Echinococcus multilocularis in the red fox Vulpes vulpes from the East Carpathian region of Poland and the Slovak Republic

The occurrence of Echinococcus multilocularis in the Poland-Slovak frontier zone of the East Carpathian region was assessed, for comparison with that in adjacent regions in both countries. A total of 392 red foxes from Poland and 427 red foxes from the Slovak Republic were examined from 2001 to 2004. Significant differences in prevalences were observed in foxes captured from the borderland and adjacent zones in both countries. The mean prevalence of E. multilocularis in the Polish borderland reached 45.7+/-18.6% and in the Slovak border 35.0+/-10.7%. In both countries, the prevalence of E. multilocularis in red foxes from adjacent districts, outside the frontier Carpathian region, was considerably lower (18.9+/-9.2% in Poland and 20.8+/-9.0% in Slovakia). These differences are probably due to geomorphological and ecological factors, which contribute to the survival of the tapeworm eggs and the subsequent spread of infection. The Carpathian regions of northeast Slovakia and southeast Poland are characterized by specific climatic conditions such as low mean annual air temperatures, low temperatures in active soil surfaces, high soil humidities and a high mean annual rainfall.     

Craniometric variation in the Eurasian badger, Meles meles

A multivariate examination of cranial variation within and between European populations of Meles meles (L.) revealed that populations from Ireland, Great Britain, Norway, and the Slovak Republic could be differentiated both by cranial form and by the degree of sexual dimorphism exhibited. Irish material was characterized by low sexual dimorphism, particularly when compared to Slovak specimens. Badgers from the British Isles had larger skulls than other samples and were more similar to each other than they were to badgers from mainland Europe. Size played a greater role in differentiating samples of female badgers than it did in males. Significant variation occurred within the British Isles, with individual samples being highly differentiable. There was, however, little relationship between morphological similarity and geographic proximity. We contend that macrogeographical (between-country) variation in the species is primarily determined by historical factors and adaptation to current conditions, while microgeographic (within-country) variation is a result of selectively neutral processes.     

The genetic structure of raccoon introduced in Central Europe reflects multiple invasion pathways

Invasions of non-native species are of great concern as they have a devastating impact on native biodiversity and can also affect the economy of a region. Multiple introductions in several locations of a new range greatly promote the success of non-native species. The raccoon (Procyon lotor) is an omnivore whose native distribution extends from southern Canada to Panama. It has been successfully introduced in many European countries. We examined the microsatellite and mitochondrial diversity of raccoon populations in Central Europe (Germany, Poland, and Czech Republic) in order to determine their introduction sources and pathways as well as the factors affecting genetic structure in this invasive species. We found low diversity of the mtDNA control region and moderate diversity of microsatellite markers. Raccoon showed three hierarchic levels of genetic structure which separate at different levels sampled from Czech Republic, Germany and raccoon inhabiting two different habitats in Poland. In Poland the raccoon population was established through migration from Germany to Czech Republic. Analysis of the intensity of migration between two different habitat types indicated source-sink dynamics in the Polish populations of raccoons. Our results confirm the high intensity of the raccoon invasion in Central Europe and point to specific measures needed as part of an effective management strategy.     

Constant multiple birth rates in the Czech Republic and the Slovak Republic until recently, 1972-1995.

Using vital statistics, yearly changes in the twinning and triplet rates by zygosity were investigated in the Czech Republic and the Slovak Republic during the period 1972-1995. Monozygotic (MZ) twinning rates in both countries had remained nearly constant (about 3 per 1000 total births) during that period. With a few exceptions, the dizygotic (DZ) twinning rates remained constant from 1972 to 1994, and increased in 1995 for both countries. MZ twinning rates for both countries were the lowest in Europe. As for triplet rates, overall rates increased significantly year by year in the Czech Republic, but not in the Slovak Republic. The triplet rate was significantly higher in 1995 than in the period 1972-1982 for both countries. The MZ triplet rate remained constant during that period in the Czech Republic. The trizygotic (TZ) triplet rates increased 3-fold for the Czech Republic and 4-fold for the Slovak Republic in 1972-1976 and 1992-1995. In the later period, the TZ rate was 1.5-fold higher in the Czech Republic than in the Slovak Republic. The quadruplet rate increased 2.3-fold from 2.9 per million births in 1982-1986 to 6.7 in 1992-1995 in the Czech Republic. The corresponding values were 2.7, 2.20 and 5.9-fold in the Slovak Republic. Both the Czech and the Slovak Republics were not affected by fertility drugs and assisted reproductive techniques until recently.     

Comparative analysis of morphometric characters of juvenile sterlet Acipenser ruthenus L. from natural population and aquaculture

A comparative analysis was performed on sixteen morphometric characters, in three different juvenile sterlet Acipenser ruthenus L. populations. Specimens were collected from a wild population in the Serbian part of the Danube River (n = 46), from aquaculture stocks in the Czech Republic originating from Russia (n = 40), and aquaculture stocks in the Slovak Republic originating from Slovakian part of the Danube River (n = 28). Average values for total length were 29·9 ± 3·9 cm, 29·1 ± 3·7 cm and 27·3 ± 7·7 cm for Serbia, Czech Republic and Slovak Republic, respectively. Populations were compared using t‐test and sequential Bonferroni correction for multiple comparisons was applied in order to determine significant differences between them. Results of analysis showed that all three populations differed in prebarbel length, interocular distance and maximum head width. Although all these characters are head‐related, head length itself was very uniform among all populations. The natural population from the Serbian part of the Danube River differed from the populations reared in aquaculture in seven morphometric characters. The two populations reared in aquaculture consistently showed lower morphological variability than the wild population, even though they had different genetic backgrounds (Russia and Slovakia). Future genetic studies will show if this tendency is caused by a reduction in genetic variability.     

Population genetic structure of the European ground squirrel in the Czech Republic

The European ground squirrel (Spermophilus citellus) is considered an endangered species with declining numbers throughout Europe, most pronounced at the western margin of its distribution area. Being extinct in Germany and Poland, the western margin of its distribution is in the Czech Republic. Here, landscape fragmentation has restricted the ground squirrels into few and very isolated localities where local extinctions still occur. In the present study we analysed European ground squirrels from six Czech and one Slovak localities using five microsatellite loci as genetic marker. The results show a strong genetic differentiation among the investigated populations (mean value of F _ST = 0.16) and high levels of inbreeding (values of F _IS ranged from 0.34 to 0.90). High level of inbreeding is generally considered to affect the viability of each population, which could lead to extinction. One of the most important factors is the lack of migration due to the large distances between the populations and the presence of migration barriers. Based on the results obtained we recommend a few suggestions for a conservation management of this species.     

Population dynamics of fisheries stock enhancement

A comparative analysis was performed on sixteen morphometric characters, in three different juvenile sterlet Acipenser ruthenus L. populations. Specimens were collected from a wild population in the Serbian part of the Danube River ( n  = 46), from aquaculture stocks in the Czech Republic originating from Russia ( n  = 40), and aquaculture stocks in the Slovak Republic originating from Slovakian part of the Danube River ( n  = 28). Average values for total length were 29·9 ± 3·9 cm, 29·1 ± 3·7 cm and 27·3 ± 7·7 cm for Serbia, Czech Republic and Slovak Republic, respectively. Populations were compared using t ‐test and sequential Bonferroni correction for multiple comparisons was applied in order to determine significant differences between them. Results of analysis showed that all three populations differed in prebarbel length, interocular distance and maximum head width. Although all these characters are head‐related, head length itself was very uniform among all populations. The natural population from the Serbian part of the Danube River differed from the populations reared in aquaculture in seven morphometric characters. The two populations reared in aquaculture consistently showed lower morphological variability than the wild population, even though they had different genetic backgrounds (Russia and Slovakia). Future genetic studies will show if this tendency is caused by a reduction in genetic variability.     

A comparative study of zygotic twinning and triplet rates in eight countries, 1972-1999

Annual changes in twinning and triplet rates by zygosity were investigated in eight countries during the period 1972-1999 using vital statistics. The monozygotic (MZ) twinning rates in Denmark, Switzerland and the Slovak Republic remained more or less constant throughout this period, whereas those in England and Wales, the Federal Republic of Germany (Germany), the Netherlands, the Czech Republic and Japan increased significantly year by year. With the exception of the Slovak Republic, the dizygotic (DZ) twinning rate increased significantly year by year in each country. It was 2.9 times higher in Denmark and 1.5 times higher in Germany in 1999 than in 1972, and within the same range in the other countries. With two exceptions, the MZ triplet rates remained more or less constant in each country. On the other hand, the DZ and trizygotic (TZ) triplet rates increased significantly year by year in each country. The TZ rate increased 30-fold in Germany, 16.6-fold in Japan, 11.7-fold in Switzerland, 9.7-fold in the Czech Republic, 8.7-fold in the Netherlands, 6.4-fold in Denmark, 5.6-fold in England and Wales and 3.5-fold in the Slovak Republic. The higher DZ twinning rate and higher DZ and TZ triplet rates since 1983 have been attributed to the higher proportion of mothers being treated with ovulation-inducing hormones and in vitro fertilization (IVF) in Denmark, England and Wales, Germany, the Netherlands, Switzerland and Japan. After the introduction of fertility drugs and IVF, variations in the DZ twinning and triplet rates and the TZ triplet rates were not only due to biological factors, but also depended on the popularity of fertility drugs and IVF in each country. In the Slovak Republic, where human fertility might not be affected by some adverse environmental factors, the DZ:MZ ratio remained constant during the period 1972-1999.     

The status of the Badger Meles meles (L., 1758) (Carnivora, Mustelidae) in Europe

The Eurasian Badger occurs throughout the Palaearctic, and in all states of Europe west of the border with the former Soviet Union. Within this area it is absent only from the arctic zones, high-altitude areas, and some islands. The Badger is currently a protected species in the UK, the Irish Republic, Spain, Portugal, Italy, Belgium, the Netherlands, Albania, Greece, Estonia, Luxembourg and Hungary, but Luxembourg and Hungary are to reconsider protected status. Elsewhere, the species is either considered as small-game or as a pest, hunting being regulated by closed seasons. At present Finland and Burgenland (Austria) afford protection to breeding females, whilst Bulgaria, Macedonia and the Austrian Bundesländer of Steiermark and Salzburg permit Badger hunting throughout the year. Where the species is protected, provisions usually exist for the removal or culling of ‘pest’ individuals. The official European game-bag currently totals about 118,000 Badgers, but poaching is common, particularly in the UK and Ireland. Published population estimates, coupled with national population minima obtained by extrapolation from game-bag statistics, indicate a minimum European Badger population of 1,220,000; the true figure may exceed this considerably. Badger populations appear to be either stable or increasing throughout much of Europe, although no data are currently available for the populations of Greece, Italy, Spain, Croatia, Bosnia-Hercegovina and Portugal. Badgers are uncommon in the Netherlands, Estonia, Belgium, the Slovak Republic, and possibly Poland. Only the populations of Albania and possibly of some parts of the former Yugoslavia appear to be decreasing. The Dutch population remains at considerable risk, despite modest recent increases. The population status of the endemic sub-species of Crete and Rhodes remain uncertain and require urgent clarification. A series of management recommendations are proposed to improve the status of the Badger in Europe.     

Larval trematodes (Digenea) of the great pond snail, Lymnaea stagnalis (L.), (Gastropoda, Pulmonata) in Central Europe: a survey of species and key to their identification

A survey of cercariae and metacercariae (Trematoda, Digenea) from the great pond snail (Lymnaea stagnalis) in Central Europe (Austria, Czech Republic, South-East Germany, Poland and Slovak Republic) is presented, based on a study of 3,628 snails examined from 1998 to 2005. A total of 953 (26.3%) L. stagnalis were infected with 24 trematode species comprising 19 species of cercariae and 11 species of metacercariae (six species occurred both as cercarie and metacercarie) of eight families. The dominant cercariae were those of Opisthioglyphe ranae (159 hosts infected), Plagiorchis elegans (141) (both family Plagiorchiidae) and Echinoparyphium aconiatum (153) (Echinostomatidae); 14 double infections were found. The most frequent metacercariae were those of Neoglyphe locellus (71) (Omphalometridae), E. aconiatum (66), Echinostoma sp. (59) and Moliniella anceps (48) (Echinostomatidae). In the previous studies carried out in Central Europe, a very similar spectrum of nine trematode families of 22 cercariae determined to species level and 43 types of cercariae reported under generic or provisional names, which can be in many cases conspecific with the previous taxa, were found. A simple key to identification of cercariae and metacercariae, together with their illustrations, is provided.     

The codon 408 mutation associated with haplotype 2 is predominant in Polish families with phenylketonuria

Summary The incidence of phenylketonuria (PKU) in the western part of Poland is 1 in 5000 live births. Restriction fragment length polymorphism (RFLP) haplotypes at the phenylalanine hydroxylase locus have been analysed in 46 Polish families with PKU. Among 43 fully-informative families 16 RFLP haplotypes were identified. Haplotype 2 is the most frequently (62%) associated with Polish PKU alleles, and the codon 408 mutation is in complete linkage disequilibrium with this haplotype in Poland. This finding is in agreement with observations in other eastern European countries (German Democratic Republic, Czechoslovakia, and Hungary) and in contrast to the genotype distribution observed in western European countries. The present observation suggests the spread of classical PKU, due to the codon 408 mutation associated with haplotype 2, from east to west in European populations. Perhaps more important for genetic counselling, 62% of all PKU chromosomes in the Polish population can now be detected using only one mutantspecific oligonucleotide probe.     

Genomic characterization of Pinzgau cattle: genetic conservation and breeding perspectives

A genome-wide scan of Slovak Pinzgau cattle was prepared for the first time in order to estimate their genetic diversity at a more detailed level compared to previously published studies. The aim of this study was to describe the genetic diversity based on the runs of homozygosity (ROH_s), linkage disequilibrium (LD) and effective population size (N_eLD) using genome-wide data. Moreover, Bayesian clustering algorithms and multivariate methods were used to detect the population structure, potential admixture level and relationship between Austrian and Slovak Pinzgau cattle with respect to a large meta-population consisting of 15 European cattle breeds. The proportion of ROH segments ranged from 0.43 to 1.91% in Slovak Pinzgau, depending on the minimum size of an ROH. The genomic inbreeding coefficients were higher than the pedigree ones possibly due to the limited number of available generations in pedigree data. The observed N_eLD was close to the limit value characterizing the endangerment status, based both on genomic and pedigree data. Population structure within analyzed breeds based on the Wright’s F_ST index, Nei’s genetic distances, and unsupervised as well as supervised analysis has been established. Overall, these analyses clearly distinguished populations based on their origin. A detailed analysis of the introgression of each breed into the Pinzgau breeds prepared using a Bayesian approach showed that the contribution of Holstein cattle in Austrian as well as Slovak Pinzgau was larger than contribution of beef breeds. A possible reason is the recent usage of Holstein sires to increase milk production. There are considerable differences between well-defined regions that clearly distinguish Austrian and Slovak Pinzgau, despite their close common history. Generally, the breeding program of Austrian Pinzgau is more focused on meat production than Slovak Pinzgau, which was clearly reflected in the obtained autozygosity islands. Considering the genetic establishment of Slovak Pinzgau population the genetic potential of the breed is insufficiently used. On a long term, more global breeding program including very close populations will be more efficient providing higher genetic progress and diversity. Established methodology how to distinguish genealogically close populations on high-throughput molecular information based of Slovak and Austrian Pinzgau can be proposed as general for analysis of differences in all highly related breeds.     

Direct comparison of assessment methods using benthic macroinvertebrates: a contribution to the EU Water Framework Directive intercalibration exercise

The aim of the intercalibration exercise presently performed by the EU is to identify and resolve significant inconsistencies between the ecological quality classifications of EU Member States and the normative definitions of the EU Water Framework Directive. Based on benthic macroinvertebrate data of two European stream types (small siliceous mountain streams and medium-sized lowland streams in Central and Western Europe) we correlated the indices of 10 river quality assessment methods (ASPT, BMWP, DSFI, German Multimetric Index, Saprobic Indices) applied in Austria, Czech Republic, Denmark, Germany, Poland, Slovak Republic, Sweden and United Kingdom. National class boundaries were compared via regression analysis. Assessment methods of the same type (Saprobic Indices, BMWP/ASPT scores) showed best correlation results (R²>0.7). The good quality status boundaries of the national methods deviated up to 25%; thus indicating the necessity to harmonize the national classification schemes. Prerequisites of the presented intercalibration approach are (1) a sufficiently large and consistent dataset representative of the respective common intercalibration types and (2) agreement on common type specific reference conditions.     

Allozyme diversity in relation to geographic distribution and population size inLathyrus vernus (L.) Bernh. (Fabaceae)

Lathyrus vernus (L.) Bernh. is a diploid, long-lived perennial and insect-pollinated herb with no special adaptation to long-distance dispersal. It occurs on neutral soil in deciduous forests throughout western Eurasia. Due to specific habitat preferences,L. vernus has a fragmented distribution with isolated populations. We investigated allozyme variation at eleven loci in 20 populations ofL. vernus from one geographically central region (the Czech Republic and the Slovak Republic) and two geographically marginal regions (southern and central Sweden) in the species present-day distribution. There was a clear differentiation between the three regions and the genetic distance between the populations was highly correlated with geographic distance. The total genetic diversity (H_T) was 0.354. The proportion of genetic diversity due to differentiation between regions, and to differentiation between populations within regions, accounted for 10% each. There was no difference in level of genetic diversity between the three regions. No significant difference in level of genetic diversity was found between small and large populations. The genetic diversity inL. vernus may either be a result of the long generation-time of the species or peculiarities in the post-glacial migration species, e.g. survival only in refugia far east of the sampled populations and/or migration as a continuous process not involving founder-events.     

Comparison of Y-STR polymorphisms in three different Slovak population groups

Eleven Y-chromosomal microsatellite loci included in the Powerplex Y multiplex kit were analyzed in different Slovak population samples: Habans (n = 39), Romanies (n = 100) and Slovak Caucasian (n = 148) individuals, respectively, from different regions of Slovakia. The analysis of molecular variance between populations indicated that 89.27% of the haplotypic variations were found within populations and only 10.72% between populations (Fst = 0.1027; p = 0.0000). The haplotype diversities were ranging from 0.9258 to 0.9978, and indicated a high potential for differentiating between male individuals. The study reports differences in allele frequencies between the Romanies, Habans and Slovak Caucasian men. Selected loci showed that both the Romany and Haban population belonged to endogamous and relatively small founder population groups, which developed in relatively reproductive isolated groups surrounded by the Slovak Caucasian population.     

Genetic diversity and the phylogeography of parthenogenesis: comparing bisexual and thelytokous populations of Nemasoma varicorne (Diplopoda: Nemasomatidae) in Denmark

The millipede, Nemasoma varicorne, represents a textbook example of geographic parthenogenesis with thelytokous populations being distributed north, east, south and west of the distribution of the bisexual ancestor in the deciduous forests of central Europe. We here describe variation in amplified fragment length polymorphisms (AFLP's) in sympatric bisexual and thelytokous populations of N. varicorne in Denmark and compare the relationships of Danish populations with animals from The Czech Republic, England and Poland. Thelytokes from Denmark, England and Poland form a monophyletic cluster that differs from bisexuals from Denmark and Czechia for about 30% of the fragments. A single clone is widely spread over Denmark (34 of 38 localities), with rare clones being detected at four other localities. The phylogenetic pattern implies strongly that thelytoky evolved prior to the post-glacial colonization of northern Europe. This further suggests that the two forms have interacted extensively during this colonization and that the thelytokes have been excluded from older forests by competition with the bisexual forms. Our results further suggest that the success of the thelytokous form, at least in Denmark, is not due to abundant clonal diversity as hypothesized by the frozen niche variation model.     

Genetic diversity of a relict plant species, Ligularia sibirica (L.) Cass. (Asteraceae)

Rare plant species can be divided into naturally, ‘old rare’ species and anthropogenically, ‘new rare’ species. Many recent studies explored genetic diversity of ‘new rare’ species. Less is, however, known about genetic diversity of ‘old rare’ species. We examined isozyme genetic variability of 20 populations of an ‘old rare’ plant species, Ligularia sibirica (Asteraceae) in the Czech and Slovak Republic. It is a long-lived perennial herb with mixed-mating breeding system, widely distributed from East Asia to European Russia, with few isolated relict populations in the remaining part of Europe.The results showed high genetic diversity within populations (80.8%) and a low level of genetic differentiation (F_ST = 0.179). Genetic distance between populations correlated significantly with geographic distance. There was also a significant positive correlation between genetic diversity and population size. This is probably caused by destruction of habitats in last centuries and subsequent decrease of population size. Patterns of genetic diversity suggest that the recent distribution is a result of stepwise postglacial migration of the species and subsequent natural fragmentation.We conclude that L. sibirica populations preserve high levels of genetic diversity and are not yet threatened by genetic factors. However, this may change if changes in habitat conditions continue.     

Examining <Emphasis Type="Italic">Ambrosia</Emphasis> pollen episodes at Poznań (Poland) using back-trajectory analysis

The pollen grains of Ambrosia spp. are considered to be important aeroallergens in parts of southern and central Europe. Back-trajectories have been analysed with the aim of finding the likely sources of Ambrosia pollen grains that arrived at Poznań (Poland). Temporal variations in Ambrosia pollen at Poznań from 1995–2005 were examined in order to identify Ambrosia pollen episodes suitable for further investigation using back-trajectory analysis. The trajectories were calculated using the transport model within the Lagrangian air pollution model, ACDEP (Atmospheric Chemistry and Deposition). Analysis identified two separate populations in Ambrosia pollen episodes, those that peaked in the early morning between 4 a.m. and 8 a.m., and those that peaked in the afternoon between 2 p.m. and 6 p.m.. Six Ambrosia pollen episodes between 2001 and 2005 were examined using back-trajectory analysis. The results showed that Ambrosia pollen episodes that peaked in the early morning usually arrived at Poznań from a southerly direction after passing over southern Poland, the Czech Republic, Slovakia and Hungary, whereas air masses that brought Ambrosia pollen to Poznań during the afternoon arrived from a more easterly direction and predominantly stayed within the borders of Poland. Back-trajectory analysis has shown that there is a possibility that long-range transport brings Ambrosia pollen to Poznań from southern Poland, the Czech Republic, Slovakia and Hungary. There is also a likelihood that Ambrosia is present in Poland, as shown by the arrival of pollen during the afternoon that originated primarily from within the country.     

Historical sketch of Slovak Haban (Hutterite) population based on autosomal STR analysis

According to the Hutterite chronicles, the Habans arrived from Austrian Tyrol, Switzerland, and northernmost Italy and stayed in four regions of Slovakia (Soboti?te, Vel'ké Leváre, Moravsky Sv?ty Ján, Tren?ín). There are some communities in western Slovakia that retained their Haban cultural identity and still identify themselves as descendents of the Hutterite population with their own specific customs. Slovak Habans are typical founder population with significant social isolation for which high degree of inbreeding is typical. Present study investigated STR polymorphisms as a powerful genetic tool for population genetic studies. The aim was to perform a comparative, population genetic study based on 15 STR loci widely used in forensic genetics, of the Haban population, the Slovak majority population and the population of Tyrol. We analyzed allele frequencies and other statistical parameters in three selected populations in order to identify groups of specific ethnic origin and establish their genetic relationship. The data set included 110 unrelated Habans and 201 unrelated individuals from the Slovak majority population, as well as allelic frequencies for the population of Austrian Tyrol available in the literature. Population pairwise FST values used as a short term genetic distance between populations showed significant differentiation between the Habans and both reference populations (FST=0.0025 and 0.0042 for comparison with the Slovaks and Austrians, respectively; p<10(-3)). The Slovak Hutterites were demonstrated to be genetically distinct and more closely related to their geographic neighbors than to their historical ancestral population, which may be at least partially explained by gene flow between neighboring Haban and Slovak populations.     

Effect of salt stress on fatty acid alterations in some strains of Dipodascopsis and Dipodascus spp.

The effect of salt stress (8% w/v NaCl) on fatty acid composition of eight strains of Dipodascus and Dipodascopsis spp. varied from being of slight influence only (Dipodascopsis uninucleata), to decreasing the content of 18:2 (D. reesii, D. tetrasperma and D. australiensis) and to decreasing both 18:1 and 18:2 (D. tothii and D. aggregatus) with a concomitant rise of 14:1 and 16:1. With the exception of D. aggregatus, NaCl inhibited lipid accumulation in all strains. Only trace amounts of fatty acids over C₁₈ in chain length were found.J. ajbidor, M. Lamaka, A. Chrastina and P. Pokreisz are with the Department of Biochemical Technology, Slovak Technical University, Radlinskeho 9, 812 37, Bratislava, Slovak Republic; E. Breirerová is with the Institute of Chemistry, Slovak Academy of Science, Dúbravská cesta 9, Slovak Republic. M. Certík is with the Research Institute of Gerontology, Zámocká ul, P.O. Box 25, 901 01 Malacky, Slovak Republic.