A contact anti-aphrodisiac pheromone supplied by the spermatophore in the rove beetle Aleochara curtula: mode of transfer and evolutionary significance.

By reducing the attractiveness of their mating partner via an anti-aphrodisiac pheromone, males can prevent a remating of the female and thus reduce the risk of sperm competition. For females, the main benefit from allowing the chemical manipulation of their attractiveness is probably the avoidance of sexual harassments from rival males. While mating plugs generally constitute a physical barrier which hinders male mating attempts, chemical manipulations must trustfully inform the responding male of the female's reluctance to mate; otherwise, it would be beneficial to ignore the repellent information. In our experiments, males of the polyandrous rove beetle Aleochara curtula chemically manipulated the attractiveness of their mating partner. Coincident with the deposition of a spermatophore into the female genital chamber, an anti-aphrodisiac pheromone was transferred and readily spread onto the female surface, where it was subsequently perceived by rival males via parameres, the claspers of the male genitalia. Males aborted contact with the mated female to avoid further time- and energy-consuming elements of the mating sequence. The chemical mode of action was demonstrated inter alia by spicing virgin females with spermatophore extracts. The action of the anti-aphrodisiac correlated with the persistence of the spermatophore in the female genital chamber and corresponded to the length of stay of the mated female at a carcass, where the density of rival males is highest. The ensuing benefits for all three parties involved in this communication system, which render this post-copulatory mate guarding strategy evolutionary stable, are discussed.

Interaction of liquid epicuticular hydrocarbons and tarsal adhesive secretion in Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae).

Species of various insect orders possess specialised tarsal adhesive structures covered by a thin liquid film, which is deposited in the form of footprints. This adhesive liquid has been suggested to be chemically and physiologically related to the epicuticular lipid layer, which naturally covers the body of insects and acts as the prime barrier to environmental stresses, such as desiccation. The functional efficiency of the layer, however, is jeopardised by partial melting that may occur at physiological temperatures. In this study, light microscopic images of elytral prints show that the epicuticular lipid layer of the Colorado potato beetle Leptinotarsa decemlineata actually is partially liquid and chemical investigations reveal the high similarity of the epicuticular hydrocarbon pattern and the tarsal liquid. By means of chemical manipulation of the surface hydrocarbon composition of live beetles, the substance exchange between their tarsal adhesive hairs and the body surface is monitored. Histological sections of L. decemlineata tarsi, furthermore, reveal glandular cells connected to individual adhesive setae and departing from these results, an idea of a general mechanism of tarsal secretion is developed and discussed in a functional-ecological context.

Impact of chemical manipulation of tarsal liquids on attachment in the Colorado potato beetle, Leptinotarsa decemlineata.

Insect tarsal attachment forces are thought to be influenced by the viscosity and surface tension of a thin film of adhesive liquid (wet adhesion). In beetles, this fluid has been shown to be composed mainly of lipophilic substances that are similar to the cuticular lipids. In this study we investigate whether and how the chemical composition of footprint lipids affects attachment forces in the Colorado potato beetle, Leptinotarsa decemlineata. After application of standardised mixtures of synthetic n-alkanes or alkenes, or a concentrated hydrocarbon extract to the surface of the elytra, we tested the beetles' attachment performance using a beam force transducer. The results show that only the unsaturated components, but not the straight-chained alkanes reduced friction forces, confirming that attachment performance is influenced by the chemical composition of the adhesive secretion. We estimated the volume of footprint droplets and calculated a mean thickness of the liquid layer of 0.04 microm. The measured friction exceeded the viscous and capillary force expected for a film of this thickness. Therefore, alternative mechanisms (i.e. shear-thinning and solid-like behaviour) for the generation of attachment forces and their dependence on the chemical composition of the liquid are discussed.

A sex pheromone in the desert tenebrionid beetle Parastizopus armaticeps.

Males of the desert beetle Parastizopus armaticeps (Pér.) (Coleoptera: Tenebrionidae) exhibit a characteristic calling behavior that attracts females by raising the tip of the abdomen, exposing the aedeagus, and remaining in this posture for a few seconds while emitting a pheromone. We collected the pheromone by holding a solid phase microextraction fiber (100 mum polydimethylsiloxane) close to the aedeagus for 5 s and analyzed the volatiles collected by gas chromatography/mass spectrometry. The volatiles consisted of 3-methylphenol (52%), ethyl-1,4-benzoquinone (48%), and 3-ethylphenol (2%). The pheromone originated from the aedeagal glands. In the gland reservoirs, these compounds (2.1%) were mixed with ethyl, isopropyl, and propyl esters of fatty acids (24.2%), and a mixture of hydrocarbons (69.1%). The mean amount of volatiles extracted from gland reservoirs was 0.92 +/- 0.83 microg. Chemo-orientation experiments with a servosphere show that females responded only to the ternary volatile mixture. Females stopped walking, elevated the front parts of their bodies with erected antennae, turned slowly on their own axis, and walked upwind toward the odor source. Single components or binary mixtures did not elicit responses from females. Males did not respond to the pheromone. Evolutionary aspects of this pheromone system are discussed.

The chemistry of the postpharyngeal gland of female European beewolves.

Females of the European beewolf, Philanthus triangulum, possess a large glove-shaped gland in the head, the postpharyngeal gland (PPG). They apply the content of the PPG to their prey, paralyzed honeybees, where it delays fungal infestation. Here, we describe the chemical composition of the gland by using combined GC-MS, GC-FTIR, and derivatization. The PPG of beewolves contains mainly long-chain unsaturated hydrocarbons (C23-C33), lower amounts of saturated hydrocarbons (C14-C33), and minor amounts of methyl-branched hydrocarbons (C17-C31). Additionally, the hexane-soluble gland content is comprised of small amounts of an unsaturated C25 alcohol, an unknown sesquiterpene, an octadecenylmethylester, and several long-chain saturated (C25, C27) and unsaturated (C23-C27) ketones, some of which have not yet been reported as natural products. Surprisingly, we found a dimorphism with regard to the major component of the PPG with some females having (Z)-9-pentacosene, whereas others have (Z)-9-heptacosene as their predominant component. The biological relevance of the compounds for the prevention of fungal growth on the prey and the significance of the chemical dimorphism are discussed.

Ethyl 4-methyl heptanoate: a male-produced pheromone of Nicrophorus vespilloides.

Sexually mature male beetles of the genus Nicrophorus (Coleoptera: Silphidae) exhibit a conspicuous behavior, recognized as pheromone-releasing activity. Laboratory and field studies demonstrated that females are attracted to males that exhibit this behavior, both on or off reproductive resources. Here, we report the results of a study in which volatile chemicals released by calling Nicrophorus vespilloides were collected by solid-phase microextraction and analyzed by using coupled gas chromatography-mass spectrometry. These analyses revealed that ethyl 4-methyl heptanoate and (E)-geranylacetone are emitted by males that engage in the behavior. In the field, traps baited with racemic ethyl 4-methyl heptanoate caught roughly equal numbers of male and female N. vespilloides. Some male and female Nicrophorus vespillo and male Nicrophorus humator were also caught in traps baited with this compound. Traps baited with (E)-geranylacetone did not catch significant numbers of beetles.

1-Tridecene--male-produced sex pheromone of the tenebrionid beetle Parastizopus transgariepinus.

Males of the genus Parastizopus (Coleoptera: Tenebrionidae) exhibit a special pheromone-emitting behaviour. They do a headstand, expose the aedeagus and remain in this posture for a few seconds. The pheromone emitted by P. transgariepinus was collected by solid-phase micro-extraction (100 microm polydimethylsiloxane fibre) and identified as 1-tridecene by gas chromatography/mass spectrometry. Presumably, this compound originates from the aedeagal gland, a special feature in Parastizopus, as 1-tridecene is the main compound in the gland reservoirs (23.6+/-3.8%), accompanied by various less volatile fatty acid esters (25.2+/-2.0%) and hydrocarbons (51.2+/-5.7%). 1-Tridecene is also part of the pygidial defensive secretion of both sexes, together with other 1-alkenes, monoterpene hydrocarbons and 1,4-benzoquinones, but as none of these other compounds was detected during calling, the pygidial gland could be ruled out as pheromone source. Extracts of the aedeagal gland reservoirs and the pygidial defensive secretion contained comparable amounts of 1-tridecene, 1.24+/-0.41 and 1.88+/-0.54 microg/male, respectively. Chemo-orientation experiments using a servosphere showed that 1 microg of 1-tridecene was attractive to females but not to males.

The smell of parents: breeding status influences cuticular hydrocarbon pattern in the burying beetle Nicrophorus vespilloides.

The waxy layer of the cuticle has been shown to play a fundamental role in recognition systems of insects. The biparental burying beetle Nicrophorus vespilloides is known to have the ability to discriminate between breeding and non-breeding conspecifics and also here cuticular substances could function as recognition cue. However, it has not yet been demonstrated that the pattern of cuticular lipids can reflect the breeding status of a beetle or of any other insect. With chemical analysis using coupled gas chromatography-mass spectrometry, we showed that the chemical signature of N. vespilloides males and females is highly complex and changes its feature with breeding status. Parental beetles were characterized by a higher amount of some unusual unsaturated hydrocarbons than beetles which are not caring for larvae. The striking correlation between cuticular profiles and breeding status suggests that cuticular hydrocarbons inform the beetles about parental state and thus enable them to discriminate between their breeding partner and a conspecific intruder. Furthermore, we found evidence that nutritional conditions also influence the cuticular profile and discuss the possibility that the diet provides the precursors for the unsaturated hydrocarbons observed in parental beetles. Our study underlines the fact that the cuticular pattern is rich of information and plays a central role in the burying beetles' communication systems.

A review of myrmecophily in ant nest beetles (Coleoptera: Carabidae: Paussinae): linking early observations with recent findings.

Myrmecophily provides various examples of how social structures can be overcome to exploit vast and well-protected resources. Ant nest beetles (Paussinae) are particularly well suited for ecological and evolutionary considerations in the context of association with ants because life habits within the subfamily range from free-living and predatory in basal taxa to obligatory myrmecophily in derived Paussini. Adult Paussini are accepted in the ant society, although parasitising the colony by preying on ant brood. Host species mainly belong to the ant families Myrmicinae and Formicinae, but at least several paussine genera are not host-specific. Morphological adaptations, such as special glands and associated tufts of hair (trichomes), characterise Paussini as typical myrmecophiles and lead to two different strategical types of body shape: while certain Paussini rely on the protective type with less exposed extremities, other genera access ant colonies using glandular secretions and trichomes (symphile type). We compare these adaptations with other taxonomic groups of insects by joining contemporary research and early sources and discuss the possibility of an attracting or appeasing effect of the secretion. Species that are ignored by their host ants might use chemical mimicry instead. Furthermore, vibrational signals may contribute to ant-beetle communication, and chemical signals have proven to play a role in host finding. The powerful defense chemistry of paussines as "bombardier beetles" is not used in contact with host ants. We attempt to trace the evolution of myrmecophily in paussines by reviewing important aspects of the association between paussine beetles and ants, i.e. morphological and potential chemical adaptations, life cycle, host specificity, alimentation, parasitism and sound production.

Queue up, please! Spermathecal filling in the rove beetle Drusilla canaliculata (Coleoptera, Staphylinidae).

Most studies on insect sperm motility have been conducted in vitro using artificial environments outside the animal's body. Only little is known about the function of motile insect sperm at different sites within the male or female genital tracts. We dissected genital tracts of female rove beetles (Drusilla canaliculata) to show that spermatozoa use their own motility to migrate from the spermatophore into the spermatheca. Our dissection method allowed direct observation and filming of the spermathecal filling process inside the female's genital tract. Spermatozoa were found to enter the spermatheca individually, sometimes in groups of two or three. Although exhibiting only weak motility and no progressive motion in buffer solution, the spermatozoa inside the female show vigorous lashing and reach an average velocity of 47.5 microm s(-1). To gain mobility and speed, the spermatozoa likely utilize the relatively small diameter of the spermathecal duct to push themselves off the duct walls, rather than swimming freely in seminal fluid. The spermatozoa (approximately 1,250 microm) are considerably longer than the distance they have to travel along the spermathecal duct (approximately 800 microm). Our study provides the first direct observation of active sperm migration within the female of an insect stressing the importance of the genital tract as a prerequisite for functional sperm motility.

Food wrapping with the postpharyngeal gland secretion by females of the European beewolf Philanthus triangulum.

Ground-nesting animals share their habitat with countless microorganisms that can play important roles as pathogens or competitors for food resources. Thus, species that store food in the soil, either for themselves or for their progeny, must protect these resources against microbial degradation. Females of the European beewolf, Philanthus triangulum, hunt honeybees as provisions for their brood and store the paralyzed prey in their subterranean nests. A previous study had shown that females lick the surface of prey before oviposition and that this licking treatment delays mold growth. Here, we showed that females apply large amounts of a secretion from their postpharyngeal glands onto the surface of their prey during the licking behavior. Inhibition-zone assays showed that comparatively large amounts of the gland secretion had no direct antimycotic effect. We discuss our findings with regard to other possible mechanisms of the postpharyngeal gland secretion against fungal growth.

On-line monitoring and identification of bioaerosols.

Fast analysis of bioaerosols in clean room environments is necessary in order to prevent contamination of pharmaceutical products, minimize machine downtimes, or both. The detection and identification of microbes will be carried out in several steps: After impaction of the aerosol on a surface, the particles are presorted with glancing light illumination and fluorescence imaging in order to distinguish between abiotic and biotic particles. Since only the biotic particles are of interest, the analysis time can be minimized due to reduction of the data set. The biotic particles are then analyzed further with Raman spectroscopy and identified with a support vector machine.

Chemotaxonomic identification of single bacteria by micro-Raman spectroscopy: application to clean-room-relevant biological contaminations.

Microorganisms, such as bacteria, which might be present as contamination inside an industrial food or pharmaceutical clean room process need to be identified on short time scales in order to minimize possible health hazards as well as production downtimes causing financial deficits. Here we describe the first results of single-particle micro-Raman measurements in combination with a classification method, the so-called support vector machine technique, allowing for a fast, reliable, and nondestructive online identification method for single bacteria.

Ultrastructure and motility pattern of the spermatozoa of Aleochara curtula (Coleoptera, Staphylinidae).

Ultrastructure and motility pattern of spermatozoa of the rove beetle Aleochara curtula were examined using electron and light microscopic methods. The spermatozoon is about 100 microm long and filiform. The head piece comprises a 5 microm long triple layered acrosome and 10 microm long nucleus. The flagellum consists of a 9+9+2 axoneme, two accessory bodies and two mitochondrial derivatives about equal in size but of different shape in their cross sections. In both derivatives there are paracrystalline inclusions. The flagellum is attached to the head by a 2 microm long centriole adjunct which is characterized by its electron dense material that forms a three layered folded lamellar structure. When liberated in buffer solution the sperm flagella assume a coiled hook-like form with the excentric stiff head protruding in front. The spermatozoa are driven through the medium by a small helicoidal wave of high frequency superimposed to the bent flagella. The maximum speed measured was 15.2 microm/s. The sperm architecture of A. curtula is similar to that of other Aleochara species but differs in total length and dimensions of the mitochondrial derivatives. For that reason Aleochara sperm can certainly prove useful to study the effect of the mitochondrial derivatives on sperm motility.