Multisource noninvasive genetics of brown bears (Ursus arctos) in Greece reveals a highly structured population and a new matrilineal contact zone in southern Europe

In human‐dominated landscapes, connectivity is crucial for maintaining demographically stable mammalian populations. Here, we provide a comprehensive noninvasive genetic study for the brown bear population in the Hellenic Peninsula. We analyze its population structuring and connectivity, estimate its population size throughout its distribution, and describe its phylogeography in detail for the first time. Our results, based on 150 multilocus genotypes and on 244‐bp sequences of the mtDNA control region, show the population is comprised by three highly differentiated genetic clusters, consistent with geographical populations of Pindos, Peristeri, and Rhodope. By detecting two male bears with Rhodopean ancestry in the western demes, we provide strong evidence for the ongoing genetic connectivity of the geographically fragmented eastern and western distributions, which suggests connectivity of the larger East Balkan and Pindos‐Dinara populations. Total effective population size (Ne) was estimated to be 199 individuals, and total combined population size (NC) was 499, with each cluster showing a relatively high level of genetic variability, suggesting that migration has been sufficient to counteract genetic erosion. The mtNDA results were congruent with the microsatellite data, and the three genetic clusters were matched predominantly with an equal number of mtDNA haplotypes that belong to the brown bear Western mitochondrial lineage (Clade 1), with two haplotypes being globally new and endemic. The detection of a fourth haplotype that belongs to the Eastern lineage (Clade 3a1) in three bears from the western distribution places the southernmost secondary contact zone between the Eastern and Western lineages in Greece and generates new hypotheses about postglacial maxima migration routes. This work indicates that the genetic composition and diversity of Europe's low‐latitude fringe population are the outcome of ancient and historical events and highlight its importance for the connectivity and long‐term persistence of the species in the Balkans

Frugivory and seed dispersal by carnivores in the Rhodopi mountains of northern Greece

The frugivory and seed dispersal by carnivores were studied using a combination of vegetation surveys, fruit production counts and analysis of faeces distribution and content. The study site is situated close to the Greek-Bulgarian border within a protected area. The habitat comprises of a mixed forest of beech, pine, oak and spruce which is occasionally interrupted by patches of fruiting trees. Faecal samples were collected on five permanent transects which were sampled monthly between May and October of 1993 and 1994.Fruiting plant density was found to be slightly higher in the forest than along forest roads, however the species diversity was much higher on the latter. In some cases, immature fruiting plants were found on transects with no mature plants in the vicinity. Availability of ripe fruit was found to increase steadily between May and September. There was a significant difference between the numbers of faeces deposited by the carnivores, with fox being the most numerous, followed by marten, bear and wolf. There was spatial and temporal variation in the number of faeces deposited. Nevertheless, there was no variation between different altitudinal zones. Martens were found to defecate more often on stones when compared with the other carnivores. The analysis of fruit consumption revealed that bears were the most frugivorous carnivores followed by foxes, wolves and martens on the basis of frequency of consumption. The temporal availability of each ripe fruit species coincided with their consumption by the carnivores in most of the cases. A number of seed species were deposited at altitudes where the plants do not normally grow. Foxes dispersed the highest numbers of seeds in the study area and bears were second as they deposited large-size faeces which contained many seeds. Of the dispersed seeds, those of Rubus sp., Rosa sp. and Fragaria vesca were deposited in the highest numbers. Only a small number fraction of seeds were damaged through handling by carnivores

Survey tools to predict and monitor the presence of brown bears (Ursus arctos) in Greece: A case study integrating spatial analysis and invertebrate-derived DNA

Increasing populations of brown bear (Ursus arctos) in Greece have resulted in recolonisation of areas previously unoccupied for decades. Large carnivore recolonisation often threatens hard-established human-wildlife coexistence efforts, therefore the need to monitor and predict bear presence and movement becomes more apparent as the animals make use of corridors existing between their current range and other suitable habitats. The challenge of carrying out surveys in infrequently used areas, such as wildlife corridors, is compounded by limited funds for wildlife conservation. This calls for the exploration of alternative monitoring techniques that are more cost and time-efficient than the standard methods such as scat surveys. This study explored the use of habitat suitability modelling techniques in mapping brown bear ecological networks in Greece throughout their current range and sites of potential future recolonisation. Using these suitability maps as a guide, the study utilised an innovative non-invasive genetic monitoring technique, invertebrate-derived DNA (iDNA), to survey the species in the field. A single-species targeted qPCR approach was used and method development experiments were conducted to form a protocol to optimise its performance in the field. Next, iDNA surveys combined with scat surveys were conducted to model bear distribution in northern Greece using occupancy modelling. These models describe the probability of bear detection in a landscape that incorporates a core brown bear habitat, a presumed corridor and a recently recolonised area. A review of the laboratory experiments and a comparison of the resulting occupancy models indicated that iDNA can effectively monitor the presence of a species as well as be subsequently used in occupancy modelling analyses. iDNA can be seen as an effective and complementary method of assessing brown bear distribution to inform conservation strategies and has the potential to assist with the conservation monitoring of other bear species

Bioavailability and bioaccumulation characterization of essential and heavy metals contents in various plant food from polluted and referent areas

Bioavailability of metals occurring in soil is the basic source of its accumulation in plants. The impact of soil pollution (due to urban and mining areas) on the food chain presents a challenge for many investigations. Bioavailability of metals in a potentially polluted soil and their possible transfer and bioaccumulation in several vegetable species and herbs was examined. Microwave digestion was applied for total digestion of the plant tissues, while on the soil samples open wet digestion with a mixture of acids was applied. Three extraction methods were implemented for determination of bioavailable metals in the soil. Atomic emission spectrometry with inductively coupled plasma was used for determination of the total contents of 21 elements. Significant enrichments in agricultural soil for As, Pb and Zn (in urban area), Cd, Cu and Ni (in a copper mine area), compared with the respective values from European standards were detected. On the basis of three different extraction methods, higher availability was assumed for both lithogenic and anthropogenic elements. Translocation factors higher than 1 were obtained for As, Cd, Cu, Ni, Pb and Zn. Higher root to shoot translocation of these metals indicated that plants species have vital characteristics to be used for phytoextraction of these metals. The obtained data also suggested that Spinacia oleracea and Rumex acetosa were singled out to have a phytostabilization potential for Cd, Cu, Ni and Pb, while U. dioica only for Cu. R. acetosa has a potential for phytoextraction of Cd in urban and copper polluted areas

Between a rock and a hard place : site selection for Brush-tailed Rock-wallaby (Petrogale penicillata) translocations using an explicit decision-making process

Translocation programs are used in species conservation, but their success rates, particularly in Australia, are low. The poor outcomes for translocation programs are commonly attributed to a lack of information about habitat requirements, especially for rare and endangered species. To address this, the identification of suitable release sites using analytical methods such as Habitat Suitability Models (HSM), has become common. Since biodiversity losses are predicted to increase in the future, effective strategies to reduce species decline are required. Evidence-based decisions and scientific data to support conservation decision-making is paramount in conservation efforts. Once widely distributed across the mountainous country of south-eastern Australia, the distribution of the Brush-tailed Rock-wallaby (BTRW) has contracted particularly within the southern extent of their range. Considered critically endangered in Victoria and extinct in the Australian Capital Territory (ACT), it is more important than ever that the habitat requirements of the BTRW are understood to inform future translocations and the protection of this species in the wild. In this study, an explicit decision-making process was combined with Habitat Suitability Modelling (HSM) and ground-truthing methods to inform potential Brush-tailed Rock-wallaby (BTRW) translocations sites within Victoria and the ACT. Although this study is not the first of its kind to use HSM to predict BTRW habitat suitability, it is the first of its kind to apply an explicit decision-making process at two key stages: 1. to inform the selection of key physical and management attributes to be used in a HSM; and 2. to prioritise sites, within experts’ local knowledge areas, for ground-truthing By combining HSM and an explicit decision-making process, this study has been able to identify six suitable and two highly suitable habitat for BTRW across Victoria and the ACT. The use of an explicit decision-making process and ground-truthing methods to validate the model outputs and prioritise sites for ground-truthing has ensured that the final ranking of potential candidate sites can be reliably used by the Southern Brush-tailed Rock-wallaby Recovery Team to inform future translocations.Masters by Researc

Neolithic land-use in the Dutch wetlands: estimating the land-use implications of resource exploitation strategies in the Middle Swifterbant Culture (4600-3900 BCE)

The Dutch wetlands witness the gradual adoption of Neolithic novelties by foraging societies during the Swifterbant period. Recent analyses provide new insights into the subsistence palette of Middle Swifterbant societies. Small-scale livestock herding and cultivation are in evidence at this time, but their importance if unclear. Within the framework of PAGES Land-use at 6000BP project, we aim to translate the information on resource exploitation into information on land-use that can be incorporated into global climate modelling efforts, with attention for the importance of agriculture. A reconstruction of patterns of resource exploitation and their land-use dimensions is complicated by methodological issues in comparing the results of varied recent investigations. Analyses of organic residues in ceramics have attested to the cooking of aquatic foods, ruminant meat, porcine meat, as well as rare cases of dairy. In terms of vegetative matter, some ceramics exclusively yielded evidence of wild plants, while others preserve cereal remains. Elevated δ15N values of human were interpreted as demonstrating an important aquatic component of the diet well into the 4th millennium BC. Yet recent assays on livestock remains suggest grazing on salt marshes partly accounts for the human values. Finally, renewed archaeozoological investigations have shown the early presence of domestic animals to be more limited than previously thought. We discuss the relative importance of exploited resources to produce a best-fit interpretation of changing patterns of land-use during the Middle Swifterbant phase. Our review combines recent archaeological data with wider data on anthropogenic influence on the landscape. Combining the results of plant macroremains, information from pollen cores about vegetation development, the structure of faunal assemblages, and finds of arable fields and dairy residue, we suggest the most parsimonious interpretation is one of a limited land-use footprint of cultivation and livestock keeping in Dutch wetlands between 4600 and 3900 BCE.NWOVidi 276-60-004Human Origin

Taphonomy, environment or human plant exploitation strategies?: Deciphering changes in Pleistocene-Holocene plant representation at Umhlatuzana rockshelter, South Africa

The period between ~40 and 20 ka BP encompassing the Middle Stone Age (MSA) and Later Stone Age (LSA) transition has long been of interest because of the associated technological change. Understanding this transition in southern Africa is complicated by the paucity of archaeological sites that span this period. With its occupation sequence spanning the last ~70,000 years, Umhlatuzana Rock Shelter is one of the few sites that record this transition. Umhlatuzana thus offers a great opportunity to study past environmental dynamics from the Late Pleistocene (MIS 4) to the Late Holocene, and past human subsistence strategies, their social organisation, technological and symbolic innovations. Although organic preservation is poor (bones, seeds, and charcoal) at the site, silica phytoliths preserve generally well throughout the sequence. These microscopic silica particles can identify different plant types that are no longer visible at the site because of decomposition or burning to a reliable taxonomical level. Thus, to trace site occupation, plant resource use, and in turn reconstruct past vegetation, we applied phytolith analyses to sediment samples of the newly excavated Umhlatuzana sequence. We present results of the phytolith assemblage variability to determine change in plant use from the Pleistocene to the Holocene and discuss them in relation to taphonomical processes and human plant gathering strategies and activities. This study ultimately seeks to provide a palaeoenvironmental context for modes of occupation and will shed light on past human-environmental interactions in eastern South Africa.NWOVidi 276-60-004Human Origin