The European Stag Beetle (Lucanus cervus) Monitoring Network: International Citizen Science Cooperation Reveals Regional Differences in Phenology and Temperature Response.

To address the decline in biodiversity, international cooperation in monitoring of threatened species is needed. Citizen science can play a crucial role in achieving this challenging goal, but most citizen science projects have been established at national or regional scales. Here we report on the establishment and initial findings of the European Stag Beetle Monitoring Network (ESBMN), an international network of stag beetle (Lucanus cervus) monitoring schemes using the same protocol. The network, started in 2016, currently includes 14 countries (see results) but with a strong variation in output regarding the number of transects (148 successful transects in total) and transect walks (1735). We found differences across European regions in the number of stag beetles recorded, related to phenology and temperature, but not for time of transect start. Furthermore, the initial experiences of the ESBMN regarding international cooperation, citizen science approach, and drop-out of volunteers is discussed. An international standardised protocol that allows some local variation is essential for international collaboration and data management, and analysis is best performed at the international level, whereas recruiting, training, and maintaining volunteers is best organised locally. In conclusion, we appeal for more joint international citizen science-based monitoring initiatives assisting international red-listing and conservation actions.

Genetic structure of recently fragmented suburban populations of European stag beetle.

Habitat loss and fragmentation due to urbanization can negatively affect metapopulation persistence when gene flow among populations is reduced and population sizes decrease. Inference of patterns and processes of population connectivity derived from spatial genetic analysis has proven invaluable for conservation and management. However, a more complete account of population dynamics may be obtained by combining spatial and temporal sampling. We, therefore, performed a genetic study on European stag beetle (Lucanus cervus L.) populations in a suburban context using samples collected in three locations and during the period 2002-2016. The sampling area has seen recent landscape changes which resulted in population declines. Through the use of a suite of F ST, clustering analysis, individual assignment, and relatedness analysis, we assessed fine scale spatiotemporal genetic variation within and among habitat patches using 283 individuals successfully genotyped at 17 microsatellites. Our findings suggested the three locations to hold demographically independent populations, at least over time scales of relevance to conservation, though with higher levels of gene flow in the past. Contrary to expectation from tagging studies, dispersal appeared to be mainly female-biased. Although the life cycle of stag beetle suggests its generations to be discrete, no clear temporal structure was identified, which could be attributed to the varying duration of larval development. Since population bottlenecks were detected and estimates of effective number of breeders were low, conservation actions are eminent which should include the establishment of suitable dead wood for oviposition on both local and regional scales to increase (re)colonization success and connectivity among current populations.

Supporting dataset and methods for body sizes and concentrations of chemical elements measured in elytra and abdomens of Stag Beetles Lucanus cervus.

The dataset presented in this data paper supports "Breaking down insect stoichiometry into chitin-based and internal elemental traits: Patterns and correlates of continent-wide intraspecific variation in the largest European saproxylic beetle" (Orlowski et al. 2020). Here we present the supplementary data and description of methods on the following: (1) mass of elytra and abdomens across 28 local Stag Beetle Lucanus cervus populations in Europe. (2) Population origin and coverage of six major land-cover types, including transport infrastructure, measured in three radii (500 m, 1000 m and 5000 m) around the sampling sites of these populations. (3) The relationship between the mass and concentrations of elements measured in abdomens and elytra in 28 Stag Beetle populations and major land-cover types around the sampling sites.

Breaking down insect stoichiometry into chitin-based and internal elemental traits: Patterns and correlates of continent-wide intraspecific variation in the largest European saproxylic beetle.

Stoichiometric, trophic and ecotoxicological data have traditionally been acquired from patterns of variation in elemental traits of whole invertebrate bodies, whereas the critical issue of the extracellular origin of some portion of elements, such as those present in ingested food and internal organs, has been ignored. Here we investigated an unexplored, yet crucial, question relating to whether, and to what degree, metals from two major body fractions: exoskeleton (elytra) and internal (body organs with gut material present in abdomens), are correlated with each other in wild populations of the largest European saproxylic insect, the Stag Beetle Lucanus cervus, and how metals from these two fractions vary with insect size and local habitat conditions. We examined the continent-wide variation in the concentrations of 12 chemical elements (Ca, Mg, K, Na, Mn, Fe, Zn, Cu, As, Cd, Pb and Ni) measured in the elytra and abdomen of specimens from 28 populations inhabiting an urban-woodland habitat gradient across the species' entire distributional range from Spain to Russia. Across populations, elemental concentrations (except Ni and Pb) were 2-13 times higher in abdominal samples than in elytra, and the magnitude of these differences was related to both insect size and local habitat conditions. Smaller individuals from both woodland and urban habitat tended to have higher concentrations of trace elements (Zn, As, Cd, Pb and Ni). The concentration of only six elements (Mg, K, Na, Mn, Cd and Ni) was correlated in the elytra and abdomen at the individual and population levels, implying a limitation to the broader applicability of elytra as a surrogate for internal elemental pools. We highlight that in non-feeding adult saproxylic beetles, minerals, acquired during the larval stage, may be concentrated in the large quantities of residual body fat.

A database of threat statuses and life-history traits of Red List species in Flanders (northern Belgium).

BACKGROUND: Red Lists estimate the extinction risk of species at global or regional levels and are important instruments in conservation policies. Global Red List assessments are readily available via the IUCN website (https://www.iucnredlist.org) and are regularly updated by (taxonomic) experts. Regional Red Lists, however, are not always easy to find and often use local criteria to assess the local extinction risk of species. NEW INFORMATION: Here, we publish a database with the outcome of 38 Red List assessments in Flanders (northern Belgium) between 1994 and 2018. In total, the database contains 6,224 records of 5,039 unique taxa pertaining to 24 different taxonomic groups. Using a quality control procedure, we evaluated the criteria used, the number of records, the temporal and spatial distribution of the data and the up-to-dateness of the Red Lists. This way, nineteen Red Lists were approved as being of sufficient high quality (i.e. validated) and nineteen others were not. Once validated, Red Lists are approved by the regional Minister of Environment and published in the Belgian Official Gazette acquiring legal status. For the validated Red Lists, we additionally compiled (life-history) traits that are applicable to a wide variety of species groups (taxonomic kingdom, environment, biotope, nutrient level, dispersal capacity, lifespan and cuddliness). The publication of this dataset allows comparison of Red List statuses with other European regions and countries and permits analyses about how certain (life-history) traits can explain the Red List status of species. The dataset will be regularly updated by adding new Red List (re)assessments and/or additional (life-history) traits.

Phylogeographic structure and ecological niche modelling reveal signals of isolation and postglacial colonisation in the European stag beetle.

Lucanus cervus (L.), the stag beetle, is a saproxylic beetle species distributed widely across Europe. Throughout its distribution the species has exhibited pronounced declines and is widely considered threatened. Conservation efforts may be hindered by the lack of population genetic data and understanding of the spatial scale of population connectivity. To address this knowledge gap this research details the first broad scale phylogeographic study of L. cervus based on mitochondrial DNA (mtDNA) sequencing and microsatellite analysis of samples collected from 121 localities across Europe. Genetic data were complemented by palaeo-distribution models of spatial occupancy during the Last Glacial Maximum to strengthen inferences of refugial areas. A salient feature of the mtDNA was the identification of two lineages. Lineage I was widespread across Europe while lineage II was confined to Greece. Microsatellites supported the differentiation of the Greek samples and alongside palaeo-distribution models indicated this area was a glacial refuge. The genetic endemism of the Greek samples, and demographic results compatible with no signatures of spatial expansion likely reflects restricted dispersal into and out of the area. Lineage I exhibited a shallow star like phylogeny compatible with rapid population expansion across Europe. Demographic analysis indicated such expansions occurred after the Last Glacial Maximum. Nuclear diversity and hindcast species distribution models indicated a central Italian refuge for lineage I. Palaeo-distribution modelling results also suggested a western refuge in northern Iberia and south-west France. In conclusion the results provide evidence of glacial divergence in stag beetle while also suggesting high, at least on evolutionary timescales, gene flow across most of Europe. The data also provide a neutral genetic framework against which patterns of phenotypic variation may be assessed.

Testing the performance of a fragment of the COI gene to identify western Palaearctic stag beetle species (Coleoptera, Lucanidae).

THE TAXONOMY OF STAG BEETLES (COLEOPTERA: Lucanidae) remains challenging, mainly due to the sexual dimorphism and the strong allometry in males. Such conjecture confounds taxonomic based conservation efforts that are urgently needed due to numerous threats to stag beetle biodiversity. Molecular tools could help solve the problem of identification of the different recognized taxa in the "Lucanus cervus complex" and in some related Palaearctic species. We investigated the potential use of a 670 bp region at the 3' end of the mitochondrial cytochrome c oxidase subunit I gene (COI) for barcoding purposes (different from the standard COI barcoding region). Well resolved species and subspecies were L. tetraodon, L. cervusakbesianus, L. c. laticornis, as well as the two eastern Asian outgroup taxa L. formosanus and L. hermani. Conversely, certain taxa could not be distinguished from each other based on K2P-distances and tree topologies: L. c. fabiani / L. (P.) barbarossa, L. c. judaicus / an unknown Lucanus species, L. c. cervus / L. c. turcicus / L. c. pentaphyllus / L. (P.) macrophyllus / L. ibericus. The relative roles of phenotypic plasticity, recurrent hybridisation and incomplete lineage sorting underlying taxonomic and phylogenetic discordances are discussed.