Mercury content in fish from drinking-water reservoirs in the Morava River Basin (Czech Republic).

This study focused on the total mercury content in fish from seven drinking-water reservoirs located in the Morava River Basin: Bojkovice, Boskovice, Hubenov, Karolinka, Landstejn, Ludkovice and Nova Rise in the Czech Republic. A total of 308 fish were collected for the analysis. The content of total mercury was measured in the muscle tissue of bream, roach and perch using atomic absorption spectrometry and varied from 0.057±0.009 to 0.440 mg kg-1 in bream, from 0.030±0.005 to 0.393±0.138 mg kg-1 in roach and from 0.092±0.007 to 0.638±0.042 mg kg-1 in perch. The highest total mercury content was found in perch from Landstejn and the lowest was measured in roach from Ludkovice. A positive statistically significant relationship was found between fish weight and total mercury content in fish muscle for almost all species and all sampling sites, except for roach from Ludkovice. A total of 19 samples exceeded the maximum mercury level set by legislation on food contaminants-0.5 mg kg-1 in freshwater fish.

Low-level pathogen transmission from wild to farmed salmonids in a flow-through fish farm.

While the potential effects of pathogens spread from farmed fish to wild populations have frequently been studied, evidence for the transmission of parasites from wild to farmed fish is scarce. In the present study, we evaluated natural bacterial and parasitic infections in brown trout (Salmo trutta m. fario) collected from the Cerná Opava river (Czech Republic) as a potential source of infections for rainbow trout (Oncorhynchus mykiss) reared in a flow-through farm system fed by the same river. The prevalence of bacterial and protozoan infections in farmed fish was comparable, or higher, than for riverine fish. Despite this, none of the infected farmed fish showed any signs of severe diseases. Substantial differences in metazoan parasite infections were observed between wild and farmed fish regarding monogeneans, adult trematodes, nematodes, the myxozoan Tetracapsuloides bryosalmonae found in riverine fish only, and larval eye-fluke trematodes sporadically found in farmed fish. The different distribution of metazoan parasites between brown and rainbow trout most probably reflects the availability of infected intermediate hosts in the two habitats. Despite the river being the main water source for the farm, there was no significant threat of parasite infection to the farmed fish from naturally infected riverine fish.

Myxozoan hidden diversity: the case of Myxobolus pseudodispar Gorbunova, 1936.

Myxobolus pseudodispar Gorbunova, 1936 (Myxozoa) was originally described as a parasite of common roach, Rutilus rutilus (Linnaeus), with developing stages in muscles and spores disseminated in macrophage centres of different organs and tissues. Later, this parasite was described from several other cyprinids, but with relatively large intraspecific differences based on SSU rDNA gene sequences. Within our long-term study on myxozoan biodiversity, we performed a broad microscopic and molecular screening of various freshwater fish species (over 450 specimens, 36 species) from different localities. We investigated the cryptic species status of M. pseudodispar. Our analysis revealed four new unique SSU rDNA sequences of M. pseudodispar as well as an infection in new fish host species. Myxobolus pseudodispar sequence analysis showed clear phylogenetic grouping according to fish host criterion forming 13 well-recognised clades. Using 1% SSU rDNA-based genetic distance criterion, at least ten new species of Myxobolus Bütschli, 1882 may be recognised in the group of M. pseudodispar sequences. Our analysis showed the paraphyletic character of M. pseudodispar sequences and the statistical tests rejected hypothetical tree topology with the monophyletic status of the M. pseudodispar group. Myxobolus pseudodispar represents a species complex and it is a typical example of myxozoan hidden diversity phenomenon confirming myxozoans as an evolutionary very successful group of parasites with a great ability to adapt to a new hosts with subsequent speciation events.

Morphometric and genetic evidence for cryptic diversity in Gyrodactylus (Monogenea) infecting non-native European populations of Ameiurus nebulosus and A. melas.

Gyrodactylid parasites were observed on non-native populations of North-American freshwater catfishes, Ameiurus nebulosus and Ameiurus melas (Siluriformes: Ictaluridae), at several sites in the Elbe River basin, Czech Republic, Europe. Using a combination of morphological and genetic analyses, the parasites infecting A. nebulosus were determined to be Gyrodactylus nebulosus, a North American parasite co-introduced to Europe along with its Ameiurus fish hosts. Subtle morphometrical differences, as well as seasonal variations, were observed among parasites collected from A. nebulosus and A. melas. The host-related variation was further supported through genetic analysis of the partial 18S rDNA, ITS1-5.8S-ITS2 and COI, showing 0.2, 3.0 and 4.8% divergence, respectively. Consistent genetic differences indicated there were two distinct genotypes. Subtle morphological differences associated with the shape of sickle toe, anchor root and ventral bar membrane, according to host species, also supported the description of a new cryptic species, Gyrodactylus melas n. sp., infecting A. melas. Multivariate morphometrical analysis of haptoral hard parts showed significant differences between the anchor lengths of G. nebulosus and G. melas n. sp. However, the measurements of the haptoral hard structures partially overlapped between species, limiting the usage of these parameters for species delineation.

Concentrations of Thirteen Trace Metals in Scales of Three Nototheniid Fishes from Antarctica (James Ross Island, Antarctic Peninsula).

In this study, we assessed concentrations of 13 trace metals in the scales of Notothenia coriiceps, Trematomus bernacchii and Gobionotothen gibberifrons caught off the coast of James Ross Island (Antarctic Peninsula). Overall, our results for scales broadly match those of previous studies using different fish and different organs, with most metals found at trace levels and manganese, aluminium, iron and zinc occurring at high levels in all species. This suggests that scales can serve as a useful, non-invasive bioindicator of long-term contamination in Antarctic fishes. High accumulation of manganese, aluminium, iron and zinc is largely due to high levels in sediments associated with nearby active volcanic sites. Manganese, vanadium and aluminium showed significant positive bioaccumulation in T. bernacchii (along with non-significant positive accumulation of iron, zinc, cobalt and chromium), most likely due to greater dietary specialisation on sediment feeding benthic prey and higher trophic species. Levels of significance in bioaccumulation regressions were strongly affected by large-scale variation in the data, driven largely by individual differences in diet and/or changes in habitat use and sex differences associated with life stage and reproductive status. Increased levels of both airborne deposition and precipitation and meltwater runoff associated with climate change may be further adding to the already high levels of manganese, aluminium, iron and zinc in Antarctic Peninsula sediments. Further long-term studies are encouraged to elucidate mechanisms of uptake (especially for aluminium and iron) and possible intra- and interspecific impacts of climate change on the delicate Antarctic food web.

Non-native parasitic copepod Neoergasilus japonicus (Harada, 1930) utilizes non-native fish host Lepomis gibbosus (L.) in the floodplain of the River Dyje (Danube basin).

The parasitic copepod Neoergasilus japonicus (Harada, 1930) (Ergasilidae), native to east Asia, is widely distributed in Asia, Europe, and North and Central America. Recently, this species appeared in lentic water bodies of the River Dyje floodplain (Danube basin, Czech Republic). It was first recorded in 2015 and in 2 years it reached a 100% prevalence in recently expanding non-native fish host, Lepomis gibbosus (Linnaeus, 1758) (Centrarchidae, native to North America) at two borrow pits. Abundance of N. japonicus increased with fish length, with maximum intensity of infection reaching 99 parasites per fish. The parasite was most frequently found attached to the dorsal and anal fins of fish, while preference for the dorsal fin was more evident with lower infection intensities. Utilization of expanding fish hosts in water bodies that are regularly interconnected via natural or managed flooding may support the rapid dispersal of this non-native parasite.

Methodological issues affecting the study of fish parasites. III. Effect of fish preservation method.

The aim of this study was to evaluate the influence of preservation method on the results of parasite community studies. Two host species, European perch Perca fluviatilis and European bitterling Rhodeus amarus, were examined for parasites after having been subjected to 4 different storage treatments: freezing, preservation in 4% formaldehyde or 70% ethanol and transportation of live (fresh) fish as a control. Preservation prior to dissection resulted in a loss of information, leading to incomplete quantitative data (all preservation treatments), qualitative data (ethanol and formaldehyde preservation) and a lowered ability to determine parasites to species level based on morphology compared to dissecting fresh fish. Of the more abundant taxa, only crustaceans and acanthocephalans provided relatively even results between treatments. We conclude that preservation media, such as ethanol or formaldehyde, significantly affects the ability to obtain precise parasite community data; hence, we recommend the use of freshly sacrificed fish for parasite community studies whenever possible. Alternatively, freezing may prove acceptable for evaluating parasite community taxonomic composition.

European distribution for metacercariae of the North American digenean Posthodiplostomum cf. minimum centrarchi (Strigeiformes: Diplostomidae).

Metacercariae of a North American digenean Posthodiplostomum cf. minimum centrarchi (Strigeiformes: Diplostomidae), have been reported from seven localities in Europe, with cysts recorded in the mesentery and internal organs of two invasive non-indigenous fishes (Lepomis gibbosus and Micropterus salmoides) from sites in Bulgaria, the Czech Republic and Portugal. Analysis of rDNA locus ITS1-ITS2-28S confirmed a closer relationship to the American Posthodiplostomum species than the common European species P. cuticola or P. brevicaudatum. Our data indicate limited potential of this parasite for switch to local fish fauna and confirm the occurrence at distant sites across Europe, suggesting that birds as definitive parasite hosts may play an important role for parasite dispersal. Further detailed studies are needed to confirm the actual means of introduction to Europe.

Methodological issues affecting the study of fish parasites. II. Sampling method affects ectoparasite studies.

In this study, we assessed the impact of sampling method on the results of fish ectoparasite studies. Common roach Rutilus rutilus were sampled from the same gravel pit in the River Dyje flood plain (Czech Republic) using 3 different sampling methods, i.e. electrofishing, beach seining and gill-netting, and were examined for ectoparasites. Not only did fish caught by electrofishing have more of the most abundant parasites (Trichodina spp., Gyrodactylus spp.) than those caught by beach seining or gill-netting, they also had relatively rich parasite infracommunities, resulting in a significantly different assemblage composition, presumably as parasites were lost through handling and 'manipulation' in the net. Based on this, we recommend electrofishing as the most suitable method to sample fish for parasite community studies, as data from fish caught with gill-nets and beach seines will provide a biased picture of the ectoparasite community, underestimating ectoparasite abundance and infracommunity species richness.

Methodological issues affecting the study of fish parasites. I. Duration of live fish storage prior to dissection.

We tested the ability of parasite species to respond quickly to artificial conditions (e.g. by changing abundance or even decreasing to extinction) while host fish species were being held alive prior to dissection. Prussian carp Carassius gibelio were sampled by electrofishing from 2 ponds alongside the River Dyje (Czech Republic) during 'cold' and 'warm' seasons. All fish were transported to the laboratory in aerated pond water and kept in a 1 m3 outdoor basin with aged tap water for 6 d. Twenty fish were dissected on consecutive days (total 120 fish for each site). Our results indicated that there was little change in parasite loading over the first 3 d of holding, suggesting no impact on parasitological studies undertaken over this period. From the fourth day, however, overall parasite abundance increased due to rapid reproduction of some parasite species, especially gyrodactylids in the cold season and dactylogyrids in the warm season. Parasite diversity appeared less stable in the warm season, with significant differences being registered as early as the second day. In addition to holding period, environmental conditions during fish holding will also play an important role in parasite community shifts.

Non-native gobies facilitate the transmission of Bucephalus polymorphus (Trematoda).

BACKGROUND: Introduced species can modify local host-parasite dynamics by amplifying parasite infection which can 'spill-back' to the native fauna, whether they are competent hosts for local parasites, or by acting as parasite sinks with 'dilution' of infection decreasing the parasite burden of native hosts. Recently infection by the trematode Bucephalus polymorphus has increased in several European rivers, being attributed to the introduction of intermediate host species from the Ponto-Caspian region. Using a combination of field and experimental data, we evaluated the competence of non-native and native fish as intermediate hosts for B. polymorphus and its role for parasite development in a definitive host. METHODS: The density of 0+ juvenile fish (the second intermediate hosts for B. polymorphus) was measured in the River Morava, Czech Republic and fish were screened for natural metacercariae infection. The stomach contents of predatory fish that are definitive hosts of B. polymorphus were examined to assess the importance of non-native gobies for parasite transmission. In semi-natural conditions, parasite establishment, initial survival, and maturity rates in experimentally infected definitive hosts pikeperch Sander lucioperca were measured in flukes recovered from native white bream Abramis bjoerkna and non-native tubenose goby Proterorhinus semilunaris and round goby Neogobius melanostomus. Adult fluke size and egg production was also measured to evaluate the potential effect of intermediate host species on parasite fitness. RESULTS: We detected high natural infection parameters of B. polymorphus in native cyprinids and non-native gobies compared to data from the period prior to goby establishment. Both fish groups are consumed by predatory fish and represent a major component of the littoral fish community. Parasite establishment and adult size in definitive hosts was equivalent among the second intermediate host species, despite a lower size of metacercariae recovered from round gobies. However, development in the definitive host of flukes recovered from gobies was reduced, showing higher mortality, delayed maturity and lower egg production, in comparison with parasites from native hosts. CONCLUSIONS: Substantial 'spill-back' of B. polymorphus due to higher transmission rates after establishment of non-native gobies was partially buffered by decreased fitness of B. polymorphus that underwent development in gobies.

Hidden diversity and evolutionary trends in malacosporean parasites (Cnidaria: Myxozoa) identified using molecular phylogenetics.

Malacosporeans represent a small fraction of myxozoan biodiversity with only two genera and three species described. They cycle between bryozoans and freshwater fish. In this study, we (i) microscopically examine and screen different freshwater/marine fish species from various geographic locations and habitats for the presence of malacosporeans using PCR; (ii) study the morphology, prevalence, host species/habitat preference and distribution of malacosporeans; (iii) perform small subunit/large subunit rDNA and Elongation factor 2 based phylogenetic analyses of newly gathered data, together with all available malacosporean data in GenBank; and (iv) investigate the evolutionary trends of malacosporeans by mapping the morphology of bryozoan-related stages, host species, habitat and geographic data on the small subunit rDNA-based phylogenetic tree. We reveal a high prevalence and diversity of malacosporeans in several fish hosts in European freshwater habitats by adding five new species of Buddenbrockia and Tetracapsuloides from cyprinid and perciform fishes. Comprehensive phylogenetic analyses revealed that, apart from Buddenbrockia and Tetracapsuloides clades, a novel malacosporean lineage (likely a new genus) exists. The fish host species spectrum was extended for Buddenbrockia plumatellae and Buddenbrockia sp. 2. Co-infections of up to three malacosporean species were found in individual fish. The significant increase in malacosporean species richness revealed in the present study points to a hidden biodiversity in this parasite group. This is most probably due to the cryptic nature of malacosporean sporogonic and presporogonic stages and mostly asymptomatic infections in the fish hosts. The potential existence of malacosporean life cycles in the marine environment as well as the evolution of worm- and sac-like morphology is discussed. This study improves the understanding of the biodiversity, prevalence, distribution, habitat and host preference of malacosporeans and unveils their evolutionary trends.

No relationship found between mercury and lead concentrations in muscle and scales of chub Squalius cephalus L.

We examined the relationship between muscle and scale mercury (Hg) and lead (Pb) concentrations in chub Squalius cephalus L. from six riverine sites in the Czech Republic in order to determine whether scale analysis alone could provide a nonlethal and convenient method for prediction of heavy metal concentration in muscle tissue. Our results confirm tissue-specific heavy metal accumulation in chub, with Hg tending to accumulate primarily in muscle tissue and Pb in scales. We found no significant relationship, however, for concentrations of either Pb or Hg between muscle tissue and scales of chub. Our results indicate that scales cannot be used for predicting heavy metal contamination in muscle of chub and we recommend, therefore, that muscle biopsy methods continue as the preferred method of analysis.

Spatiotemporal trends of heavy metal concentrations in fish of the River Morava (Danube basin).

The aim of this study was to assess mercury (Hg), cadmium (Cd) and lead (Pb) contamination in muscle tissue of fishes over the longitudinal profile of the River Morava (Czech Republic, Danube basin) and to detect any temporal trends over the past 18 years. Fish samples were collected in 1992, 1998, 2000, 2003 and 2009 at 6 study sites situated just downstream of important pollution sources. Chub (Leuciscus cephalus L.) were selected as indicator species at 5 sites, and brown trout (Salmo trutta m. fario L.) at the uppermost site where chub do not occur. In total, muscle tissue of 175 specimens of chub and 19 specimens of brown trout were analysed. Concentrations of heavy metals ranged as follows: mercury 0.015-0.369 mg/kg; cadmium 0.001-0.254 mg/kg and lead 0.006-1.505 mg/kg. Mercury levels did not exceed the maximum allowed concentration in the Czech Republic (0.5 mg/kg). Content of cadmium and lead in fish muscle exceeded the maximum allowed levels (0.05 and 0.3 mg/kg respectively) in 11 and 4 samples, respectively. On average, the order of metal concentration in fish muscle was: Hg>Pb>Cd. No significant differences were found between sites along the longitudinal profile of the river. Significant differences were found, however, for the interannual comparison of cadmium and lead (but not mercury) at different sites (P < 0.05). A catastrophic flood in 1997 resulted in an increase in metal concentrations, especially cadmium and lead, in the following 1998 season. Our results indicate that the Morava river basin does not represent a threatening source of mercury, cadmium or lead for the River Danube downstream.

Assessing relationships between chemical exposure, parasite infection, fish health, and fish ecological status: a case study using chub (Leuciscus cephalus) in the Bílina River, Czech Republic.

Multiple stressor scenarios, as they are relevant in many watersheds, call for approaches extending beyond conventional chemical-focused approaches. The present study, investigated the fish population, represented by chub (Leuciscus cephalus), in the Bílina River (Czech Republic), which is impacted by various pollution sources and might pose a risk on the fish population. To confirm or reject this hypothesis it was examined whether there exists an association between abundance of chub and exposure to toxic chemicals as well as natural stressors, represented by parasites, and whether health-related suborganismal traits, namely, organ indices, tissue histopathology, and immune parameters, would help in revealing relationships between stressor impact and population status. Toxic pressure was assessed by the toxic unit approach, which gives an integrative estimate of toxic effect concentrations and by measuring the biomarkers cytochrome P4501A and vitellogenin, which indicate exposure to bioavailable arylhydrocarbon- or estrogen receptor ligands. Parasite pressure was estimated by determining abundance and species composition of ecto- and endoparasites of chub. Chub abundance was high upstream in the Bílina, low to zero in the middle stretches, and increased again downstream. Toxic pressure increased in the downstream direction, while parasite intensity decreased in this direction. Health status of chub did not differ clearly between up-, middle-, and downstream sites. Thus, it appears that neither toxic pressure nor parasite pressure nor their combination translates into a change of chub health status. By using varied assessment tools, this study provides evidence against a presumed causative role of toxicants impairing the fish ecological status of the Bílina River.

The interaction between an introduced fish host and local parasite fauna: Neogobius kessleri in the middle Danube River.

Parasite communities of introduced fish Neogobius kessleri Günther (Gobiidae) were studied at five localities in the Slovak section of the Danube River during 2002-2005. Thirty-three metazoan parasite species were identified. All fish were infected with at least two parasite species; most of the parasite species were generalists. At all sampling sites, high susceptibility to local parasites was observed. The parasite community was dominated by three parasite species: glochidia of Anodonta anatina, larval or subadult acanthocephalan Pomphorhynchus laevis, and larval nematode Raphidascaris acus. The infection of both A. anatina and P. laevis was affected by season and habitat type, with higher abundance in spring and more frequent occurrence of A. anatina in side channels and P. laevis in main river channels. At both the component and infracommunity levels, a more diverse parasite community was found in side channels. This habitat was dominated by actively transmitted parasites, whilst endoparasites were more abundant in fish from the main river channel. Larval stages of parasites dominated the endoparasite community at all sampling sites. The introduced N. kessleri was used as intermediate host for most of the recorded parasites, in some cases also as a paratenic host. Finally, the importance of gobies as suitable hosts for local non-native parasite species (Anguillicoloides crassus, Anodonta woodina, Hydrozetes lacustris) is discussed.

Parasite assemblages of European bitterling (Rhodeus amarus), composition and effects of habitat type and host body size.

Parasite community composition of European bitterling (Rhodeus amarus), the only bitterling species occurring on the European continent, was investigated in 16 different localities from four European sea drainages during 1998-2007. A total of 41 species of metazoan parasites was identified. Nine parasite species are new records for European bitterling, namely Dactylogyrus rarissimus, D. suecicus, D. yinwenyingae, Gyrodactylus vimbi, Sphaerostomum globiporum, Petasiger sp., Paryphostomum radiatum, Ichthyocotylurus variegatus and Posthodiplostomum brevicaudatum. The specialist Gyrodactylus rhodei was the most widely distributed and one of the most prevalent species. The most frequent digenean species, represented by larval stages, was Metorchis xanthosomus. The parasite community of European bitterling was characterised by the dominance of generalists and parasites with autogenic life cycles. The rare occurrence of strictly endoparasitic species reflected the specific diet of the fish host. The character of the habitat significantly affected the parasite assemblages of bitterling. The greatest similarity was associated with lentic habitats (gravel pits and oxbows) and the lowest similarity between gravel pits and rivers. Juvenile bitterling from 8mm in length upwards were colonised by metazoan parasites, firstly by the monogenean G. rhodei. Host body size was positively correlated with parasite species richness, but the variability explained by length was low.

Parasitism, life history traits and immune defence in cyprinid fish from Central Europe.

BACKGROUND: The main prediction of life-history theory is that optimal energy allocated among the traits is related to the growth, maintenance and survival. It is hypothesized that the optimal resource allocated to immune function, which generates resistance towards parasites and reduce the fitness losses caused by parasitism, is depending on other requirements for energetic resource and the benefits associated with them. The aims of this study are to investigate in a comparative way (1) how parasitism is related to fish life history traits (fecundity, longevity, mortality), (2) whether there is a trade-off between reproduction and immune investments in fish females (i.e. energetic hypothesis) and in males (i.e. immunohandicap hypothesis), (3) whether parasitism influences host immunity (spleen size) and reproduction (gonad size) in females and males. RESULTS: Data on metazoan parasites of 23 cyprinid fish species from Central Europe were used for the analyses as well as new data collected from a field study. Ectoparasite species richness was negatively correlated with the fish mortality estimated by the k-value and positively correlated with fish body size, suggesting that parasite diversity increases with fish longevity. A negative relationship between spleen size and gonad size, controlling for fish body size, was found in females but not in males. Moreover, parasite abundance was positively correlated with fish spleen size and negatively with fish gonad size in females. CONCLUSION: The comparative analyses using cyprinid fish species demonstrated that natural mortality could be considered as a factor contributing to the variability of parasite species richness and moreover, parasite species benefit from long-lived fish. The results obtained from the analyses investigating the potential trade-off between reproduction and immunity could be interpreted as an energetic trade-off between female reproduction and immune function. The lack of negative relationship between gonad size and spleen size in males did not support our prediction based on the immunohandicap hypothesis.

Seasonal changes of immunocompetence and parasitism in chub (Leuciscus cephalus), a freshwater cyprinid fish.

Seasonal variation of water characteristics, predominantly temperature, is considered to strongly affect fish physiology and immunology. In nature, this variation directly influences the life cycle especially in fish parasites, but the infection of parasites is also altered by the host immune response. This study is aimed to analyze the seasonal changes in selected physiological and immunological parameters, the latter a potential measure of fish immunocompetence. Moreover, the seasonal changes in metazoan parasite infection were investigated, and the potential associations between fish physiology, immunocompetence, and parasitism were estimated. No differences in gender were recognized for immunological parameters. The significant differences in the spleen-somatic index were found among fish samples of different seasons indicating higher investment in spleen size in April (after winterizing) and August (post-breeding with low gonado-somatic index [GSI]). The significant seasonal differences in erythrocyte and leukocyte cell counts, as well as in phagocyte count affecting respiratory burst, were recorded. The general trend of leukocyte composition was similar in all seasons investigated; however, the changes in proportion of different neutrophilic cells were reported. The parasite diversity and the infection parameters in parasite communities were highest in spring and early summer. When comparing parasite abundance in infracommunities among seasons, the statistically highest values were observed in April and June for Monogenea, in April and November for Acanthocephala, and in April for Cestoda. The positive correlations between GSI and the parasite abundance of groups with higher infection parameters were found in males. Moreover, the positive association between Monogenea as the dominant parasite group and respiratory burst was observed. The higher investment in condition and the seasonal variation in GSI were associated with a decrease immune function measured by spleen size or leukocyte cell count especially for males suggesting the seasonal energy allocation between immune function and somatic or reproductive investment.

MODELKEY. Models for assessing and forecasting the impact of environmental key pollutants on freshwater and marine ecosystems and biodiversity.

BACKGROUND: Triggered by the requirement of Water Framework Directive for a good ecological status for European river systems till 2015 and by still existing lacks in tools for cause identification of insufficient ecological status MODELKEY (http:// www.modelkey.org), an Integrated Project with 26 partners from 14 European countries, was started in 2005. MODELKEY is the acronym for 'Models for assessing and forecasting the impact of environmental key pollutants on freshwater and marine ecosystems and biodiversity'. The project is funded by the European Commission within the Sixth Framework Programme. OBJECTIVES: MODELKEY comprises a multidisciplinary approach aiming at developing interlinked tools for an enhanced understanding of cause-effect-relationships between insufficient ecological status and environmental pollution as causative factor and for the assessment and forecasting of the risks of key pollutants on fresh water and marine ecosystems at a river basin and adjacent marine environment scale. New modelling tools for risk assessment including generic exposure assessment models, mechanistic models of toxic effects in simplified food chains, integrated diagnostic effect models based on community patterns, predictive component effect models applying artificial neural networks and GIS-based analysis of integrated risk indexes will be developed and linked to a user-friendly decision support system for the prioritisation of risks, contamination sources and contaminated sites. APPROACH: Modelling will be closely interlinked with extensive laboratory and field investigations. Early warning strategies on the basis of sub-lethal effects in vitro and in vivo are provided and combined with fractionation and analytical tools for effect-directed analysis of key toxicants. Integrated assessment of exposure and effects on biofilms, invertebrate and fish communities linking chemical analysis in water, sediment and biota with in vitro, in vivo and community level effect analysis is designed to provide data and conceptual understanding for risk arising from key toxicants in aquatic ecosystems and will be used for verification of various modelling approaches. CONCLUSION AND PERSPECTIVE: The developed tools will be verified in case studies representing European key areas including Mediterranean, Western and Central European river basins. An end-user-directed decision support system will be provided for cost-effective tool selection and appropriate risk and site prioritisation.