Population genetic studies in the Balkans. II. DNA-STR-systems

Within a study of the genetics of Southeastern European populations four DNA-STR-systems (D21S11, FGA, TH01, VWA) were examined in seven samples (samples of three Aromuns and four other Balkan populations). The results have been compared to data from four samples from literature (Austrians, Germans, Hungarians, Slovenians). The results show three clusters: a) the Aromuns from Albania (Andon Poci) and Macedonia (Stip region), b) the Romanian Aromuns (Kogalniceanu), Romanians (Constanta, Ploiesti) and Albanians (Tirana) und c) the data from literature. A sample of Northeastern Greece clearly differs from these three clusters. Including seven serum protein polymorphisms (without the populations from literature) results in two clusters: a) the three Aromun populations and b) Albanians and Romanians. Again the sample of Northeastern Greece clearly differs from these clusters.     

Allele frequencies of D21S11, FGA, TH01, and VWA in populations of the Balkans

This study is part of an extensive investigation of the genetic relationship between Balkan populations, especially the Aromuns. Allele frequencies of four STRs (D21S11, FGA, TH01, VWA) from Macedonians (Skopje), Gramostian Aromuns from the Stip region (Macedonia), Moskopolian Aromuns from Krusevo (Macedonia), and Musequiar Aromuns from Dukasi (Albania) are presented.     

Dermatoglyphic studies in eastern and south-eastern Europe

In nine population samples from Bulgaria, Croatia, Greece, Hungary, Republic of Macedonia (Skopje and Aromuns from Stip region), Romania, Serbia and Slovakia 12 dermatoglyphic variables have been studied. There are distinct differences between the populations and between males and females. The Macedonian Aromuns are clearly separated from the other populations.     

Phenotype and allele frequencies of some serum protein polymorphisms in populations of the Balkans

Within a study of the genetics of Southeastern European populations seven serum protein polymorphisms (AMY2, BF, C3, CP, GC, HPA, TF) were examined in two samples of Aromuns and one reference sample (Musequiar-Aromuns from Dukasi in Albania, Moskopolian-Aromuns from Krusevo, Republic of Macedonia, and Macedonians from Skopje). The neighbor joining tree as well as the principal component analysis show results which do not correspond well to the geographic and historic background. This indicates that in the present case the serum protein polymorphisms give no clearly defined information about the relationships between the Balkan populations and to the origin of Aromuns.     

DNA-PCR systems TH01 and VWA31: population data from Albania and Romania

The microsatellite systems TH01 and VWA31 have been typed and analyzed for three populations from Albania (city population of Tirana, and Aromuns from a small-sized village, named Andon Poci, near Gjirokaster) and Romania (city population of Constanta). The genetic data have been compared with other, particularly South-Eastern European population.     

Isonymy, consanguinity and repeated pairs of surnames in Aromun populations

The Aromuns represent a small and almost unknown people that live scattered over the Balkan Peninsula. Due to their language, that is very similar to classical Latin, they are in a special position. The Aromuns settled only in more recent times. Until now they lived as shepherds, as caravan guides and merchants and lead a semi-nomadic life. We are currently carrying out studies to determine the genetic structure of this population. To facilitate the interpretation of these data, we are also trying to obtain other important parameters that pertain to migration processes and the genealogical structure of this populations. The data arise from three areas in Albania, the Republic of Macedonia and Romania. The inbreeding coefficient and the proportion of repeated pairs of surnames was calculated through the use of genealogies and the isonymy method. The difference between these three populations are due primarily to confounding by selection of mates and family composition.     

Population genetic studies of the pancreatic amylase (AMY2, E.C. 3.2.1.1) in Bulgaria

The pancreatic amylase (AMY2, E.C. 3.2.1.1) polymorphism has been studied in 2346 individuals from south-central and south-eastern Bulgaria. The allele frequencies have been determined as AMY2*1 = 0.9520 and AMY2*2 = 0.0480. The neighbor joining tree of seven subpopulations revealed only small genetic distances. Compared with other populations, the Bulgarian sample clustered with samples from Romania, Hungary, Germany and Switzerland, with larger distances to Albania, Greece and Macedonia.     

Variability of blood group frequencies in Romania

This paper presents the results of a study of two population samples from Dobruja and Muntenia (Romania) as well as data from the literature (AB0, MN, Rhesus (D/d) and HPA). The data have been pooled according to the different regions of Romania. Interestingly, the Carpathian Mountains do not form a genetical barrier: there are clear genetical similarities between the provinces east and west as well as between the provinces south and north from the mountains. The differences and similarities between Romania as a whole and other populations in this region reflect the historical processes of the last centuries.     

Subgeneric division of the genus Orcula Held?1837 with remarks on Romanian orculid data (Gastropoda,Pulmonata, Orculidae)

The genital anatomy of Orcula jetschini (Romania), Orcula zilchi (Bulgaria), and Orcula wagneri (Albania) is described. Based on anatomical features (morphology of the penial caecum) shell characters (sculpture and shape) and unpublished molecular data the genus Orcula is subdivided into three subgenera. Orcula zilchi was classified within the monotypic subgenus Orcula (Hausdorfia) subgen. n.; Orcula jetschini, Orcula wagneri, and Orcula schmidtii were classified to Orcula (Illyriobanatica) subgen. n. (type species: Pupa schmidtii) whereas the other Orcula species remain in the nominotypical subgenus. Orcula (Hausdorfia) is known from South-Eastern Bulgaria and North-Western Turkey, Orcula (Illyriobanatica) inhabits Western Romania, North-Western Greece, Albania, Macedonia, Kosovo, and Montenegro. The nine species of Orcula (Orcula) are known mainly from the Alps and the Western Carpathians (from Eastern France to Eastern Hungary and Slovakia).The occurrence of only one Orcula species namely Orcula jetschini is verified from Romania. Available information suggests that data on the Romanian occurrence of Orcula dolium and Orcula gularis were based on wrongly identified specimens. Sphyradium dobrogicum (=Orcula dobrogica) is considered as a synonym of Sphyradium doliolum.     

Genomic characterization of human and environmental polioviruses isolated in Albania.

Between April and December 1996, a serious outbreak of poliomyelitis occurred in Albania; almost 140 subjects were involved, and the episode presented an unusually high mortality rate (12%). During the outbreak, water samples from the Lana River in Tirana, Albania, and stool samples from two cases of paralytic poliomyelitis were collected and analyzed for the presence of polioviruses. Six polioviruses were isolated from the environmental and human samples, according to standard methods. All the samples were characterized by partial genomic sequencing of 330 bases across the 5' untranslated region (5'-UTR) (nucleotide positions 200 to 530) and of 300 bases across the VP1 region (nucleotide positions 2474 to 2774). Comparison of these sequences with those present in data banks permitted the identification of environmental isolates Lana A and Lana B as, respectively, a Sabin-like type 2 poliovirus and an intertypic recombinant poliovirus (Sabin-like type 2/wild type 1), both bearing a G instead of an A at nucleotide position 481. The two other environmental polioviruses were similar to the isolates from the paralytic cases. They were characterized by a peculiar 5'-UTR and by a VP1 region showing 98% homology with the Albanian epidemic type 1 isolates reported by other authors. This study confirms the environmental circulation in Albania of recombinant poliovirus strains, likely sustained by a massive vaccination effort and by the presence in the environment of a type 1 poliovirus, as isolated from the Lana River in Tirana about 2 months before the first case of symptomatic acute flaccid paralysis was reported in this town.     

Clonal population structure of the chestnut blight fungus in expanding ranges in southeastern Europe

Expanding populations are often less genetically diverse at their margins than at the centre of a species' range. Established, older populations of the chestnut blight fungus, Cryphonectria parasitica, are more variable for vegetative compatibility (vc) types than in expanding populations in southeastern Europe where C. parasitica has colonized relatively recently. To test whether vc types represent clones, we genotyped 373 isolates of C. parasitica from southern Italy, Romania, Bulgaria, Macedonia, Greece and Turkey using 11 sequence-characterized amplified region (SCAR) markers. Ten SCAR loci and six vegetative incompatibility (vic) loci were polymorphic in these samples. These populations are clonal by all criteria tested: (i) among 373 isolates, we found only eight multilocus haplotypes, and the same haplotypes were found in multiple countries, sometimes separated in time by as much as 12 years; (ii) the number of haplotypes observed was significantly less than expected under random mating; (iii) populations are in linkage disequilibrium; (iv) the two sets of independent markers, SCARs and vc types, are highly correlated; and (v) sexual structures of C. parasitica were found only in Bulgaria and Romania. One mating type (MAT-1) was found in 98% of the isolates sampled. In contrast, a population in northern Italy, in the central part of the range in Europe, had 12 multilocus haplotypes among 19 isolates. The spread of a few clones could be the result either of founder effect and restricted migration, or these clones have greater fitness than others and spread because they are better adapted to conditions in southeastern Europe.     

Jumping into the grids: mapping biodiversity hotspots in groundwater habitat types across Europe

Biodiversity hotspots are routinely identified by grid-based analyses, despite grids encompassing different habitats, thus hindering the potential to assess which habitat type accounts for the conservation priority assigned to a grid. In this study, we aimed at identifying the main hotspots for the conservation of the European stygobitic Crustacea Copepoda Harpacticoida at the groundwater habitat scale. A multi-metric approach was used, based on six biodiversity indicators: species richness, endemicity, evolutionary origin, phylogenetic rarity, taxonomic distinctness, habitat specificity. The Hot Spot Analysis, based on the statistics Getis-Ord Gi*, was used to compare the local to the global average values of each indicator to identify hotspots of conservation. The operational units used to perform the analyses were the groundwater habitat types, in order to gather all the possible patterns of spatial occupancy in terms of habitat variability. Eight biodiversity hotspots of stygobitic Crustacea Harpacticoida were highlighted: 1) the Pyrenees (Spain and France), 2) the Jura Massif (France), 3) the Alpine arc (France, Switzerland and Italy) embracing southward the River Po alluvial plain and the Slovenian External Dinarides, 4) the Central Apennines (Italy), 5) the Carpathian and Balkan mountains in Romania and at the boundary between western Bulgaria and north-west Macedonia, 6) the Dinaric Alps (from Croatia to Albania), 7) the Sardinia Island, 8) an area in central-northern Europe embracing Denmark, the Netherlands and Germany. The hotspots showed a clear spatial distribution in southern Europe where they were distributed predominantly south to the 45th parallel, in line to what reiteratively observed in previous studies. Many hotspots embraced more than one habitat type. The adoption of discrete groundwater habitat types as working spatial units rather than grids provided a higher resolution of where the stygobitic harpacticoid species effectively live, with the possibility of intervening more precisely to preserve them and their habitats.     

Expression of stress inducible protein 1 (Stip1) in the mouse testis

Phthalate esters are considered endocrine disruptors that interfere with the endocrine balance and development of the mammalian testis. Mono-2-ethylhexyl phthalate (MEHP), the active metabolite of the ubiquitously used plasticizer di-2-ethylhexyl phthalate (DEHP), acts upon Sertoli cells as initial target. By subtractive cDNA libraries we identified genes deregulated as response to MEHP in primary cultures of mouse Sertoli cells. The expression of mouse stress inducible protein 1 (Stip1) was detected as upregulated as a result of MEHP exposure. Stip1 is a cochaperone protein that is homologous to the human heat shock cognate protein 70 (hsc70)/heat shock protein 90 (hsp90)-organizing protein (Hop). To assess the presence and localization of Stip1 in mouse testis and its potential role in stress defense, we studied the expression pattern of the Stip1 protein by immunohistochemistry and of the mRNA by in situ hybridization. Both the protein and the mRNA of Stip1 were mainly found in the cytoplasm of all types of spermatogonia and spermatocytes up till zygotene, the expression decreased during late pachytene and was very weak in diplotene spermatocytes and round spermatids. Interestingly, this expression pattern resembled the pattern of stress sensitivity of spermatogenic cells in that the most sensitive cell types show the weakest expression of Stip1. This suggests an important role for Stip1 in the ability of germ cells to survive in stress conditions including high temperatures.     

The location of the<Emphasis Type="Italic">Mus musculus/M. domesticus</Emphasis> hybrid zone in the Balkans: clues from morphology

Three morphological characters were used to depict the position of the hybrid zone between two species of house mice,M. musculus Linnaeus, 1758 andM. domesticus Schwarz et Schwarz, 1943, across a vast area covering countries of the former Yugoslavia, Albania, Bulgaria and Greece. Quantitative approach based on a morphological index (MI), resembling the hybrid index widely used in allozyme-based genetic studies, was used. The zone crosses Slovenia south of the Sava River, and then follows the Dinaric Mts to Montenegro and northern Albania. Contrary to many previously published results, the zone was found to run parallel with northern borders of Albania and the former Yugoslavian Macedonia, about 150 km north of the Greek border, thus giving its course rather “shallow” appearance at this part of the Balkan Peninsula.     

Distribution and genetic status of brown bears in FYR Macedonia: implications for conservation

Conservation and management of large carnivores is often hampered by the lack of information of basic biological parameters. This is particularly true for brown bears (Ursus arctos) in the Former Yugoslav Republic (FYR) of Macedonia. The bear population in this country is important, as it links bear populations of the central part of the Dinaric–Pindos population and the endangered population to the south in Greece. The aim of this study was to assess bear presence in FYR Macedonia and to provide the first evaluation of the genetic status of the species in this country. Bear presence was assessed through a questionnaire and sign surveys, while the genetic status of the species was evaluated through noninvasive genetic sampling from power poles and microsatellite analysis. The results of the study indicate the continuous and permanent presence of brown bears in FYR Macedonia from the border to Kosovo in the northwest, along the border to Albania and Greece in the south; bear presence around Mount Ko?uf in the south of the country was seasonal. High levels of genetic diversity were recorded, and it appears that this bear population is currently not threatened by low genetic variability. Cross-border movements of bears between FYR Macedonia and Greece were documented, indicating the presence of an interconnected population and outlining the necessity for a coordinated international approach in the monitoring and conservation of the species in southeastern Europe.     

Genomic Characterization of Human and Environmental Polioviruses Isolated in Albania

Between April and December 1996, a serious outbreak of poliomyelitis occurred in Albania; almost 140 subjects were involved, and the episode presented an unusually high mortality rate (12%). During the outbreak, water samples from the Lana River in Tirana, Albania, and stool samples from two cases of paralytic poliomyelitis were collected and analyzed for the presence of polioviruses. Six polioviruses were isolated from the environmental and human samples, according to standard methods. All the samples were characterized by partial genomic sequencing of 330 bases across the 5′ untranslated region (5′-UTR) (nucleotide positions 200 to 530) and of 300 bases across the VP1 region (nucleotide positions 2474 to 2774). Comparison of these sequences with those present in data banks permitted the identification of environmental isolates Lana A and Lana B as, respectively, a Sabin-like type 2 poliovirus and an intertypic recombinant poliovirus (Sabin-like type 2/wild type 1), both bearing a G instead of an A at nucleotide position 481. The two other environmental polioviruses were similar to the isolates from the paralytic cases. They were characterized by a peculiar 5′-UTR and by a VP1 region showing 98% homology with the Albanian epidemic type 1 isolates reported by other authors. This study confirms the environmental circulation in Albania of recombinant poliovirus strains, likely sustained by a massive vaccination effort and by the presence in the environment of a type 1 poliovirus, as isolated from the Lana River in Tirana about 2 months before the first case of symptomatic acute flaccid paralysis was reported in this town.     

The bryophyte association Solorino saccatae–Distichietum capillacei Reimers 1940 on the Mediterranean mountains: A tool for evaluating natural and conservative environments

This study focuses on Solorino saccatae–Distichietum capillacei, a terri-saxicolous bryophyte association belonging to the alliance Distichion capillacei of the class Ctenidietea mollusci. With this research, we extend the Mediterranean distribution area including eastern Sicily, southern Italy, Albania and Macedonia. The results of the research demonstrate that this association is linked to natural, lasting and conservative habitat of which it can be considered a good bioindicator. This is revealed by the bryovegetational analysis, based on biological and ecological parameters (life form, life strategy and hemerobic gradient), which can be interpreted to provide useful information about the environmental quality.     

First outbreak of norovirus in Albania

Aims: Noroviruses (NoVs) represent the most important enteric viruses responsible for acute gastroenteritis world‐wide. This study objective is to characterize the first outbreak of NoV that occurred in Ballsh, a small city in Albania. Methods and Results: Stool specimens were collected from people attending to the hospital. Samples were also collected from the aqueduct for bacteriological and virological tests. Overall 33 stools and five drinking water samples were collected, respectively, from the hospital in Ballsh and from the municipal aqueduct. No water samples were scored positive whereas ten stool samples (30·3%) were scored GGII NoV positive. All the GGII isolates were identified as GGII·4 genotype, and no GGI was identified. The alignment and protein analysis were performed using, respectively, Clustal V and the mega 4 software. Conclusions: This is the first report of NoV GGII·4 in Albania causing an outbreak. The genetic analysis showed several point mutations and amino acid substitutions with respect to the international strains. Significance and Impact of Study: Over the last decades, Albania has suffered from different outbreaks as cholera, poliomyelitis, hepatitis A and now, for the first time, it has been documented an outbreak of NoV.