Binary mixtures of feeding deterrents mitigate the decrease in feeding deterrent response to antifeedants following prolonged exposure in the cabbage looper, <Emphasis Type="Italic">Trichoplusia ni</Emphasis> (Lepidoptera: Noctuidae)

Summary. The effect of rearing larvae of Trichoplusia ni on individual feeding deterrents or on binary mixtures of deterrents on their subsequent gustatory sensitivity was measured in paired choice leaf disc bioassays. Our working hypothesis was that mixtures of antifeedants (pure allelochemicals) would mitigate decreased feeding deterrent response following prolonged exposure in this generalist herbivore. Neonate larvae were reared on cabbage leaves treated with individual feeding deterrents (digitoxin, thymol, toosendanin or xanthotoxin), or with binary mixtures of these until the third instar. Feeding deterrent responses to each antifeedant or mixture was then determined in a leaf disc choice bioassay. All of the mixtures produced additive deterrence when presented to naïve larvae. Larvae reared on individual antifeedants showed a significantly decreased feeding deterrent response (except to digitoxin), whereas larvae reared on binary mixtures of antifeedants did not show a decreased feeding deterrent response to any of them. Such mixtures were synergistic in terms of their feeding deterrence to experienced larvae. Our experiment supports the hypothesis (Jermy 1986) that mixtures of deterrents can prevent decreased feeding deterrent response following prolonged exposure, and provides one explanation for the multiplicity of chemical defenses found in many plants.     

Bark salicylates and condensed tannins reduce vole browsing amongst cultivated dark-leaved willows (<Emphasis Type="Italic">Salix myrsinifolia</Emphasis>)

Summary. Vole feeding amongst herbal willows that have a high concentration of salicylates in their bark and leaves, and may therefore be cultivated for use as raw material for herbal medicine was tested in the field and in laboratory conditions. Eight clones of dark-leaved willow (Salix myrsinifolia Salisb.) were cultivated for two years with six different methods combining three fertilisation levels (none, low and high), black plastic mulch applied for suppressing weed competition and unmulched control. Samples for the laboratory feeding trial were taken from the unfertilised plants during willow winter dormancy and twigs were fed to 16 voles as a multi-choice experiment. The bark area removed was calculated from image analysis of the material left by the voles. The diameter and the bark thickness of the twigs were measured. Concentrations of salicin, salicortin, HCH-salicortin, picein, triandrin, triandrin derivative, gallocatechin, (+)-catechin, luteolin-7-glucoside, hyperin, total condensed tannins and total nitrogen were measured from the twigs fed to voles in the laboratory. Browsing by a natural population of voles amongst winter-dormant willows was measured in the field. In the laboratory, voles browsed on 80% of the twigs and in the field voles browsed on 33% of the twigs. Vole feeding followed similar patterns in the field and in the laboratory experiment; feeding was clearly higher amongst the plants grown in unmulched control compared to those in plastic mulch. The same clones, 1, 2 and 6 were preferred in both experiments. Voles preferred thin twigs to thick ones. Feeding correlated negatively with concentrations of salicylates and tannins. As vole feeding seems to be highly affected by willow cultivation method and plant genotype, careful selection of cultivated clones and cultivation methods can enhance the reliability of herbal willow cultivation.     

The iridoid glycoside,catalpol, as a deterrent to the predatorCamponotus floridanus (Formicidae)

Summary We investigated the role of the iridoid glycoside, catalpol, as a deterrent to the predator,Camponotus floridanus. Four laboratory colonies of this ant were offered buckeye caterpillars (Junonia coenia: Nymphalidae) raised on diets with and without catalpol. The same colonies were offered sugar-water solutions containing varying concentrations of catalpol, in both no-choice and choice tests. Regardless of diet, buckeye caterpillars appeared to be morphologically protected from predation by the ants, possibly because of their large spines or tough cuticle. However, buckeyes raised on diets with catalpol had high concentrations of catalpol in their hemolymph; extracts of this high-catalpol hemolymph proved to be an effective deterrent to the ants. When starved ants were not given the choice of food items, they were more likely to consume sucrose solutions that contained 5 mg catalpol/ml or 10 mg catalpol/ml than they were to consume solutions with 20 mg catalpol/ml. When they were given a choice of sugar solution or a sugar solution containing catalpol, the ants avoided solutions with catalpol at any of these concentrations. Ant colony responses to catalpol in sucrose solutions varied considerably over time and among colonies.     

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.     

Male endogenous pheromonal component of Bactrocera carambolae (Diptera: Tephritidae) deterred gecko predation

Summary. Geckos consumed immature Bactrocera carambolae males readily, but their consumption was reduced significantly when offered sexually mature males. They showed a similar aversion for sexually mature males previously fed with methyl eugenol. When male flies at different ages were offered to starved geckos, geckos’ consumption showed an age-related anti-feeding effect, which was concomitant with the production trend of 6-oxo-1-nonanol, a major endogenous component synthesized in the male rectal gland. The production of 6-oxo-1-nonanol was also age-related and corresponded with sexual maturity as reported previously. Topical application of the authentic chemical of 6-oxo-1-nonanol onto the thorax of female fruit flies, at the amount normally detected in the rectal gland of a sexually mature male, had the same deterrent effect against geckos.     

A stingless bee (Melipona panamica) indicates food location without using a scent trail

The study of location specification in recruitment communication by bees has focused on two dimensions: direction and distance from the nest. Yet the third dimension, height above ground, may be significant in the tall and dense forest habitats of stingless bees. Foragers of the stingless bee Scaptotrigona postica recruit to a specific three-dimensional location by laying a scent trail. Stingless bees in the genus Melipona are thought to have a more sophisticated recruitment system that communicates distance through sounds inside the nest and direction through pointing zig-zag flights outside the nest. However, prior research on Melipona has not examined height communication or even established that foragers can recruit newcomers to a specific location. We used identical paired feeders to investigate recruitment to food in M panamica on Barro Colorado Island, Panama. We trained foragers from an observation hive to one feeder and monitored both feeders for the subsequent arrival of newcomers. We changed the relative positions of the feeders to test for correct direction, distance, and canopy-level communication. A 40-m canopy tower located inside the forest enabled us to examine canopy-level communication. We found that M. panamica foragers can recruit to a specific (1) direction, (2) distance, and (3) canopy level. To test the possibility that foragers accomplish this by means of a scent trail, we placed the colony on one shore of a small cove and trained bees over 116 m of open water to a feeder located on the opposite shore. We also placed a second feeder on this shore, equidistant from the colony but 20 m from the first feeder. Significantly more newcomers consistently arrived at the feeder visited by the foragers. Thus foragers evidently do not need a scent trail to communicate direction. Inside the nest, a forager produces pulsed sounds while visibly vibrating her wings after returning from a good food source. She is attended by other bees who cluster and hold their antennae around her, following her as she rapidly spins clockwise and counterclockwise. Locational information may be encoded in this behavior. However, foragers may also directly lead newcomers to the food source. Further experiments are planned to test for such piloting and other communication mechanisms.     

Quantitative composition of the defensive secretion ofBledius species (Coleoptera: Staphylinidae: Oxytelinae) is adapted to naturally occurring predators

Summary The adaptation of defensive secretions to their target organisms was examined for the abdominal gland secretions ofBledius furcatus, B. spectabilis andB. arenarius. Therefore the target organisms of the secretion of theseBledius species (i.e. their predators) had to be identified. At the collection sites examined these were the earwigLabidura riparia, the antCataglyphis bicolor, the flyLispe candicans, different carabids of the generaPogonus, Dichirotrichus, Dyschirius, Bembidion andCalathus and the wading birdsHaematopus ostralegus andCalidris alba. The secretion of the abdominal glands contains the toxin ptoluquinone dissolved in either-dodecalactone and 1-undecene (B. furcatus andB. spectabilis) or in octanoic acid and octyloctanoate (B. arenarius). The ratio of these solvents is species-specific. Application experiments using some of the natural insect predators (L. riparia, C. bicolor, Pogonus, Di. gustavii, Dyschirius) revealed that these solvent ratios provided a more effective deterrent than other possible ratios. Thus by combining the solvents in certain ratios, the capability of cuticular penetration and therefore the effectiveness of the defensive secretions are adapted to their natural targets.     

Scent mediation of intraspecific,interspecific, and intergeneric agonistic behavior among sympatric species of voles (Microtinae)

Summary The hypothesis that the posterolateral scent glands of male microtine rodents (Muridae) direct the attacks of other males was tested.Montane voles (Microtus montanus) and sagebrush voles (Lagurus curtatus) were established as residents in terraria, and conspecific and heterospecific males, from which posterolateral scent glands had been unilaterally removed, were introduced. More attacks by residents were aimed at the glandular sides than at the glandless sides. Male M. montanus also attacked the unilateral hip gland of longtail voles (M. longicaudus), the growth of which had been induced by exogenous hormone inasmuch as this species does not normally possess these glands. Resident male M. longicaudus did not similarly attack the induced glands of conspecific males.The findings are related to demographic patterns, mterspecific relationships, and the evolutionary development of posterolateral scent glands among microtine rodents.     

Toxicity of the venom in three neotropical Crematogaster ants (Formicidae: Myrmicinae)

Bioassays were performed to investigate and compare the toxic properties of the contact venoms of three Neotropical Crematogaster species (C. sp. prox. abstinens, C. distans and C. brevispinosa rochai), whose main venom compounds are chemically different (long chain derivatives linked to an aldehyde or a primary acetate, and furanocembrenoid diterpenes, respectively). Different quantities of venom were topically applied on the bodies of three target insect species (workers of the termite Nasutitermes ephratae, workers of the ant Solenopsis sp., “media” and “major” workers of the ant Cephalotes pusillus). The toxicity of the venom greatly varied according to the Crematogaster species, the venom of C. b. rochai showing the highest toxic activity towards all target species. The sensitivity of the target species also greatly varied, the workers of N. ephratae showing a great sensitivity to the venoms, while the heavily armoured workers of the ant C. pusillus, particularly “major” ones, were resistant. Cuticle thickness was found to be a key factor in the resistance to the toxic effects of Crematogaster venom. Sensitivity of the target workers was also directly related to the amount of venom applied to their cuticle. Workers of C. distans were resistant to topical application of their own venom, while C. sp. prox. abstinens and C. b. rochai were not. In all cases, the venoms were slow acting, as several hours to one or even several days were needed to obtain a significant mortality. The main biological activities of the Crematogaster venom could be related to more immediate repellent properties, as recently shown with the European species C. scutellaris.     

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.     

Leaf surface compound fromBrassica oleracea (Cruciferae) induces oviposition byPieris brassicae (Lepidoptera: Pieridae)

Summary Chemicals present on the surface of cabbage (Brassica oleracea L.) leaves were extracted by dipping these leaves for 3 s in dichloromethane followed by a 3 s dip in methanol. When offered in dual choice bioassays using green paper cards as a substrate, the methanol extract stimulated oviposition activity byPieris brassicae L. (Lepidoptera: Pieridae) females. The oviposition stimulant was isolated using medium pressure liquid chromatography, reversed-phase HPLC, ion-pair HPLC and ion exchange chromatography. Using¹H-NMR spectroscopy, the stimulant could be identified as glucobrassicin (3-indolyl-methyl-glucosinolate). When pure glucobrassicin was offered at a dose identical to that in the crude methanol extract, butterflies did not discriminate between these two substrates in a dual choice test. It is argued that a high sensitivity for indole glucosinolates as host recognition factors may confer an adaptive value for these specialist crucifer feeders. The nutritional significance of their precursor tryptophan and the non-volatile nature of the aglycones formed upon enzymic hydrolysis in damaged tissues are proposed as properties of indole glucosinolates that contribute to this possible adaptive advantage.     

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.     

Attraction,deterrence or intoxication of bees (Apis mellifera) by plant allelochemicals

The influence of 63 dietary allelochemicals (alkaloids, terpenes, glycosides,etc.) on the feeding behaviour of bees (Apis mellifera) was tested in terms of deterrency and attraction. For 39 compounds a deterrent (mostly alkaloids, coumarins and saponins) and for 3 compounds an attractive response (mostly terpenes) was obtained in choice tests, which allowed the calculation of respective ED₅₀-values. Under no-choice conditions, 17 out of 29 allelochemicals caused mortality at concentrations between 0.003 and 0.6%. Especially toxic were alkaloids, saponins, cardiac glycosides and cyanogenic glycosides. These data show that bees which are confronted with plant allelochemicals in nectar and pollen, are not especially adapted (i.e. insensitive) to the plants' defence chemistry. GLC and GLS-MS data are given on the alkaloid composition of nectar and pollen ofBrugmansia aurea, Atropa belladonna andLupinus polyphyllus.     

Local and systemic induced responses to cabbage root fly larvae (Delia radicum) in Brassica nigra and B. oleracea

Summary. Feeding by belowground herbivores may induce systemic changes in shoot defence levels that affect the performance of above ground herbivores and higher trophic levels. In this paper two wild Brassica species, B. nigra and B. oleracea were experimentally infested with 10 larvae of the cabbage root fly, Delia radicum. Plant dry masses and glucosinolate levels in shoots, main roots, and fine roots were determined at 3, 7, 12 and 14 days after infestation and compared to those of control plants. The systemic response in the leaves differed between plant species. In B. nigra shoot glucosinolate levels in D. radicum infested plants steadily increased with time until they were almost twice those of controls 14 days after infestation. B. oleracea plants infested with D. radicum did not show significant changes in shoot glucosinolate levels within 14 days, which may be due to the unexpected poorer performance of D. radicum on this species. Both plant species showed a local increase in indole glucosinolates in the main roots, which are the preferred feeding site of D. radicum larvae. B. oleracea plants however showed a stronger (1.9 – 4.7 times) increase in indole glucosinolate levels than B. nigra (1.5 – 2.6 times). The increase in indole glucosinolates in B. nigra main roots, was counterbalanced by a significant decrease in aromatic glucosinolate levels. These differences in local responses to D. radicum feeding between the two species may have contributed to the slower growth rates of the larvae on B. oleracea. D. radicum feeding did not result in altered glucosinolate levels in the fine roots in either plant species. The differences in glucosinolate induction patterns between the summer annual B. nigra and the perennial B. oleracea are discussed in the light of their different life histories.     

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.     

Trichinella spiralis infection in voles alters female odor preference but not partner preference

Females may choose mates based on secondary sex traits that reflect disease resistance. Accordingly, females should be able to distinguish between unparasitized and parasitized males, and should prefer to mate with unparasitized individuals. Mate and odor preferences for uninfected males or males infected with the nematode, Trichinella spiralis, were examined among prairie voles (Microtus ochrogaster) and meadow voles (M. pennsylvanicus). In a 15-min odor preference test, only female meadow voles distinguished between bedding from parasitized and unparasitized conspecific males, and preferred to spend time with bedding from unparasitized males. Although T. spiralis infection influenced odor preference in female meadow voles, there was no effect of infection status on mate preference among either species. Testosterone and corticosterone concentrations were not different between parasitized and unparasitized males. However, among prairie voles, males that spent an increased amount of time with females during the mate preference test had elevated testosterone concentrations. Taken together, these data suggest that (1) female meadow voles can discriminate between unparasitized and parasitized males, (2) the effects of infection on steroid hormone concentrations may be masked by the effects of social interactions, and (3) parasites may represent a selective constraint on partner preference in voles; however, the life cycle of parasites may influence female preference and should be considered in studies of female preference. Received: 23 April 1998 / Accepted after revision: 25 October 1998     

Net energetic advantage drives honey bees (Apis mellifera L) to nectar larceny in Vaccinium ashei Reade

Carpenter bees (Xylocopa spp.) act as primary nectar thieves in rabbiteye blueberry (Vaccinium ashei Reade), piercing corollas laterally to imbibe nectar at basal nectaries. Honey bees (Apis mellifera L) learn to visit these perforations and thus become secondary nectar thieves. We tested the hypothesis that honey bees make this behavioral switch in response to an energetic advantage realized by nectar-robbing flower visits. Nectar volume and sugar quantity were higher in intact than perforated flowers, but bees (robbers) visiting perforated flowers were able to extract a higher percentage of available nectar and sugar so that absolute amount of sugar (mg) removed by one bee visit is the same for each flower type. However, because perforated flowers facilitate higher rates of bee flower visitation and the same or higher rates of nectar ingestion, they are rendered more profitable than intact flowers in temporal terms. Accordingly, net energy (J) gain per second flower handling time was higher for robbers on most days sampled. We conclude that the majority evidence indicates an energetic advantage for honey bees that engage in secondary nectar thievery in V. ashei.Communicated by R. Page     

Behaviour and choice of refuge by voles under predation risk

Animals often show a strong antipredatory response when they are exposed to their most deadly predator. In northern vole populations, the least weasel, Mustela nivalis nivalis, is probably the most important predator of voles. Because of its small size and slender body shape, the least weasel is capable of hunting voles in their burrows. However, small voles can potentially escape weasel predation by selecting holes smaller than those weasels can enter. We studied the choice of nest holes and refuges by two species of voles under simulated predation risk. In a laboratory experiment, voles were provided with four nest boxes as refuges, with individually adjusted entrance sizes. When exposed to the odour of a weasel, voles did not choose the smallest opening; they rather seemed to trade full protection for easy and immediate access by choosing the nest box with an intermediate entrance size. When outside the nest at the time when a weasel entered the arena, voles avoided the refuges with the smallest holes. In addition to using refuges on ground level, voles climbed on top of the boxes as an escape reaction, as well as exhibiting a variety of behavioural responses, such as fast running, freezing and sneaking.Communicated by P.A. Bednekoff     

The antifungal activity of the leaf gland volatile oil of sweet gale (Myrica gale) (Myricaceae)

Summary The antifungal activity of volatile oil obtained from the leaves of sweet gale (Myrica gale) has been investigated. Fungi, isolated from the leaves ofM. gale, were grown in liquid medium containing the oil and its effect on biomass production was gauged. The oil inhibited growth of all species of fungi to greater or lesser extent. Sesquiterpenes were more active than monoterpenes, and among the sesquiterpenes germacrone was marginally more active than -elemenone. These results support the proposition that the volatile oil ofM. gale has a role in resistance to fungal pathogens.     

Susceptibility of Eucalyptus globulus to Mnesampela privata defoliation in relation to a specific foliar wax compound

Summary. Increased abundance of several aliphatic benzyl and phenylethyl alkanoates were previously associated with reduced defoliation of Eucalyptus globulus Labill. by the autumn gum moth (Mnesampela privata Guenée) in two clonally replicated F₂ families. We examine the robustness of this association by comparing the abundance of the implicated foliar wax compounds with defoliation levels in three plantation sites and a genetics trial in northern Tasmania. At all locations, the aliphatic alkanoate, benzyl n-tetracosanoate (C₂₄), was found in significantly higher abundance in the foliar wax of E. globulus trees that had displayed low M. privata damage (designated resistant) compared to trees that had displayed high damage (designated susceptible). Further, when sites were combined, benzyl n-tetracosanoate (C₂₄) accounted for 16 % of variation in field defoliation. Field defoliation was also positively correlated to M. privata oviposition both in the field and on foliage sprigs in a cage. In the cage bioassay, oviposition accounted for 20 % of variation in field defoliation making it a better predictor of tree defoliation in the field than either field oviposition, which accounted for 9 % of variation, or screening foliage for the abundance of benzyl n-tetracosanoate (C₂₄). Despite both benzyl n-tetracosanoate (C₂₄) and oviposition being related to field defoliation, there was no statistically significant relationship between the abundance of benzyl n-tetracosanoate (C₂₄) and oviposition in the field or cage bioassay. Further work is therefore required to determine if increased levels of this compound in the foliar wax operates as a direct deterrent to M. privataoviposition.