A fish-parasite sentinel system in an assessment of the spatial distribution of polychlorinated biphenyls.

The spatial distribution of polychlorinated biphenyls (PCBs), in the Zemplínska Sírava water reservoir and adjacent tributaries in the Bodrog River Basin were investigated using a fish-parasite sentinel system. PCB concentrations were detected in various fish matrices (dorsal and abdominal muscles, liver and intestine) of the Wels catfish (Silurus glanis) and its intestinal cestode Glanitaenia osculata. PCB concentrations in the fish from the water reservoir, located closest to the chemical plant, the primary source of the PCB pollution, were the highest. The analysis of these contaminants in catfish matrices showed the highest concentrations in the abdominal muscle, followed by the dorsal muscle, liver and intestine. Concentrations of ∑PCBs exceeding the limits for food set by European regulations were measured in the muscle tissue of catfish at all sites, even in the Bodrog River, 60 km away from the primary source of contamination, posing a significant risk to humans in the Zemplín region. For the first time, the ability of cestode G. osculata to accumulate higher amounts of PCBs compared to fish matrices has been demonstrated. Due to the enormous ability of the parasites to accumulate PCBs, we recommend this approach for alternative biomonitoring of PCBs in contaminated aquatic environments.

New cytogenetic data on Caryophyllaeus laticeps and Paracaryophyllaeus gotoi, parasites of evolutionary interest.

Caryophyllideans are intestinal parasites of freshwater fishes, occupying a basal position among the 'true' tapeworms. We performed detailed cytogenetic analyses of the well-known caryophyllidean species Caryophyllaeus laticeps. For comparison, we also examined for the first time the chromosomes of Paracaryophyllaeus gotoi, a specific parasite of loaches in China. Both species showed a diploid chromosome number of 2n = 20, n = 10m. Chromomycin A3 (CMA3)/diamidino-2-phenylindole (DAPI) staining performed for the first time in the class Cestoda revealed CMA3+/DAPI− bands in the pericentromeric regions of the short arms of chromosome pair no. 7 in the karyotype of C. laticeps. Fluorescence in situ hybridization with the 18S rDNA probe confirmed the presence of a single cluster of major rDNA near the centromere on a pair of small chromosomes in both species. These findings support the hypothesis that the ancestral state in the family Caryophyllaeidae is a single interstitial cluster of major rDNA genes and thus one nucleolar organizer region per haploid genome. Our results, which we presented together with literature data plotted on a phylogenetic tree, show stability of caryophyllidean karyotypes at the genus level, but showed differences between genera without a clear phylogenetic signal. The data allowed us to at least formulate a hypothesis about the ancestral haploid chromosome number of n = 10 for the family Caryophyllaeidae and possibly for the sister family Capingentidae. In addition, we compared two populations of C. laticeps from water bodies with different levels of polychlorinated biphenyl contamination, showing a slightly increased incidence of chromosomal abnormalities at the contaminated site.

Hazardous pollutants in the environment: Fish host-parasite interactions and bioaccumulation of polychlorinated biphenyls.

The present paper reports on the interrelationships of fish, parasites and the bioaccumulation of hazardous organic compounds in the Zemplínska Sírava water reservoir in eastern Slovakia, which is heavily polluted with polychlorinated biphenyls (PCBs). The concentrations of these contaminants were measured in various fish matrices (dorsal and abdominal muscle tissues, hepatopancreas, intestine wall and adipose tissue) of the freshwater bream, Abramis brama (Cyprinidae), and in its intestinal parasite Caryophyllaeus laticeps (Cestoda), which was used for the first time as a model for a PCB bioaccumulation study. Regarding the fish, the highest concentrations of PCBs were found in the intestine, followed by hepatopancreas and muscle tissues. The amounts of PCBs were higher in abdominal muscles than in their dorsal parts. Concentrations of ∑PCBs above the limits set by European regulations were detected in both muscle parts in the fish, confirming the persistent unfavorable conditions in this locality and high risk for biota and humans. Based on bioconcentration factor values (BCFs), PCBs reached much higher levels in cestodes compared to bream matrices. Some significant differences in PCB amounts between infected and uninfected bream were determined. Fulton's condition factor (CF) significantly differed in infected and non-infected fish (p ˂ 0.05), with CF values surprisingly lower in fish free of parasites compared to parasitized fish, which suggests a "mutualistic" relationship between the parasite and its host.

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.

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.

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.

Ultrastructural features of the spermatozoon and its differentiation in Brandesia turgida (Brandes, 1888) (Digenea, Microphalloidea, Pleurogenidae).

Spermatological characters of the digenean Brandesia turgida (Brandes, 1888), an intestinal parasite of the frog Pelophylax ridibundus (Pallas, 1771), have been investigated by means of transmission electron microscopy for the first time. The process of the spermatozoon formation begins with the appearance of the differentiation zone bordered by cortical microtubules and containing two centrioles associated with striated rootlets and with an intercentriolar body. The intercentriolar body is made up of seven distinct electron-dense plates, two less electron-dense, and four electron-lucent zones. The orthogonal development of the two flagella is followed by a flagellar rotation and their proximodistal fusion with the median cytoplasmic process. This process is accompanied by an extension of both the mitochondrion and nucleus into the median cytoplasmic process. The mature spermatozoon of B. turgida contains two parallel axonemes of unequal lengths with the 9 + "1" trepaxonematan pattern, mitochondrion, nucleus, parallel cortical microtubules, four electron-dense attachment zones, an external ornamentation of the plasma membrane, and electron-dense glycogen granules. The anterior extremity of the male gamete contains one complete centriole, a small component of the central element of the second centriole, and peripheral cortical microtubules (up to 45). The posterior extremity of the mature spematozoon exhibits tubular elements of the disorganized axoneme. The present study provides the first data on spermiogenesis within the family Pleurogenidae. Variations of the spermatozoa ultrastructural characters within Digenea, in particular, between different families of the superfamily Microphalloidea, are discussed.

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.