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.     

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

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

Tropane and pyrrolizidine alkaloids in the ithomiinesPlacidula euryanassa andMiraleria cymothoe (Lepidoptera: Nymphalidae)

Summary Larvae of the ithomiine butterflyPlacidula euryanassa sequester tropane alkaloids (TAs) from the host plantBrugmansia suaveolens and pass them through the pupae to freshly emerged adults. Wild caught adults also show in their tissues, variable amounts of pyrrolidizine alkaloids (PAs), probably sequestered from variable plant sources and subject to dynamics of incorporation, accumulation and utilization of PAs by ithomiine butterflies. The ratio TAs/PAs is also variable between different populations.Miraleria cymothoe, another ithomiine that feeds onB. suaveolens as larvae, does not sequester TAs from the host-plant, but sequesters PAs from plant sources visited by the adult butterflies. The main alkaloid found in both butterflies is lycopsamine, which also is the principal PA found in all genera of Ithomiinae.     

Phenological fate of plant-acquired pyrrolizidine alkaloids in the polyphagous arctiid Estigmene acrea

Summary. The alkaloid profiles of the life history stages of the highly polyphagous arctiid Estigmene acrea were established. As larvae individuals had free choice between a plain diet (alkaloid-free) and a diet that was supplemented with Crotalaria-pumila powder with a known content and composition of pyrrolizidine alkaloids (PAs). Idiosyncratic retronecine esters (insect PAs) accounted for approximately half of the PAs recovered from the larvae. These alkaloids were synthesized by the larvae through esterification of dietary supinidine yielding the estigmines, and esterification of retronecine yielding the creatonotines. The retronecine is derived from insect-mediated degradation of the sequestered pumilines (macrocyclic PAs of the monocrotaline type). With one exception, the PA profiles established for larvae were found almost unaltered in all life-stages as well as larval exuviae and pupal cocoons. The exception is the males, which in comparison to pupae and adult females, showed a significantly decreased quantity of the creatonotines and pumilines. These data support the idea that the creatonotines are direct precursors of the PA-derived male courtship pheromone, hydroxydanaidal. Crosses of PA-free males with PA-containing females and vice versa confirmed an efficient trans-mission of PAs from males to females and then from females to eggs. In single cases a male bestowed almost his total PA load to the female, and a female her total load to the eggs. The results are discussed with respect to pheromone formation, PA transmission between life-stages, and the defensive role of PAs against predators and parasitoids     

Larval chemical defence and evolution of host shifts in Chrysomela leaf beetles

Summary. Larvae of Chrysomela leaf beetles release for defence volatile compounds belonging to various chemical families. This study focuses on the defensive strategy based on the esterification of isobutyric acid and 2-methylbutyric acid with a wide variety of alcohols taken up from the host plant. To date, only two species are known to produce these repellents C. interrupta, which is associated with Betulaceae and C. lapponica which occurs either on Betulaceae or Salicaceae.? In order to know if other species have developed this chemical defence and how the food plant influences the secretion of these toxins, we targeted by mass spectrometry the presence of iso- and 2-methylbutyric acids and esters of them in the defensive secretions of Chrysomela larvae exclusively associated with Betulaceae or Salicaceae. ?Screening analyses reveal that the synthesis of these compounds is a common character restricted to all the members belonging to the C. interrupta group sensu Brown (1956) regardless of the host-plant family. These results suggest that the biochemical mechanism leading to the synthesis of these compounds could be considered as a synapomorphy meaning that the group is probably monophyletic. ?Defensive secretions of the members of the interrupta group are quantitatively assayed for iso- and 2-methylbutyric acids and their (Z)-3-hexenyl esters. Results reveal a chemical plasticity developed by Chrysomela species associated with Salicaceae. The amounts of iso- and 2-methylbutyric acids derivatives and of salicylaldehyde in their larval secretions depend on the food plant and on its content in phenolglucosides. Received 5 October 1998; accepted 25 November 1998.     

Sequestration of plant pyrrolizidine alkaloids by chrysomelid beetles and selective transfer into the defensive secretions

Summary Oreina cacaliae andO. speciosissima (Coleoptera, Chrysomelidae) sequester in their elytral and pronotal defensive secretions pyrrolizidine alkaloids (PAs) as Noxides (PA N-oxides). The PA N-oxide patterns found in the beetles and their host plants were evaluated qualitatively and quantitatively by capillary gas chromatography/mass spectrometry (GC-MS). Of the three host plantsAdenostyles alliariae (Asteraceae) is the exclusive source for PA N-oxide sequestration in the defensive secretions of the beetles. With the exception of O-acetylseneciphylline the N-oxides of all PAs ofA. alliariae, i.e. senecionine, seneciphylline, spartioidine, integerrimine, platyphylline and neoplatyphylline were identified in the secretion. PA N-oxides typical ofSenecio fuchsii (Asteraceae) were detected in the bodies of the beetles but not in their secretion. No PAs were found in the leaves of the third host plant,Petasites paradoxus (Asteraceae). The results suggest the existence of two distinctive storage compartments for PA N-oxides in the beetle: (1) the defensive secretion, containing specifically PA N-oxides acquired fromA. alliariae; (2) the body of the beetle, sequestering additionally but less selectively PA N-oxides from other sources,e.g. S. fuchsii or monocrotaline N-oxide fed in the laboratory. The concentration of PA N-oxides in the defensive secretion is in the range of 0.1 to 0.3 mol/1, which is more than 2.5 orders of magnitude higher than that found in the body of the beetle. No significant differences exist in the ability of the two species of beetles to sequester PA N-oxides fromA. alliariae, althoughO. speciosissima, but notO. cacaliae, produces autogenous cardenolides. A negative correlation seems to exist between the concentrations of plant-derived PA N-oxides andde novo synthesized cardenolides in the defensive secretion ofO. speciosissima.     

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.     

Chemical defense and evolutionary trends in biosynthetic capacity among dorid nudibranchs (Mollusca: Gastropoda: Opisthobranchia)

Summary. An evolutionary scenario incorporating recent advances in phylogenetic research begins with an opisthobranch-pulmonate common ancestor that was herbivorous and had some diet-derived chemical defense. The Nudibranchia and their closest relatives, the Notaspidea, form a lineage the ancestors of which had switched to feeding upon sponges and deriving protection from metabolites contained in them. Subsequently there have been repeated shifts in food and defensive metabolites, and trends are evident in the ability to detoxify, sequester and utilize metabolites from food, as well as to synthesize defensive compounds de novo. The Notaspidea display a minor adaptive radiation that foreshadows a more extensive one in the various lineages of nudibranchs. This review emphasizes changes that have occurred within the Holohepatica, or dorid nudibranchs (order Doridacea). Their sister-group, the Cladohepatica, consists of three other orders, Dendronotacea, Arminacea, and Aeolidiacea, in which there has been a shift from sponges to Cnidaria as food. The Dendronotacea often feed upon Octocorallia, which combine spicules, chemical defense, and stinging capsules and thereby suggest a transition from feeding on sponges. A previous diet of Octocorallia is suggested by the defensive use of prostaglandins in the dendronotacean Tethys fimbria, which eats crustaceans. A shift to bryozoans in some Arminacea is accompanied by use of different metabolites. Dorid nudibranchs evidently began as sponge-feeders, but some lineages have shifted to a variety of other food organisms, and others have specialized in the kind of sponges they feed on and how they do it. There have been shifts to bryozoans (Ectoprocta) and ascidians (Chordata: Urochordata) that track metabolites rather than the taxonomy of the food. There is a crude correlation between the genealogy and the defensive metabolites of the sponge-feeding dorids. De novo synthesis is well documented in this order and the metabolites are appropriately positioned so as to have an adaptive effect. The hypothesis that the capacity for de novo synthesis was acquired by gene transfer across lineages is rejected, partly on the basis of different chirality of metabolites in the nudibranchs and their food organisms. Instead it is proposed that there has been a preadaptive phase followed by evolution in a retrosynthetic mode, with selection favoring enzymes that enhance the yield of end products that are already present in the food. Received 5 February 1999; accepted 26 July 1999     

Why do plants ‘talk’?

Summary Plant defence can be induced by herbivory. This is true for both direct and indirect plant defence. Induced direct defence has been the most studied of the two. However, in most cases induced direct defence does not appear to be a water-tight defence option. In contrast, induced indirect defence through the production of herbivore-induced carnivore attractants can be a decisive factor in the extermination of herbivore populations. In this paper the main characteristics of induced attraction of carnivores by plants are reviewed. This includes the similarities and dissimilarities among tritrophic systems. There are two main patterns of induced carnivore attraction. (1) Through the emission of the same bouquet as that emitted by mechanically damaged plants, but in larger quantities and for a longer period of time after damage. (2) Through emission of large amounts of new volatiles that are synthesizedde novo in response to herbivore feeding andnot in response to mechanical wounding.Herbivore populations may be decimated by carnivores. Therefore it should be realized that herbivoreinduced carnivore attractants are essential in an important step in carnivore foraging,i.e. long-distance herbivore location. Once herbivores have started feeding on a plant and direct defence is not effective, induced indirect defence may be decisive for plant survival. Therefore, it is concluded that indirect defence is an essential aspect of induced plant defence directed at herbivorous arthropods.     

Precopulatory sexual interaction in an arctiid moth (Utetheisa ornatrix): Role of a pheromone derived from dietary alkaloids

Summary Males of Utetheisa ornatrix have a pair of brushlike glandular structures, the coremata, which they evert from the abdomen during close-range precopulatory interaction with the female. Males experimentally deprived of coremata are less acceptable to females. The principal chemical associated with the coremata, identified as a pyrrolizine (hydroxydanaidal), has a proven pheromonal role: males raised under conditions where they fail to produce hydroxydanaidal are also less likely to succeed in courtship, and the compound itself, as its (-)-isomer, is capable of inducing the principal receptive response (wing raising) of the female. Evidence is presented indicating that Utetheisa derive hydroxydanidal from defensive pyrrolizidine alkaloids that they sequester from their larval foodplants (Crotalaria spp.). It is proposed that in addition to signalling male presence to the female, hydroxydanaidal may provide the means whereby the female assesses the alkaloid content of the male and therefore his degree of chemical protectedness. The argument is made that such pheromonal assessment of defensive capacity may occur also in other insects, including danaid butterflies, many of which share with Utetheisa a dependence on pyrrolizidine alkaloids for sex-pheromone production.     

Storage and metabolism of radioactively labeled pyrrolizidine alkaloids by butterflies and larvae of Mechanitis polymnia (Lepidoptera: Nymphalidae,Ithomiinae)

Summary. Tracer feeding studies with radioactively labeled pyrrolizidine alkaloids (PAs) were performed to attain experimental information about the specificity and mechanisms of uptake, metabolism and storage of PAs in the alkaloid sequestering ithomiine butterfly Mechanitis polymnia. Adult butterflies easily ingest the tracers offered dissolved in a saturated sugar solution. Feeding of [¹⁴C]rinderine (free base) confirmed that M. polymnia is well adapted to sequester and maintain PAs of the lycopsamine type. Approximately 80% of the ingested radioactivity can be recovered in methanol extracts of the butterflies over a period of at least 6 hours. Labeled rinderine is efficiently N-oxidized and transformed into a metabolite of still unknown structure. These two metabolites are formed in almost equal amounts and account for more than 90% of total radioactivity. After four hours the toxic free base is only detectable in traces. Radioactively labeled senecioylretronecine (free base), a PA that often accompanies PAs of the lycopsamine type in plants, is metabolized in a different manner. The toxic free base disappeared as fast as the tertiary rinderine, but the final products which accumulated in a stable ratio after 12 hours were mainly two polar metabolites of unknown structure; senecioylretronecine N-oxide accounts for less than 10% of total PAs. Labeled senecionine a macrocyclic PA, which never has been found in wild caught M. polymnia is only slowly N-oxidized. In females ca 50% of the ingested senecionine is still present as free base after 24 hours, whereas under the same conditions in males this percentage is only ca 20%. This difference in N-oxidation was the only significant sex-specific difference observed in various experiments. Larvae of M. polymnia which feed on Solanum tabacifolium, a plant that does not contain PAs, are able to sequester and partly N-oxidize labeled senecioylretronecine and senecionine. However, the storage is not very efficient; with the two tracers less than 5% of radioactivity remained in the bodies after 24 hours. Received 19 October 1999; accepted 24 November 1999     

Faecal shield chemical defence is not important in larvae of the tortoise beetle Chelymorpha reimoseri (Chrysomelidae: Cassidinae: Stolaini)

The most frequently investigated defence mechanism among larvae of tortoise beetles is protection by faecal shields, which generally present chemicals that are directly sequestered and/or modified from larval host-plants. In the present work we investigate the tortoise beetle Chelymorpha reimoseri that feeds on the leaves of Ipomoea carnea fistulosa (Convolvulaceae), seeking for the importance of this mechanism to their larvae. We show that 4th instar larvae suffer low predation regardless of the presence of shields in field and laboratory bioassays with ants and chicks. Chloroform extract from larvae without shields provided high protection against predation as shown in bioassays in the field, as well as against chicks, suggesting that C. reimoseri does not rely on the shield for protection. The aqueous extract of the shields did not show activity in such bioassays. The compounds responsible for this protection have yet to be identified, and it remains an open question as to whether they are sequestered from the host plant or de novo biosynthesized. This is the first record of chemical defence in cassidine beetles without the need for faecal shields. These findings indicate that more attention should be paid to chemicals present in the tissues of larvae and/or adults of tortoise beetles; the protective compounds sequestered from host plants or de novo biosynthesized can provide an alternative or complementary strategy against predation in these insects.     

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.     

Effects of fertilization and elevated CO<Subscript>2</Subscript> on larval food and butterfly nectar amino acid preference in<Emphasis Type="Italic"> Coenonympha pamphilus</Emphasis> L.

The effects of larval diet on the nutritional preferences of butterflies has rarely been examined. This study investigates whether alterations in the larval diet result in changes in adult preferences for nectar amino acids. Larvae of Coenonympha pamphilus were raised on fertilized or unfertilized Festuca rubra, grown under ambient (350 ppm) or elevated (750 ppm) atmospheric CO ₂environments. Fertilization led to marked increases in leaf nitrogen concentration. In plants grown under elevated CO ₂conditions, leaf water and nitrogen concentrations were significantly lower, and the C/N-ratio increased significantly. Fertilization of the host plant shortened the development time of C. pamphilus larvae, and pupal weight increased. In contrast, larvae of C. pamphilus developed significantly slower on F. rubra grown under elevated CO ₂, but adult emergence weight was not affected by CO ₂treatment of the plant. C. pamphilus females showed a clear preference for nectar mimics containing amino acids, whereas males, regardless of treatment, either preferred the nectar mimic void of amino acids or showed no preference for the different solutions. Female butterflies raised on fertilized plants showed a significant decline in their preference for nectar mimics containing amino acids. A slight, but not significant, trend towards increased nectar amino acid preference was found in females raised on plants grown under elevated CO ₂. We clearly demonstrate that alterations in larval host quality led to changes in butterfly nectar preferences. The ability of the butterfly to either rely less on nectar uptake or compensate for poor larval conditions represents a trade-off between larval and adult butterfly feeding.     

Cardenolide content,emetic potency,and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus,and their larval host-plant milkweed,Asclepias humistrata,in Florida

Summary This paper is the fourth in a series on cardenolide fingerprints of monarch butterflies (Danaus plexippus, Danainae) and their host-plant milkweeds (Asclepiadaceae) in the eastern United States. Cardenolide concentrations ofAsclepias humistrata plants from north central Florida ranged from 71 to 710 µg/0.1 g dry weight, with a mean of 417 µg/0.1 g. Monarchs reared individually on these plants contained cardenolide concentrations ranging from 243 to 575 µg/0.1 g dry weight, with a mean of 385 µg/0.1 g. Cardenolide uptake by butterflies was independent of plant concentration, suggesting that sequestration saturation occurs in monarchs fed cardenolide-rich host plants. Thinlayer chromatography resolved 19 cardenolides in the plants and 15 in the butterflies. In addition to humistratin,A. humistrata plants contained several relatively non-polar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. These produced a butterfly cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. In emetic assays with the blue jay,Cyanocitta cristata, the 50% emetic dose (ED₅₀) per jay was 57.1 µg, and the average number of ED₅₀ units per butterfly was 13.8, establishing that this important south eastern milkweed produces highly emetic, chemically defended monarchs. Our data provide further support for the use of cardenolide fingerprints of wild-caught monarchs to make ecological predictions concerning defence against natural enemies, seasonal movement and larval host-plant utilization by monarch butterflies during their annual cycle of migration, breeding and overwintering.     

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.     

Methylalkylpyrazines in aposematic insects,their hostplants and mimics

Summary Adults of forty-five aposematic insect species from four orders, and/or, in eight cases, their early stages, were examined for the presence of 2-methoxy-3-alkylpyrazines by coupled gas chromatography/mass spectrometry. The survey also included eight cryptic and related species and six unrelated mimics, twelve selected hostplants of those herbivores that had proved positive for such pyrazines, and nine well known toxic plants. Positive results were as follows: aposematic adult species, 26 (58 %); obscurely coloured species, 1 (12.5%); mimics, 2 (33%); host-plants, 5 (42%); well-known toxic plants, 4 (44%). Among the taxonomic groups, these pyrazines were found in the lepidopteran families and subfamilies Papilionidae, Danainae, Heliconiinae, Acraeinae, Zygaenidae and Arctiidae, and among the Coleoptera, in the Lycidae, Cantharidae, Endomychidae, Coccinellidae, Oedemeridae and Meloidae. Single cases of the occurrence of these pyrazines were also detected in the Hemiptera (Cercopidae) and Orthoptera (Pyrgomorphidae). Mimics were found with and without the pyrazines. Among the plants they were detected in Aristolochiaceae, Asclepiadaceae, Asteraceae, Papaveraceae, Passifloraceae, Solanaceae and Urticaceae. Some aposematic and phytophagous insects apparently sequester pyrazines from their hostplants whereas others, particularly the predatory species, manufacture these substancesde novo. Alerting signals in general are discussed.     

Responses of three mouse species to deterrent chemicals in the monarch butterfly. I. Taste and toxicity tests using artificial diets laced with digitoxin or monocrotaline

Summary Of three common mouse species at the Mexican overwintering sites of the monarch butterfly, onlyPeromyscus melanotis eats monarchs. We hypothesized thatP. aztecus andReithrodontomys sumichrasti reject monarchs because they are more sensitive to the bitter taste and/or toxic effects of the cardiac glycosides (CGs) and pyrrolizidine alkaloids (PAs) in the butterflies. Two-choice preference tests revealed no difference in taste avoidance thresholds to free base and N-oxide forms of the PA, monocrotaline, but very different avoidance thresholds to the CG, digitoxin. Avoidance thresholds forR. sumichrasti andP. aztecus were, in respective order, 1020 and 34 times less than that forP. melanotis. We also tested the toxic sensitivity of juvenile mice by chronically feeding diets containing digitoxin or monocrotaline at concentrations similar to those used in the preference tests. No species developed CG toxicity, but bothP. melanotis andP. aztecus developed moderate PA toxicity (R. sumichrasti was not tested for PA toxicity).P. aztecus grew more slowly and manyP. melanotis had hepatic metabolic lesions. Thus, the three mouse species responded very differently to the taste and toxic properties of CGs and PAs at ecologically relevant concentrations: 1) CGs were taste rejected by all species exceptP. melanotis, while PAs were not; and 2) PAs were toxic, while CGs were not.     

Rodent predation on hibernating peacock and small tortoiseshell butterflies

Insects that hibernate as adults have a life span of almost a whole year. Hence, they must have extraordinary adaptations for adult survival. In this paper, we study winter survival in two butterflies that hibernate as adults and have multimodal anti-predator defences—the peacock, Inachis io, which has intimidating eyespots that are effective against bird predation, and the small tortoiseshell, Aglais urticae, which does not have an effective secondary defence against birds. We assessed predation on wild butterflies hibernating in the attic of an unheated house, as well as survival of individually marked butterflies placed by hand on different sites in the attic. Our objectives were to assess (1) the number of butterflies that were killed during hibernation, (2) whether survival differed between butterfly species, and (3) how predation was related to hibernation site and the identity of the predator. There was a strong pulse of predation during the first 2 weeks of hibernation: 58% of A. urticae and 53% of I. io were killed during this period. Thereafter, predation decreased and butterfly survival equalled 98% during the final 16 weeks of hibernation. There was no difference in survival between the two butterfly species, but predation was site-specific and more pronounced under light conditions in locations accessible to a climbing rodent, such as the common yellow-necked mouse, Apodemus flavicollis. We contend that small rodents are likely important predators on overwintering butterflies, both because rodents are active throughout winter when butterflies are torpid and because they occur at similar sites.     

Larval hemolymph feeding in the ant Leptanilla japonica by use of a specialized duct organ,the “larval hemolymph tap” (Hymenoptera: Formicidae)

Summary The larvae of the migratory Japanese ant Leptanilla japonica Baroni Urbani have a specialized duct organ on each side of the 4th abdominal segment. Behavioral and histological studies have shown that the adult ants are able to imbibe hemolymph directly from the larval body cavity through these organs, each of which is referred to here as a larval hemolymph tap. Laboratory observations further confirm that larval hemolymph feeding (LHF) is the sole source of nutrient for the queens. L. japonica is cyclical in its brood production. All larvae in a colony develop in concert, and when they are mature, the queen performs much active LHF on them. Nourished in this way, she achieves full physogastry within a period of only a few days and then lays a batch of 100–200 eggs. LHF thus facilitates synchronization of brood maturation and concentrated production of eggs by the queen. The larval hemolymph tap has probably evolved de novo in Leptanilla, in relation to cyclical brood production by species with small colony populations. Obviously, the function of this larval organ is non-selfish, contributing to overall colony-level functioning. The larvae of Leptanilla are thus properly qualified as a distinct class of caste in ant societies.