A comparison of tarsal morphology and traction force in the two burying beetles Nicrophorus nepalensis and Nicrophorus vespilloides (Coleoptera, Silphidae).

Our aim was to compare friction and traction forces between two burying beetle species of the genus Nicrophorus exhibiting different attachment abilities during climbing. Specifically, the interaction of adhesive hairs and claws during attachment with respect to various surface properties was investigated by using a 2 × 3 experimental design. Traction force was measured for two different surface energies (hydrophilic vs hydrophobic) varying in roughness from smooth to micro-rough to rough. Nanotribometric tests on single legs were also performed. The external morphology of the attachment devices investigated by scanning electron microscopy suggested higher intra-specific (intersexual) than inter-specific differences. Whereas differences between the two species in traction force were high on smooth surfaces, no differences could be detected between males and females within each species. With claws intact, both species showed the highest forces on rough surfaces, although N. nepalensis with clipped claws performed best on a smooth surface. However, N. nepalensis beetles outperformed N. vespilloides, which showed no differences between smooth and rough surfaces with clipped claws. Both species demonstrated poor traction forces on micro-rough surfaces. Results concerning the impact of surface polarity were inconclusive, whereas roughness more strongly affected the attachment performance in both species. Nanotribometric analyses of the fore tarsi performed on micro-rough and rough surfaces revealed higher friction in the proximal (pull) direction compared with the distal (push) direction. In these experiments, we detected neither differences in friction performance between the two species, nor clear trends concerning the influence of surface polarity. We conclude that the investigated morphological traits are not critical for the observed interspecific difference in attachment ability on smooth surfaces. Furthermore, interspecific differences in performance are only clear on smooth surfaces and vanish on micro-rough and rough surfaces. Our results suggest that even subtle differences in the adhesion-mediating secretion in closely related species might result in qualitative performance shifts.

Author Correction: Volatiles Emitted by Calling Males of Burying Beetles and Ptomascopus morio (Coleoptera: Silphidae: Nicrophorinae) Are Biogenetically Related.

The original version of this article unfortunately contained a mistake. The name of the main component released by Nicrophorus defodiens in Fig. 1a should read "Fuscumyl acetate", not "Fuscumol".

Volatiles Emitted by Calling Males of Burying Beetles and Ptomascopus morio (Coleoptera: Silphidae: Nicrophorinae) Are Biogenetically Related.

In burying beetles, Nicrophorus spp. (Coleoptera: Silphidae: Nicrophorinae) mate finding is mediated by male produced volatile compounds. To date, pheromone components of only two species have been identified. In an attempt to better understand the evolution of male pheromone signaling in burying beetles, we investigated the male released volatiles of ten Nicrophorus species and one closely related nicrophorine species, Ptomascopus mori. Volatiles emitted by calling males were collected in the laboratory by means of solid phase micro extraction and analyzed using gas chromatography coupled with mass spectrometry. Identified volatiles included short chain esters of 4-methylcarboxylic acids, terpenoids, and some other aliphatic compounds. The long-range volatile signals of the burying beetle species included in this study are blends of two to seven components. We found that methyl or ethyl esters of 4-methylheptanoic acid and 4-methyloctanoic acid are produced by eight of the ten investigated Nicrophorus species. These esters may play a key role in chemical communication. Their widespread occurrence suggests that these compounds did not evolve recently, but appeared relatively early in the phylogeny of the genus. Although Ptomascopus is considered the sister genus of Nicrophorus, P. morio males do not produce any of the Nicrophorus compounds, but release 3-methylalkan-2-ones, which are absent in Nicrophorus. A better understanding of the evolution of burying beetle pheromones, however, will only be possible once more species have been studied.

Phoretic Poecilochirus mites specialize on their burying beetle hosts.

Recurring species interactions can cause species to adapt to each other. Specialization will increase the fitness of symbionts in the coevolved association but may reduce the flexibility of symbiont choice as it will often decrease fitness in interactions with other than the main symbiont species. We analyzed the fitness interactions between a complex of two cryptic mite species and their sympatric burying beetle hosts in a European population. Poecilochirus mites (Mesostigmata, Parasitidae) are phoretic on burying beetles and reproduce alongside beetles, while these care for their offspring at vertebrate carcasses. While Poecilochirus carabi is typically found on Nicrophorus vespilloides beetles, P. necrophori is associated with N. vespillo. It has long been known that the mites discriminate between the two beetle species, but the fitness consequences of this choice remained unknown. We experimentally associated both mite species with both beetle species and found that mite fitness suffered when mites reproduced alongside a nonpreferred host. In turn, there is evidence that one of the beetle species is better able to cope with the mite species they are typically associated with. The overall fitness effect of mites on beetles was negative in our laboratory experiments. The Poecilochirus mites studied here are thus specialized competitors or parasites of burying beetles.

Too fresh is unattractive! The attraction of newly emerged Nicrophorus vespilloides females to odour bouquets of large cadavers at various stages of decomposition.

The necrophagous burying beetle Nicrophorus vespilloides reproduces on small carcasses that are buried underground to serve as food for their offspring. Cadavers that are too large to bury have previously been postulated to be important food sources for newly emerged beetles; however, the attractiveness of distinct successive stages of decomposition were not further specified. Therefore, we investigated the potential preference of newly emerged N. vespilloides females for odour bouquets of piglet cadavers at specific stages of decomposition. Analyses of walking tracks on a Kramer sphere revealed a significantly higher mean walking speed and, consequently, a higher mean total track length when beetles were confronted with odour plumes of the decomposition stages 'post-bloating', 'advanced decay' or 'dry remains' in comparison with the solvent control. Such a change of the walking speed of newly emerged N. vespilloides females indicates a higher motivation to locate such food sources. In contrast to less discriminating individuals this behaviour provides the advantage of not wasting time at unsuitable food sources. Furthermore, in the advanced decay stage, we registered a significantly higher preference of beetles for upwind directions to its specific odour plume when compared with the solvent control. Such a change to upwind walking behaviour increases the likelihood that a large cadaver will be quickly located. Our findings are of general importance for applied forensic entomology: newly emerged N. vespilloides females on large cadavers can and should be regarded as potential indicators of prolonged post mortem intervals as our results clearly show that they prefer emitted odour bouquets of later decomposition stages.

Fitness costs associated with chemical signaling.

The production, maintenance and transmission of chemical signals often entail costs. Costs can arise, for instance, if signal production depends on the availability of limited nutritional resources or if signal transmission leads to attraction of predators. Many species effectively reduce these costs by signaling at specific times or in certain contexts only. We previously reported that breeding burying beetle females facultatively adjust their pheromone emission in response to their social environment, emitting high amounts of their chemical signal in the presence of a male partner, but not when providing uniparental care. Here we present data showing that chemical signaling is costly, and that higher investments in signaling result in reduced clutch sizes, but not a shorter life span, in the burying beetle Nicrophorus vespilloides.

Social environment determines degree of chemical signalling.

Few studies have attempted to distinguish between cues and signals in the context of chemical communication. A number of chemical substances have been shown to vary with physiological state, such as stage of oestrus cycle, fertility, dominance status or nutritional condition, but little is known about whether this variation is incidental or adaptive. Here, we provide evidence of a substance whose emission varies with breeding state, but is not merely an incidental by-product of physiological state, but rather, an evolved signal. Breeding females of the facultative biparental burying beetle, Nicrophorus vespilloides, release methyl geranate, a substance that helps males to identify breeding status and to distinguish between their female partners and non-breeding intruders. We demonstrate that females respond flexibly to their social environment and emit high amounts of methyl geranate only in the presence of a male partner, i.e. a receiver. In contrast, cuticular hydrocarbons, which also have been shown to change with breeding status, are not modulated and do not differ between single and paired breeding females. Receiver-dependent chemical signalling is expected to evolve when costs are involved in the production or transmission of the signal; such signal modulation might be more common than previously thought.

Starving the competition: a proximate cause of reproductive skew in burying beetles (Nicrophorus vespilloides).

Proximate mechanisms underlying reproductive skew are obscure in many animals that breed communally. Here, we address causes of reproductive skew in brood-parasitic associations of burying beetles (Nicrophorus vespilloides). Male and female burying beetles feed and defend their larvae on buried carcasses. When several females locate the same small carcass, they engage in violent physical altercations. The subordinate then acts as an intraspecific brood parasite, laying eggs, but not providing care. The dominant female largely monopolizes access to the carcass; she alone provides parental care and her share of the brood is much larger than the subordinate's. On larger carcasses, subordinates have greater access to the carcass than on small ones, and reproductive skew is reduced. Differential fecundity, ovicide and larvicide have been suggested as causes of skew on small carcasses. Here, we report the results of the experiments pertaining to the first two of these potential mechanisms. Ovicide did not significantly contribute to reproductive skew on small carcasses, but differential fecundity did. Fecundity differences were due to dominance status, not body size per se. Fecundity differences disappeared when supplemental food was available, suggesting that reduced access to the carcass limits fecundity by causing nutritional deficiencies. Supplemental food prevented such nutritional deficiencies and allowed subordinates to produce as many eggs as dominants. Apparently, aggressive behaviour by dominants functions in the context of reproductive competition, limiting subordinate reproduction by preventing food intake on the carcass.

The Coolidge effect, individual recognition and selection for distinctive cuticular signatures in a burying beetle.

The ability to recognize individuals is an important aspect of social interactions, but it can also be useful to avoid repeated matings with the same individual. The Coolidge effect is the progressive decline in a male's propensity to mate with the same female combined with a heightened sexual interest in new females. Although males that recognize previous partners and show a preference for novel females should have a selective advantage as they can distribute sperm evenly among the females they encounter, there are few invertebrate examples of the Coolidge effect. Here we present evidence for this effect in the burying beetle Nicrophorus vespilloides and examine the mechanism underlying the discrimination between familiar and novel mates. Burying beetles feed and reproduce on vertebrate carcasses, where they regularly encounter conspecifics. Males showed greater sexual interest in novel females (virgin or mated) than in females they had inseminated before. The application of identical cuticular extracts allowed us to experimentally create females with similar odours, and male responses to such females demonstrated that they use female cuticular patterns for discrimination. The chemical analysis of the cuticular profile revealed greater inter-individual variation in female than in male cuticular patterns, which might be due to greater selection on females to signal their individual identity.

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.

Maternal nutritional condition and genetic differentiation affect brood size and offspring body size in Nicrophorus.

In most animal species, brood size and body size exhibit some variation within and between populations. This is also true for burying beetles (genus Nicrophorus), a group in which the body size of offspring depends critically on the number of offspring competing for food due to the discrete nature of resource used for larval nutrition (vertebrate carcasses). In one species, brood size and body size are correlated with population density, and appear to be phenotypically plastic. We investigated potential proximate causes of between-population variation in brood size and body size in two species, Nicrophorus vespilloides and Nicrophorus defodiens. Our first experiment supported the notion that brood size is phenotypically plastic, because it was affected by environmental variation in adult nutritional condition. We found that the pre-breeding nutritional status of female N. vespilloides affected the number of eggs they laid, the number of surviving larvae in their broods, and the body size of their offspring. We do not know whether this plasticity is adaptive because greater offspring body size confers an advantage in contests over breeding resources, or whether starved females are constrained to produce smaller clutches because they cannot fully compensate for their poor pre-breeding nutritional status by feeding from the carcass. Our second experiment documents that brood size, specifically the infanticidal brood-size adjustment behavior, has undergone genetic differentiation between two populations of N. defodiens. Even under identical breeding conditions with identical numbers of first-instar larvae, females descended from the two populations produced broods of different size with corresponding differences in offspring body size.

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.

The dispersal behaviour of the phoretic mite Poecilochirus carabi (Mesostigmata, Parasitidae): adaptation to the breeding biology of its carrier Necrophorus vespilloides (Coleoptera, Silphidae).

When the phoretic mite Poecilochirus carabi reproduces in the brood chamber of its carrier Necrophorus vespilloides, a beetle with biparental brood care, the first deuteronymphs of the new mite generation aggregate on the male beetle. They do not use sex-specific traits to discriminate between male and female beetles in the brood chamber, but traits that are related to the beetles' behaviour and may be displayed by both parent beetles. When the male beetle departs, it carries virtually all deuteronymphs then present in the brood chamber. Deuteronymphs that develop later congregate on the female, which leaves the crypt some days after the male. Only those deuteronymphs that miss the female's departure disperse on the beetle larvae, meaning they have to wait in their pupal chambers until the beetles have completed their development. On average, 86% of the deuteronymphs leave the brood chamber on the parent beetles, thereby gaining the advantage of an early departure. As soon as their carrier arrives at one of the beetles' meeting places, the deuteronymphs can transfer between the beetles present. Choice experiments revealed that the deuteronymphs tend to even out density differences between congregating carriers, and prefer sexually mature to immature beetles. Therefore, transferring between beetles results in a dispersion of deuteronymphs on the sexually mature beetles of the population.