Transfer of Heavy Metals Through Three Components: Sediments, Plants and Fish in the Area with Previous Mining Activity.

This study aims to evaluate the ecological risk and distribution of heavy metals in sediment, plants and fish in a seriously polluted water reservoir in Krompachy, Slovakia. Special attention was given to the different food web positions of individual fish species (predators, omnivores) and their size. The degree of heavy metal contamination in sediments decreased in the order Cu > Pb > Cr > Hg > Cd, and their mutual proportion was largely consistent with concentrations found in aquatic plants, i.e. water sedges (Carex acutiformis). Of the seven fish species investigated, piscivorous perch (Perca fluviatilis) accumulated higher quantities of metal than fish situated at lower trophic levels. Interestingly, co-equal levels of heavy metals to those found in perch (P. fluviatilis) also occurred in rudd (Scardinius erythrophthalmus). The Hg values in some fish muscles exceeded the maximum permissible limits suggesting a persistent problem of old environmental burden from former mining activities.

Morphological abnormalities in fish parasites: a potential tool for biomonitoring natural contaminants?

A comparative study on the strobilar morphology of the tapeworm Proteocephalus percae (Müller, 1780) (Cestoda), a parasite of the perch Perca fluviatilis (L.), showed a high percentage of abnormally developed parasite individuals. The evaluation of biological samples showed seven types of morphological abnormalities, mostly related to reproductive organs of the model tapeworm species. The most commonly identified deformity was an incomplete segmentation of the strobila. A malformed ovary, which is a structural anomaly linked with proglottization and maturation of the strobila, was also shown to be rather frequent. Offish hosts (P. percae) were collected from two localities with different levels of heavy metal pollution, the highly contaminated water reservoir Ruzín and a control locality, the water reservoir Palcmanská Masa, which belongs to the European network of protected areas in Slovakia. Tapeworm abnormalities occurred more frequently in individuals from the contaminated environment (29.9%) compared with individuals from the control site (4.9%). The concentrations of heavy metals found in the parasites and their fish hosts from the heavily polluted reservoir support our assumption that the occurrence of abnormalities could be linked with the destructive effect of toxic substances. The present study also demonstrates that the enumeration of body deformities exceeding the common level of phenotypic variability of particular parasitic species could potentially be used as an indicator of environmental problems.

Variability in responses of soil nematodes to trace element contamination.

Free-living soil nematodes and plant parasites were previously repeatedly tested for their use as bioindicators of soil contamination. We investigated soil nematode communities from grasslands in four different industrial areas in Slovakia to estimate their responses to heavy metals. We considered differences or similarities in behavioural reactions of nematodes to different quantities of selected heavy metals in the soil. Using the CCA analysis and t-value biplot diagrams with van Dobben circles, we split individual nematode genera by their tolerance/intolerance to the particular heavy metal into several clusters. Our results showed that (a) representatives of nematode genera showed concurrently positive and negative associations with two or more heavy metals, (b) most nematode genera expressed a strong positive relation with Ni and to a lesser extend with Cr and Zn, (c) taxonomically closely related nematodes did not demonstrated similar responses to the same pollutant. We assume that nematode genera with a higher level of tolerance to trace elements should be prioritized in the heavy metal bioindication instead of sensitive species, due to their longer persistence in the stressed environment and better indicative ability. We pointed nematode genera, which according to received results may serve as suitable sentinels for specific soil pollutants.

A synoptic review of Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes.

Tapeworms of the genus Caryophyllaeus Gmelin, 1790 (Caryophyllidea: Caryophyllaeidae), common parasites of cyprinid fishes, are reviewed and taxonomic status of 42 nominal taxa that have been placed in the genus during its long history is clarified. The following seven species occurring in the Palaearctic Region are recognised as valid: C. laticeps (Pallas, 1781), C. auriculatus (Kulakovskaya, 1961), C. balticus (Szidat, 1941) comb. n. (syn. Khawia baltica Szidat, 1941), C. brachycollis Janiszewska, 1953, C. fimbriceps Annenkova-Chlopina, 1919, C. syrdarjensis Skrjabin, 1913, and newly described Caryophyllaeus chondrostomi sp. n. (= C. laticeps morphotype 4 of Bazsalovicsová et al., 2014) from common nase, Chondrostoma nasus (Linnaeus), found in Austria and Slovakia. The new species differs by the paramuscular or cortical position of preovarian vitelline follicles, a large, robust body (up to 64 mm long), conspicuously long vas deferens, flabellate scolex with small wrinkles on the anterior margin, and anteriormost testes located in a relatively short distance from the anterior extremity. Caryophyllaeus kashmirenses Mehra, 1930 and Caryophyllaeus prussicus (Szidat, 1937) comb. n. are considered to be species inquirendae, C. truncatus von Siebold in Baird, 1853 and C. tuba von Siebold in Baird, 1853 are nomina nuda. Data on the morphology, host spectra, distribution and known life-cycles of valid species are provided. Phylogenetic interrelations of four species of the genus including its type species and newly described C. chondrostomi were assessed based on an analysis of sequences of lsrDNA and cox1. A key to identification of all valid species of Caryophyllaeus is also provided.

The effects of vegetation cover on soil nematode communities in various biotopes disturbed by industrial emissions.

Better understanding of interactions among belowground and aboveground components in biotopes may improve our knowledge about soil ecosystem, and is necessary in environment assessment using indigenous soil organisms. In this study, we proposed that in disturbed biotopes, vegetation play important role in the buffering of contamination impact on soil communities and decrease the ecological pressure on soil biota. To assess the effects of these interactions we compared nematode communities, known for their bioindication abilities, from four types of disturbed and undisturbed biotopes (coniferous forest, permanent grassland, agricultural field, clearings), where the main stress agent was represented by long-term acidic industrial emissions containing heavy metals (As, Cd, Cu, and Pb). To understand the ecological interactions taking place in studied biotopes, we studied abiotic factors (soil properties) and biotic factors (vegetation, nematode communities). Except significant increase in metals total and mobile concentrations in disturbed biotopes soil, we found acidification of soil horizon, mainly in the clearings (pH=3.68), due to SO2 precipitation. These factors has caused in clearings degradation of native phytocoenoses and decrease in decomposition rate characterized by high amount of organic matter (Cox=4.29%). Nematodes reacts to these conditions by shifts in trophic structure (bacteriovores to fungal feeders), increase in c-p 2 genera (Aphelenchoides, Acrobeloides, and Cephalobus), absence of sensitive groups (c-p 3-5, omnivores, predators), and decrease in ecological indices (SI, MI, MI2-5, H'). Similar contamination was found in forest biotope, but the nematodes composition indicates more suitable conditions; more complex community structure (presence of sensitive trophic and higher c-p groups), higher abundance and indices values, comparable with less stressed field and grassland biotopes. As showed our results, the vegetation undoubtedly plays an important role not only as a resource of services indispensable for the ecosystem, but also as a significant buffer of negative impacts acting within.

Host-parasite relationships as determinants of heavy metal concentrations in perch (Perca fluviatilis) and its intestinal parasite infection.

The concentrations of As, Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn and their bioconcentration factors (BCFs) were determined in two intestinal parasites, an acanthocephalan, Acanthocephalus lucii, a tapeworm, Proteocephalus percae, present in the same host, the European perch (Perca fluviatilis, L.), in the heavily polluted Ruzín reservoir in eastern Slovakia. The bioaccumulation of heavy metals in the fish organs and parasites was studied for acanthocephalan and tapeworm monoinfections or mixed infections by the two parasites and for the size of their parasitic infrapopulations. Bioconcentration factors (c[parasite]/c[muscle tissue]) showed that the concentrations of As, Ni, Pb and Zn were higher in mixed infections than in monoinfections. Negative correlations between heavy metal concentrations in perch organs and the parasites were found. For example, higher concentrations of Ni and Zn in both parasite species corresponded with lower metal concentrations in perch and hard roe. Likewise, significant negative relationships between metal concentrations in fish organs and number of parasites were noticed with lower levels of Pb in fish harbouring higher numbers of tapeworms. Similarly, in both parasite species the concentrations of some essential elements (Cr, Mn) were lower at high infection intensities compared to low intensities. Our study revealed that the differential concentration of heavy metals in perch organs was affected by the type of infection (mono- or mixed-infection), and needs to be considered in field ecotoxicological and parasitological studies as a potentially important factor influencing the pollutant concentrations in fish.

Asian fish tapeworm, Khawia japonensis (Yamaguti, 1934), has expanded its European invasive range.

The invasive fish tapeworm, Khawia japonensis (Yamaguti, 1934) originally described in Japan, is reported for the first time in Slovakia. The tapeworm was found in farmed common carp (Cyprinus carpio L.) from East-Slovakian breeding fish pond. This finding is registered 4 years after the first announcement of this parasite in Europe (Po River Basin in Italy) in 2010 and increases its distribution area within this continent. Morphological characterization of K. japonensis supplemented with DNA sequences of cox1 and ribosomal lsrDNA genes is provided. Specimens from Slovakia phenotypically corresponded with those from feral and farmed carps from China, Vietnam, and Italy. Moreover, 100 and 98.7% identity of partial ribosomal lsrDNA gene and mitochondrial cox1 genes, respectively, were detected with K. japonensis from Japan. The invasive and pathogenic potential of K. japonensis in commercial breeding fisheries and its possible further spread in natural habitats is difficult to estimate for now. As yet, K. japonensis appears to be without a major impact on commercial breeding fisheries, but calls for more attention to the problem of biological invasions.

Morphological polymorphism in tapeworms: redescription of Caryophyllaeus laticeps (Pallas, 1781) (Cestoda: Caryophyllidea) and characterisation of its morphotypes from different fish hosts.

Recent morphological and molecular data have shown that one of the most common parasites of freshwater fish in the Palaearctic Region, the cestode Caryophyllaeus laticeps (Pallas, 1781) (Eucestoda: Caryophyllidea), is highly polymorphic. Five distinct morphotypes of C. laticeps, largely corresponding to different fish hosts and representing separate, yet closely related genetic lineages, have been recognised and they are characterised in the present paper. Morphotype 1 from breams, Abramis brama (L.) (type-host) and Ballerus spp., corresponds to the original Taenia laticeps Pallas, 1781 and its neotype (paragenophore ex A. brama in Russia) is designated. This morphotype is characterised by a slender body and flabellate scolex. Morphotype 2 was found in the Macedonian vimba Vimba melanops (Heckel) and the vimba bream V. vimba (L.); it is typified by a more robust body, with most anterior extent of the vitelline follicles near the scolex and the cirrus-sac situated more anteriorly than in other morphotypes. Morphotype 3 is represented by worms from the common carp Cyprinus carpio L. that possess a cuneicrispitate scolex (having the form of a wedge with shallow indentations on anterior margin). Morphotype 4 from the common nase Chondrostoma nasus (L.) has a large, robust body and a wide scolex with numerous superficial grooves (wrinkles) in its anterior part. Morphotype 5 is represented by worms from the white-eye bream Ballerus sapa (Pallas); its typical characteristics are a festoon-like anterior margin of the scolex, the absence of vitelline follicles posterior to the cirrus-sac and the absence of a well-developed internal seminal vesicle. Discriminant analysis of 15 morphometric variables readily separated Morphotypes 3, 4 and 5 and confirmed the key discriminating power of traits related to the reproductive system, especially the terminal reproductive organs. Morphological polymorphism and the genetic divergence of different morphotypes of C. laticeps correspond to its wide spectrum of fish definitive hosts and a large distribution area that includes Europe, most of Palaearctic Asia and northern Africa.

Development of microsatellite markers in Caryophyllaeus laticeps (Cestoda: Caryophyllidea), monozoic fish tapeworm, using next-generation sequencing approach.

Caryophyllaeus laticeps (Pallas 1781) (Cestoda: Caryophyllidea) is a monozoic tapeworm of cyprinid fishes with a distribution area that includes Europe, most of the Palaearctic Asia and northern Africa. Broad geographic distribution, wide range of definitive fish hosts and recently revealed high morphological plasticity of the parasite, which is not in an agreement with molecular findings, make this species to be an interesting model for population biology studies. Microsatellites (short tandem repeat (STR) markers), as predominant markers for population genetics, were designed for C. laticeps using a next-generation sequencing (NGS) approach. Out of 165 marker candidates, 61 yielded PCR products of the expected size and in 25 of the candidates a declared repetitive motif was confirmed by Sanger sequencing. After the fragment analysis, six loci were proved to be polymorphic and tested for heterozygosity, Hardy-Weinberg equilibrium and the presence of null alleles on 59 individuals coming from three geographically widely separated populations (Slovakia, Russia and UK). The number of alleles in particular loci and populations ranged from two to five. Significant deficit of heterozygotes and the presence of null alleles were found in one locus in all three populations. Other loci showed deviations from Hardy-Weinberg equilibrium and the presence of null alleles only in some populations. In spite of relatively low polymorphism and the potential presence of null alleles, newly developed microsatellites may be applied as suitable markers in population genetic studies of C. laticeps.

Conflict between morphology and molecular data: a case of the genus Caryophyllaeus (Cestoda: Caryophyllidea), monozoic tapeworms of cyprinid fishes.

Molecular analysis of an extensive specimen collection of morphologically distinct tapeworms of the genus Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes in the Palaearctic Region, brought evidence of host-related plasticity in critical morphological characters widely used for species circumscription and classification of these tapeworms. The results obtained here do not support the traditionally used morphological concept of species-defining characters of the order Caryophyllidea Carus, 1863, especially due to high morphological plasticity of the scolex and the anterior end of the body. Several morphotypes within both Caryophyllaeus laticeps (Pallas, 1781) and Caryophyllaeus brachycollis Janiszewska, 1953 generally corresponding to different hosts and geographical regions, were recognised likely suggesting host- and geography-related intraspecific morphological variability. Genetic data confirmed euryxenous host specificity of both species, most profoundly C. laticeps, but did not support the existence of cryptic species. In fact, some of the fish hosts may harbour both of the congeneric species including several of their respective morphotypes. The pattern of morphological and genetic divergence observed in both cestode species studied indicates a scenario of possible host range extensions and subsequent parasite diversification. If molecular sequence variability and host-related morphological polymorphism are confirmed in other groups of monozoic cestodes, it may complicate species identification and straightforward classification of these parasites.

Diversity and food web structure of nematode communities under high soil salinity and alkaline pH.

A long-term and intensive magnesium (Mg) ore processing in Slovenské Magnezitové Závody a.s. in Jelsava has resulted in a high Mg content and alkaline pH of the soil environment, noticeable mainly in the close vicinity of the smelter. Nematode communities strongly reacted to the contamination mostly by a decrease in abundance of the sensitive groups. Nematodes from c-p 1 group and bacterivores, tolerant to pollution played a significant role in establishing the dominance at all sites. With increasing distance from the pollution source, the nematode communities were more structured and complex, with an increase in proportion of sensitive c-p 4 and 5 nematodes, composed mainly of carnivores and omnivores. Various ecological indices (e.g. MI2-5, SI, H') indicated similar improvement of farther soil ecosystems.

Phenotypic plasticity in Caryophyllaeus brachycollis Janiszewska, 1953 (Cestoda: Caryophyllidea): does fish host play a role?

Recent molecular phylogenetic studies on fish tapeworms of the genus Caryophyllaeus Gmelin, 1790 (Cestoda: Caryophyllidea), parasites of cyprinid fishes in the Palaearctic Region, have revealed unexpected phenotypic plasticity that seems to be related to definitive hosts. In the present paper, Caryophyllaeus brachycollis Janiszewska, 1953 is redescribed and its two morphotypes are circumscribed on the basis of newly-collected specimens. Morphotype 1 from barbels [Barbus spp. including the type-host Barbus barbus (L.); Barbinae] and chubs (Squalius spp.; Leuciscinae) is characterised by a more robust body with spatulate scolex, which is only slightly wider than a very short neck region, and the anterior position of the testes and vitelline follicles, which begin immediately posterior to the scolex. Specimens of Morphotype 2 from breams (Abramis spp., Ballerus spp. and Blicca spp.; Abraminae), which have been previously misidentifed as Caryophyllaeus laticeps (Pallas, 1781), possess a more slender body with a flabellate scolex, which is much wider than a long neck, and the first testes begin at a considerable distance posterior to the first vitelline follicles. Despite conspicuous differences in the scolex morphology and the anterior extent of the testes and vitelline follicles, both morphotypes are identical in the morphology of the posterior end of the body, in particular that of the cirrus-sac, which is large, thick-walled, elongate-pyriform, and contains a long cirrus, and in the distribution of the vitelline follicles, which surround medially vas deferens near the cirrus-sac. A specimen of Morphotype 1 from B. barbus from the Argens River, France, is designated as neotype of C. brachycollis. The presence of phenotypic plasticity in morphological characteristics previously used for differentiation of species of Caryophyllaeus may confound species identification, which is crucial for biodiversity, ecological and evolutionary studies. To avoid these potential problems, combination of morphological and molecular data is strongly recommended.

Ultrastructure and chemical composition of the proboscis hooks of Acanthocephalus lucii (Müller, 1776) (Acanthocephala: Palaeacanthocephala) using X-ray elemental analysis.

The ultrastructure and chemical composition of the proboscis hooks and surrounding tegument of Acanthocephalus lucii (Müller, 1776), a parasite of European perch, Perca fluviatilis Linnaeus, were examined using scanning (SEM) and transmission (TEM) electron microscopy and X-ray microanalysis (EDXA). The blade of middle hooks consists of three layers: an outer homogeneous layer, an inner heterogeneous layer and a central core. TEM observation revealed the presence of hollow tubes, which spaced the central core; fibrous inner hook layer surrounded by an electron-dense margin and the basal tegumental layer filled with electron-dense bodies and outer layer. We found for the first time that the so-called 'epidermal covering' surrounding of the exposed hook blade (outer hook layer) is a modified striped portion of the tegumental layer and there are no special contact sites between these two morphologically different structures, i.e. striped layer of the syncytial tegument and following proper outer hook layer, which is a homogeneous, moderately electron-dense layer of -0.3 µm in thickness. The hook root is embedded into subtegumental fibrous layer. X-ray microanalysis of both the surface and internal parts of A. lucii hooks demonstrated the presence of calcium, magnesium, phosphorus and sulphur. The highest concentration of sulphur was recorded at the tip of hooks, whereas the middle part of the hooks was most rich in calcium, phosphorus and magnesium. The proximal part of the hooks contained lower concentrations of sulphur, calcium and phosphorus. In the proboscis tegument, only two elements, calcium and silicon, were found. The differences observed in the chemical composition of the hook 'epidermal covering' and the proboscis tegument support our ultrastructural findings that the hook tegumental covering is a modified structure compared with that of the general proboscis tegument.

Nematodes as bioindicators of soil degradation due to heavy metals.

The effect of distance from a heavy metal pollution source on the soil nematode community was investigated on four sampling sites along an 4 km transect originating at the Kovohuty a.s. Krompachy (pollution source). The soil nematode communities were exposed to heavy metal influence directly and through soil properties changes. We quantified the relative effects of total and mobile fraction of metals (As, Cd, Cr, Cu, Pb, and Zn) on soil ecosystem using the nematode community structure (trophic and c-p groups,) and ecological indices (Richness of genera, H', MI2-5, etc.). Pollution effects on the community structure of soil free living nematodes was found to be the highest near the pollution source, with relatively low population density and domination of insensitive taxa. A decrease in heavy metals contents along the transect was linked with an increase in complexity of nematode community. The majority of used indices (MI2-5, SI, H') negatively correlated (P < 0.05 or P < 0.01) with heavy metals content and were sensitive to soil ecosystem disturbance. Contamination by heavy metals has negatively affected the soil environment, which resulted in nematode community structure and ecological indices changes. Results showed that the free-living nematodes are useful tools for bioindication of contamination and could be used as an alternative to the common approaches based on chemical methods.

Perch and its parasites as heavy metal biomonitors in a freshwater environment: the case study of the Ruzín water reservoir, Slovakia.

Heavy metal concentrations were determined in 43 perches (Perca fluviatilis) and in two of its most common parasites, the acanthocephalan Acanthocephalus lucii and the cestode Proteocephalus percae, collected in the period 2009-2010 from Ruzín, a seriously polluted water reservoir in Slovakia. Samples of muscle, liver, kidney, brain, male and female reproductive organs and adipose tissue of fish and both parasites were analyzed for As, Cd, Cr, Cu, Hg, Mn, Ni, Pb and Zn, by ICP-MS. Mean concentrations of individual heavy metals in all fish samples decreased in the order zinc > copper > manganese > mercury > arsenic > chromium > cadmium > nickel > lead. Zinc was found to be the dominant element and its antagonistic interaction with copper was confirmed. The kidney was a key target organ receiving the highest mean concentrations of all analyzed metals, but some metals showed specific affinity for particular tissues. In terms of human health, concentration of Hg in fish muscle, which exceeded more than two-times its maximum level admitted in foodstuffs in European countries, is of great importance and should be taken into account. Bioaccumulation factors (C([parasite])/C([fish tissue])) calculated for all elements indicated much higher detection skills of A. lucii and P. percae parasites than fish organs and hence, present results allow proposing both parasite models as useful tools to monitor aquatic environmental quality. Acanthocephalans, however, seem to be superior for heavy metal monitoring, also demonstrated under experimental conditions. Present results also indicate the decreasing heavy metal burden of the reservoir and its gradual recovery in the course of time.

Bioaccumulation of six PCB indicator congeners in a heavily polluted water reservoir in Eastern Slovakia: tissue-specific distribution in fish and their parasites.

Concentrations of six indicator PCB congeners (IUPAC nos. 28, 52, 101, 138, 153, and 180) were measured in several organs and adipose tissue of a freshwater predatory fishes (European perch, northern pike, pike perch, wels catfish) as well as in nonpredators (common carp, freshwater bream, goldfish, white bream) and in acanthocephalan Acanthocephalus lucii from the water reservoir Zemplínska sírava (Eastern Slovakia), which is considered to be one of the most PCB-contaminated places in Europe. Concentration of PCBs was determined by capillary gas chromatography in samples from May to September 2009. The two-way main-effect ANOVA confirmed that feeding habits of fish (P < 0.00001) and peculiarity of individual fish organs (P < 0.01) affect PCB bioaccumulation. The total amount of PCBs was significantly higher (P < 0.05) in predators compared to nonpredators. Tissue-specific differences were found in PCB accumulation in both fish groups. PCBs were predominantly accumulated in the liver and hard roe. Individual congeners were not distributed homogeneously within the investigated organs and adipose tissue. PCB 153 was present in higher concentrations than the other congeners in all fish organs as well as in adipose tissue comprising an average 31 and 34 % of ΣPCB in predators and nonpredators, respectively. Acanthocephalans, attached to the intestine of perch, absorbed significantly higher concentrations of PCBs (P < 0.001) than the muscles, liver, kidney, brain, and adipose tissue of their host. About 20 times lower amount of PCBs was detected in the liver and almost 3 times in muscles of infected perch. Data on PCB accumulation in perch infected with acanthocephalans demonstrated a decline of PCB values in all organs as well as in adipose tissue compared to noninfected fish. About 20 times lower amount of PCBs was detected in the liver and almost 3 times in muscles of infected perch. Present results could indicate that some parasitic organisms may influence positively their hosts in PCB-contaminated environment.

Biomonitoring of polychlorinated biphenyls (PCBs) in heavily polluted aquatic environment in different fish species.

The distribution and concentrations of polychlorinated biphenyls (PCBs) were determined in fish species (European perch Perca fluviatilis, northern pike Esox lucius, pike perch Sander lucioperca, wels catfish Silirus glanus, common carp Cyprinus carpio, European eel Anguilla anguilla, freshwater bream Abramis brama, goldfish Carassius auratus, and roach Rutilus rutilus) in a heavily polluted water reservoir Zemplínska sírava (Slovakia). The study performed at two different time points 5 years apart (2004 and 2009) revealed serious PCB contamination of fish muscle tissue and significant interspecies as well as tissue-specific differences in PCB uptake by fish. Total PCBs broadly correlated with the trophic position of individual fish species within a food chain (P < 0.01). The concentrations were particularly high in predatory fish species, perch, pike, and pike perch (108.0, 90.1, and 113.0 mg kg(-1) lipid wt, respectively), but comparable PCB values were also found in non-predatory detrivorous freshwater bream (128.0 mg kg(-1) lipid wt). The lowest PCB values were surprisingly assessed in European eel (17.1 mg kg(-1) lipid wt). Tissue analysis showed the highest storage capacity of the liver (hepatopancreas in cyprinids) with maximum concentrations recorded found in northern pike (214.0 mg kg(-1) lipid wt) and freshwater bream (163.0 mg kg(-1) lipid wt). Negative correlations, mostly not significant, between the total PCB concentrations and fish weight were observed (P > 0.05). The study has shown that the kind of fish, its feeding habit, and specific conditions of the habitat are mutually interrelated factors that are responsible for significant variations in fish body burdens. A tendency to PCB biomagnification was also proved in some fish species of this water reservoir.

Revision of Khawia spp. (Cestoda: Caryophyllidea), parasites of cyprinid fish, including a key to their identification and molecular phylogeny.

Monozoic cestodes of the genus Khawia Hsü, 1935 (Caryophyllidea: Lytocestidae), parasites of cyprinid fish in Europe, Asia, Africa and North America, are revised on the basis of taxonomic evaluation of extensive materials, including recently collected specimens of most species. This evaluation has made it possible to critically assess the validity of all 17 nominal species of the genus and to provide redescriptions of the following seven species considered to be valid: Khawia sinensis Hsü, 1935 (type species); K. armeniaca (Cholodkovsky, 1915); K. baltica Szidat, 1941; K. japonensis (Yamaguti, 1934); K. parva (Zmeev, 1936); K. rossittensis (Szidat, 1937); and K. saurogobii Xi, Oros, Wang, Wu, Gao et Nie, 2009. Several new synonyms are proposed: Khawia barbi Rahemo et Mohammad, 2002 and K. lutei Al-Kalak et Rahemo, 2003 are synonymized with K. armeniaca; K. coregoni Kritscher, 1990 with Caryophyllaeus laticeps (Pallas, 1781) (family Caryophyllaeidae); K. cyprini Li, 1964 and K. iowensis Calentine et Ulmer, 1961 with K. japonensis; K. dubia (Szidat, 1937) (syn. Bothrioscolex dubius Szidat, 1937) with K. rossittensis; and Tsengia neimongkuensis Li, 1964 and T. xiamenensis Liu, Yang et Lin, 1995 with K. sinensis. Khawia prussica (Szidat, 1937) (syn. Bothrioscolex prussicus Szidat, 1937) is considered to be species incertae sedis, but its morphology indicates it may belong to Caryophyllaeus Gmelin, 1790 (Caryophyllaeidae). The molecular analysis of all seven valid species, based on comparison of sequences of two nuclear ribosomal and two mitochondrial genes, has shown that the species form three major groups clustered according to their fish hosts. Five species from common and crucian carp and goldfish were grouped together, whereas K. armeniaca from barbels (Barbinae) and K. baltica from tench (Tinca) formed separate clades. In contrast, geographical distribution does not seem to play a crucial role in grouping of individual taxa. A phylogenetic tree based on morphological characters was incongruent with that inferred from molecular data, which indicates that some morphological traits may be homoplastic. A key to identification of all species of Khawia based on morphological characteristics is provided.

Nematode community structure in the vicinity of a metallurgical factory.

Soil nematode communities (taxa composition, trophic structure, ecological indices) in the area of metallurgical factory (Oravské ferozliatinárske závody) in Siroká, Northern Slovakia were investigated in 2009. The factory belongs to main sources of emissions originated by ferroalloy production in this region. Four sites (meadows) were selected in a downwind direction from the factory: site A was located 0.85 km far from the factory, and the other sites were maintained in approximately 2-km intervals from each other. Chemical analysis of soil samples showed low concentrations of heavy metals (As, Cd, Cr, Cu, Ni, Pb and Zn), with all values being under Slovak limit concentrations of heavy metals in soils. Only the Cd content in the soil sample from site A slightly exceeded the allowable threshold, but it was decreasing with the distance from the factory, similarly as remaining metals except Cr, with slightly increasing trend of concentration. Within 64 identified nematode genera, the Helicotylenchus, Paratylenchus, Pratylenchus, Acrobeloides, Cephalobus and Rhabditis were most common and eudominant. This was clearly reflected on the trophic structure of nematode communities, where plant feeding nematodes and bacteriovorous prevailed. Significant negative correlation (P < 0.05) was observed between the abundance of bacteriovores and the concentration of Cu in the soil. On the other hand, fungivores showed significant correlation with Ni and Cr (P < 0.05) as well as predators with Cd, Pb and Zn contents in the soil (P < 0.01). The highly significant correlation (P < 0.05; P < 0.01) was found between As, Cd, Ni, Pb and Zn and Maturity Index 2-5. A negative relationship was detected between Maturity Index and the concentration of Cr in the soil (P < 0.01). On the other hand, Cu was in positive correlation with MI values. The MI, reflecting the degree of disturbances and changes in the structure and function of the soil ecosystem, was found to be the most sensitive indicator among all used indices.

Population study of Atractolytocestus huronensis (Cestoda: Caryophyllidea), an invasive parasite of common carp introduced to Europe: mitochondrial cox1 haplotypes and intragenomic ribosomal ITS2 variants.

The invasive monozoic tapeworm Atractolytocestus huronensis, a specific parasite of common carp, was originally found and described in the North American continent. It has been introduced to Europe and reported in several countries in the last 15 years, as well. In the current study, tapeworms from one North American (USA) and five European localities (United Kingdom/UK, Slovakia, Hungary, Croatia, and Romania) were subjected to molecular analyses in order to determine the level of intrapopulation and intraspecific molecular variation and to assess interrelationships among American and European populations of the parasite. Partial sequences (672 bp) of the mitochondrial cytochrome c oxidase subunit I (cox1) revealed the presence of only two cox1 haplotypes, in accordance with the nonnative character of the populations. The first haplotype was common for all tapeworms from the Continental Europe (Slovakia, Hungary, Croatia, and Romania); no differences were determined either within or among respective A. huronensis populations. The second cox1 haplotype was characterized in all individuals from the USA and UK, indicating their close genetic relationship. Both haplotypes differed in three nucleotide positions (99.6% identity) which did not change the amino acid sequence. The cox1 data imply that introduction of the parasite to Europe was probably the result of two independent events directed to the UK and Continental Europe. The very close genetic relationship between British and American A. huronensis was reflected also by similar ribosomal internal transcribed spacer 2 (ITS2) sequence structure; considerable intragenomic ITS2 variability was detected in all individuals of both geographic populations. Divergent ITS2 copies were mostly induced by different numbers of short repetitive motifs within the sequences, allowing their assortment into two ITS2 variants.