Defensive volatile secretions of two diplopod species attract the carrion ball roller scarab <Emphasis Type="Italic">Canthon morsei</Emphasis> (Coleoptera: Scarabaeidae)

Summary. We report field and laboratory evidence indicating that the defensive compounds secreted by the millipedes Amplinus bitumidus (Diplopoda: Polydesmida) and Anadenobolus putealis (Diplopoda: Spirobolida) attract the carrion scarab Canthon morsei (Scarabaeidae: Scarabaeinae). In a Mexican tropical rain forest, C. morsei was mostly caught in pitfall traps baited with the carcasses of both millipede species. Only a few beetles were attracted to fish carrion. Experimental evidence indicates that C. morsei is attracted to freshly dead millipedes. Benzaldehyde and HCN were identified from Amplinus bitumidus; but 2-methyl-1,4-benzoquinone (toluquinone) and 2-methoxy- 3-methyl-1,4-benzoquinone from Anadenobolus putealis. Benzaldehyde attracted significantly more C. morsei individuals than fish meat, or millipedes of both species did after rinsing in hexane. This is the first report of Polydesmida attracting dung beetles.     

Chemical defense of an earwig (Doru taeniatum)

Summary. The earwig Doru taeniatum (Dermaptera, Forficulidae) has a pair of defensive glands, opening on the 4th abdominal tergite, from which it discharges a spray when disturbed. It aims the discharges by revolving the abdomen, a maneuver that enables it simultaneously to use its pincers in defense. The secretion contains two quinones (methyl-1,4-benzoquinone and 2,3-dimethyl-1,4-benzoquinone) present in the glands as a crystalline mass, together with pentadecane and a (presumably) aqueous phase. The gland openings are minute, with the result that virtually no quinone crystals are expelled with the spray. Only the two liquid phases are discharged, together with the ca. 1% quinone they carry in solution. Such a solute-economizing discharge mechanism appears to be without parallel among insect defensive glands. Received 14 February 2000; accepted 21 February 2000     

Chemical composition and pheromonal function of the defensive secretions in the subtribe Stizopina (Coleptera, Tenebrionidae, Opatrini)

The chemical composition of the defensive secretions of 52 species from 15 genera of the tenebrionid subtribe Stizopina was analyzed. The secretions of all species contained 1,4-benzoquinones, 1-alkenes, and monoterpene hydrocarbons, only one species was lacking the latter. Methyl- and ethyl-1,4-benzoquinone were ubiquitous, mostly accompanied by smaller amounts of 1,4-benzoquinone as well as isopropyl- and propyl-1,4-benzoquinone. 1-Alkenes were dominated by 1-undecene with varying admixtures of other 1-alkenes. The monoterpene hydrocarbons always consisted of a mixture of α-pinene, camphene, β-pinene and limonene, but also p-cymene, α-terpinene or α-phellandrene were found in some species. Furthermore, some species synthesized additional compounds such as phenols, ketones, 2,5-dihydroxy-6-methylbenzoate, 2-hydroxy-4-methoxyacetophenone and naphthoquinones. Bioassays showed that the defensive secretion co-functioned as an aggregation pheromone in the subtribe Stizopina. All nine tested species from six genera were attracted to defensive secretion of Stizopina species, but they did not distinguish between defensive secretions of different Stizopina species. This lack of discrimination might be the initial step for the formation of interspecific aggregations and the evolution of cleptoparasitism within the Stizopina.     

Chemical defense of a primitive Australian bombardier beetle (Carabidae):Mystropomus regularis

Summary The Australian bombardier beetle,Mystropomus regularis, sprays a mixture of quinones (1,4-benzoquinone, 2-methyl-1,4-benzoquinone, 2-ethyl-1,4-benzoquinone) and hydrocarbons (principallyn-pentadecane). The defensive fluid ist generated explosively in two-chambered glands, and is ejected audibly and hot (maximal recorded temperature = 59°C).Mystropomus is a member of the paussoid lineage of bombardiers. In common with other members of the group, it has a pair of elytral flanges (flanges of Coanda), associated with the gland openings, that serve as launching guides for anteriorly-aimed ejections of spray. It is argued thatMystropomus may be the least derived of flanged paussoids, and the closest living relative of the most primitive of extant bombardiers (Metriini).Paper no. 101 of the series Defense Mechanisms of Arthropods; no. 100 is Attygalleet al. J Chem Ecol 17: 805 (1991)     

The use of the tergal gland defensive secretion in male intrasexual aggression of the rove beetle,Aleochara curtula (Coleoptera: Staphylinidae), measured by closed-loop-stripping-analyses and tandem bioassay — mass fragmentography

Summary 2-methyl-1,4-benzoquinone as a target compound of the tergal gland secretion of the rove beetle,Aleochara curtula, was quantified from the reservoirs of individual beetles. Males store less secretion than females, and they evaporate more of the secretion as measured by adsorption from the air (closed-loop-stripping-analysis). The amount of emitted quinone is increased during aggressive interactions of rival males. The pulsed emission of the secretion during aggression is demonstrated by a novel technique combining observation of behavior with the on-line measurement of target fragment ions by mass-spectrometry (tandem bioassay — mass fragmentography). The emission of the secretion is used as a weapon in combats between males and may result in the repulsion of subordinate males from the mating site, but may also serve to mimic females chemically in order to avoid aggressive encounters.     

Age- and sex-related variation in defensive secretions of adult <Emphasis Type="Italic">Chlaenius cordicollis</Emphasis> and evidence for their role in sexual communication

Defensive secretions of adult Chlaenius cordicollis (Coleoptera: Carabidae) from Manitoba and Pennsylvania were analyzed across the life cycle in a multi-year study using solid phase microextraction and gas chromatography-mass spectrometry. In concurrent experiments using Y-tube olfactometry, we observed the response of each sex when male or female source beetles were induced to discharge defensive secretions. Almost all defensive secretion samples contained five alkylphenolic compounds, dominated by 3-methylphenol; frequently, two alkoxyphenolic compounds and one ester were also detected. As beetles aged, the composition of their defensive secretion shifted, showing an exponential decrease in the relative proportion of 2,3-dimethylphenol as a function of day of year. Previously reported differences observed in autumn between secretions of beetles from Manitoba and Pennsylvania are attributable to different times of onset of the decline of 2,3-dimethylphenol in the two locales. Regardless of airborne odor, responding beetles exhibited upwind anemotaxis in the olfactometer and, outside of the reproductive period, avoidance was the most frequent response to defensive secretions. Sexual dimorphism was evident in the major components of the defensive secretions during the reproductive period and resulted from an increase over time in 2,5-dimethylphenol in male beetles. In the reproductive period, neither sex avoided odor from male source beetles, females preferred clean air to odor from source females, and males were attracted to females that discharged defensive secretion but showed no preference when females had not discharged the secretion. The defensive secretion appears to function as a sex attractant during the reproductive period.     

Production of semiochemical and allelobiotic agents as a consequence of aphid feeding

Aphids have an extremely intimate relationship with their plant hosts. Although this might suggest that aphid infestation would be largely cryptic, there are a wide range of changes that can be detected behaviourally, chemically and also at the molecular genetic level. Colonisation by aphids can cause release of semiochemicals characteristic of the aphid species and besides reducing acceptability to incoming aphids; these can enable recruitment of specific parasitoids. Some semiochemicals involved in these processes can also influence the defence status of neighbouring intact plants through air or the rhizosphere. Electrophysiological and behavioural studies on the aphids, their parasitoids and other organisms facilitate the identification of compounds having direct effects on plant defence. New developments from work on semiochemicals such as (Z)-jasmone and 6-methyl-5-hepten-2-one, and allelobiotic agents such as 6-hydroxy-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one, as well as new work on these compounds and others as potential phytopheromones are discussed.     

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.     

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.     

Inter-individual variation in sequestration (as measured by energy dispersive spectroscopy) predicts efficacy of defensive secretion in lubber grasshoppers

Summary. We tested whether inter-individual variation in concentration of sequestered compounds predicts defensive efficacy of the defensive secretion of onion-fed Eastern lubber grasshoppers. When fed onion, lubbers have the ability to sequester sulfur compounds into their defensive secretion, and the secretion from onion-fed lubbers is more deterrent of ants than the secretion from lubbers fed other diets (Jones et al. 1989). To test three hypotheses, we developed a method for measuring total sequestered sulfur in the secretion using energy dispersive spectroscopy (EDS). First, we found that lubbers fed an ad libitum, monophagous diet of onion produce more secretion and have higher concentrations of total sulfur in their secretions than lubbers fed other diets. These sulfur concentrations varied three-fold; we believe that this variation in the composition of defensive secretions stems from physiological variation. Second, our method was sensitive enough to detect sulfur in the secretion of some lubbers that had been fed onion only once. Third, the sulfur concentration of a secretion sample significantly predicted its ability to deter ants from foraging. Hence, our results suggest that inter-individual variation in defensive chemistry in insects offered the same diet may be ecologically important. Received 25 November 1997; accepted 5 February 1998.     

Synthesis and in vitro anti-inflammatory activity of novel benzo[d] imidazolyl dihydrospiro[indoline-3,1′-isoindole]-2,4′,7′-triones

Synthesis of novel benzo[d]imidazolyldihydrospiro[indoline-3,1′-isoindole]-2,4′,7′-triones has been described. The key intermediate 2-[(2-sulfanyl-1H-benzo[d]imidazol-5-yl)amino]acetic acid is obtained by reacting 5-amino-2-mercapto benzimidazole and chloroacetic acid. The three-component reaction of 2-[(2-sulfanyl-1H-benzo[d]imidazol-5-yl)amino]acetic acid with isatin and 1,4-benzoquinone in the presence of 10 mol% of L-proline at ambient temperature afforded 2′-(2-mercapto-1H-benzo [d] imidazol-5-yl)-3′,3a′-dihydrospiro[indoline-3,1′-isoindole]-2,4′,7′(2′H,7a′H)-triones. All the synthesized compounds were evaluated for in vitro anti-inflammatory activity.     

On the enigmatic scent glands of dyspnoan harvestmen (Arachnida,Opiliones): first evidence for the production of volatile secretions

While considerable knowledge on the chemistry of the scent gland secretions from the opilionid suborders Laniatores and Cyphophthalmi has been compiled, it is the Palpatores (Eupnoi and Dyspnoi) where chemical data are scarce. In particular, the Dyspnoi have remained nearly unstudied, mainly due to their reported general reluctance to release secretions as well as to the phenomenon of production of insoluble—and inaccessible—solid secretion. We here show that at least certain nemastomatid Dyspnoi, namely all three species of genus Carinostoma, indeed produce a volatile secretion, comprising octan-3-one, 6-methyl-5-hepten-2-one and acetophenone in species-specific combinations. In all Carinostoma spp., these volatiles are embedded in a semi-volatile, naphthoquinone matrix (mainly 1,4-naphthoquinone and 6-methyl-1,4-naphthoquinone). In detail, acetophenone and traces of naphthoquinones characterize the secretions of Carinostoma carinatum. A mixture of octan-3-one, 6-methyl-5-hepten-2-one and large amounts of naphthoquinones were found in C. elegans, and 6-methyl-5-hepten-2-one together with small amounts of naphthoquinones in the secretions of C. ornatum. So far, exclusively naphthoquinones had been reported from a single dyspnoan hitherto studied, Paranemastoma quadripunctatum.     

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.     

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.     

Mimicry: imitative depiction of discharged defensive secretion on carapace of an opilionid

Summary. The opilionid, Parampheres ronae, has a pair of orange markings on its carapace at a location where some other opilionids discharge yellowish, quinone-containing defensive fluid. P. ronae itself produces a defensive secretion, but the fluid is quinone-free and nearly translucent. We suggest that the orange markings in P. ronae are aposematic in the sense that they are imitative of the glandular emissions of quinone-producing opilionids.     

Volatile substances in the pygidial secretion of gyrinid beetles (Coleoptera: Gyrinidae)

Summary Gyrinid beetles are common in freshwater habitats. They have paired pygidial glands with a secretion that contains high molecular weight substances rendering them toxic for predatory fish. In this paper we report on a laboratory study on volatile components released by three different gyrinid species when irritated. The volatile pattern had a clear difference between the different species.Gyrinus substriatus andG. aeratus, both produced 3-methyl-1-butanal and 3-methyl-1-butanol when irritated, but in quite different amounts. The third tested species,G. minutus, did not produce any substances above the detection level. It is suggested that the volatile compounds may be part of the beetles' communication and/or defence system.     

How <Emphasis Type="Italic">Doratomyces stemonitis</Emphasis> copes with Benzoxazolin-2(3<Emphasis Type="Italic">H</Emphasis>)-one (BOA), its derivatives and detoxification products

Summary. Doratomyces stemonitis (Hyphomycetales, Dematiaceae) is a saprotrophic fungus belonging to the mycobiota of the cereal rhizosphere. The fungus is able to metabolize benzoxazolin-2-(3H)-one and a variety of its derivatives including higher plant detoxification products, microbial degradation products and the chemically rather stable 2-amino-(3H)-phenoxazin-3-one. D. stemonitis can use all of these compounds as sole C-sources but their utilization, especially that of microbial degradation products and 2-amino-(3H)-phenoxazin-3-one, seems to be highly energy consuming, resulting in slow mycelium growth and a change of colony morphology. Benzoxazolin-2-(3H)-one derived compounds induce the synthesis of different isoforms of a glycosylated protein with sequence homologies to the endo-1,3-β-glucanase Asp f2, an allergen from Aspergillus fumigatus and other Asp f2-like proteins e.g., from Verticillium dahliae or PRA1 antigen from Candida albicans. The induction of the protein is regarded as a stress response.     

Quinones in cockchafers: additional function of a sex attractant as an antimicrobial agent

Summary. Females of the forest cockchafer, Melolontha hippocastani Fabr. (Coleoptera: Scarabaeidae, Melolonthinae), are known to produce toluquinone and 1,4-benzoquinone. The latter component has been shown in our earlier studies to act as a sex attractant that enhances synergistically the attraction of males towards plant volatiles induced by feeding females. Analyses of whole body extracts from adults and grubs (L3) of M. hippocastani by coupled gas chromatography--mass spectrometry revealed the presence of 1,4-benzoquinone and toluquinone also in males and larvae. Quantitative analyses showed that extracts from females contained significantly more of 1,4-benzoquinone and toluquinone than extracts from males. Remarkably, extracts from grubs contained significantly higher amounts of both compounds than extracts from females. Inhibition assays using Escherichia coli, Saccharomyces cerevisiae, and the entomopathogenic fungi Metarhizium anisopliae and Beauveria brongniartii revealed inhibitory effects of 1,4-benzoquinone and toluquinone against all tested microorganisms. However, the amounts necessary for inhibition of entomopathogenic fungi and S. cerevisiae in the laboratory assay were higher than those found in the extracts from M. hippocastani.     

Spray mechanism of crepidogastrine bombardier beetles (Carabidae; Crepidogastrini)

Summary.   The defensive glandular apparatus of primitive bombardier beetles of the tribe Crepidogastrini (Carabidae) is described for the first time. As exemplified by two African species (Crepidogaster ambreana and C. atrata), the apparatus conforms to the basic bombardier plan, in that the glands are bicompartmented and the secretion is quinonoid (it contains 1,4-benzoquinones and hydrocarbons), hot, and discharged audibly. In a number of morphological respects the crepidogastrine apparatus resembles that of the classical bombardiers of the tribe Brachinini (rather than that of bombardiers of the paussoid lineage), reinforcing the view, already held on taxonomic grounds, that the Crepidogastrini and Brachinini are closely related. That the Crepidogastrini may be primitive relative to Brachinini is underscored by the finding that, unlike brachinines, crepidogastrines do not pulse their secretory emissions. Moreover, they discharge their secretion as a mist, rather than forcibly in the form of jets. Received 22 May 2001; accepted 29 May 2001.     

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.