Factors influencing carrion communities are only partially consistent with those of deadwood necromass.

Research on decomposer communities has traditionally focused on plant litter or deadwood. Even though carrion forms highly nutrient-rich necromass that enhance ecosystem heterogeneity, the factors influencing saprophytic communities remain largely unknown. For deadwood, experiments have shown that different drivers determine beetles (i.e., decay stage, microclimate, and space), fungi (i.e., decay stage and tree species) and bacteria (decay stage only) assemblages. To test the hypothesis that similar factors also structure carrion communities, we sampled 29 carcasses exposed for 30 days that included Cervus elaphus (N = 6), Capreolus capreolus (N = 18), and Vulpes vulpes (N = 5) in a mountain forest throughout decomposition. Beetles were collected with pitfall traps, while microbial communities were characterized using amplicon sequencing. Assemblages were determined with a focus from rare to dominant species using Hill numbers. With increasing focus on dominant species, the relative importance of carcass identity on beetles and space on bacteria increased, while only succession and microclimate remained relevant for fungi. For beetle and bacteria with focus on dominant species, host identity was more important than microclimate, which is in marked contrast to deadwood. We conclude that factors influencing carrion saprophytic assemblages show some consistency, but also differences from those of deadwood assemblages, suggesting that short-lived carrion and long-lasting deadwood both provide a resource pulse with different adaptions in insects and microbes. As with deadwood, a high diversity of carcass species under multiple decay stages and different microclimates support a diverse decomposer community.

Dung-visiting beetle diversity is mainly affected by land use, while community specialization is driven by climate.

Dung beetles are important actors in the self-regulation of ecosystems by driving nutrient cycling, bioturbation, and pest suppression. Urbanization and the sprawl of agricultural areas, however, destroy natural habitats and may threaten dung beetle diversity. In addition, climate change may cause shifts in geographical distribution and community composition. We used a space-for-time approach to test the effects of land use and climate on α-diversity, local community specialization (H 2') on dung resources, and γ-diversity of dung-visiting beetles. For this, we used pitfall traps baited with four different dung types at 115 study sites, distributed over a spatial extent of 300 km × 300 km and 1000 m in elevation. Study sites were established in four local land-use types: forests, grasslands, arable sites, and settlements, embedded in near-natural, agricultural, or urban landscapes. Our results show that abundance and species density of dung-visiting beetles were negatively affected by agricultural land use at both spatial scales, whereas γ-diversity at the local scale was negatively affected by settlements and on a landscape scale equally by agricultural and urban land use. Increasing precipitation diminished dung-visiting beetle abundance, and higher temperatures reduced community specialization on dung types and γ-diversity. These results indicate that intensive land use and high temperatures may cause a loss in dung-visiting beetle diversity and alter community networks. A decrease in dung-visiting beetle diversity may disturb decomposition processes at both local and landscape scales and alter ecosystem functioning, which may lead to drastic ecological and economic damage.

Linking bacteria, volatiles and insects on carrion: the role of temporal and spatial factors regulating inter-kingdom communication via volatiles.

Multi-kingdom community complexity and the chemically mediated dynamics between bacteria and insects have recently received increased attention in carrion research. However, the strength of these inter-kingdom interactions and the factors that regulate them are poorly studied. We used 75 piglet cadavers across three forest regions to survey the relationship between three actors (epinecrotic bacteria, volatile organic compounds (VOCs) and flies) during the first 4 days of decomposition and the factors that regulate this interdependence. The results showed a dynamic bacterial change during decomposition (temperature-time index) and across the forest management gradient, but not between regions. Similarly, VOC emission was dynamic across a temperature-time index and the forest management gradient but did not differ between regions. However, fly occurrence was dynamic across both space and time. The strong interdependence between the three actors was mainly regulated by the temperature-time index and the study regions, thereby revealing regulation at temporal and spatial scales. Additionally, the actor interdependence was stable across a gradient of forest management intensity. By combining different actors of decomposition, we have expanded our knowledge of the holistic mechanisms regulating carrion community dynamics and inter-kingdom interactions, an important precondition for better describing food web dynamics and entire ecosystem functions.

Carcasses at Fixed Locations Host a Higher Diversity of Necrophilous Beetles.

In contrast to other necromass, such as leaves, deadwood, or dung, the drivers of insect biodiversity on carcasses are still incompletely understood. For vertebrate scavengers, a richer community was shown for randomly placed carcasses, due to lower competition. Here we tested if scavenging beetles similarly show a higher diversity at randomly placed carcasses compared to easily manageable fixed places. We sampled 12,879 individuals and 92 species of scavenging beetles attracted to 17 randomly and 12 at fixed places exposed and decomposing carcasses of red deer, roe deer, and red foxes compared to control sites in a low range mountain forest. We used rarefaction-extrapolation curves along the Hill-series to weight diversity from rare to dominant species and indicator species analysis to identify differences between placement types, the decay stage, and carrion species. Beetle diversity decreased from fixed to random locations, becoming increasingly pronounced with weighting of dominant species. In addition, we found only two indicator species for exposure location type, both representative of fixed placement locations and both red listed species, namely Omosita depressa and Necrobia violacea. Furthermore, we identified three indicator species of Staphylinidae (Philonthus marginatus and Oxytelus laqueatus) and Scarabaeidae (Melinopterus prodromus) for larger carrion and one geotrupid species Anoplotrupes stercorosus for advanced decomposition stages. Our study shows that necrophilous insect diversity patterns on carcasses over decomposition follow different mechanisms than those of vertebrate scavengers with permanently established carrion islands as important habitats for a diverse and threatened insect fauna.

Temporal variability of the rove beetle (Coleoptera: Staphylinidae) community on small vertebrate carrion and its potential use for forensic entomology.

Rove beetles (Coleoptera: Staphylinidae) are one of the most important colonizers of vertebrate cadavers. We have previously investigated carrion-associated rove beetle communities across various forests and demonstrated that the study regions are the main drivers of the local rove beetle species pool that is, in turn, able to colonize a carcass. Nevertheless, little is known about their temporal variation in community composition during the decomposition process. The aim of our study has been to analyze the temporal changes of the composition of the rove beetle community and to identify new, potentially useful candidate species for forensic entomological evaluations. We determined 80 rove beetle species that were attracted to 60 piglet cadavers across various forest stands in Germany. Both the abundance and the community composition changed over the decomposition process, independently of the species-specific geographical variation across study regions. In the region Schorfheide-Chorin, species from the genus Philonthus proved to be a suitable group for future forensic entomological investigations. They appeared in markedly high numbers at piglet cadavers from the bloated stage until the advanced decay stage. For the study region Schwäbische Alb, we newly describe the species Omalium septentrionis as a member of the carrion-associated rove beetle fauna. Based on the geographical variation in rove beetle community compositions, we have filtered out several species that might contribute to advances in postmortem interval estimations or other applied fields in forensic entomological contexts.

Local and Landscape Effects on Carrion-Associated Rove Beetle (Coleoptera: Staphylinidae) Communities in German Forests.

Intensification of anthropogenic land use is a major threat to biodiversity and thus to essential ecosystem services provided by insects. Rove beetles (Coleoptera: Staphylinidae), which react sensitively to habitat changes, are species-rich colonizers of vertebrate cadavers and contribute to the important ecosystem service of carrion decomposition. The unveiling of anthropogenic and environmental drivers that modify carrion-associated rove beetle communities should improve our understanding of the plasticity of cadaver decay. We report the presence of 80 rove beetle species on 65 decomposing piglet cadavers at forest sites characterized by a gradient of management intensity across three geographic regions in Germany. Local and landscape drivers were revealed that shape beetle abundance, diversity, and community composition. Forest management and regions affect rove beetle abundance, whereas diversity is influenced by local habitat parameters (soil pH, litter cover) and regions. The community composition of rove beetles changes with management intensification by promoting generalist species. Regarding single species, Philonthus decorus and Anotylus mutator are linked to unmanaged forests and Ontholestes tessellatus to highly used forest stands. The spatial information provided about carrion-associated rove beetle communities in German forests is not only of carrion-ecological but also of forensic entomological interest.

The Attraction of the Dung Beetle Anoplotrupes stercorosus (Coleoptera: Geotrupidae) to Volatiles from Vertebrate Cadavers.

During decomposition, vertebrate carrion emits volatile organic compounds to which insects and other scavengers are attracted. We have previously found that the dung beetle, Anoplotrupes stercorosus, is the most common dung beetle found on vertebrate cadavers. Our aim in this study was to identify volatile key compounds emitted from carrion and used by A. stercorosus to locate this nutritive resource. By collecting cadaveric volatiles and performing electroantennographic detection, we tested which compounds A. stercorosus perceived in the post-bloating decomposition stage. Receptors in the antennae of A. stercorosus responded to 24 volatiles in odor bouquets from post-bloating decay. Subsequently, we produced a synthetic cadaver odor bouquet consisting of six compounds (benzaldehyde, DMTS, 3-octanone, 6-methyl-5-hepten-2-ol, nonanal, dodecane) perceived by the beetles and used various blends to attract A. stercorosus in German forests. In field assays, these beetles were attracted to a blend of DMTS, 3-octanone, and benzaldehyde. Generalist feeding behavior might lead to the super-dominant occurrence of A. stercorosus in temperate European forests and have a potentially large impact on the exploitation and rapid turnover of temporally limited resources such as vertebrate cadavers.

The Impact of Environmental Factors on the Efficacy of Chemical Communication in the Burying Beetle (Coleoptera: Silphidae).

There is growing evidence that a wide range of insect sex pheromones are condition dependent and play a fundamental role in mate choice. However, the effectiveness of pheromonal communication might not only depend on internal factors of the sender, but also on attributes of the microhabitat, in which the signaler chooses to emit its chemical signal. For example, the degree of anthropogenic land use might affect how successful the signal is transmitted, as land use has been shown to affect animal communities and the complexity of biotic interactions. To test the hypothesis that parameters of the microenvironment determine males' ability to attract females via their sex pheromone, we used the burying beetle Nicrophorus vespilloides Herbst (Coleoptera: Silphidae) as our model system. We exposed 144 males across differently managed forest stands and analyzed the impact of 29 environmental parameters. Our data revealed that human land use intensity had no effect on a male's attractiveness. However, the harvested tree biomass positively affected the proportion of competitors attracted. Furthermore, we found that soil characteristics were important factors determining the amount and body size of females a male was able to attract. Consequently, we present evidence that the environmental context of a signaling male influences the effectiveness of chemical signaling either because it affects the transmission process or the prevailing abundance of potential signal receivers. Thus, our results demonstrate that males need to make careful decisions about the location where they emit their pheromone, as this choice of microhabitat has an impact on their fitness.

Forest habitat parameters influence abundance and diversity of cadaver-visiting dung beetles in Central Europe.

Dung beetles provide crucial ecosystem services and serve as model organisms for various behavioural, ecological and evolutionary studies. However, dung beetles have received little attention as consumers of large cadavers. In this study, we trapped copronecrophagous dung beetles on above-ground exposed piglet cadavers in 61 forest plots distributed over three geographically distinct regions in Germany, Central Europe. We examined the effects of land use intensity, forest stand, soil characteristics, vascular plant diversity and climatic conditions on dung beetle abundance, species richness and diversity. In all three regions, dung beetles, represented mainly by the geotrupid species Anoplotrupes stercorosus and Trypocopris vernalis, were attracted to the cadavers. High beetle abundance was associated with higher mean ambient temperature. Furthermore, A. stercorosus and T. vernalis were more abundant in areas where soil contained higher proportions of fine sand. Additionally, an increased proportion of forest understorey vegetation and vascular plant diversity positively affected the species richness and diversity of dung beetles. Thus, even in warm dry monocultured forest stands exploited for timber, we found thriving dung beetle populations when a diverse understorey was present. Therefore, forestry practices that preserve the understorey can sustain stable dung beetle populations and ensure their important contribution to nutrient cycles.

Effects of abiotic environmental factors and land use on the diversity of carrion-visiting silphid beetles (Coleoptera: Silphidae): A large scale carrion study.

Anthropogenic land use causes global declines in biodiversity. Despite the knowledge that animal carrion is the most nutrient-rich form of dead organic matter, studies on landscape and local scales determining whether and the means by which land use intensity influences the diversity of the carrion-associated insect fauna are globally scarce. We investigated the effects of land use intensity and abiotic and biotic environmental factors on the abundance, species richness, and diversity of the important ecosystem-service-providing silphid beetle taxon (carrion beetles) in three regions of Germany. In 61 forest stands distributed over three geographically distinct regions in Central Europe, we trapped silphid beetles on exposed piglet cadavers during late summer. In all three regions, higher ambient temperatures and higher fine sand contents were associated with the abundance of the silphid beetle taxa. The carrion community silphid diversity was negatively affected by an increase in mean ambient temperature in all three regions. Although management intensity in forests did not affect the overall abundance of Silphidae, the abundance of Nicrophorus humator decreased significantly with higher forest management intensity across all three regions. Unmanaged and age-class forests showed a higher abundance of N. humator compared with extensively managed forest stands. These findings indicate that N. humator has potential as an indicator species for anthropogenic disturbances in forests. Overall, the direct responses of the silphid beetle community to diverse soil characteristics underline soil as an important factor determining the abundance and diversity of necrophagous carrion beetles in Central Europe. To protect these valuable ecosystem-service providers, forest-management-induced soil modifications need to be paid close attention.

Function of bacterial community dynamics in the formation of cadaveric semiochemicals during in situ carcass decomposition.

The decomposition of dead mammalian tissue involves a complex temporal succession of epinecrotic bacteria. Microbial activity may release different cadaveric volatile organic compounds which in turn attract other key players of carcass decomposition such as scavenger insects. To elucidate the dynamics and potential functions of epinecrotic bacteria on carcasses, we monitored bacterial communities developing on still-born piglets incubated in different forest ecosystems by combining high-throughput Illumina 16S rRNA sequencing with gas chromatography-mass spectrometry of volatiles. Our results show that the community structure of epinecrotic bacteria and the types of cadaveric volatile compounds released over the time course of decomposition are driven by deterministic rather than stochastic processes. Individual cadaveric volatile organic compounds were correlated with specific taxa during the first stages of decomposition which are dominated by bacteria. Through best-fitting multiple linear regression models, the synthesis of acetic acid, indole and phenol could be linked to the activity of Enterobacteriaceae, Tissierellaceae and Xanthomonadaceae, respectively. These conclusions are also commensurate with the metabolism described for the dominant taxa identified for these families. The predictable nature of in situ synthesis of cadaveric volatile organic compounds by epinecrotic bacteria provides a new basis for future chemical ecology and forensic studies.

Volatile Organic Compounds of Decaying Piglet Cadavers Perceived by Nicrophorus vespilloides.

In the necrophagous burying beetle Nicrophorus vespilloides (Coleoptera: Silphidae), cadaver preference appears to depend on cadaver size and on the maturity of the beetle. We previously showed that newly emerged females with immature ovaries prefer later stages of decomposition of large cadavers. Our present aim is the determination of specific chemical compounds involved in the discrimination of cadaveric odor bouquets and the recognition of specific stages of decomposition. We used headspace samples of maggot-infested piglet cadavers at various decomposition stages and performed gas chromatography coupled with electroantennography (GC-EAD) to record 45 EAD-active compounds. Using GC coupled with mass spectrometry, we identified 13 of the EAD-active compounds. The headspace of the fresh decomposition stage was characterized mainly by high relative amounts of trimethylpyrazine. High relative amounts of dimethyl trisulfide were characteristic of bloated, post-bloating, and advanced decay stages. The advanced decay and dry remains stages were dominated by high relative amounts of phenol. Statistically, this compound had the highest impact on discrimination between the fresh decomposition stage, which is important for mature burying beetles for reproduction, and the advanced decay stage, which is nutritionally more attractive for newly emerged beetles. Phenol might, therefore, function as a key substance for newly emerged female burying beetles, so that they can locate suitable cadavers for feeding to maturation.

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.

The attraction of virgin female hide beetles (Dermestes maculatus) to cadavers by a combination of decomposition odour and male sex pheromones.

INTRODUCTION: The hide beetle Dermestes maculatus (Coleoptera: Dermestidae) feeds as an adult and larva on decomposing animal remains and can also be found on human corpses. Therefore, forensic entomological questions with regard to when and how the first receptive females appear on carcasses are important, as the developmental stages of their larvae can be used to calculate the post-mortem interval. To date, we know that freshly emerged males respond to the cadaver odour of post-bloated carcasses (approximately 9 days after death at Tmean = 27°C), being attracted by benzyl butyrate. This component occurs at its highest concentration at this stage of decay. The aim of our study was to determine the principle of attraction of virgin females to the feeding and breeding substrate. For this purpose, we tested the response of these females to headspace samples of piglet cadavers and male sex pheromones [(Z9)-unsaturated fatty acid isopropyl esters] in a Y-olfactometer. Because we expected that such an odour combination is of importance for virgin female attraction, we tested the following two questions:1) Are virgin female hide beetles attracted by a combination of cadaver odour and male sex pheromones?2) During which decomposition stage do the first virgin females respond to cadaver odour when combined with male sex pheromones? RESULTS: We found that young virgin females were attracted to the cadaver by a combination of cadaver odour and male sex pheromones. Neither cadaver odour alone nor male sex pheromones alone was significantly more attractive than a solvent control. Our results also gave a weak indication that the first young virgin females respond as early as the post-bloating stage to its associated decomposition odour when combined with male sex pheromones. CONCLUSIONS: Our results indicate that freshly emerged males possibly respond to cadaver odour and visit carcasses before virgin females. Being attracted to cadavers when male sex pheromone is perceived as well, virgin females can optimise their reproductive possibilities.