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1. Introduction. Multiple cytokines have been studied for their role in the propagation of the inflammatory process related to inflammatory bowel diseases (IBD), but the role of interleukin-4 remains controversial. The aim of this study was to evaluate the role of two IL-4 gene single nucleotide polymorphisms (SNPs) in disease susceptibility and phenotypic expression. 2. Materials and Methods. A group of 160 patients with IBD (86CD/74UC) and 160 healthy controls were genotyped for IL-4 rs2243250/−590C/T and rs2070874/−34C/T using real-time polymerase chain reaction with TaqMan assay. 3. Results. The analysis of IBD patients and controls revealed a significantly reduced frequency of the minor allele T of both SNPs in CD patients (p = 0.03, OR 0.55 and p = 0.02, OR 0.52) and for the entire IBD group (p = 0.01, OR 0.57 and p = 0.01, OR 0.55). Haplotype analysis identified the most frequent haplotype (rs2243250/rs2070874 CC) associated with a high risk for developing IBD (either UC or CD) (p = 0.003). IBD patients with extraintestinal manifestations had significantly increased frequency of the minor alleles T. We also found an association between the presence of allele C of rs2070874 and response to antiTNF treatment. 4. Conclusions. This is the first study to investigate the IL-4 gene’s relation to IBD susceptibility conducted in Romania. Both SNPs were found to be associated with disease susceptibility and phenotypic features, such as extraintestinal manifestations and response to antiTNF agents.

In the present study 423 Lucanus cervus individuals belonging to the entomological collection of the” Grigore Antipa” National Museum of Natural History (MNINGA) were checked for the presence of morphological differences and anomalies. Various malformations were identified in 16 specimens. This study also presents the first cases of bilateral symphysocery 2 + 3 and unilateral symphysocery 2 + 3 + 4 at the antennal clubs for L. cervus, as revealed by scanning electron microscopy. The teratological cases were more prevalent on the left rather than the right antenna (36.3% vs. 18.18%). The normal structural variations in L. cervus were classified as follows: different number of lamellate antennomeres of the antennal club, different number of pores on the last lamella and different shapes of the clypeus. More than 94% of the examined individuals had four antennomeres at the antennal club for both left and right antennae. The rounded shape (72%) with a uniporous sensilla (98%) of the last lamellate antennomere was the most common morph in the analysed samples. Among variations, V-shaped clypeus form was prevalent (37.3%) in L. cervus analysed males. Some structural modifications could have been caused by environmental factors producing alterations in the pupation or eclosion stages. For a better understanding of the causes of teratology and the presence of morphological changes in populations of L. cervus, further investigations are needed.

Dr. Dumitru Murariu was born on September 21, 1940 in Ungureni, Botoşani County, as the first child in a peasant family. He attended primary and secondaryschool in his home village. Since the beginning he proved himself to be one of the brightest students in the class and, as a consequence, the school teachers advised his parents to have him continue his education. At the end of secondary school, the young Dumitru Murariu enrolled at the “August Treboniu Laurian” theoretical 280 Popa & Popa high school in the city of Botoşani. During these years (1955–1957), the teacher of “the Fundamentals of Darwinism” made a strong impression on the future scientist, with practical lessons, in a small garden, on the correlation between the natural selection and the variability of organisms. On the way from the main building to the above‑mentioned garden, the professor taught his pupils how to identify the trees on the sidewalks and from the “Public Garden”. This teacher’s name was Remus Cehovschi - former Assistant Professor at the University of Chernivtsi (Cernăuți – North Bukovina), from where he took refuge to Botoșani in 1944. After graduating high school, Dumitru Murariu returned to his home village, where he occupied a position of unqualified teacher in the village school. In the fall of 1958, he was drafted for the mandatory military service until mid-January 1961. Returning home, he resumed his school position but in the autumn of the same year, he successfully passed the admission exams at the Faculty of Biology-Geography, the Department of Biology-Zoology at the “Al. I. Cuza” University of Iași. Based on his academic excellence he received a scholarship until graduating in 1966.

The unique aquatic Pontocaspian (PC) biota of the Black Sea Basin (BSB) is in decline. The lack of detailed knowledge on the status and trends of species, populations, and communities hampers a thorough risk assessment and precludes effective conservation. This paper reviews PC biodiversity trends in the BSB (Bulgaria, Romania, Moldova, Ukraine, and Russia) using endemic mollusks as a model group. We aim to assess changes in PC habitats, community structure, and species distribution over the past century and to identify direct anthropogenic threats. The presence/absence data of target mollusk species were assembled from literature, reports, and personal observations. Pontocaspian biodiversity trends in the northwestern BSB coastal regions were established by comparing 20th‐ and 21st‐century occurrences. The direct drivers of habitat and biodiversity change were identified and documented. We found that a pronounced decline of PC species and communities is driven by (a) damming of rivers, (b) habitat modifications that disturbed previous natural salinity gradients and settings in the studied area, (c) pollution and eutrophication, (d) invasive alien species, and (e) climate change. Four out of the 10 studied regions, namely, the Danube Delta–Razim Lake system, Dniester Liman, Dnieper–Bug estuary, and Taganrog Bay–Don Delta, contain favorable ecological conditions for PC communities and still host threatened endemic PC mollusk species. Distribution data are incomplete, but the scale of deterioration of PC species and communities is evident from the assembled data, as are major direct threats. Pontocaspian biodiversity in the BSB is profoundly affected by human activities. Standardized observation and collection data as well as precise definition of PC biota and habitats are necessary for targeted conservation actions. This study will help to set the research and policy agenda required to improve data collection to accommodate effective conservation of the unique PC biota., Distribution data of unique, endemic Pontocaspian (PC) biodiversity of the Black Sea Basin is incomplete, but the scale of deterioration of PC species and communities is evident from the assembled data, as are major direct threats. This review will help to set the research and policy agenda required to improve data collection to accommodate effective conservation of the unique PC biota.

Simple Summary During the last decade, our understanding of the biogeography of Western-Palaearctic leeches (genus Hirudo) has begun to unravel, unveiling their diversity in practically all of Europe, except for its westernmost peninsula, Iberia. We discovered for the first time H. verbana in Spain and conducted an integrative approach (combining morphology, anatomy, ecology with genetics) to characterize the newly recorded Iberian populations. We found two endemic and geographically separated Iberian lineages of H. verbana. One of them is easily distinguished by its distinctive colour-pattern and is described as H. verbana bilineata ssp. nov. We established its phylogenetic relationships with other European Hirudo spp. and confirmed the presence of a second species in Iberia, H. troctina. The provided distribution pattern of medicinal leeches contributes to a better understanding of the complexity of the Iberian Peninsula as a glacial refugium/cradle for endemisms, sheltering species populations that began to settle throughout the Pleistocene. Finally, we highlight the urgent need for implementing conservation measures to reverse the evident decline of Iberian populations of medicinal leeches. Abstract Until the beginning of the 21st century, the famous medicinal leech was thought to be represented by only one species, Hirudo medicinalis. However, recent publications have demonstrated that under that name, at least five different species of medicinal leeches were hidden. During the last decade, the biogeography of Western-Palaearctic leeches has begun to unravel, untangling their diversity in practically all of Europe, except for its westernmost peninsula, Iberia. Hirudo medicinalis has been repeatedly reported from Iberia, but those records were considered questionable. We discovered H. verbana in northern Spain, constituting its first record in Iberia. Using an integrative approach (combining morpho-anatomical data and molecular analyses using three genes, COI,12S rRNA, and ITS2), two endemic and geographically separated Iberian lineages have been found. One of them is easily distinguished by its distinctive colour-pattern and is described as H. verbana bilineata ssp. nov. We characterized the new subspecies morphologically, ecologically, and genetically. We also established its phylogenetic relationships with other European Hirudo spp. and confirm the presence of H. troctina in Iberia, occurring as far as 43° lat. N. Iberian H. verbana records constitute its westernmost known distribution to date. The provided distribution pattern of H. verbana contributes to a better understanding of the complexity of Iberia as a glacial refugium/cradle for endemisms, harbouring populations with a high degree of genetic structure that began to settle throughout the Pleistocene. Iberian Hirudo populations are declining in recent decades and there is an urgent need to assess their conservation status and to initiate conservation measures to reverse their decline.

A new species of the feather mite genusTrouessartia(Trouessartiidae) is described from the Large NiltavaNiltavagrandis(Blyth) (Passeriformes, Muscicapidae) in Northeast India (Meghalaya, Jaintia Hills, Shnongrim village).TrouessartianiltavaeConstantinescu,sp. n.is morphologically closely related (no phylogenetic meaning) toT.bulligeraGaud, 1968 fromClytorhynchushamlini(Mayr) (Passeriformes: Monarchidae), sharing in males a unique character within the genus, by having setaeeon legs IV hemispheroid, with spine-shaped apex. Males of the new species have the prodorsal shield without ornamentation, the prohysteronotal shield and lobar shield connected, and the terminal cleft parallel sided. Females have the posterior half of the hysteronotal shield ornamented with large ovate lacunae in central area and small elliptical lacunae marginally. To the morphological description of this new feather mite species we added sequence data on the mitochondrial cytochrome c oxidase subunit I gene fragment (COI). The phylogenetic relationships betweenTrouessartiaspecies are briefly discussed.

A new species of the feather mite genus Proterothrix (Proctophyllodidae: Pterodectinae) is described from the Large Niltava Niltava grandis (Blyth) (Passeriformes, Muscicapidae) in northeast India (Meghalaya, Jaintia Hills, Shnongrim village). Proterothrix chachulae Constantinescu, sp. n. differs from all known species of the genus by having in males the aedeagus with bilobate tip. The morphological description is supplemented with molecular characterisation of a fragment f near the 5` terminus of the mitochondrial COI gene.

Psoriatic arthritis (PsA) is a chronic inflammatory disorder that belongs to the group of spondyloarthritis (SpA). It was found that single nucleotide polymorphisms (SNPs) of endoplasmic reticulum aminopeptidase (ERAP1 and ERAP2) genes influence the risk of ankylosing spondylitis, the most common form of SpA and the risk of psoriasis. Our purpose was to investigate the possible association of ERAP1 and ERAP2 gene SNPs with psoriatic arthritis susceptibility in Romanian population. Subsequent analyses included patients' subgroups according to HLA-B27 status. Psoriatic arthritis patients (N = 98) and random healthy controls (N = 139) were genotyped for ERAP1/2 genes SNPs rs30187, rs27044, rs2910686, and rs2248374 by TaqMan Allelic Discrimination Assays. An additional control group (N = 108; 100% HLA-B27 positive) was used for subsequent analyses. The results showed the association of rs2248374 SNP of ERAP2 gene with the risk of PsA, especially for HLA-B27 negative disease (p = 0.02; OR 1.59). ERAP2 haplotype GT (rs2248374/rs2910686) was significantly under-represented in PsA patients than in controls (43 vs. 55%; p = 0.02). The analysis of ERAP1 SNPs in HLA-B27 positive controls and PsA subgroup showed strong evidence of association for rs30187 (p = 0.005; OR 2.73) and for CC rs30187/rs27044 haplotype (47% in patients vs. 70.5% in controls; p = 0.006). In conclusion, we found a significant association of ERAP2 with PsA and HLA-B27 negative PsA, while ERAP1 association was restricted only to HLA-B27 positive disease. To our knowledge, this is the first study that investigated ERAP2 polymorphisms in relation to PsA susceptibility.

Objective. Our aim was to investigate whether two ERAP2 single nucleotide polymorphisms (rs2910686 and rs2248374) influence spondyloarthritis (SpA) susceptibility in Romanians. Methods. The case control study included 139 controls and 192 SpA patients. The two polymorphisms were genotyped by real time polymerase chain reaction (RT-PCR). The association tests for allele, genotype and haplotype frequencies for each polymorphism were performed with PLINK 1.9. Results. The genotypes and allele frequencies of the two SNPs in general SpA group vs. controls showed no association except for a possible marginal one for the minor allele A of rs2248374 (p = 0.08). In HLA-B27 negative SpA cohort the minor allele (A) frequency (55.4%) of SNP rs2248374 was significantly higher than the one in HLA-B27 controls (44.5%) (p = 0.012). HLA-B27 negative carriers of minor allele A present a higher risk of developing SpA (p = 0.015). Also for the second ERAP2 gene variant investigated (rs2910686) the minor allele T frequency was significantly higher (46.5%) in HLA-B27 negative SPA patients when compared with HLA-B27 negative controls (36%) (p = 0.02). The haplotype of the minor alleles (AC) is a risk factor for HLA-B27 negative SpA (p = 0.019), while the haplotype of the major alleles (GT) is a protective one against the disease in HLA-B27 negative cohorts (p = 0.009). Conclusions. Both ERAP2 gene polymorphisms investigated, especially rs2248374, influence SpA susceptibility, but this influence is limited only to the HLA-B27 negative individuals.

Funding: Grantová Agentura České Republiky (GrantNumber(s): 13-05872S) Romanian Ministry of Education (GrantNumber(s): CNCS UEFISCDI PN II-RU-PD-2012-3-0479). Understanding the invasive potential of species outside their native range is one of the most pressing questions in applied evolutionary and ecological research. Admixture of genotypes of invasive species from multiple sources has been implicated in successful invasions, by generating novel genetic combinations that facilitate rapid adaptation to new environments. Alternatively, adaptive evolution on standing genetic variation, exposed by phenotypic plasticity and selected by genetic accommodation, can facilitate invasion success. We investigated the population genetic structure of an Asian freshwater mussel with a parasitic dispersal stage, Sinanodonta woodiana, that has been present in Europe since 1979 but which has expanded rapidly in the last decade. Data from a mitochondrial marker and nuclear microsatellites have suggested that all European populations of S. woodiana originate from the River Yangtze basin in China. Only a single haplotype was detected in Europe, in contrast to substantial mitochondrial diversity in native Asian populations. Analysis of microsatellite markers indicated intensive gene flow and confirmed a lower genetic diversity of European populations compared to those from the Yangtze basin, though that difference was not large. Using an Approximate Bayesian Modelling approach, we identified two areas as the probable source of the spread of S. woodiana in Europe, which matched historical records for its establishment. Their populations originated from a single colonization event. Our data do not support alternative explanations for the rapid recent spread of S. woodiana; recent arrival of a novel (cold‐tolerant) genotype or continuous propagule pressure. Instead, in situ adaptation, facilitated by repeated admixture, appears to drive the ongoing expansion of S. woodiana. We discuss management consequences of our results. Publisher PDF

A new species of the feather mite genus Trouessartia (Trouessartiidae) is described from the Large NiltavaNiltavagrandis (Blyth) (Passeriformes, Muscicapidae) in Northeast India (Meghalaya, Jaintia Hills, Shnongrim village). Trouessartianiltavae Constantinescu, sp. n. is morphologically closely related (no phylogenetic meaning) to T.bulligera Gaud, 1968 from Clytorhynchushamlini (Mayr) (Passeriformes: Monarchidae), sharing in males a unique character within the genus, by having setae e on legs IV hemispheroid, with spine-shaped apex. Males of the new species have the prodorsal shield without ornamentation, the prohysteronotal shield and lobar shield connected, and the terminal cleft parallel sided. Females have the posterior half of the hysteronotal shield ornamented with large ovate lacunae in central area and small elliptical lacunae marginally. To the morphological description of this new feather mite species we added sequence data on the mitochondrial cytochrome c oxidase subunit I gene fragment (COI). The phylogenetic relationships between Trouessartia species are briefly discussed.

The use of DNA barcoding in alien invasions has recently proved to be a powerful tool in delineating dispersal pathways and clarifying doubtful identifications. Morphological similarities between Anadara kagoshimensis (Tokunaga, 1906) and Anadara inaequivalvis (Bruguière, 1789) require the use of genetic markers in identifying the ark shell species that has recently invaded the Black Sea. The high genetic similarity (99.8-100%) hereby found between COI sequences obtained from the Black Sea samples and Japanese A. kagoshimensis confirms at a molecular level that the ark clam species invading the Romanian Black Sea belong to this taxon.

Objective. Our aim was to investigate whether two ERAP1 gene variants, rs30187 and rs27044, influence the clinical characteristics of ankylosing spondylitis (AS) (the age of onset and the type of articular manifestations - axial or mixed) in Romanian patients. Methods. We studied 94 AS patients and 139 healthy controls. The method used for genotyping the two non-synonymous single nucleotide polymorphisms (SNPs) was real-time polymerase chain reaction. Association tests were carried out using PLINK 1.07 software. We analyzed separately the subgroups of AS patients with early onset (age 30 years), as well as the subgroups of patients with axial manifestations and mixed manifestations (axial and peripheral). Results. Significant association between ERAP1 polymorphisms and AS is only present for patients who experienced an early onset (p = 0.04 for rs30187 and p = 0.007 for rs27044) and not for those with late onset (p = 0.32 for rs30187 and p = 0.29 for rs27044). Polymorphism rs30187 is associated only with the axial form of AS (p = 0.02), while rs27044 is associated only with the mixed form of the disease (p = 0.02). Conclusions. Our findings demonstrate a consistent association between the studied ERAP1 gene SNPs and certain phenotypic characteristics of AS, suggesting that these gene variants may influence the AS onset and the presence of axial or mixed manifestations of AS.

The freshwater mussel Anodonta (Sinanodonta) woodiana (Lea, 1834) (Chinese Huge Mussel or Swan Mussel) (Bivalvia: Unionidae) is the largest unionid species present in the European fauna. Its native range is in East Asia (South-Eastern Russia to Malaysia), but it has spread rapidly across Europe over the last few decades and the species is invasive also in other parts of the world ( Bogan et al., 2011 and Demayo et al., 2012). Studies of the population genetics of A. woodiana in Europe used slowly evolving markers such as allozymes and mitochondrial COI DNA sequences for the analysis of several isolated populations ( Nagel et al., 1996, Soroka, 2005 and Soroka et al., 2014). However, for the recent and rapid spread of the species across Europe, fast evolving markers, such as DNA microsatellites, are needed to understand important aspects of the population genetics of this invasive species: the route(s) of invasion, the time and number of colonization events, and other details. The first eight microsatellite markers for the species have been described in 2011 (Popa et al., 2011). This number is rather low to infer aspects of the evolutionary history of populations and additional microsatellite loci are needed to increase to power of future genetic studies of this species (Koskinen et al., 2004). In this paper we describe the development of nine new polymorphic microsatellite loci for A. woodiana. We also combined new and previously described loci into three multiplex sets allowing reducing the time and money costs of genotyping, as well as decreasing the risk for samples mishandling.

The Transylvanian bush-cricket, Pholidoptera transsylvanica, is a protected mesophytic species endemic to the Carpathian Mountains grasslands. We developed 14 polymorphic microsatellite markers in order to investigate the population genetic structure of this species. These markers were tested on a population sample and the average number of alleles was 10.929 ± 0.730, while the mean of the observed and expected heterozygosities was 0.820 ± 0.020 and 0.814 ± 0.019 respectively. These markers can become a useful tool in genetic studies important in conservation strategies for this species.

Background The cytokines IL12 and IL23 have been recently implicated in the pathogenesis of psoriatic arthritis (PsA). In this study we investigated the genetic variations in the genes coding for IL12, IL23 and IL23 receptor as a plausible source of susceptibility and modification of clinical symptoms of PsA in Romanian population. Methods Twenty five SNPs mapping to IL12A, IL12B, IL23A, IL23R and IL12RB1 genes were genotyped in 94 PsA patients and 161 healthy controls of Romanian ethnicity using the Sequenom genotyping platform. Results The exonic SNP rs11171806 from IL23A gene was significantly underrepresented in patients versus controls (p = 0.03, OR 0.391) and the carriers of rs11171806/rs2066808 AC haplotype had decreased risk for PsA (p = 0.03). The two SNPs of the highly conserved gene IL23A are in complete LD in our population. Genetic variants of IL12B gene were associated with polyarticular subtype of PsA. No associations were found between SNPs from IL12A, IL23R and IL12RB1 genes and susceptibility to PsA and its phenotypes. Conclusion We confirm the previously described association of rs2066808 variant with psoriasis and PsA and we show evidence of an extended genomic region inside IL23A gene as carrier of true disease susceptibility factors. These data suggest a role for IL23 in the PsA pathogenesis in Romanians.

We investigated two nonsynonymous variants (rs30187 and rs27044) of ERAP1 gene in HLA-B27 positive individuals (150 spondyloarthritis and 108 controls) and in general ankylosing spondylitis (AS) patients (n = 137) vs random controls (n = 139). Both single nucleotide polymorphisms (SNPs) were associated with the risk of spondyloarthritis [odds ratio (OR) 1.80, 95% confidence interval (CI) 1.24-2.62, P = 0.001 for rs30187, OR 1.58, 95% CI 1.07-2.34, P = 0.02 for rs27044]. The CC haplotype was a protective factor (P = 0.002), while the TG haplotype was a risk factor (P = 0.01) for spondyloarthritis. The SNP rs30187 was also associated with the risk of HLA-B27+ AS. For the general group of AS, the carriers of minor alleles showed an increased risk for the disease (OR 1.92, 95% CI 1.17-3.13 for rs30187, OR 1.74, 95% CI 1.08-2.80 for rs27044). This is the first study that shows the association of ERAP1 gene variants and haplotypes with HLA-B27 positive spondyloarthritis.

Ten microsatellite loci were isolated and characterized for Stys's bush-cricket, Isophya stysi Cejchan (Orthoptera: Tettigoniidae), an endemic Orthoptera species to the Carpathian Basin, using an enriched genomic library procedure. The polymorphism of these loci were tested in two populations of I. stysi, and the number of alleles per locus varied from 4 to 16. The expected and observed heterozygosities ranged from 0.612 to 0.925 and from 0.625 to 1.000, respectively. The interspecific applicability of these microsatellites was evaluated by amplification in 20 related species: Isophya camptoxypha, Isophya sicula, Isophya ciucasi, Isophya pienensis, Isophya harzi, Isophya kraussii, Isophya zubovskii, Isophya rectipennis, Isophya modesta, Isophya longicaudata, Isophya dobrogensis, Isophya hospodar, Isophya speciosa, Isophya modestior, Poecilimon fussii, Poecilimon affinis, Polysarcus denticauda, Barbitistes constrictus, Leptophyes discoidalis, Phaneroptera falcata. All primer pairs for the 10 loci yielded successful amplifications in at least one other taxon from the Isophya genus. This set of microsatellite loci would be useful for genetic studies in I. stysi and other species of the genus Isophya.

Tumor necrosis factor alpha (TNF-alpha) is an important pro-inflammatory cytokine implicated in the pathogenesis of psoriatic arthritis. We have performed a case-control association study of three TNF-alpha gene polymorphisms in a group of Romanian psoriatic arthritis patients versus ethnically matched controls. A second group of patients with undifferentiated spondyloarthritis was used in order to look for similarities in the genetic background of the two rheumatic disorders. The −857C/T polymorphism was associated with susceptibility to psoriatic arthritis in our population at the individual level (p = 0.03, OR 1.65, 95% CI 1.05–2.57) and in combined haplotypes with the −238G/A and −308G/A SNPs. Regarding the investigated polymorphisms and derived haplotypes, no potential association was found with the susceptibility to undifferentiated spondyloarthritis in Romanian patients.

Sinanodonta woodiana (Lea, 1834) is a large Unionid species with a real invasion success. It colonized Europe, Central America, the Indonesian Islands and recently North America. The species life cycle involves a larval parasitic stage on freshwater fish species which contributes to the spread of the mussel. In this paper we describe, for the first time, eight polymorphic microsatellite loci for the species Sinanodonta woodiana. The genetic screening of individuals confirmed that all loci were highly polymorphic. The number of alleles per locus ranged from 7 to 14 and the observed heterozygosity ranged from 0.650 to 0.950. These loci should prove useful to study the species population genetics which could help to infer important aspects of the invasion process.

Infestation of the pumpkinseed Lepomis gibbosus (Teleostei: Cyprinidae) by the copepod Lernaea Cyprinacea (Crustacea) - some ecological aspects The copepod Lernaea cyprinacea Linnaeus, 1758 is known by the large spectrum of fish hosts which it infests, and to which it can cause haemorrhages and ulcerations. The paper focuses on some aspects of the relation between the Pumpkinseed sunfish and this copepod: infestation parameters, preferred attaching situs, relationship between the host size and parasitisation degree, as well as the visible effects of this parasite left on the fish body, in the Moara Domnească Lake, Romania.

In this study, we used data from morphology and three DNA markers to assess the taxonomic status of the putative bivalve species Hypanis colorata and Hypanis angusticostata in a Black Sea lagoon, the Razelm Lake in Romania. The morphological data (the shape of shell ribs and the multivariate analysis of morphometric variance of three variables constructed as the ratios between the main dimensions of the shell) confirmed that the two analyzed species are distinct morphological entities. Three molecular markers, one from the nuclear genome (18S rRNA) and two from the mitochondrial genome (16S rRNA and COI), showed extremely reduced sequence divergence (0–0.1%) between the two putative species. Based on these results, we suggest that H. angusticostata and H. colorata are morphotypes of a single species.

We determined the distribution of human leukocyte antigen-C (HLA-C) allelic groups in a cohort of psoriatic arthritis (PsA) patients and a control population of Romanian ethnicity. A nominal association of HLA-C*06 with susceptibility to PsA was observed [P = 0.014, p(corr) > 0.05, odds ratio (OR) 2.1, 95% confidence interval (CI) 1.08-4.46]. When subanalyzing data according to PsA clinical phenotypes, association was noticed between HLA-C*06 and PsA with psoriasis onset before 40 years (p(corr) = 0.013, OR 3.7, 95% CI 1.58-9). This first report from Romania confirmed the association of HLA-C*06 with type I psoriasis in PsA patients. Other study findings, such as the relationship between HLA-C*06 and spondylitis or the protective effect of HLA-C*07 for the polyarthritis clinical phenotype of PsA, are of preliminary character and require verification.

Hypanis colorata (Eichwald, 1829) (Cardiidae: Lymnocardiinae) is a bivalve relict species with a Ponto-Caspian distribution and is under strict protection in Romania, according to national regulations. While the species is depressed in the western Black Sea lagoons from Romania and Ukraine, it is also a successful invader in the middle Dniepr and Volga regions. Establishing a conservation strategy for this species or studying its invasion process requires knowledge about the genetic structure of the species populations. We have isolated and characterized nine polymorphic microsatellite markers in H. colorata. The number of alleles per locus ranged from 4 to 28 and the observed heterozygosity ranged from 0.613 to 1.000. The microsatellites developed in the present study are highly polymorphic and they should be useful for the assessment of genetic variation within this species.

We describe here eight polymorphic microsatellite markers for Nannospalax leucodon, an exclusively subterranean and highly specialized rodent species which exhibits range contractions and population declines in Europe. The level of polymorphism was tested on 36 individuals of blind mole rat from Doubroudja region, in South-Eastern part of Romania. The number of alleles across all loci ranged from 7 to 12, with an average of 6.1 ± 0.571 alleles per locus. Observed heterozygosity ranged from 0.735 to 1. These loci will be a useful tool for the study of genetic structure and diversity of blind mole rat populations, whit immediate consequences in conservation strategy for these species.

Morphometric analysis of some populations of lymnocardiid species (Mollusca: Bivalvia) from Razelm Lake Complex (Romania) In this paper we report the morphometric analysis of some populations of Lymnocardiid species from the lakes Razelm and Goloviţa. We used three measurements ratios to perform a discriminant analysis and a multivariate analysis of variance (MANOVA), in order to compare the species Hypanis colorata vs. Hypanis angusticostata in the two lakes, H. colorata in Razelm vs. Goloviţa, and H. angusticostata in Razelm vs. Goloviţa. From this analysis we concluded that the multivariate means of the morphological variables used in this study were highly significantly different (p=3.2e-05) between the two species. Concerning the geographical variability, in both species, the analysis showed no significant difference between the populations in the two investigated lakes. We also determined from a fitting curve analysis, that the growth pattern of both species shows length-height isometry and width-length and width-height negative allometry.

This paper is a synthesis of the faunistic data present in the specialized literature on the alien decapod species identified in the Romanian fauna between 1951 and 2010: Orconectes limosus (Rafinesque, 1817), Callinectes sapidus Rathbun, 1896, Rhithropanopeus harrisii (Gould, 1841), Eriocheir sinensis H. Milne Edwards, 1853, Hemigrapsus sanguineus (de Haan, 1835), Palaemon macrodactylus Rathbun, 1902, Dyspanopeus sayi (Smith, 1869). Resume. Ce travail presente une synthese des donnees faunistiques de la litterature specialisee sur les especes allochtones decapodes identifie dans la faune de Roumanie entre 1951 et 2010: Orconectes limosus (Rafinesque, 1817), Callinectes sapidus Rathbun, 1896, Rhithropanopeus harrisii (Gould, 1841), Eriocheir sinensis H. Milne Edwards, 1853, Hemigrapsus sanguineus (de Haan, 1835), Palaemon macrodactylus Rathbun, 1902, Dyspanopeus sayi (Smith, 1869).

Objectives This work aimed to describe the genetic landscape of the Balkan Peninsula, as revealed by STR markers commonly used in forensics and spatial methods specifically developed for genetic data. Methods We generated and analyzed 16 short tandem repeats (STRs) autosomal genotypes in 287 subjects from ten administrative/geographical regions of Eastern Europe (Romania and the Republic of Moldova). We report estimates of the allele frequencies in these sub-populations, their fixation indexes, and use these results to complement previous spatial analyses of Southern Europe. Results In seven out of ten analyzed regional samples the heterozygosity, averaged across loci, was lower than expected. The average Fis was 0.011. Among the 16 loci, five returned a significant fixation index Fst. The composite Fst across the 16 loci, among the 10 regional samples, was 0.00417, a figure twice as large as that obtained with the same markers across the entire Northern Mediterranean. The first spatial principal component (sPC1) returned the picture of a Central-European pattern of frequencies for the Carpathians, which extended to the Southern boundary of the Balkan Peninsula. However, the 8 alleles extracted by sPC1 returned a picture of a strong reduction of the migration rate in the Carpathian region, mostly between the inner locations. Conclusions Our results revealed an unexpected heterogeneity in the area. We believe that populations from some regions will require treatment as distinct entities when considered in forensic applications.

The analysis of 310 Romanian spondyloarthritides patients confirmed the association of the HLA-B27 marker with the susceptibility to different diseases of this group. For ankylosing spondylitis, the HLA-B27 frequency in Romanian patients (72.1%) was similar to that found in several regions in the Mediterranean area.

To analyze the contribution of the Czech population to the Y-chromosome diversity landscape of Europe and to reconstruct past demographic events, we typed 257 males from five locations for 21 UEPs. Moreover, 141 carriers of the three most common haplogroups were typed for 10 microsatellites and coalescent analyses applied. Sixteen Hg's characterized by derived alleles were identified, the most common being R1a-SRY(10831) and P-DYS257*(xR1a). The pool of haplogroups within I-M170 represented the third most common clade. Overall, the degree of population structure was low. The ages for Hg I-M170, P-DYS257*(xR1a), and R1a-SRY(10831) ap peared to be comparable and compatible with their presence during or soon after the LGM. A signal of population growth beginning in the first millennium B.C. was detected. Its similarity among the three most common Hg's indicated that growth was characteristic for a gene pool that already contained all of them. The Czech population appears to be influenced, to a very moderate extent, by genetic inputs from outside Europe in the post-Neolithic and historical times. Population growth postdated the archaeologically documented introduction of Neolithic technology and the estimated central value coincides with a period of repeated changes driven by the development of metal technologies and the associated social and trade organization.

About 30% of couple infertilities are of male origin, some of them caused by genetic abnormalities of the Y chromosome. Deletions in AZF region can cause severe spermatogenic defects ranging from non‐obstructive azoospermia to oligospermia. The intracytoplasmatic sperm injection technique (ICSI) is rapidly becoming a versatile procedure for human assisted reproduction in case of male infertility. The use of ICSI allows Y chromosome defects to be passed from father. The goal of our study is to evaluate the frequency of microdeletions in the long arm of Y chromosome, within the AZF regions, in these cases of infertilities, using molecular genetics techniques. Thirty infertile men with azoospermia or oligozoospermia, determined by spermogram, were studied after exclusion of patients with endocrine or obstructive causes of infertility. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the Y chromosome AZF zones. Each case was checked by multiplex PCR through coamplification with the SRY marker. Three men with microdeletions of the long arm of the Y chromosome were diagnosed among the 30 patients, corresponding to a proportion of 10%. The relatively high proportion of microdeletions found in our population suggest the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions. The molecular diagnostics was performed according to the current European Academy of Andrology laboratory guidelines for molecular diagnosis of Y chromosomal microdeletions.

In 2006 and 2007 living individuals of Sinanodonta woodiana (Lea 1834) were collected from the eastern part of Romania. These samples represent the first report of this invasive species in the region and indicate that this alien species has colonized the main aquatic basins of Romania.

The occurence of Dreissena bugensis (Andrusov 1897) from the Romanian Danube is reported. This represents the most westward occurrence point of quagga mussel in Europe.

In this study we analyzed at a submeso-geographic scale (2 km) the genetic diversity of two sub-populations of Platynereis dumerilii and correlated this with the physical characteristics of the marine currents along the western Black Sea coast. For this purpose, we developed a set of 13 new polymorphic microsatellite markers and used them to assess the genetic differentiation, as well as the bi-directional migration rates between the studied sub-populations. We also computed the Peclet number (Pe) as an indicator of the relative effect of advection and eddy diffusion on larval dispersion for the specific conditions of the Black Sea study area. The results indicated no genetic structure in P. dumerilii sub-populations which indicates that the longitudinal alongshore currents dominate in the population structuring of this species. This finding is important, because with the average current speeds of 5 cm/sec on the Black Sea coast during May-August we might have expected a certain population structuring to occur. In accordance with the periodical change of direction of the longitudinal current (either form North to South, or form South to North) the gene flow was found to be bi-directional, with the same intensity.

The invasive softshell clam (Mya arenaria Linnaeus, 1758) is native to the northwestern region of the Atlantic Ocean. This species has been introduced in the northeast Pacific and along the European coasts, due to intense naval transports and aquaculture, and it is now present in all the European seas. In this paper we describe seven new microsatellite loci for Mya arenaria. The isolated loci are polymorphic with a number of alleles per locus between 6 and 14. The observed and expected heterozygosities ranged from 0.417 to 0.951, and from 0.643 to 0.895, with an average of 0.716 and 0.775, respectively. These microsatellite markers should be useful in analyzing this species' genetic diversity, which could explain various processes of its invasion history.

In this paper we describe a multiplex PCR useful to as-sign some major Y chromosome tree branches (DE, G2, I, J and J2) (for the nomenclature of the Y chromosome tree branches, see YCC 2002). These are prev alent in all Southern European populations, accoun ting between 51% and 77% of the total male population (Di Giacomo et al. 2003 and personal data). The identification of the allelic state at every locus in the study is done either directly (presence/a bsenc e of the specific PCR fragment), or can be simply obtained with further sequential steps of dot-blot hybrid ization with specific probes. In the last years a large number of single nucleotide polymorphic (SNP) binary markers have been identified in the non-recombining portion of the human Y chrom o-some (NRY) (YCC 2002). SNPs are ideal genetic mar k-ers for med ical and population genetic studie s, since they are widespread in the human genome and are easily de-tected by PCR based methods. In general, the SNPs ex-hibit only two, evolutionarily stable, alleles. This ad van t-age is often counterbalanced by the necessity of typing large numbers of them in many individuals. Genotyping for these markers is of great importance in forensics and in association studies. Moreover, since the NRY lacks genetic recombination during meioses, it is a pow erful tool in phylogenetic studies. In order to increase the pro-ductivity of such studies, the first choice will be to study simultaneously more SNPs in a single multiplex PCR reaction. Two papers concerning the use of mult iplex PCR in the study of the human Y chromosome SNP variation have been published recently. One of them uses a mini-sequencing/capillary electrophoresis method for Y chro-mosome typing in forensic studies (Sanchez et al. 2003); the other one uses a primer extension/mass spectroscopy method for high throughput SNP genotyping of the Y chromosome (Paracchini et al. 2002). The method pre-sented here provides a reliable effort-effective, time-effec -tive and cost-effective method for the early stages of human Y chromosome genotyping, being particularly useful when little information or no information at all is available about the genetic structure of the population under study.

Deletions of the Y chromosome are a significant cause of spermatogenic failure. Three major deletion intervals have been defined and termed AZFa, AZFb and AZFc. Here, we report an unusual case of a proximal AZFb deletion that includes the Y chromosome palindromic sequence P4 and a novel heat shock factor (HSFY). This deletion neither include the genes EIF1AY, RPS4Y2 nor copies of the RBMY1 genes. The individual presented with idiopathic azoospermia. We propose that deletions of the testis-specific HSFY gene family may be a cause of unexplained cases of idiopathic male infertility. This deletion would not have been detected using current protocols for Y chromosome microdeletion screens, therefore we recommend that current screening protocols be extended to include this region and other palindrome sequences that contain genes expressed specifically in the testis.

In order to attain a finer reconstruction of the peopling of southern and central-eastern Europe from the Levant, we determined the frequencies of eight lineages internal to the Y chromosomal haplogroup J, defined by biallelic markers, in 22 population samples obtained with a fine-grained sampling scheme. Our results partially resolve a major multifurcation of lineages within the haplogroup. Analyses of molecular variance show that the area covered by haplogroup J dispersal is characterized by a significant degree of molecular radiation for unique event polymorphisms within the haplogroup, with a higher incidence of the most derived sub-haplogroups on the northern Mediterranean coast, from Turkey westward; here, J diversity is not simply a subset of that present in the area in which this haplogroup first originated. Dating estimates, based on simple tandem repeat loci (STR) diversity within each lineage, confirmed the presence of a major population structuring at the time of spread of haplogroup J in Europe and a punctuation in the peopling of this continent in the post-Neolithic, compatible with the expansion of the Greek world. We also present here, for the first time, a novel method for comparative dating of lineages, free of assumptions of STR mutation rates.

In this study we analyzed comparatively the host-parasite associations between two fish host species invasive in Europe (Lepomis gibbosus and Pseudorasbora parva) and one known generalist parasite species, the copepod Lernaea cyprinacea. We used a fragment of the hypervariable region D1-D2 of the 28S rRNA to confirm that the copepod specimens collected on both host species in our study are indeed conspecific. The prevalence of infection was significantly different between the two host species in all three aquatic ecosystems. Two populations of L. gibbosus exhibited a positive correlation coefficient between the standard body length and infection intensity, while a negative correlation coefficient was observed in one population of P. parva. This is one of the few studies providing parasitological parameters of infections of Lernaea cypriancea in Lepomis gibbosus and Pseudorasbora parva.

The major aim of this study is to describe the first microsatellite loci for the stone-crayfish (Austropotamobius torrentium), by cross-species amplification. Austropotamobius torrentium is a priority species in the EU Habitats Directive and it needs effective conservation management efforts throughout Europe. We tested cross-species amplification of 55 decapod microsatellite primer pairs in A. torrentium and only ten of these loci, from relatively close related species, yielded PCR products of expected sizes. Five of the ten microsatellites proved to be polymorphic (allele numbers ranging from 4 to 14 in a set of 35 individuals). Three of the loci exhibited departure from Hardy-Weinberg equilibrium which could be explained by the presence of null alleles. A forth locus exhibiting HWE deviation, but no null alleles, suggest the possible presence of population substructure of the species in the investigated area. These microsatellite markers are useful for population genetic studies of stone-crayfish.

Croatian Orthoptera were never systematically studied in the past, there are only data dispersed in a lot of publications by foreign authors. In the last three years of intensive research of Croatian Orthoptera, we have added more than thirty species to the national fauna and omitted about twenty species which were included in the former lists. For example: Rammeihippus dinaricus was found for the very first time since the description of the species, western border of the Leptophyes discoidalis distribution was reported and some of the recent problems of European Tetrigidae taxonomy were solved and a new subspecies – Tetrix transsylvanica hypsocorypha was described. For the first time, exhaustive faunistical research was performed on Papuk Mtand Dinara Mountain. We found a new species from the genus Gryllotalpa, a new species from the Poecilimon elegans group and Balkan species of the genus Psorodonotus are being reviewed. One of the oldest mysteries of Croatian and European orthopterology, the Platycleis kraussi case has just been solved (Skejo et al. in prep.), Stenobothrus croaticus was re- discovered, as well as Rhacocleis buchichii, Barbitistes kaltenbachi and Paramogoplistes novaki – all of them in new and interesting localities. There are a lot of ongoing projects concerning the Chorhippus biguttulus aggregate, the Platycleidini tribe, the description of a new species of Leptophyes from the Dinaric Alps (Szövényi & Puskás in litt.), new cryptic species of Oedipoda (Hochkirch in litt.), the discovery of Xya variegata (and checking out for some new localities) and our major project: the first annotated checklist and catalogue of Croatian grasshoppers. There are a lot of novelties concerning molecular, morphological, morphometrical, ecological and bioacoustical data.

Groundhoppers are one of the less studied Orthopteran groups in the world, mostly because of their small body size, lack of audible song and unusual ecology. Altough European list of groundhoppers’ species (Caelifera: Tetrigidae) counts only 2 genera, 15 species and 3 subspecies, taxonomical situation of the family in Europe is still not steady. There are at least six problems that are to be solved: (i) new genus should be erected for Tetrix nodulosa incertae sedis, (ii) the subspecies Tetrix ceperoi brachyptera, firstly described as a variety of T. ceperoi should be considered as invalid according to the International code of zoological nomenclature, (iii) the (sub)specific status of Tetrix transsylvanica hypsocorypha needs to be reevaluated on the larger series, (iv) the (sub)specific status of T. undulata gavoyi is to be re-evaluated: are the differences between this and the nominal subspecies enough to consider T. undulata gavoyi as the valid taxon, (v) Tetrix kraussi vs. Tetrix bipunctata issues – there are a few ideas how to solve this problem: Mayr’s species concept – it is best for the solution of the problem and (vi) T. wagai needs to be synonymized with T. tuerki. Besides these six taxonomical problems there will be discussed how well do we know the distribution of groundhoppers in Europe: the Balkans Peninsula – the least studied area in Europe.