Evidence of a phylogeographic break in the Romanian brown bear (Ursus arctos) population from the Carpathians

We analysed 33 brown bears from the Romanian Carpathians and the Italian Apennines at sequences of the mitochondrial control region and nine polymorphic microsatellite loci with regard to genetic variability and haplotype distribution. The Italian brown bears were monomorphic for mtDNA sequences. The Romanian bears yielded the highest variability found so far in this species. Haplotypes of both previously identified mtDNA lineages (western and eastern) were found in Romania. In the eastern part of the Carpathians western and eastern haplotypes occurred sympatrically, the bears from the western part of the mountain range only exhibited western-type sequences. This pattern provides evidence of a mitochondrial phylogeographic break in the distribution of the eastern lineage within the Romanian Carpathians. Conservation implications of this finding are discussed.     

Isotopic insights on cave bear palaeodiet

Abstract

More than 300 cave bear bones from all over Europe have carbon and nitrogen isotopic composition that match overwhelmingly a diet based on plants, except for samples from two caves in Romania, for which high nitrogen-15 amounts have been interpreted as reflecting an omnivorous diet. This paper aims at deciphering the various factors influencing the carbon and nitrogen isotopic composition of a potential omnivorous species like cave bear, those linked to trophic levels and variations among plants and those caused by physiological factors. The comparison of European cave bears with coeval Late Pleistocene large mammals with different diets clearly shows that all the cave bear populations, including those from Romania, present isotopic values overlapping with herbivores, not with carnivores. Therefore omnivory is very unlikely for cave bears. Consumption of plants with high δ¹⁵N values, such as graminoids, forbs and possibly fungi, could explain in part the observed isotopic pattern. In addition, the variations in δ¹⁵N values through ontogeny support the hypothesis of a different hibernation pattern for the Romanian cave bears with high δ¹⁵N values. Future investigations using new isotopic approaches, especially nitrogen isotopic composition of collagen amino acids, should contribute to decipher the paleoecology of these Romanian cave bears.     

Conflict Bear Translocation: Investigating Population Genetics and Fate of Bear Translocation in Dachigam National Park,Jammu and Kashmir,India

The Asiatic black bear population in Dachigam landscape, Jammu and Kashmir is well recognized as one of the highest density bear populations in India. Increasing incidences of bear-human interactions and the resultant retaliatory killings by locals have become a serious threat to the survivorship of black bears in the Dachigam landscape. The Department of Wildlife Protection in Jammu and Kashmir has been translocating bears involved in conflicts, henceforth ‘conflict bears’ from different sites in Dachigam landscape to Dachigam National Park as a flagship activity to mitigate conflicts. We undertook this study to investigate the population genetics and the fate of bear translocation in Dachigam National Park. We identified 109 unique genotypes in an area of ca. 650 km² and observed bear population under panmixia that showed sound genetic variability. Molecular tracking of translocated bears revealed that mostly bears (7 out of 11 bears) returned to their capture sites, possibly due to homing instincts or habituation to the high quality food available in agricultural croplands and orchards, while only four bears remained in Dachigam National Park after translocation. Results indicated that translocation success was most likely to be season dependent as bears translocated during spring and late autumn returned to their capture sites, perhaps due to the scarcity of food inside Dachigam National Park while bears translocated in summer remained in Dachigam National Park due to availability of surplus food resources. Thus, the current management practices of translocating conflict bears, without taking into account spatio-temporal variability of food resources in Dachigam landscape seemed to be ineffective in mitigating conflicts on a long-term basis. However, the study highlighted the importance of molecular tracking of bears to understand their movement patterns and socio-biology in tough terrains like Dachigam landscape.     

Low genetic variability of <Emphasis Type="Italic">Rhinolophus mehelyi</Emphasis> (Mehely's horseshoe bat) in Romania

Rhinolophus mehelyi (Mehely's horseshoe bat) is a vulnerable species with an increasingly fragmented distribution. In Romania, populations of R. mehelyi have experienced a dramatic decline over the past 50 years, and the current population size is estimated at only 100 adult individuals inhabiting almost exclusively the Limanu cave. In the present study, we investigated the genetic consequences of population decline for the viability of the remaining population of R. mehelyi in Romania. We sequenced and analyzed a 359-bp fragment of the mitochondrial control region from the only known Romanian population and compared it with two geographically close colonies from Bulgaria. A single haplotype was found in the Romanian population compared to 10 in the Bulgarian population, suggesting genetic isolation.     

Predominance of sexual reproduction in Romanian populations of the aphid Sitobion avenae inferred from phenotypic and genetic structure

Models of coexistence of sexual and asexual lineages in aphids assume that obligate parthenogenetic lineages predominate in areas with mild winter climate because of their high reproductive output, while sexual lineages predominate in areas with severe winter because they produce eggs resistant to frost. To validate this hypothesis in natural conditions, the reproductive mode of populations of the aphid Sitobion avenae was assessed in two very contrasting climatic situations, Romania (severe winter) and Western France (mild winter). To achieve this, reproductive modes were inferred from both (1) the population composition in sexual and asexual forms in autumn, and (2) the genetic structure of Romanian and French populations of S. avenae using microsatellite markers. Romanian populations encompassed a high proportion of sexual forms and were characterised by a very high genotypic diversity and low linkage disequilibrium. In constrast, the French population showed frequent linkage disequilibria, low genetic diversity, and high level of clonal amplification with two asexual genotypes representing over 60% of the sample. In agreement with the model's predictions, these results clearly indicate that sexual reproduction in S. avenae is predominant under the continental climate of Romania, while asexual lineages prevail under the oceanic climate of Western France.     

Conservation genetics of the European brown bear - a study using excremental PCR of nuclear and mitochondrial sequences

In the Brenta area of northern Italy, a brown bear Ursus arctos population is rapidly going extinct. Restocking of the population is planned. In order to study the genetics of this highly vulnerable population with a minimum of stress to the animals we have developed a PCR-based method that allows the study of mitochondrial and nuclear gene sequences from droppings collected in the field. This method is generally applicable to animals in the wild. Using excremental as well as hair samples, we show that the Brenta population is monomorphic for one mitochondrial lineage and that female as well as male bears exist in the area. In addition, 70 samples from other parts of Europe were studied. As others have previously reported, the mitochondrial gene pool of European bears is divided into two major clades, one with a western and the other with an eastern distribution. Whereas populations generally belong to either one or the other mitochondrial clade, the Romanian population contains both clades. The bears in the Brenta belong to the western clade. The implications for the management of brown bears in the Brenta and elsewhere in Europe are discussed.     

Evaluation of Holocene pollen records from the Romanian Plain

This study is a critical review of pollen analyses carried out on Holocene sequences from 15 sites in and near the Romanian Plain. Three sites come from natural sediments, 10 sites are from anthropogenic deposits and two are from both anthropogenic and natural settings. The general reconstruction is of a steppe-forest-steppe vegetation through the Holocene. The nature of the deposits, however, casts doubts on this reconstruction. Deposits of archaeological sites generally yield pollen spectra that are influenced by human activities and thus unsuitable for vegetation reconstructions. Loess deposits are also unfavorable for pollen preservation because of high pH and porosity. Consequently, pollen spectra from loess deposits are strongly biased by selective pollen destruction. Research and experiments carried out by several authors suggest that spectra dominated by Asteraceae, Poaceae, Chenopodiaceae or Pinus pollen in soils and loess are a result of selective pollen destruction, especially if low pollen concentrations, progressive pollen deterioration or high frequencies of deteriorated or unidentifiable pollen are evidenced. The fact that pollen records from the Romanian Plain come from loess, alkaline peat or archaeological sites reduces their reliability for reconstructions of vegetation. The vegetation history of similar regions in Hungary, Bulgaria and Turkey suggests that early Holocene steppe vegetation was gradually replaced by forest or forest-steppe vegetation in the late Holocene. Records from lake sediments are required to find out whether the Holocene vegetation history of the Romanian Plain was similar.     

Genotypic variation in safflower (Carthamus spp.) cadmium accumulation and tolerance affected by temperature and cadmium levels

Soil pollution is a world-wide problem, with heavy metals being a major part of the concern. To investigate the effect of temperature on cadmium (Cd) uptake and translocation, as well as Cd tolerance in wild and cultivated species of safflower, a hydroponic experiment was conducted under controlled conditions. The responses of four wild genotypes (Isfahan, Arak, Azari, and Shiraz) and four cultivated genotypes (AC-Sterling, 2811, Saffire, and C111) of safflower to nine levels of CdCl₂ (0, 0.5, 1, 5, 10, 20, 50, 100, and 500 μM) in solution were examined under two temperatures (18 and 23 °C). Cadmium sensitivity was determined using the Weibull model on the total dry weight of the plants. Cadmium uptake and translocation were analyzed on 1 μM Cd treated plants. Results revealed that safflower genotypes differed in terms of uptake, translocation, and tolerance to Cd, with AC-Sterling and Arak indicating the most and the least tolerance to Cd, respectively. Relative Cd uptake and Cd concentration in roots and shoots increased with an increase in temperature in all genotypes, with the exception of AC-Sterling. Net accumulation of Cd via root increased with an increase in temperature for the wild Azari and the cultivated 2811, Saffire, and C111, though it decreased for the rest of genotypes. Cadmium translocation to shoots significantly increased with increased temperature in all genotypes. Cadmium translocation from roots to shoots in cultivated genotypes was significantly greater than in wild genotypes. Root Cd concentration in wild genotypes was significantly greater than in cultivated genotypes. It seems that wild and cultivated species of safflower differ in their response to Cd. Furthermore, temperature may affect the plant's tolerance to Cd, probably through accompanying changes in Cd uptake and translocation from root to shoot.     

The power of genetic monitoring for studying demography,ecology and genetics of a reintroduced brown bear population

Genetic monitoring has rarely been used for wildlife translocations despite the potential benefits this approach offers, compared to traditional field‐based methods. We applied genetic monitoring to the reintroduced brown bear population in northern Italy. From 2002 to 2008, 2781 hair and faecal samples collected noninvasively plus 12 samples obtained from captured or dead bears were used to follow the demographic and geographical expansion and changes in genetic composition. Individual genotypes were used to reconstruct the wild pedigree and revealed that the population increased rapidly, from nine founders to >27 individuals in 2008 (λ = 1.17–1.19). Spatial mapping of bear samples indicated that most bears were distributed in the region surrounding the translocation site; however, individual bears were found up to 163 km away. Genetic diversity in the population was high, with expected heterozygosity of 0.74–0.79 and allelic richness of 4.55–5.41. However, multi‐year genetic monitoring data showed that mortality rates were elevated, immigration did not occur, one dominant male sired all cubs born from 2002 to 2005, genetic diversity declined, relatedness increased, inbreeding occurred, and the effective population size was extremely small (Ne = 3.03, ecological method). The comprehensive information collected through genetic monitoring is critical for implementing future conservation plans for the brown bear population in the Italian Alps. This study provides a model for other reintroduction programmes by demonstrating how genetic monitoring can be implemented to uncover aspects of the demography, ecology and genetics of small and reintroduced populations that will advance our understanding of the processes influencing their viability, evolution, and successful restoration.     

Microsatellite and Mitochondrial DNA Study of Native Eastern European Cattle Populations: The Case of the Romanian Grey

The Eastern European Grey cattle are regarded as the direct descendants of the aurochs (Bos taurus primigenius). Nowadays in Romania, less than 100 Grey animals are being reared and included in the national gene reserve. We examined the genetic diversity among Romanian Grey, Brown, Spotted and Black and White cattle breeds, with a particular focus on Romanian Grey through the use of (i) 11 bovine specific microsatellite markers on 83 animals and (ii) 638 bp length of mitochondrial DNA (mtDNA) D-loop region sequence data from a total of 81 animals. Both microsatellite and mtDNA analysis revealed a high level of genetic variation in the studied breeds. In Romanian Grey a total of 100 alleles were found, the mean number of observed alleles per locus was 9.091; the average observed heterozygosity was 0.940; the Wright’s fixation index (F_IS) was negative (-0.189) and indicates that there is no inbreeding and no selection pressure. MtDNA analysis revealed 52 haplotypes with 67 variable sites among the Romanian cattle breeds without any insertion or deletion. Haplotype diversity was 0.980 ± 0.007 and ranged from 0.883 ± 0.056 (Brown) to 0.990 ± 0.028 (Spotted and Black and White). The highest genetic variability of the mtDNA was recorded in the Grey breed, where 18 haplotypes were identified. The most frequent mtDNA D-loop region belonged to T3 haplogroup (80.247%), which was found across all studied breeds, while T2 haplotypes (16.049%) was only found in Grey, Spotted and Black and White genotypes. The T1 haplotypes (3.704%) were found in the Grey and Spotted. The current results contribute to the general knowledge on genetic diversity found in Eastern European cattle breeds and could prove a valuable tool for the conservation efforts of animal genetic resources (FAnGR).     

Genetic variation of the East Balkan Swine (Sus scrofa) in Bulgaria,revealed by mitochondrial DNA and Y chromosomal DNA

East Balkan Swine (EBS) Sus scrofa is the only aboriginal domesticated pig breed in Bulgaria and is distributed on the western coast of the Black Sea in Bulgaria. To reveal the breed's genetic characteristics, we analysed mitochondrial DNA (mtDNA) and Y chromosomal DNA sequences of EBS in Bulgaria. Nucleotide diversity (π_n) of the mtDNA control region, including two newly found haplotypes, in 54 EBS was higher (0.014 ± 0.007) compared with that of European (0.005 ± 0.003) and Asian (0.006 ± 0.003) domestic pigs and wild boar. The median‐joining network based on the mtDNA control region showed that the EBS and wild boar in Bulgaria comprised mainly two major mtDNA clades, European clade E1 (61.3%) and Asian clade A (38.7%). The coexistence of two mtDNA clades in EBS in Bulgaria may be the relict of historical pig translocation. Among the Bulgarian EBS colonies, the geographical differences in distribution of two mtDNA clades (E1 and A) could be attributed to the source pig populations and/or historical crossbreeding with imported pigs. In addition, analysis of the Y chromosomal DNA sequences for the EBS revealed that all of the EBS had haplotype HY1, which is dominant in European domestic pigs.     

Changes in soluble amino-N,soluble proteins and free amino acids in leaves and roots of salt-stressed maize genotypes

Abstract

The effects of salt stress on the contents of organic solutes and on the pattern of free amino acids were studied in leaves and roots of two maize genotypes, BR5033 (salt-tolerant) and BR5011 (salt-sensitive). In leaves and roots of salt-stressed plants, soluble amino-N increased with time when compared to the controls. Salt stress increased the soluble protein content only in leaves of BR5011. Salinity increased the content of the majority of the free amino acids in leaves and roots of genotypes studied. Results suggest the hypothesis of disturbances in translocation of N-containing compounds from shoot to root in the salt-sensitive genotype. Results also suggest that the accumulation of organic solutes, mainly in roots of BR5033, may have an important role in the tolerance of this genotype to salt stress.     

Diagnosing Mechanisms of Decline and Planning for Recovery of an Endangered Brown Bear (Ursus arctos) Population

Background

The usual paradigm for translocations is that they should not take place in declining populations until the causes(s) of the decline has been reversed. This approach sounds intuitive, but may not apply in cases where population decline is caused by behavioral or demographic mechanisms that could only be reversed by translocation itself.

Methodology/Principal Findings

We analyzed a decade of field data for Pyrenean brown bears (Ursus arctos) from two small populations: the growing Central population - created from a previous translocation and the endemic Western population - believed to be declining because of excessive human-caused mortality. We found that adult survival rates for both populations were as high as those observed for most other protected brown bear populations. However, the Western population had much lower reproductive success than the Central population. Adult breeding sex ratio was male-biased in the Western population and female-biased in the Central population. Our results exclude high anthropogenic mortality as a cause for population decline in the West but support low reproductive success, which could result from sexually selected infanticide induced by a male-biased adult sex ratio or inbreeding depression. Using a stochastic demographic model to compute how many bears should be released to ensure viability, we show that the Western population could recover provided adequate numbers of new females are translocated.

Conclusions/Significance

We suggest that a translocation could take place, even if the decline has not yet been reversed, if the translocation itself removes the biological mechanisms behind the decline. In our case, the ultimate cause of low reproductive success remained unknown (infanticide or inbreeding), but our proposed translocation strategies should eliminate the proximate cause (low reproductive success) of the decline and ensure population recovery and viability.     

Evaluating the role of environmental familiarity and behaviour in the success of wildlife translocation: A grizzly bear case study using agent-based modelling

Human-carnivore systems are built on multi-scalar complex processes often resulting in conflicts that force wildlife managers to address what are conceived as problem individuals. In North America, the grizzly bear (Ursus arctos) is often involved in human-bear conflict with management measures such as translocations, in which problem individuals are moved to new areas, being used to reduce conflict risk. While translocations offer a non-lethal alternative to managing conflict animals, they show varying levels of success. Our objective was to perform a novel assessment of grizzly bear translocation success through agent-based simulation by evaluating how familiarity with landscape features coupled with behavioral traits affects the way individuals use resources in a new environment. Our results showed that bears translocated to familiar habitat used high-quality habitat more than bears moved to areas with unfamiliar landscape characteristics. Increased exploration led to greater use of high-quality habitat in the long run but resulted in reduced use of high-quality habitat during the first two years following a translocation. Habitat quality use depended on scale, with bears translocated to less familiar environments accessing higher quality areas at a finer scale than bears translocated to familiar habitats. We emphasize the need to account for wildlife behavioral traits and habitat characteristics at multiple scales when selecting suitable translocation locations. Understanding the role of factors such as these on translocation outcome will help ensure the success of translocations not only as a method for managing problem wildlife, but also for population restoration, species reestablishment, and conservation translocations across the globe.     

Marker assisted detection of gene (1Dx5) and translocation (1B/1R) in wheat genotypes

Detection of 1Dx5 gene and presence of 1B/1R wheat rye translocation were studied in nineteen elite Indian wheat genotypes using AS-PCR and STS markers, respectively. Fifteen genotypes had 1B/1R translocation whereas ten showed presence of 1Dx5 gene. More than 50 per cent of the genotypes tested were found positive for both 1Dx5 and 1B/1R translocation. The results are in conformity with HMW glutenin SDS-PAGE profile for 1Dx5 and cytological observations for 1B/1R translocation.     

Primates Face to Face: Conservation Implications of Human–Nonhuman Primate Interconnections Edited by Agustín Fuentes and Linda D. Wolfe

ABSTRACT

Sterilization of companion animals is in most cases an expression of the caretaker's will. However, little is known about caretakers' attitudes toward pet sterilization in general—these can influence their decision to sterilize their own animals. Three hundred and twelve Romanian caretakers of non-sterilized companion animals (194 women and 118 men) were surveyed before the implementation of a pet management program in Transylvania, the North-Western part of Romania. The caretakers were administered a questionnaire that gathered demographic data and measured their general attitude toward pet sterilization and their attitude toward the sterilization of their own pet (male or female). Questionnaires were filled in by caretakers of 202 dogs and 110 cats. Romanian pet owners showed an overall level of slight disagreement toward the sterilization of companion animals. Gender-biased attitudes of the Romanian caretakers toward the sterilization of their own pets emerged only when the sex of the animals was taken into consideration, regardless of the type of pet. Romanian men disagreed with neutering male pets while Romanian women were more likely to agree with neutering male pets. No gender difference was detected regarding the spaying of female pets, with both genders showing a slight level of disagreement. The results are discussed from sociological, anthropological, and evolutionary perspectives.     

North-South Differentiation and a Region of High Diversity in European Wolves (Canis lupus)

European wolves (Canis lupus) show population genetic structure in the absence of geographic barriers, and across relatively short distances for this highly mobile species. Additional information on the location of and divergence between population clusters is required, particularly because wolves are currently recolonizing parts of Europe. We evaluated genetic structure in 177 wolves from 11 countries using over 67K single nucleotide polymorphism (SNP) loci. The results supported previous findings of an isolated Italian population with lower genetic diversity than that observed across other areas of Europe. Wolves from the remaining countries were primarily structured in a north-south axis, with Croatia, Bulgaria, and Greece (Dinaric-Balkan) differentiated from northcentral wolves that included individuals from Finland, Latvia, Belarus, Poland and Russia. Carpathian Mountain wolves in central Europe had genotypes intermediate between those identified in northcentral Europe and the Dinaric-Balkan cluster. Overall, individual genotypes from northcentral Europe suggested high levels of admixture. We observed high diversity within Belarus, with wolves from western and northern Belarus representing the two most differentiated groups within northcentral Europe. Our results support the presence of at least three major clusters (Italy, Carpathians, Dinaric-Balkan) in southern and central Europe. Individuals from Croatia also appeared differentiated from wolves in Greece and Bulgaria. Expansion from glacial refugia, adaptation to local environments, and human-related factors such as landscape fragmentation and frequent killing of wolves in some areas may have contributed to the observed patterns. Our findings can help inform conservation management of these apex predators and the ecosystems of which they are part.     

Control of late maturity alpha-amylase in wheat by the dwarfing gene Rht-D1b and genes on the 1B/1R translocation

The occurrence of late maturity alpha-amylase (LMA) was investigated using two doubled haploid wheat populations segregating for the dwarfing gene Rht-D1b and the 1B/1R translocation. Genotypes were assessed in the field and in controlled environments where a cold-shock treatment was used to induce LMA. Results from field-grown genotypes from the cross Spark × Rialto suggest that the absence of Rht-D1b or the presence of the 1B/1R translocation increases the expression of LMA.These two genetic factors were found to act independently and to have a positive interaction (complementary epistasis). In Option × Potent genotypes fixed for Rht-D1b, the 1B/1R effect was similar to that seen in the equivalent Spark × Rialto genotypes. Under controlled environment conditions, genotypes with the 1B/1R translocation showed a higher occurrence of LMA under both control and cold-shock conditions. 1B/1R was present in the majority of genotypes expressing LMA under control and cold-shock conditions. The results point to the novel finding that the 1B/1R translocation increases the expression of alpha-amylase in LMA-prone germplasm independently of effects of Rht-D1b, whereas previously it had been thought to act by a modification of the Rht-D1b effect.     

Nitrate Uptake and Partitioning by Corn Root Systems : Differential Effects of Ammonium among Genotypes and Stages of Root Development

The relative effects of ammonium on nitrate uptake and partitioning during induction were compared among decapitated seedlings of three corn (Zea mays L.) genotypes at two developmental stages. This study tested the hypothesis that root systems efficient at translocating products of ammonium assimilation away from sites of nitrate uptake or reduction would exhibit less inhibition of nitrate uptake by ammonium compared to root systems with inefficient N translocation efficiency. Inhibition of nitrate uptake by ammonium was relatively slight at day 5 ranging from 0% to 20% among the three genotypes, as compared to greater inhibition, from 20% to 37%, at day 8. Five-day-old roots exhibited negligible xylem translocation capacity in comparison with those grown for 8 days. Thus, although the capability to translocate ammonium assimilates out of the root increased between days 5 and 8, inhibitory effects of ammonium also increased. In the absence of ammonium, nitrate uptake per unit root mass increased between days 5 and 8. This increased activity of the uptake system was proportionally more sensitive to ammonium.

Partitioning of entering nitrate into the reduction process was positively correlated with lateral root development of the inbred root systems at 5 and 8 days. This is supportive of a localization of a major portion of nitrate reduction occurring in root apical regions. Nitrate reduction was the partitioning process most severely inhibited by ammonium in all cases, ranging from 39% to 55% inhibition. In contrast, ammonium-inhibition of nitrate accumulation in the root tissue and translocation via xylem vessels varied with genotype and root age.

Two mechanisms of ammonium-inhibition of nitrate are implicated, one which directly affects nitrate reduction and the uptake system associated with it, and another which may involve potassium as an intermediate regulator of nitrate accumulation in the root tissue and nitrate translocation out of the root tissue.