edulis was investigated. Laboratory experiments revealed that ~ 90% of bivalve larvae offered to mussels was ingested and apparently fully digested. The shell of the bivalve larvae offered no protection against digestive processes, resulting in high larval mortality once inside the stomach. Stomach content analysis (September 2001–January 2003) showed that bivalve larvae were ingested by farmed mussels year-round, with the exception of March 2002. Numbers of ingested larvae were highest in October 2001 and May 2002, which coincides with known spawning times of farmed mussels in Ireland. Mussels ingested a large size-range of bivalve larvae, suggesting that all stages of the bivalve life cycle are vulnerable to predation. It is suggested that adult bivalves routinely filter larvae from the surrounding water and that, given the high biomass of mussels present in mussel farms, filtration by adult bivalves significantly reduces numbers of bivalve larvae in nearby waters.Communicated by J.P. Thorpe, Port Erin     

Transformation of soft coral (Coelenterata: Octocorallia) terpenes by Ovula ovum (Mollusca: Prosobranchia)

The faecal pellets from specimens of the prosobranch mollusc Ovula ovum found feeding on the soft coral Sarcophyton sp. at Eclipse Island, Palm Island Group (18°46S; 146°33E) in November 1980 were analysed. The only terpene present in the faeces, 7,8-deoxysarcophytoxide, differed from the major constituent of the soft coral, sarcophytoxide, suggesting that the latter had been transformed into the former within the cowrie. This transformation is not trivial, and could not be produced simply by acid catalysis. Subsequent analysis of tissues dissected from different regions of O. ovum indicates that the transformation is probably effected by enzymes in the digestive diverticula stomach region of the prosobranch. The transformed compound is significantly less toxic to the mosquito fish Gambusia affinis Baird and Girard than the ingested compound.     

Effects on larval crabs of exposure to algal toxin via ingestion of heterotrophic prey

We tested whether ingesting toxic algae by heterotrophic prey affected their nutritional value to crab larval predators, using toxic algal strains that are either ingested directly by larval crabs or rejected by them. Ingestion of toxic strains of the dinoflagellates Alexandrium andersoni and A. fundyense by the rotifer Brachionus plicatilis was confirmed. Rotifers having ingested either algal type for five days were fed to freshly hatched larvae of three crab species, with larval survival and stage durations determined. For both algal/rotifer treatments in all three crab species, larvae fed algae directly died during the first zoeal stage, while those fed rotifers that had been fed either algal strain survived to the experiment’s end (zoeal stage 3). Survival was lower, and stage duration longer, for larvae fed rotifers cultured on toxic algae when compared to those fed non-toxic algae. The role of toxic algae in the planktonic food web may be influenced by its direct or indirect ingestion by larval crabs.     

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     

Differential responses in American (<Emphasis Type="Italic">Castanea dentata</Emphasis> Marshall) and Chinese (<Emphasis Type="Italic">C. mollissima</Emphasis> Blume) chestnut (Falales: Fagaceae) to foliar application of jasmonic acid

Summary.  Tannins are plant defense compounds that exhibit antibiotic (e.g. toxic) and antixenotic (e.g. repellent) effects against an array of plant pests. They are broadly divided into two major groups, hydrolysable tannins (gallotannins and ellagitannins) and proanthocyanidins, each with an undetermined number of compounds. We investigated constitutive levels of hydrolysable tannins and proanthocyanidins in the leaves and stems of American (Castanea dentata Marshall) (Fagales: Fagaceae) and Chinese (C. mollissima Blume) chestnut. American chestnut contained more proanthocyanidins in leaves and stems than Chinese chestnut, but Chinese chestnut contained more foliar hydrolysable tannins. Regardless of these differences, gypsy moth (Lymantria dispar L, Lepidoptera: Lymantriidae) performance did not differ when fed American and Chinese chestnuts. We also investigated the effects of jasmonic acid (JA) treatment on differential tannin induction in American and Chinese chestnut leaves and stems. JA treatment increased proanthocyanidins in American chestnut stems and hydrolysable tannins in both tissue types of American chestnut, but did not influence tannin concentrations in Chinese chestnut leaves or stems. HPLC-ESI-MS analysis of pooled samples suggested that hydrolysable tannins in each tissue were qualitatively comprised primarily of ellagitannins, and JA generally increased the number of hydrolysable tannins that could be detected by ESI-MS. Third, we investigated the performance of gypsy moths on JA treated and untreated American and Chinese chestnut. Caterpillar relative growth was not influenced by JA treatment on Chinese chestnut, but decreased in response to JA application on American chestnut. Our results indicate that JA-dependent defenses differ between these chestnut species. This study improves our understanding of ecologically important differences in tannin induction and herbivore susceptibility in Castanea, and has important implications in efforts for American chestnut restoration and commercial chestnut production.     

Resistance of the generalist moth Trichoplusia ni (Noctuidae) to a novel chemical defense in the invasive plant Conium maculatum

Summary. Conium maculatum is an apiaceous species native to Eurasia that is highly toxic to vertebrates due to the presence of piperidine alkaloids, including coniine and γ-coniceine. More than 200 years after invading the United States this species remains mostly free from generalist insect herbivores. The presence of novel chemical defenses in the introduced range could provide invasive species with a competitive advantage relative to native plants. The cabbage looper (Trichoplusia ni) is a generalist lepidopteran found throughout the US that occasionally feeds on C. maculatum. We evaluated the toxicity of piperidine alkaloids to T. ni and determined putative resistance mechanisms, both behavioral and physiological, that allows this insect to develop successfully on C. maculatum foliage. T. ni larvae raised on diets enriched with coniine and γ-coniceine showed a decrease in consumption and longer development time, but no effects on growth were found at any alkaloid concentration. In a diet choice experiment T. ni larvae showed no avoidance of alkaloid-enriched diets, suggesting that the deterrence produced by alkaloids was related to a post-ingestive metabolic response. The ability of T. ni to consume diets high in alkaloid content could be due to at least three different mechanisms: 1) a decreased consumption rate, 2) efficient excretion of at least 1/3 of ingested alkaloids unmetabolized in frass, and 3) partial detoxification of alkaloids by cytochrome P450 s, as shown by the decreased larval growth in the presence of piperonyl butoxide, a P450 inhibitor. Even though T. ni tolerates C. maculatum alkaloids, the use of this species as a host plant could be ecologically disadvantageous due to prolonged larval growth and thus increased exposure to predators. Novel plant secondary compounds do not guarantee increased resistance to generalist herbivores.     

Communication ecology of webbing clothes moth: 4. Identification of male- and female-produced pheromones

Summary. We investigated the hypothesis that aggregation signals produced by male webbing clothes moths (WCM), Tineola bisselliella (Hum.) (Lepidoptera: Tineidae), and close-range male attractant signals produced by females have a pheromonal basis, at least in part. Gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of bioactive methanolic extracts of male WCM disclosed three candidate pheromone components: hexadecanoic acid methyl ester (16:Ester), (Z)-9-hexadecenoic acid methyl ester (Z9—16:Ester), and octadecanoic acid methyl ester (18:Ester). In bioassay experiments in a large Plexiglas™ arena, a blend of synthetic 16:Ester plus Z9—16:Ester was attractive to male and virgin (but not mated) female WCM; the 18:Ester was inactive. GC-EAD analyses of pheromone gland extracts from female WCM revealed (E,Z)-2,13-octadecadienal (E2Z13—18:Ald) and (E,Z)-2,13-octadecadienol (E2Z13—18:OH) as candidate sex pheromone components. In arena bioassay experiments, 1—5 female equivalents of synthetic E2Z13—18:Ald (0.2 ng) and E2Z13—18:OH (0.1 ng) were more attractive to male WCM than were two virgin female WCM. We anticipate that the combination of aggregation and sex pheromones, male-produced sonic aggregation signals, and habitat-derived semiochemicals will be highly effective in attracting male and female WCM to commercial traps. Received 12 January 2001; accepted 8 June 2001.     

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.     

Characteristics of feeding on dinoflagellates by newly hatched larval crabs

The zoeal larvae of brachyuran crabs must feed soon after hatching on a diet that includes large micro- and mesozooplankton in order to satisfy nutritional requirements. However, newly hatched larvae have been shown to ingest a variety of dinoflagellates, perhaps using microbial carbon sources to sustain them until they encounter more favored prey. Ingestion of dinoflagellates by larval crabs has been documented previously under conditions in which the larvae were exposed to algae provided in monoculture or in defined mixtures of cells. We report here on experiments conducted on the hatching stage of five crab species to determine if ingestion of dinoflagellates occurred when they were provided in combination with Artemia sp. nauplii or after a period of feeding on mesozooplankton. Quantitative measurements of chl a in the larval guts provided evidence of ingestion of algal cells. Active ingestion of the dinoflagellate Prorocentrum micans at specified intervals during an extended feeding period was determined on larvae of two crab species using fluorescently labeled cells provided for brief periods at prescribed time intervals. Stage 1 larvae of four of the five crab species ingested dinoflagellates when they were provided in combination with nauplii and larvae of all five species ingested cells after feeding solely on nauplii for 24 h. Ingestion of algal cells was first evident in the larval guts after 6 h of feeding at both low (200 cell ml⁻¹) and high (1,000 cells ml⁻¹) prey densities. Higher prey densities resulted in higher gut chl a. Larvae continuously exposed to dinoflagellates actively ingested cells at every 3 h interval tested over a 36 h period. Results confirm previous studies that larvae will ingest dinoflagellates even when they are encountered in a mixed prey field or when having previously fed. Ingestion of cells may occur on a continual basis over time.     

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.     

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.