Hydroquinones from defensive secretion of a giant Pacific millipede, <Emphasis Type="Italic">Acladocricus setigerus</Emphasis> (Diplopoda: Spirobolida)

Summary. The presence of considerable quantities of hydroquinones including hydroquinone, 2-methylhydroquinone, 2,3- dimethoxyhydroquinone, 2-methoxy-3-methylhydroquinone, and 2,3-dimethoxy-5-methylhydroquinone renders the defensive secretion of Acladocricus setigerus (Silvestri, 1897) significantly different from that of other quinone-producing millipedes. In addition, two hitherto undescribed compounds, namely, 2,3-dimethoxy-5-methylhydroquinone and 2-methyl-3,4-methylenedioxyphenol, were characterized from the defensive secretion. However, it is uncertain if the latter compound is formed after the release of the secretion. The two major compounds, 2-methyl-1,4-benzoquinone and 2-methoxy-3-methyl-1,4-benzoquinone, constitute about 75% of the defensive fluid. Furthermore, hydrocarbons, which are typically present in the secretions of most other arthropods that use benzoquinones as repellents, are notably absent in the secretion of A. setigerus.     

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.     

Chemical studies on Indopacific Ceratosoma nudibranchs illuminate the protective role of their dorsal horn

Summary. Two Ceratosoma species, C. trilobatum and C. gracillimum (Mollusca: Gastropoda: Opistobranchia), collected along the South China Sea coast, have been studied in order to investigate their defensive strategies. Both species contain four typical sponge furanosesquiterpenoids, probably with a dietary origin. The tissue distribution of these compounds has been studied by ¹H-NMR analysis and preliminary bioassays have been carried out in order to evaluate the ecological role of the isolated metabolites. The main sesquiterpene component in both species was (–)-furodysinin (2), which showed significant feeding-deterrent and ichthyotoxic properties in bioassays with freshwater organisms. The work has been performed studying a very limited number of molluscs that prevented the realization of ecologically relevant experiments. However, the almost exclusive presence of compound 2 in the crude extract of the mantle glands concentrated in the dorsal horn of C. gracillimum strongly supports the defensive role of that dorsal protuberance, which probably acts as a defensive lure attracting potential predators to the part of the animal that contains the distasteful chemicals.     

Odour profile congruity in two closely related desert tenebrionid beetles: Homology as the basis for a cleptoparasitic relationship?

Summary Parastizopus armaticeps andEremostibes opacus are two closely related desert tenebrionids which also live in close association, the former having biparental brood care and the latter cleptoparasitising the brood.E. opacus is unable to discriminate between the conspecific and host odour even in the absence of physical contact. Gas chromatographic analysis of headspace volatiles of resting animals showed almost complete qualitative and quantitative odour congruity between them. Comparison of these odour profiles with those of two other tenebrionids sharing the same ecological niche,Gonopus agrestis andHerpiscius sp. (damaralis?) showed that congruity was independent of common foodplant utilisation. It is also independent of common defensive gland secretions. Parallels between resting odour spectra, defensive secretion spectra and systematic status suggest that the origins of congruity lie in odour homology, by means of which the cleptoparasite was able to exploit its host.     

Anatomical and ultrastructural studies of chemical defence in the sponge Dysidea fragilis

The marine sponge Dysidea fragilis from El Mar Menor, a hypersaline coastal lagoon (Murcia, Spain), contains the furanosesquiterpenoid ent-furodysinin as the major secondary metabolite. D. fragilis emits a defensive white fluid when it is disturbed. Electron micrographs of this fluid revealed intact vesiculated cells together with other amorphous material. Dissociated cells are more rounded in shape but maintain the same ultrastructural features as cells observed in ultra-thin sections of the whole sponge. The defensive secretion is composed mainly of sponge cells with abundant light vesicles. Sometimes these light vesicles appear to open into the intercellular space; this correlates with surface blebs on these cells observed under scanning electron microscopy. The intracellular location of ent-furodysinin was confirmed by Erlich staining. In laboratory assays, we examined the role of ent-furodysinin as a feeding deterrent to generalist fish predators. It was isolated from D. fragilis and incorporated into a carrageenan-based artificial diet. The addition of ent-furodysinin to the artificial diet reduced feeding by the fish Thalassoma pavo. Similarly, fish did not feed on artificial diet above which defensive secretion of D. fragilis had been ejected with a small syringe. Received: 4 June 1997 / Accepted: 28 January 1998     

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.     

Genetic connectivity among color morphs and Pacific archipelagos for the flame angelfish, <Emphasis Type="Italic">Centropyge loriculus</Emphasis>

Color variation is used in taxonomic classification of reef fishes, but it may not reliably indicate evolutionary divergence. In the central Pacific, there are three color morphs of the flame angelfish, Centropyge loriculus: a red morph that occurs primarily in the Hawaiian archipelago, the endemic Marquesan color morph with reduced black markings, and an orange morph that occurs throughout the rest of Oceania. The red and orange morphs co-occur at Johnston Atoll (1,300 km south of Hawai’i), but intermediate forms have not been reported. To determine whether the three color morphs represent distinct evolutionary lineages, we compared 641 base pairs of mitochondrial cytochrome b. Forty-one closely related haplotypes were observed in 116 individuals. Analysis of molecular variance (AMOVA) indicated no significant genetic structure among color morphs (Φ_ST = 0.011, P = 0.147). Likewise, there was no significant pairwise structure between sampling locations, separated by up to 5,700 km, after a Bonferroni correction (Φ_ST = 0.000–0.080, P = 0.0130–0.999). Genetic studies in conjunction with larval distribution data indicate that Centropyge species are highly dispersive. While there is a strong geographic component to the distribution of color morphs in C. loriculus, we find no evidence for corresponding genetic partitioning. We do not rule out an adaptive role for color differentiation, but our data do not support emerging species.     

Caste polyethism and collective defense in the ant,Pbeidole pallidula: the outcome of quantitative differences in recruitment

Summary During agonistic encounters, both minors and majors of the European ant P. pallidula actively cooperate in defense. Minors seize the legs of the intruder and in some cases induce the recruitment of nestmates whereas majors kill the spreadeagled alien ant. The defensive strategy of P. pallidula is very flexible and adapted to both the number of alient ants and to the intruder's superiority in fighting. On the one hand, only a massive invasion of alien minors results in a slow mobilization of resident ants to the combat area, elicited by recruiters performing weak tactile invitations and trail-laying behavior. On the other hand, the presence of 10 majors induces a fast and massive recruitment achieved by intense trail-laying and tactile invitations from the recruiters. Because of their high response threshold to invitations, resident majors are mobilized only during these intense recruitments, their exit being additionally enhanced by their preferential stimulation. The adaptiveness of this defensive strategy is discussed. It is also suggested that simple decision-making rules of recruitment and caste differences in behavioral thresholds could account for the complexity of P. pallidula defensive strategies.Senior Research Assistant at Belgian National Fund for Scientific Research Offprint requests to: C. Detrain     

Chemoecological studies of the exocrine glandular larval secretions of two chrysomelid species (Coleoptera):Phaedon cochleariae andChrysomela lapponica

Summary The exocrine glandular secretions of larvae of the subfamily Chrysomelinae are known to repel conspecific adults, other competitive phytophagous insects and natural enemies. InPhaedon cochleariae, the intraspecific activity of tlc fractions of the larval secretion was tested in order to examine the ecological significance of two fractions containing minor components and a fraction containing the major compound, the cyclopentanoid monoterpene (epi)chrysomelidial. InChrysomela lapponica, the defensive activity of the larval secretion against ants is known from specimens feeding upon willow or birch. The feeding preferences of larvae and adults ofC. lapponica from a Finnish and a Czech population were tested. The Finnish individuals significantly preferred feeding uponSalix borealis, whereas they hardly fed upon birch. The Czech specimens clearly preferred birch (Betula pubescens) to willow species. Application of salicin onto leaves of a willow species free of this phenolglycoside revealed that the Finnish individuals preferred feeding upon leaves with salicin. On the other hand, the Czech individuals avoided feeding upon leaves ofB. pubescens treated with salicin. The chemical composition of the glandular secretion of the Finnish larvae differed from the one of the Czech larvae. GC-MS-analyses of the secretions revealed that salicylaldehyde was the only major component of the secretion of Finnish larvae feeding upon the salicin-containing willowS. borealis. The glandular secretion of the Czech larvae feeding upon birch contained numerous esters of isobutyric acid and 2-methylbutyric acid. When Czech larvae had fed upon a salicin-containing willow (S. fragilis), the major compounds of their secretion were benzoic acid, salicylalcohol and benzoic acid esters; salicylaldehyde was only detected in traces. Thus,C. lapponica individuals from the Finland population adapted so closely to a salicincontaining willow that they clearly prefer this plant for food and that they obviously derive their main larval defensive compound (salicylaldehyde) from their host-plant.     

Offensive and defensive sperm competition roles in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae)

Sperm competition is predicted to generate opposing selection pressures on males. On one hand, selection should favour ‘defensive’ adaptations that protect a male’s ejaculate from displacement, while, on the other hand, selection should favour ‘offensive’ adaptations that overcome paternity assurance mechanisms of rivals. Here, we use the sterile male technique to assess sperm precedence when a male dung beetle Onthophagus taurus mates in both a defensive (first male) and an offensive (second male) role. Significant variation in a male’s sperm precedence (both P ₁ and P ₂) was detected, and an individual’s defensive (P ₁) and offensive (P ₂) abilities were positively correlated. Thus, it appears that sexual selection simultaneously selects for ‘defensive’ and ‘offensive’ adaptations in O. taurus. We discuss a variety of male traits in O. taurus that potentially contribute to a male’s ability to be successful when mating in an ‘offensive’ and a ‘defensive’ role.     

Defensive secretions from the larvae ofApatania fimbriata (Pictet) (Trichoptera: Limnephilidae)

Summary When the larvae of the caddis flyApatania fimbriata (Pictet) are threatened, drops of fluid appear on their head capsules. The secretions are produced in a gland in the dorsal part of the prothorax. The neck region contains an eversible sac with numerous single setae, groups of 3 setae, or rows of setae on the surface. The secretion is released through two paris of orifices on the lateral sides of the sac. For the most part the secretion is composed of some 30 fatty acids, with the major components having 12–14 carbon atoms and up to 4 double bonds (approx. 1–2 µg secretion per specimen). Biotests with synthetic saturated acids (C₆–C₁₂) in a stream and in the laboratory demonstrated a paralysing effect on small invertebrate predators (Rhyacophila sp.,Plectrocnemia conspersa, Hydropsyche sp., larvae, all Trichoptera). In choice experiments,Rhyacophila sp. larvae preferred larvae ofAgapetus fuscipes andDrusus annulatus (Trichoptera) as food as compared withApatania fimbriata larvae. Larger predators, such asDinocras cephalotes (Insecta, Plecoptera) and the fishCottus gobio, did not discriminate betweenApatania fimbriata and other prey species. The use of fatty acids in defensive secretions is interpreted as an adaptation to the running water environment. They are effective repellents againstRhyacophila sp. larvae, the most important predator in the natural environment ofApatania larvae.     

Complex chemical communication in the crazy ant <Emphasis Type="Italic">Paratrechina longicornis</Emphasis> Latreille (Hymenoptera: Formicidae)

Summary. The formicine ant Paratrechina longicornis is known for its extremely opportunistic foraging behaviour. Only a single trail pheromone source, the rectum, was previously described from this ant. Our detailed examination of this ant’s chemical communication system revealed the presence of at least four sources of pheromones. Rectum, poison sac, and Dufour gland contain orientation components with decreasing effectiveness and persistence (in the sequence mentioned) as well as attractants with increasing effectiveness. Furthermore, the mandibular gland contains repellents, and a releaser of defensive behaviour. This set of various signals of different strength and persistence allows an extraordinary degree of flexibility and efficiency in the collective behaviour of P. longicornis, especially food exploitation, and thus may contribute to this insect’s overall ecological success.     

Oviposition by butterflies on young leaves: Investigation of leaf volatiles

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

Effect of bombardier beetle spray on a wolf spider: repellency and leg autotomy

Summary. Data are presented on the repellency of the spray of a bombardier beetle (Pheropsophus aequinoctialis) to a lycosid spider (Lycosa ceratiola). The secretion is shown to cause the spider to desist from its assault on the beetle within, on average, 58 ms of onset of the beetle’s secretory emission, a reaction time that is at a par with latencies previously reported for startle, escape, and avoidance reactions of cockroaches, flies, and moths. Spray ejections by the beetle, are shorter in duration (43 ms, on average) than the response time of the spider, an indication that the beetle does indeed pack a formidable “punch” into its ejection. After being hit by a beetle’s spray, L. ceratiola were found occasionally to autotomize one or two of their legs. It is argued, but not proven, that this unusually severe effect from exposure to an arthropodan defensive secretion may be caused by the high temperature of the bombardier beetle spray.     

Defensive secretion of a flightless grasshopper: Failure to prevent lizard attack

Summary We tested the role of a grasshopper defensive secretion in deterring lizard predation. Adults, but not young larvae, of the chemically defended lubber grasshopperRomalea guttata (=microptera) froth a volatile secretion when attacked by predators. The lizardAnolis carolinensis failed to strike juvenile lubbers (which lack secretion) in laboratory trials. Survivorship of palatable crickets loaded with secretion offered toA. carolinensis was not significantly different from survivorship of control crickets. In experiments designed to investigate if lizards learn an aversion to the secretion, striking times forSceloporus undulatus fed wax worms coated with secretion were not significantly different over three days of trials. Three primary conclusions are drawn from these data. First, the secretion may not be necessary for lubber protection from lizards. Second, lubber secretion does not appear to deter lizards from attacking or eating prey items. Third, lizards do not appear to develop an aversion to the secretion.     

Defense mechanisms and antipredator behavior in two sympatric species of spiny lobsters, Panulirus argus and P. guttatus

The congeneric spiny lobsters Panulirus argus and P. guttatus co-occur throughout the Caribbean Sea, where they may share the coral reef habitat. Despite their phylogenetic closeness, both species have many different life-history traits that may partially explain their coexistence. However, even though both species may face the same predators, their defense mechanisms and antipredator strategies had not been compared. We compared the performance between species in 18 morphological and behavioral defense mechanisms commonly expressed by most spiny lobsters, including predator-avoidance mechanisms (activity schedule, sheltering behavior, delay to disturbance, and effect of conspecific damage-released scents on shelter choice) as well as antipredator mechanisms (body size, several parameters of the escape response and limb autospasy, clinging strength, antennal strength, and cooperative defense). As hypothesized, both species expressed all these defense mechanisms (except cooperative defense, shown only by P. argus), reflecting their phylogenetic closeness, but performed significantly differently in most, in accordance with their particular ontogenetic traits. Their comparative performance in individual defense mechanisms as well as the antipredator strategies displayed by groups of lobsters of each species in the presence of a common predator (the triggerfish Balistes vetula) showed that, in general, the defensive behavioral type of P. argus is more bold and that of P. guttatus more shy. Therefore, their distinct defensive behaviors contribute to their niche differentiation.     

Chemical defense of the ladybird beetle Epilachna paenulata

Summary. The defensive chemistry of the ladybird beetle Epilachna paenulata (Coleoptera: Coccinellidae) was characterized as a mixture of piperidine, homotropane and pyrrolidine alkaloids. Whole body extracts of adult beetles contain four major alkaloids: 1-(6-Methyl-2,3,4,5-tetrahydro-pyridin-2-yl)-propan-2-one; 1-(6-methyl-2-piperidyl)-propan-2-one; 9-aza-1-methyl-bicyclo[3.3.1]nonan-3-one and 1- (2′′- hydroxyethyl)-2-(12′-aminotridecyl)-pyrrolidine. Comparative studies of the defensive chemistry of eggs, larvae, pupae and adults showed both qualitative and quantitative differences in alkaloid composition among the four life stages, and also within adult age. Laboratory predation bioassays with wolf spiders showed that the adults are better protected than the larvae and pupae. Field tests showed the adult alkaloid extract to be deterrent to ants.     

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

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

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.     

Allelochemical effects of ergoline alkaloids fromIpomoea parasitica onHeliothis virescens

Lysergol, elymoclavine and three other van Urk-positive alkaloids were identified by 2D chromatography in the seed extract ofIpomoea parasitica. The presence of the same ergoline alkaloids was also demonstrated in vegetative tissue ofI. parasitica. Heliothis virescens larvae reared on a diet containing an alkaloid extract ofI. parasitica showed an increase in the consumption index and a reduction in the effiency of conversion of food. No difference was observed in the approximated digestibility. The percentages of pupation and emergence were reduced and the malefemale ratio was altered in insects fed on a diet containing the alkaloid extract ofI. parasitica or lysergol. The increases in the concentration of ergoline alkaloids inI. parasitica at the seedling stage and at flowering support the theory that these compounds play a defensive role against herbivory in the plant.