Ingenious Architecture and Coloration Generation in Enamel of Rodent Teeth.

Teeth exemplify architectures comprising an interplay of inorganic and organic constituents, resulting in sophisticated natural composites. Rodents (Rodentia) showcase extraordinary adaptations, with their continuously growing incisors surpassing human teeth in functional and structural optimizations. In this study, employing state-of-the-art direct atomic-scale imaging and nanoscale spectroscopies, we present compelling evidence that the release of material from ameloblasts and the subsequent formation of iron-rich enamel and surface layers in the constantly growing incisors of rodents are complex orchestrated processes, intricately regulated and independent of environmental factors. The synergistic fusion of three-dimensional tomography and imaging techniques of etched rodent́s enamel unveils a direct correlation between the presence of pockets infused with ferrihydrite-like material and the acid resistant properties exhibited by the iron-rich enamel, fortifying it as an efficient protective shield. Moreover, observations using optical microscopy shed light on the role of iron-rich enamel as a microstructural element that acts as a path for color transmission, although the native color remains indistinguishable from that of regular enamel, challenging the prevailing paradigms. The redefinition of "pigmented enamel" to encompass ferrihydrite-like infusion in rodent incisors reshapes our perception of incisor microstructure and color generation. The functional significance of acid-resistant iron-rich enamel and the understanding of the underlying coloration mechanism in rodent incisors have far-reaching implications for human health, development of potentially groundbreaking dental materials, and restorative dentistry. These findings enable the creation of an entirely different class of dental biomaterials with enhanced properties, inspired by the ingenious designs found in nature.

Molecular analysis of scats revealed diet and prey choice of grey wolves and Eurasian lynx in the contact zone between the Dinaric Mountains and the Alps.

A comprehensive understanding of the dietary habits of carnivores is essential to get ecological insights into their role in the ecosystem, potential competition with other carnivorous species, and their effect on prey populations. Genetic analysis of non-invasive samples, such as scats, can supplement behavioural or microscopic diet investigations. The objective of this study was to employ DNA metabarcoding to accurately determine the prey species in grey wolf (Canis lupus) and Eurasian lynx (Lynx lynx) scat samples collected in the Julian Alps and the Dinaric Mountains, Slovenia. The primary prey of wolves were red deer (Cervus elaphus) (detected in 96% scat samples), European roe deer (Capreolus capreolus) (68%), and wild boar (Sus scrofa) (45%). A smaller portion of their diet consisted of mesocarnivores, small mammals, and domestic animals. In contrast, the lynx diet mostly consisted of European roe deer (82%) and red deer (64%). However, small mammals and domestic animals were also present in lynx diet, albeit to a lesser extent. Our findings indicate that the dietary habits of wolves and lynx are influenced by geographical location. Snapshot dietary analyses using metabarcoding are valuable for comprehending the behaviour and ecology of predators, and for devising conservation measures aimed at sustainable management of both their natural habitats and prey populations. However, to gain a more detailed understanding of wolf and lynx dietary habits and ecological impact, it would be essential to conduct long-term genetic monitoring of their diet.

Spatial genetic structure of European wild boar, with inferences on late-Pleistocene and Holocene demographic history.

European wildlife has been subjected to intensifying levels of anthropogenic impact throughout the Holocene, yet the main genetic partitioning of many species is thought to still reflect the late-Pleistocene glacial refugia. We analyzed 26,342 nuclear SNPs of 464 wild boar (Sus scrofa) across the European continent to infer demographic history and reassess the genetic consequences of natural and anthropogenic forces. We found that population fragmentation, inbreeding and recent hybridization with domestic pigs have caused the spatial genetic structure to be heterogeneous at the local scale. Underlying local anthropogenic signatures, we found a deep genetic structure in the form of an arch-shaped cline extending from the Dinaric Alps, via Southeastern Europe and the Baltic states, to Western Europe and, finally, to the genetically diverged Iberian peninsula. These findings indicate that, despite considerable anthropogenic influence, the deeper, natural continental structure is still intact. Regarding the glacial refugia, our findings show a weaker signal than generally assumed, but are nevertheless suggestive of two main recolonization routes, with important roles for Southern France and the Balkans. Our results highlight the importance of applying genomic resources and framing genetic results within a species' demographic history and geographic distribution for a better understanding of the complex mixture of underlying processes.

Pan-European phylogeography of the European roe deer (Capreolus capreolus).

To provide the most comprehensive picture of species phylogeny and phylogeography of European roe deer (Capreolus capreolus), we analyzed mtDNA control region (610 bp) of 1469 samples of roe deer from Central and Eastern Europe and included into the analyses additional 1541 mtDNA sequences from GenBank from other regions of the continent. We detected two mtDNA lineages of the species: European and Siberian (an introgression of C. pygargus mtDNA into C. capreolus). The Siberian lineage was most frequent in the eastern part of the continent and declined toward Central Europe. The European lineage contained three clades (Central, Eastern, and Western) composed of several haplogroups, many of which were separated in space. The Western clade appeared to have a discontinuous range from Portugal to Russia. Most of the haplogroups in the Central and the Eastern clades were under expansion during the Weichselian glacial period before the Last Glacial Maximum (LGM), while the expansion time of the Western clade overlapped with the Eemian interglacial. The high genetic diversity of extant roe deer is the result of their survival during the LGM probably in a large, contiguous range spanning from the Iberian Peninsula to the Caucasus Mts and in two northern refugia.

Neutral and Selective Processes Shape MHC Diversity in Roe Deer in Slovenia.

Disease control and containment in free-ranging populations is one of the greatest challenges in wildlife management. Despite the importance of major histocompatibility complex (MHC) genes for immune response, an assessment of the diversity and occurrence of these genes is still rare in European roe deer, the most abundant and widespread large mammal in Europe. Therefore, we examined immunogenetic variation in roe deer in Slovenia to identify species adaptation by comparing the genetic diversity of the MHC genes with the data on neutral microsatellites. We found ten MHC DRB alleles, three of which are novel. Evidence for historical positive selection on the MHC was found using the maximum likelihood codon method. Patterns of MHC allelic distribution were not congruent with neutral population genetic findings. The lack of population genetic differentiation in MHC genes compared to existing structure in neutral markers suggests that MHC polymorphism was influenced primarily by balancing selection and, to a lesser extent, by neutral processes such as genetic drift, with no clear evidence of local adaptation. Selection analyses indicated that approx. 10% of amino acids encoded under episodic positive selection. This study represents one of the first steps towards establishing an immunogenetic map of roe deer populations across Europe, aiming to better support science-based management of this important game species.

Next-generation phylogeography resolves post-glacial colonization patterns in a widespread carnivore, the red fox (Vulpes vulpes), in Europe.

Carnivores tend to exhibit a lack of (or less pronounced) genetic structure at continental scales in both a geographic and temporal sense and this can confound the identification of post-glacial colonization patterns in this group. In this study we used genome-wide data (using genotyping by sequencing [GBS]) to reconstruct the phylogeographic history of a widespread carnivore, the red fox (Vulpes vulpes), by investigating broad-scale patterns of genomic variation, differentiation and admixture amongst contemporary populations in Europe. Using 15,003 single nucleotide polymorphisms (SNPs) from 524 individuals allowed us to identify the importance of refugial regions for the red fox in terms of endemism (e.g., Iberia). In addition, we tested multiple post-glacial recolonization scenarios of previously glaciated regions during the Last Glacial Maximum using an Approximate Bayesian Computation (ABC) approach that were unresolved from previous studies. This allowed us to identify the role of admixture from multiple source population post-Younger Dryas in the case of Scandinavia and ancient land-bridges in the colonization of the British Isles. A natural colonization of Ireland was deemed more likely than an ancient human-mediated introduction as has previously been proposed and potentially points to a larger mammalian community on the island in the early post-glacial period. Using genome-wide data has allowed us to tease apart broad-scale patterns of structure and diversity in a widespread carnivore in Europe that was not evident from using more limited marker sets and provides a foundation for next-generation phylogeographic studies in other non-model species.

COVID-19 related travel restrictions prevented numerous wildlife deaths on roads: A comparative analysis of results from 11 countries.

Millions of wild animals are killed annually on roads worldwide. During spring 2020, the volume of road traffic was reduced globally as a consequence of the COVID-19 pandemic. We gathered data on wildlife-vehicle collisions (WVC) from Czechia, Estonia, Finland, Hungary, Israel, Norway, Slovenia, Spain, Sweden, and for Scotland and England within the United Kingdom. In all studied countries WVC statistics tend to be dominated by large mammals (various deer species and wild boar), while information on smaller mammals as well as birds are less well recorded. The expected number of WVC for 2020 was predicted on the basis of 2015-2019 WVC time series representing expected WVC numbers under normal traffic conditions. Then, the forecasted and reported WVC data were compared. The results indicate varying levels of WVC decrease between countries during the COVID-19 related traffic flow reduction (CRTR). While no significant change was determined in Sweden, where the state-wide response to COVID-19 was the least intensive, a decrease as marked as 37.4% was identified in Estonia. The greatest WVC decrease, more than 40%, was determined during the first weeks of CRTR for Estonia, Spain, Israel, and Czechia. Measures taken during spring 2020 allowed the survival of large numbers of wild animals which would have been killed under normal traffic conditions. The significant effects of even just a few weeks of reduced traffic, help to highlight the negative impacts of roads on wildlife mortality and the need to boost global efforts of wildlife conservation, including systematic gathering of roadkill data.

Dentition and body condition: tooth wear as a correlate of weight loss in roe deer.

BACKGROUND: In many mammalian species, once the permanent teeth have erupted, the only change to dentition is a gradual loss of tooth surface/height through wear. The crown of the teeth cannot be repaired once worn. When dental crown tissue has been depleted due to wear, the animal is expected to have a suboptimal body condition. We evaluated the role of tooth wear in causing a reduction of physical condition in adult roe deer females (Capreolus capreolus). RESULTS: The progressive wearing of the lower cheek teeth was assessed in a Northern Apennines (Italy) population with a new scoring scheme based on objectively described tooth characteristics (morphotypes) being either present or absent. Eviscerated body mass and mandible length, which is a good proxy for body size in roe deer, were related to the tooth wear score by the use of linear regressions. The sum of wear scores for molariform teeth correlated most strongly with body condition (i.e., eviscerated body mass/mandible length), showing the importance of the entire chewing surface for acquiring energy by food comminution, chewing, and digestion. In comparison with individuals of comparable size experiencing minor tooth wear, the body mass of those with the most advanced stage of tooth wear was decreased by 33.7%. This method was compared to the height and the hypsodonty index of the first molar, the most commonly used indices of tooth wear. The sum of molariform wear scoring scheme resulted in a more suitable index to describe the variation in body condition of roe deer. CONCLUSIONS: Describing tooth wear patterns in hunted populations and monitoring at which tooth wear level (and therefore dental morphotype) an animal is no longer able to sustain its physical condition (i.e. when it begins to lose body mass) can be a useful tool for improving the management of the most widespread and abundant deer species in Europe. At the same time, such an approach can clarify the role of tooth wear as a proximate cause of senescence in ungulates.

Environmental factors shaping stable isotope signatures of modern red deer (Cervus elaphus) inhabiting various habitats.

Stable isotope analyses of bone collagen are often used in palaeoecological studies to reveal environmental conditions in the habitats of different herbivore species. However, such studies require valuable reference data, obtained from analyses of modern individuals, in habitats of well-known conditions. In this article, we present the stable carbon and nitrogen isotope composition of bone collagen from modern red deer (N = 242 individuals) dwelling in various habitats (N = 15 study sites) in Europe. We investigated which of the selected climatic and environmental factors affected the δ13C and δ15N values in bone collagen of the studied specimens. Among all analyzed factors, the percent of forest cover influenced the carbon isotopic composition most significantly, and decreasing forest cover caused an increase in δ13C values. The δ15N was positively related to the proportion of open area and (only in the coastal areas) negatively related to the distance to the seashore. Using rigorous statistical methods and a large number of samples, we confirmed that δ13C and δ15N values can be used as a proxy of past habitats of red deer.

First Insights into the Population Genetic Structure and Heterozygosity-Fitness Relationship in Roe Deer Inhabiting the Area between the Alps and Dinaric Mountains.

Across its pan-European distribution, the European roe deer (Capreolus capreolus) faces a wide diversity of environmental and climatic conditions; therefore, several factors, including intrinsic ones, shape life-history traits and cause significant variability in parameters of fitness. By utilizing microsatellite variations in 214 roe deer females collected throughout Slovenia, Central Europe, we determined the genetic variability and population structure of this species in the contact zone between the Alps and the Dinaric Mountains, i.e., over a wider area where data on the genetic outlook for this-the most common and widespread European wild ungulate-have been completely lacking so far. Throughout the country, we found moderate microsatellite diversity (Ho = 0.57-0.65) in relation to the observed heterozygosity reported for other roe deer populations in Europe. Spatial differences in genetic diversity of the species in Slovenia can be explained by population history linked to varying approaches to population management and/or different connectivity among subpopulations in topographically differentiated habitats. A country-wide pattern of genetic structure is clearly defined by separation of the populations into three groups present in the following regions: (i) Southern sub-Mediterranean and Karst regions, (ii) Central Slovenia, and (iii) the Sub-Pannonian Region in the north-east. This is also confirmed by evidencing a moderate isolation by distance, especially by separating southern samples (coastal Slovenia) from others. Levels of genetic differentiation vary among populations, which can be explained by the effect of natural geographical barriers or the presence of anthropogenic barriers such as urban areas and highways. In the subset of 172 yearling females, we analyzed the influence of genetic advantage (individual heterozygosity) and other genetic data (reflected in the structuring of the population) on body mass and reproductive ability. We found evidence that genetic factors influence the body mass of roe deer yearling females (explaining altogether 18.8% of body mass variance), and the level of individual heterozygosity alone also positively affected body mass, which is in accordance with the theory that heterozygosity is commonly positively correlated with fitness in wild populations. However, we did not uncover any effect of heterozygosity on two parameters of reproductive ability (fertility and potential reproductive outcome), indicating that several other factors, especially environmental ones, have a predominant effect on the parameters of fitness in roe deer.

Genetic characterisation of wild ungulates: successful isolation and analysis of DNA from widely available bones can be cheap, fast and easy.

Genetic characterisation of wild ungulates can be a useful tool in wildlife management and in obtaining a greater understanding of their biological and ecological roles in a wider spatiotemporal context. Different ways of optimising methodologies and reducing the costs of genetic analyses using widely available bone tissues collected within regular hunting allocations were examined. Successful isolation and analysis of DNA from widely available bones can be cheap, fast and easy. In particular, this study explored the possibility of using bones for extracting high quality nuclear DNA for microsatellite analysis. The utility of applying a modified demineralisation process using two commercially available DNA isolation kits, which differ significantly in price, was evaluated. The sample sets included bones and, for comparison, muscle tissues from four wild ungulate species: chamois (Rupicapra rupicapra), roe deer (Capreolus capreolus), wild boar (Sus scrofa), and Alpine ibex (Capra ibex). For the recent bones, these results confirmed that the DNA concentrations and microsatellite amplification were sufficiently high, even when using low-cost kits, after prior demineralisation. For old bones, prior demineralisation and use of a specially designed isolation kit led to a more successful extraction of DNA. Besides reducing kit-related costs, low-cost kits are much faster and therefore make genetic analysis more efficient.

Pilot Study on Genetic Associations With Age-Related Sarcopenia.

Despite strong evidence of an inheritable component of muscle phenotypes, little progress has been made in identifying the specific genetic factors involved in the development of sarcopenia. Even rarer are studies that focus on predicting the risk of sarcopenia based on a genetic risk score. In the present study, we tested the single and combined effect of seven candidate gene variants on the risk of sarcopenia. Single nucleotide polymorphisms in candidate genes were genotyped using the KASP assay. We examined 190 older adults that were classified as non-sarcopenic or sarcopenic according to the diagnostic criteria of the European Working Group on Sarcopenia in Older People. Sarcopenia was associated with Methylenetetrahydrofolate reductase, Alpha-actinin-3, and Nuclear respiratory factor 2 genotypes. The combined effect of all three polymorphisms explained 39% of the interindividual variation in sarcopenia risk. Our results suggest that the single and combined effect of Methylenetetrahydrofolate reductase, Alpha-actinin-3, and Nuclear respiratory factor 2 polymorphism is associated with sarcopenia risk in older adults. Nowadays, as the population is getting older and older, great efforts are being made to research the etiology, diagnosis and treatment of sarcopenia. At the same time, small progress has been made in understanding the genetic etiology of sarcopenia. Given the importance of research on this disease, further genetic studies are needed to better understand the genetic risk underlying sarcopenia. We believe that this small-scale study will help to demonstrate that there is still much to be discovered in this field.

Age-related effects of body mass on fertility and litter size in roe deer.

We analysed effects of females' body mass and age on reproductive capacity of European roe deer (Capreolus capreolus) in a large sample set of 1312 females (305 yearlings and 1007 adults), hunted throughout Slovenia, central Europe, in the period 2013-2015. Body mass positively affected probability of ovulation and potential litter size (number of corpora lutea), although its effect was more pronounced in yearlings than in adults. Between age groups, we found clear differences in responses of both reproductive parameters to body mass which influences primarily reproductive performance of younger, and in particular, lighter individuals: at the same body mass yearlings would at average have smaller litters than adults, and at lower body mass also young to middle-aged adults would have smaller litters than old ones. In addition, while yearlings have to reach a critical threshold body mass to attain reproductive maturity, adult females are fertile (produce ova) even at low body mass. However, at higher body mass also younger individuals shift their efforts into the reproduction, and after reaching an age-specific threshold the body mass does not have any further effects on the reproductive output of roe deer females. Increased reproductive capacity at more advanced age, combined with declining body mass suggests that old does allocate more of their resources in reproduction than in body condition.

Magnesium-Assisted Continuous Growth of Strongly Iron-Enriched Incisors.

Teeth are an excellent example where optimally designed nanoarchitectures with precisely constructed components consist of simple compounds. Typically, these simple constituent phases with insignificant properties show mechanical property amplifications when formed into composite architectures. Material properties of functional composites are generally regulated on the nanoscale, which makes their characterization particularly demanding. Using advanced analytical and imaging transmission electron microscopy techniques, we identified innovative microstructural adjustments combined with astonishing compositional adaptations in incisors of coypu. Unique constituents, recognized as an additional amorphous Fe-rich surface layer followed by a transition zone covering pigmented enamel, provide the required structural stability to withstand repeated mechanical load. The chemically diverse Fe-rich surface layer, including ferrihydrite and iron-calcium phosphates, gives the typical orange-brown coloration to the incisors. Within the spaces between elongated hydroxyapatite crystals in the pigmented enamel, only ferrihydrite was found, implying that enamel pigmentation is a very strictly controlled process. Most significantly, an unprecedentedly high amount of Mg was measured in the amorphous flake-like material within the dentinal tubules of the incisors, suggesting the presence of a (Mg,Ca) phosphate phase. This unusually high influx of Mg into the dentin of incisors, but not molars, suggests a substantial functionality of Mg in the initial formation stages and constant growth of incisors. The present results emphasize the strong mutual correlation among the microstructure, chemical composition, and mechanical properties of mineralized dental tissues.

Wild boar populations up, numbers of hunters down? A review of trends and implications for Europe.

Across Europe, wild boar numbers increased in the 1960s-1970s but stabilised in the 1980s; recent evidence suggests that the numbers and impact of wild boar has grown steadily since the 1980s. As hunting is the main cause of mortality for this species, we reviewed wild boar hunting bags and hunter population trends in 18 European countries from 1982 to 2012. Hunting statistics and numbers of hunters were used as indicators of animal numbers and hunting pressure. The results confirmed that wild boar increased consistently throughout Europe, while the number of hunters remained relatively stable or declined in most countries. We conclude that recreational hunting is insufficient to limit wild boar population growth and that the relative impact of hunting on wild boar mortality had decreased. Other factors, such as mild winters, reforestation, intensification of crop production, supplementary feeding and compensatory population responses of wild boar to hunting pressure might also explain population growth. As populations continue to grow, more human-wild boar conflicts are expected unless this trend is reversed. New interdisciplinary approaches are urgently required to mitigate human-wild boar conflicts, which are otherwise destined to grow further.

Lead and cadmium in mushrooms from the vicinity of two large emission sources in Slovenia.

Cd and Pb contents were determined in 699 samples of fruiting bodies of 55 mushrooms species, collected in the period 2000-2007 in the vicinity of the largest Slovenian thermal power plant (the Salek Valley) and near an abandoned lead smelter (the Upper Meza Valley). The present study is the first regarding lead and cadmium in mushrooms from those exposed areas. Therefore, there was a significant lack of prior data. Among 55 studied mushroom species 36 species are edible and important from an ecotoxicological perspective. However, the remaining non-edible species are important for bioindication and allowed us to compare our results with other studies carried out in other polluted areas in Europe. The highest contents of Cd were found in Agaricus arvensis Schff.: Fr. (117 mg/kg dw) and Agaricus silvicola L.: Fr. (67.9 mg/kg dw), while the highest contents of Pb were found in Macrolepiota procera (Scop.) Singer (53.8 mg/kg dw) and Lycoperdon perlatum Pers. (50 mg/kg dw), respectively. Considering the high contents of both metals in fruiting bodies of edible fungi, together with FAO/WHO directives on tolerable levels of weekly intake of Pb/Cd by humans, it is evident that consumption of some mushroom species originating from both study areas may pose a significant human health risk. A. arvensis Schff.: Fr., A. silvicola L.: Fr. and Cortinarius caperatus (Pers.) Fr. originating from the Salek Valley, and Armillaria mellea Vahl. P. Kumm., Boletus edulis Bull., L. perlatum Pers., Leccinum versipelle (Fr. & Hök) Snell, and M. procera (Scop.) Singer originating from the Upper Meza Valley should not be consumed at all. Our findings are consistent with some other studies, which emphasized that mushrooms from heavily polluted areas, such as in the vicinity of smelters, accumulate extremely high amounts of metals, and should therefore be omitted from human consumption.

Heavy metals and arsenic concentrations in ten fish species from the Salek lakes (Slovenia): assessment of potential human health risk due to fish consumption.

The study, which measured the concentrations of Hg, Pb, Cd, Zn and As in various fish tissues (muscle, gill and liver) of 10 fish species (Abramis brama danubii, Alburnus alburnus alburnus, Barbus meridionalis petenyi, Carassius auratius gibelio, Cyprinus carpio, Lepomis gibossus, Leuciscius cephalus cephalus, Perca fluviatilis fluviatilis, Rutilus rutilus, Scardinus erythrophtlalmus erythrophtlalmus) collected in the Salek lakes, is the first survey regarding metal concentrations in fish species with samples originating from Slovene lakes, while only a limited number of such studies have been carried out in southeastern Europe. Since these lakes are situated in the close vicinity of the largest Slovene thermal power plant, the study provides an insight into the potential impact of increased levels of metals in the environment as well as an estimate of the contamination of fish tissues with metals. Furthermore, it was possible to compare the results obtained with those from other studies regarding metal levels in freshwater fish species. The mean metal concentrations of different tissues irrespective of species varied in the following ranges: Zn 4.31-199 mg/kg ww, Pb 0.01-0.48 mg/kg ww, As 0.02-0.44 mg/kg ww, Hg <0.01-0.31 mg/kg ww, Cd < 0.01-0.19 mg/kg ww. In general, higher contents of Hg were found in muscles and livers than in gills and higher contents of As in gills and livers than in muscles, respectively. The accumulation of Pb and Zn was most pronounced in gills. The result obtained regarding metal concentrations in fish revealed that the ecosystems of the Salek lakes are not polluted with Hg and Pb, slightly loaded with As and Cd and moderately polluted with Zn. In addition, the potential human health risk due to fish consumption was assessed. This showed that the estimated weekly intakes for all metals were far below provisional permissible tolerable weekly intakes determined by WHO/FAO. The consumption of fish from the Salek lakes, therefore, does not pose a risk to human health.

Prevalence of mandibular osteomyelitis in roe deer (Capreolus capreolus) in Slovenia.

Mandibular osteomyelitis in free-ranging cervids is a rare, but eventually fatal, disease. We examined 41,895 defleshed mandibles of roe deer collected throughout Slovenia in 2007. Mandibles from 14,679 fawns had no signs of osteomyelitis, and were excluded from further analysis. Of the remaining 27,216 specimens, chronic osteomyelitis ("lumpy jaw") was found in 113 mandibles (4.2%; 7.0% of adults). The majority of cases were observed from the Mediterranean and subalpine regions, near larger cities and thermal power plants. There was no statistically significant correlation between severity of the mandibular osteomyelitis and body weight. Females were more frequently affected than males. Coarse and abrasive food, and to some extent dental fluorosis, are the most probable triggers for development of lesions.

Historical biomonitoring of fluoride pollution by determining fluoride contents in roe deer (Capreolus capreolus L.) antlers and mandibles in the vicinity of the largest Slovene thermal power plant.

Roe deer antlers/mandibles are a useful tool for determination of ambient fluoride pollution. Antlers have a well-defined annual cycle of growth, therefore they represent a natural standardisation of samples during winter months. On the contrary, mandibles accumulate fluorides during the whole life of an organism, thus they reflect aggregated effect of fluoride pollution trough the life-span of an organism. Both tissues are easily available; mandibles are often systematically collected with the purpose of cognitive management and control, and antlers could be gathered from private well-dated hunters' collections. Considering these benefits, fluoride contents were measured in 141 antlers (period 1960-2007) and 220 mandibles (period 1997-2009) of roe deer, shot in the vicinity of the largest Slovene Thermal Power Plant of Sostanj (STPP) as one of the major sources of fluorides in Slovenia. Fluoride contents in antlers significantly differed among age categories, and ranged from 110 to 1210 mg/kg in yearlings, 130 to 2340 mg/kg in young adults, and 250 to 2590 mg/kg in older adults, respectively. Fluoride levels in mandibles were also significantly different among age categories, and ranged from 30.0 to 227 mg/kg in fawns, 33.8 to 383 mg/kg in yearlings, and 61.5 to 1020 mg/kg in adults, respectively. Comparison of these results with previously reported fluoride contents in antlers and mandibles of roe deer from different areas of Europe revealed that the study area has never been extensively contaminated with fluorides. Moreover, trends of fluoride contents in both tissues confirmed a significant decrease of fluoride pollution in the area after the years 1995 and 2000, when flue-gas cleaning devices were constructed on the STPP. Indeed, highly positive correlations between annual emissions from the STPP and mean annual fluoride contents in antlers/mandibles confirmed that both tissues may be a useful tool for assessing temporal trends in ambient fluoride pollution.

Heavy metal concentrations in food chain of Lake Velenjsko jezero, Slovenia: an artificial lake from mining.

The concentration of metals (Pb, Cd, Zn, Hg) in different ecosystem components (lake water, sediment, plankton, macrophytes, and fish tissues) has been determined in Velenjsko jezero, an artificial lake resulting from mining activity. The risk to humans from consuming fish has been evaluated from the heavy metal load of fish muscle tissue. Heavy metals are transferred through the food chain at different levels, and Hg is found to be the element of the highest ecotoxicological concern. Although both sediment and plankton contain relatively low concentrations of Hg, this element accumulates in high levels in fish, especially in the benthivorous species Abramis brama danubii and predator species Perca fluviatilis. Moreover, Hg appears to be very mobile in the fish organism. Whereas the other metals remained mostly in liver (Cd) or gills (Zn, Pb), levels of Hg in fish muscle and liver were the same and markedly higher than in gills. However, in muscle, the average concentrations of each metal were below their maximum limits, determined either by Slovenian legislation or by the Food and Agriculture Organization.