Search

Owing to their widespread use, pharmaceuticals have become important pollutants of the aquatic environment. Pharmaceuticals and parasites can affect the immunity, physiology and behaviour of target organisms, both singly and through interaction. In this study, we related pharmaceutical concentration, parasite infection and condition in brown trout (Salmo trutta m. fario) collected at localities upstream and downstream of a sewage treatment plant (STP). The sites were separated by a weir preventing fish migration. Pharmaceutical number and concentrations differed significantly between sites. Antibiotics and antidepressants were found at the highest concentrations at both localities. Though fish condition and overall parasite abundance did not differ between sites, there was a significant difference in parasite community composition. A higher abundance of monogenean ectoparasites, along with a lower abundance of endoparasites, was observed at the downstream locality. The adverse effect of pharmaceutical load on ecto- and endoparasitic species on brown trout was further confirmed at the individual level using multivariate analysis. Both the number and concentration of pharmaceuticals in fish tissues reflected their high incidence in the environment, confirming the ability of trout to bioaccumulate pharmaceuticals. This could have important consequences on fish health, particularly in small streams where STP effluent represents a high proportion of stream flow.

Traces of psychoactive substances have been found in freshwaters globally. Fish are chronically exposed to pollution at low concentrations. The changes of aggressive behaviour of chub (Squalius cephalus) were determined under the exposure to four psychoactive compounds (sertraline, citalopram, tramadol, methamphetamine) at environmentally relevant concentrations of 1 μg/L for 42 days. We tested whether (A) the behavioural effect of compounds varies within a single species; (B) there is a correlation between the individual brain concentration of the tested pollutants and fish aggression using the novel analysis of pollutants in brain; and (C) there is detectable threshold to effective pollutant concentration in brain. Behaviour and pollutant concentrations in brain were determined repeatedly (1st, 7th, 21st, 42nd and 56th days), including a two-week-long depuration period. The effect of particular compounds varied. Citalopram and methamphetamine generally increased the fish aggression, while no such effect was found after exposure to tramadol or sertraline. The longitudinal analysis showed an aggression increase after depuration, indicating the presence of withdrawal effects in methamphetamine- and tramadol-exposed fish. The analysis of pollutant concentration in brain revealed a positive linear relationship of citalopram concentration and aggression, while no such effect was detected for other compounds and/or their metabolites. Structural break analyses detected concentration thresholds of citalopram (1 and 3 ng/g) and sertraline (1000 ng/g) in brain tissue, from which a significant effect on behaviour was manifested. While the effect of sertraline was not detected using traditional approaches, there was a reduction in aggression after considering its threshold concentration in the brain. Our results suggest that pursuing the concentration threshold of psychoactive compounds can help to reduce false negative results and provide more realistic predictions on behavioural outcomes in freshwater environments, especially in the case of compounds with bioaccumulation potential such as sertraline.

Tramadol is a widely used analgesic with additional antidepressant and anxiolytic effects. This compound has been reported in continental waters reaching concentrations of µg/L as a consequence of its inefficient removal in sewage treatment plants and increasing use over time. In this study, European chubs (Squalius cephalus) were exposed to 1 µg/L of tramadol in water for 42 days with a subsequent 14 days of depuration. Our results revealed that chubs exposed to this analgesic underwent changes in their behaviour as compared to the control group. The behavioural outcome was also influenced by the individual concentration of tramadol in brain tissue. In particular, experimental fish presented anxiolytic-like effects, characterized by less bold and less social individuals. Exposed animals were less frequently out of the shelter and moved a shorter distance, indicating that they explored the new environment less during the boldness test. In the novel object recognition experiment, although they distinguished the new item, they examined it less and displayed a reduced activity. Shoal cohesion was disrupted as observed in an increased distance between individuals. After the depuration phase, this alteration remained whereas the boldness effect disappeared. Moreover, the degree of behavioural changes was correlated with the concentration of the substance in brain. According to our findings, chronic presence of tramadol in the environment can impact the fitness of exposed aquatic fauna by altering evolutionary crucial behaviours.

To gain a better understanding of which pharmaceuticals could pose a risk to fish, 94 pharmaceuticals representing 23 classes were analyzed in blood plasma from wild bream, chub, and roach captured at 18 sites in Germany, the Czech Republic and the UK, respectively. Based on read across from humans, we evaluated the risks of pharmacological effects occurring in the fish for each measured pharmaceutical. Twenty-three compounds were found in fish plasma, with the highest levels measured in chub from the Czech Republic. None of the German bream had detectable levels of pharmaceuticals, whereas roach from the Thames had mostly low concentrations. For two pharmaceuticals, four individual Czech fish had plasma concentrations higher than the concentrations reached in the blood of human patients taking the corresponding medication. For nine additional compounds, determined concentrations exceeded 10% of the corresponding human therapeutic plasma concentration in 12 fish. The majority of the pharmaceuticals where a clear risk for pharmacological effects was identified targets the central nervous system. These include e.g. flupentixol, haloperidol, and risperidone, all of which have the potential to affect fish behavior. In addition to identifying pharmaceuticals of environmental concern, the results emphasize the value of environmental monitoring of internal drug levels in aquatic wildlife, as well as the need for more research to establish concentration-response relationships.

Illicit drug abuse presents pervasive adverse consequences for human societies around the world. Illicit drug consumption also plays an unexpected role in contamination of aquatic ecosystems that receive wastewater discharges. Here, we show that methamphetamine, considered as one of the most important global health threats, causes addiction and behavior alteration of brown trout Salmo trutta at environmentally relevant concentrations (1 µg l−1). Altered movement behavior and preference for methamphetamine during withdrawal were linked to drug residues in fish brain tissues and accompanied by brain metabolome changes. Our results suggest that emission of illicit drugs into freshwater ecosystems causes addiction in fish and modifies habitat preferences with unexpected adverse consequences of relevance at the individual and population levels. As such, our study identifies transmission of human societal problems to aquatic ecosystems.

With increasing population growth and climate change, de facto reuse practices are predicted to increase globally. We investigated a longitudinal gradient within the Uhlava River, a representative watershed, where de facto reuse is actively occurring, during Fall and Spring seasons when instream flows vary. We observed human pharmaceutical levels in the river to continuously increase from the mountainous areas upstream to downstream locations and a potable intake location, with the highest concentrations found in small tributaries. Significant relationship was identified between mass flow of pharmaceuticals and the size of human populations contributing to wastewater treatment plant discharges. Advanced ozonation and granular activated carbon filtration effectively removed pharmaceuticals from potable source waters. We observed a higher probability of encountering a number of targeted pharmaceuticals during colder Spring months when stream flows were elevated compared to warmer conditions with lower flows in the Fall despite a dilution paradigm routinely applied for surface water quality assessment and management efforts. Such observations translated to greater water quality hazards during these higher Spring flows. Future water monitoring efforts should account for periods when higher chemical uses occur, particularly in the face of climate change for regions experiencing population growth and de facto reuse.

POCIS is the most widely applied passive sampler of polar organic substances, because it was one of the first commercially available samplers for that purpose on the market, but also for its applicability for a wide range of substances and conditions. Its main weakness is the variability of sampling performance with exposure conditions. In our study we took a pragmatic approach and performed in situ calibration for a set of 76 pharmaceuticals and their metabolites in five sampling campaigns in surface water, covering various temperature and flow conditions. In individual campaigns, RS were calculated for up to 47 compounds ranging from 0.01 to 0.63 L d−1, with the overall median value of 0.10 L d−1. No clear changes of RS with water temperature or discharge could be found for any of the investigated substances. The absence of correlation of experimental RS with physical-chemical properties in combination with the lack of mechanistic understanding of compound uptake to POCIS implies that practical estimation of aqueous concentrations from uptake in POCIS depends on compound-specific experimental calibration data. Performance of POCIS was compared with grab sampling of water in seven field campaigns comprising multiple sampling sites, where sampling by both methods was done in parallel. The comparison showed that for 25 of 36 tested compounds more than 50% of POCIS-derived aqueous concentrations did not differ from median of grab sampling values more than by a factor of 2. Further, for 30 of 36 compounds, more than 80% of POCIS data did not differ from grab sampling data more than by a factor of 5. When accepting this level of accuracy, in situ derived sampling rates are sufficiently robust for application of POCIS for identification of spatial and temporal contamination trends in surface waters.

Pharmaceutically active compounds (PhACs) are ubiquitous in the aquatic environment worldwide and considered emerging contaminants. Their effects on growth, behavior, and physiological processes of aquatic organisms have been identified even at very low concentrations. Ecotoxicological investigations have primarily focused on single compound exposure, generally at a range of concentrations. In the natural environment, pollutants seldom occur in isolation, but little is known about the effects and risks of combinations of chemicals. This study aimed to investigate the effects of concurrent exposure to six psychoactive PhACs on locomotory behavior and life history traits of clonal marbled crayfish Procambarus virginalis. Crayfish were exposed to ~1 μg L−1 of the antidepressants sertraline, citalopram, and venlafaxine; the anxiolytic oxazepam; the opioid tramadol; and the widely abused psychostimulant methamphetamine. In the absence of shelter, exposed crayfish moved significantly shorter distances and at lower velocity and showed significantly less activity than controls. With available shelter, exposed crayfish moved significantly more distance, showed higher activity, and spent a significantly more time outside the shelter than controls. Molting, mortality, and spawning frequency did not vary significantly between the groups. Hemolymph glucose level did not vary among groups and was not correlated with observed behaviors. Results suggest that environmental concentrations of the tested compounds in combination can alter the behavior of non-target aquatic organisms as individual exposure of these compounds, which may lead to disruption of ecosystem processes due to their reduced caution in polluted conditions. Further research is needed using varied chemical mixtures, exposure systems, and habitats, considering molecular and physiological processes connected to behavior alterations.

Methamphetamine (METH), a central nervous system stimulant used as a recreational drug, is frequently found in surface waters at potentially harmful concentrations. To determine effects of long-term exposure to environmentally relevant levels on nontarget organisms, we analysed cardiac and locomotor responses of signal crayfish Pacifastacus leniusculus to acute stress during a 21-day exposure to METH at 1 &mu, g L&minus, 1 followed by 14 days depuration. Heart rate and locomotion were recorded over a period of 30 min before and 30 min after exposure to haemolymph of an injured conspecific four times during METH exposure and four times during the depuration phase. Methamphetamine-exposed crayfish showed a weaker cardiac response to stress than was observed in controls during both exposure and depuration phases. Similarly, methamphetamine-exposed crayfish, during METH exposure, showed lower locomotor reaction poststressor application in contrast to controls. Results indicate biological alterations in crayfish exposed to METH at low concentration level, potentially resulting in a shift in interactions among organisms in natural environment.

Recent state-of-the-art methods developed for the analysis of polar xenobiotics from different types of biological matrices usually employ liquid chromatography with mass spectrometry. However, there are limitations when a small amount of sample mass is available. For example, individual benthic invertebrates or fish tissue samples often weigh less than 100 mg (e.g., brain, liver) but are necessary to understand environmental fate and bioaccumulation dynamics. We developed ultra-fast methods based on a direct sample introduction technique. This included coupling laser diode thermal desorption with atmospheric pressure chemical ionization mass spectrometry (LDTD-APCI-MS). We then quantitated a common selective serotonin reuptake inhibitor (citalopram) in brain tissues of individual juvenile fish after in vivo exposure to environmentally relevant concentration. Two mass spectrometric methods based on low (LDTD-APCI-triple quadrupole (QqQ)-MS/MS) and high (LDTD-APCI-high-resolution product scan (HRPS)) resolutions were developed and evaluated. Individual instrument conditions were optimized to achieve an accurate and robust analytical method with minimum sample preparation requirements. We achieved very good recovery (97–108%) across the range of 1–100 ng g−1 for LDTD-APCI-HRPS. LDTD-APCI-QqQ-MS/MS showed poorer performance due to interferences from the matrix at the lowest concentration level. LDTD-APCI ionization was successfully validated for analysis of non-filtered sample extracts. Evaluation of final methods was performed for a set of real fish brain samples, including comparison of LDTD-APCI-HRPS with a previously validated LC-heated electrospray ionization-HRPS method. This new LDTD-APCI-HRPS method avoids the chromatographic step and provides important benefits such as analysis of limited sample masses, lower total sample volume (typically μL), and reduction in analysis time per sample run to a few seconds.

Aquaculture is increasing at the global scale, and beneficial reuse of wastewater is becoming crucial in some regions. Here we selected a unique tertiary treatment system for study over a one-year period. This experimental ecosystem-based approach to effluent management included a treated wastewater pond (TWP), which receives 100% effluent from a wastewater treatment plant, and an aquaculture pond (AP) that receives treated water from the TWP for fish production. We examined the fate of a wide range of pharmaceutically active compounds (PhACs) in this TWP-AP system and a control pond fed by river water using traditional grab sampling and passive samplers. We then employed probabilistic approaches to examine exposure hazards. Telmisartan, carbamazepine, diclofenac and venlafaxine, exceeded ecotoxicological predicted no effect concentrations in influent wastewater to the TWP, but these water quality hazards were consistently reduced following treatment in the TWP-AP system. In addition, both grab and passive sampling approaches resulted in similar occurrence patterns of studied compounds, which highlights the potential of POCIS use for water monitoring. Based on the approach taken here, the TWP-AP system appears useful as a tertiary treatment step to reduce PhACs and decrease ecotoxicological and antibiotic resistance water quality hazards prior to beneficial reuse in aquaculture.

Monitoring the contamination level in aquatic environments and assessing the impact on aquatic life occurs throughout the world. In the present study, an approach based on a combination of biomarkers and the distribution of various industrial and municipal pollutants was used to investigate the effect of aquatic environmental contamination on fish. Monitoring was performed in ten rivers in the Czech Republic (Berounka, Dyje, Elbe, Lužnice, Odra, Ohře, Otava, Sazava, Svratka, and Vltava rivers, with one or two locations in each river) at the same sites that were regularly monitored within the Czech National Monitoring Program in 2007–2011. Health status, hepatic ethoxyresorufin-O-deethylase (EROD) activity, total cytochrome P450 content, and the plasma vitellogenin concentration were assessed in wild chub (Squalius cephalus) males caught at the monitored sites. The contamination level was the highest in the Svratka River downstream of Brno. Among all measured persistent organic pollutants (POPs), polychlorinated biphenyls and dichlorodiphenyltrichloroethane and its metabolites were the major contributors of POPs in fish muscle. Elbe, Odra, and Svratka rivers were identified as the most polluted. Fish from these locations showed reduced gonad size, increased vitellogenin concentration in male plasma, EROD, and total cytochrome P450 content. These biomarkers can be used for future environmental monitoring assessments. Overall, this study improves our understanding of the relationship between human activities and pollutant loads and further contributes to the decision to support local watershed managers to protect water quality in this region.

Sewage treatment plants (STPs) are one of the major source of pharmaceuticals and personal care products in the aquatic environment. Generally, the effects of individual chemicals on fish are studied under laboratory conditions, which leads to results that are potentially not realistic regarding the effects of these chemicals under environmental conditions. Therefore, in this study, common carps were held in exposed pond that receive water from STP effluents for 360 days under natural conditions. Elimination of xenobiotics starts in the fish intestine, in which the microbial community strongly influences its function. Moreover, the fish intestine functions as crucial organ for absorbing lipids and fatty acids (FA), with consequent transport to the liver where their metabolism occurs. The liver is the primary organ performing xenobiotic metabolism in fish, and therefore, the presence of pollutants may interact with the metabolism of FA. The catalytic activity of CYP1A and CYP3A-like enzymes, their gene expression, FA composition and intestinal microbiome consortia were measured. The catalytic activity of enzymes and their gene and protein expression, were induced in hepatic and intestinal tissues of fish from the exposed pond. Also, fish from the exposed pond had different compositions of FA than those from the control pond: concentration of 18:1 n-9 and 18:2 n-6 were significantly elevated and the longer chain n-3 FA 20:5 n-3, 22:5 n-3 and 22:6 n-3 were significantly lowered. There were clear differences among microbiome consortia in fish intestines across control and exposed groups. Microbiome taxa measured in exposed fish were also associated with those found in STP activated sludge. This study reveals that treated STP water, which is assumed to be clean, affected measured biomarkers in common carp.

BackgroundThe aquatic environment has been contaminated with various anthropogenic pollutants, including psychoactive compounds that may alter the physiology and behavior of free-living organisms. The present study focused on the condition and related mortality of the juvenile chub (Squalius cephalus). The aim of the study was to test whether the adverse effects of the antidepressants sertraline and citalopram, the analgesic tramadol and the illicit drug methamphetamine, on fish condition exist under environmentally relevant concentrations and whether these effects persist after a depuration period. Innovative analyses of the fish brain concentrations of these compounds were performed with the aim to show relationship between compound brain tissue concentration and fish condition.MethodsThe laboratory experiment consisted of 42 days of exposure and a subsequent 14-day depuration period with regular monitoring of the condition and mortality of exposed and control fish. Identical methodology, including individual brain concentration analyses for the tested compounds, was applied for all substances. Additional study on feeding under sertraline exposure was also conducted. The feeding was measured from the 28th day of the exposure, three times in a week, by observation of food intake during 15 minutes in social environment.ResultsThe effects of particular psychoactive compounds on chub condition varied. While sertraline induced a lower condition and increased mortality, the effects of methamphetamine were inverse, and tramadol and citalopram had no significant effect at all. Individual brain concentrations of the tested compounds showed that the effects of sertraline and methamphetamine on fish condition were increased with brain concentration increases. Additionally, the food intake was reduced in case of sertraline. In contrast, there was no relationship between tramadol and citalopram brain tissue concentration and fish condition, suggesting that the concentration-dependent effect is strongly compound-specific. Methamphetamine was the only compound with a persistent effect after the depuration period. Our results demonstrate the suitability of the brain concentration evidence approach and suggest that changes in fish condition and other related parameters can be expected in freshwater ecosystems polluted with specific psychoactive compounds.

Concerns about the effect of sewage treatment plant (STP) effluent on the health of freshwater ecosystems have increased. In this study, a unique approach was designed to show the effect of an STP effluent-dominated stream on native wild brown trout (Salmo trutta L.) exposed under fully natural conditions. Zivny stream is located in South Bohemia, Czech Republic. The downstream site of Zivny stream is an STP-affected site, which receives 25% of its water from Prachatice STP effluent. Upstream, however, is a minimally polluted water site and it is considered to be the control site. Native fish were collected from the upstream site, tagged, and distributed to both upstream and downstream sites. After 30, 90, and 180 days, fish were recaptured from both sites to determine whether the downstream site of the Zivny stream is associated with the effects of environmental pollution. Several biomarkers indicating the oxidative stress and antioxidant enzyme activities, cytochrome P450 activity, xenoestrogenic effects, bacterial composition, and lipid composition were investigated. Additionally, polar chemical contaminants (pharmaceuticals and personal care products (PPCPs)) were quantified using polar organic chemical integrative samplers (POCIS). Fifty-three PPCPs were detected in the downstream site; 36 of those were constantly present during the 180-day investigation period. Elevated hepatic 7-benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylase (BFCOD) (after 90 days) and blood plasma vitellogenin concentrations in males were detected in fish downstream of the STP effluent during all sampling events. An increase in the fishes' total fat content was also observed, but with low levels of ω-3 fatty acid in muscle tissue. Two bacterial taxa related to activated sludge were found in the intestines of fish from downstream. Our results show that Prachatice STP is a major source of PPCPs in the Zivny stream, which has biological consequences on fish physiology.

The fate of fourteen target perfluoroalkyl substances (PFASs) are described within a small stream affected by a sewage treatment plant (STP) effluent. Concentrations of target PFASs in samples of water, benthic macroinvertebrates and brown trout (Salmo trutta) are presented. Two hundred brown trout individuals originating from clean sites within the same stream were tagged and stocked into an experimental site affected by the STP's effluent. As a passive sampling approach, polar organic chemical integrative samplers (POCIS) were deployed in the water to reveal the water-macroinvertebrates-fish biotransformation processes of PFASs. Bioconcentration/bioaccumulation of target compounds was monitored one, three, and six months after stocking. Twelve of the fourteen target PFASs were found in concentration above the LOQ in at least one of the studied matrices. The compound pattern varied significantly between both the studied species and water samples. Concerning the accumulation of PFASs in fish, the highest concentrations were found in the liver of individuals sampled after three months of exposure. These concentrations rapidly decreased after six months although the water concentrations were slightly increasing during experiment.

Intensive aquaculture is an important and fast-growing food production industry generating significant amounts of nutrient-rich sludge, which represents a potential environmental threat. Vermicomposting aquacultural sludge has been suggested, but remained poorly understood – only survival and growth of initial earthworm stocks have been assessed so far. The present study provides a comprehensive evaluation of the production system, examining vermicomposting of three types of sludge each at four inclusion levels and the possibility of further utilising end-products (vermicomposts and earthworms). Through an 18-week experiment, high survival of initial earthworm stocks, exceeding 90% among treatments up to week 6, was documented. Higher inclusion levels and sludge types richer in nutrients positively influenced individual weight of initial stocks and their reproduction indices (cocoon and juvenile production). The most progressive treatments sustained >300 juveniles in experimental incubators containing 200 g dw of initial substrates. Original sludge and final vermicomposts were found suitable for use in agriculture, complying with limits for heavy metals given in the most usually applied regulations. In relation to the heavy metals, earthworms were found to be a generally safe feed for fish. Only arsenic concentrations may occasionally exceed given limits. Still, observed concentrations are considered safe, presuming arsenic presence primarily in organic forms having largely reduced toxicity. Vermicomposting is recommended as a clean and sustainable technology transforming aquaculture sludge into highly valuable vermicompost and earthworm biomass.

The fungicide cyproconazole (CPZ) inhibits the biosynthesis of ergosterol, an essential sterol component in fungal cell membrane and can also affect non-target organisms by its inhibitory effects on P450 monooxygenases. The predicted environmental concentration of CPZ is up to 49.05 μg/L and 145.89 μg/kg in surface waters and sediments, respectively, and information about CPZ toxicity towards non-target aquatic organisms is still limited. This study aimed to address the lack of ecotoxicological data for CPZ, and thus, an evaluation of the lethal and sub-lethal effects of CPZ was performed using two freshwater invertebrates (the midge Chironomus riparius and the planarian Dugesia tigrina). The estimated CPZ 48 h LC50 (95% CI) was 17.46 mg/L for C. riparius and 47.38 mg/L for D. tigrina. The emergence time (EmT50) of C. riparius was delayed by CPZ exposure from 0.76 mg/L. On the other hand, planarians showed higher tolerance to CPZ exposure. Sub-lethal effects of CPZ on planarians included reductions in locomotion (1.8 mg/L), delayed photoreceptors regeneration (from 0.45 mg/L), and feeding inhibition (5.6 mg/L). Our results confirm the moderate toxicity of CPZ towards aquatic invertebrates but sub-lethal effects observed also suggest potential chronic effects of CPZ with consequences for population dynamics.

Although pharmaceuticals are frequently studied contaminants, their fate in the environment is still not completely clear. During a one year study, a complex approach including water, sediment and fish sampling was used to describe the behaviour of pharmaceuticals and their metabolites (PTMs) in the environment. Eighteen pharmaceuticals and seven of their metabolites were determined in a pond used for the tertiary treatment of wastewater effluent. A liquid chromatography-tandem mass spectrometry method was applied to determine the PTMs concentrations in all matrices. Seasonal variations in concentrations were evaluated. The partitioning of contaminants between pond compartments was estimated by means of solid water distribution coefficients (Kd) and bioaccumulation factors (BAF) for the livers of fish. Kd values were almost stable throughout the year, which may be a sign of the continuous transport of PTMs between water and sediment under the experimental conditions. Almost all of the studied compounds, with exception of sertraline (BAF of 6200), were found to not be bioaccumulative in fish livers. The pond removal efficiency was calculated for all PTMs, and favourable conditions for natural pharmaceutical removal were proposed. Further aspects regarding fish pharmaceutical exposure need to be studied.

Pharmaceutically active compounds (PhAC) have been increasingly detected in freshwater and marine waterbodies worldwide and are recognized as major emerging micropollutant threat to the aquatic environment. Despite their low concentrations in the environment, there is evidence of effects on non-target aquatic organisms in natural habitats. To assess the potential effects of PhACs on its burrowing behavior, we exposed the red swamp crayfish Procambarus clarkii to methamphetamine or tramadol at the environmentally relevant concentration of 1 μg/L. Methamphetamine-exposed females constructed burrows of lower depth and volume relative to individual weight than did controls. Tramadol-exposed females consistently exhibited a tendency for smaller burrows, but this difference was not significant. Exposed males showed a non-significant tendency to excavate larger burrows compared with the control. Control and tramadol-treated females maintained the natural tendency of constructing relatively deeper and/or larger-volume burrows compared with males. This sex-related pattern was not detected in the methamphetamine group. The rate of human therapeutic PhAC usage is relatively stable year-round, and impacts on crayfish burrowing can be particularly damaging during periods of drought, when the dilution of waste waters is reduced, and burrowing becomes a critical survival strategy. Our results suggest that an increasingly broad range of environmental impacts of PhACs on non-target organisms can be expected in natural ecosystems.

This study characterized changes in biomarker responses in common carp (Cyprinus carpio) upon exposure to effluent water discharged from a sewage treatment plant (STP) under real conditions. Fish were exposed to contamination in Cezarka pond, which receives all of its water input from the STP in the town of Vodnany, Czech Republic. Five sampling events were performed at day 0, 30, 90, 180, and 360 starting in April 2015. In total, 62 pharmaceutical and personal care products (PPCPs) were detected in the polar organic chemical integrative sampler. Compared to a control pond, the total concentration of PPCPs was 45, 16, 7, and 7 times higher in Cezarka pond at day 30, 90, 180, and 360, respectively. The result of oxidative stress and antioxidant enzyme biomarkers indicated alterations in the liver and intestine tissues of fish from Cezarka pond at day 30 and 360, respectively. High plasma vitellogenin levels were observed in both exposed females (180 and 360 days) and males (360 days) compared with their respective controls. However, only exposed female fish had higher vitellogenin mRNA expression than the control fish in these periods. Exposed female fish showed irregular structure of the ovary with scattered oocytes, which further developed to a vitellogenic stage at day 360. Low white blood cell levels were indicated in all exposed fish. Despite numerous alterations in exposed fish, favorable ecological conditions including high availability of food resulted in a better overall condition of the exposed fish after 1 year of exposure compared to the controls.

Evidence of the ecological and biological impact of pharmaceuticals in surface waters on aquatic organisms is increasing. Tramadol is a synthetic opioid analgesic used to treat chronic and acute pain. To investigate its long-term effects at environmentally relevant levels, we evaluated heart rate (HR) and locomotion of signal crayfish Pacifastacus leniusculus during a 21-day exposure to 1 μg L-1 tramadol followed by 14 days depuration. Locomotion and HR were recorded over a period 30 min before and 30 min after exposure to physiological fluids of an injured conspecific, a natural stressor, four times during the tramadol exposure and four times during depuration. A significant increase in HR following stress induction was found in the majority of tramadol-exposed and control crayfish, as well as significant group-specific HR changes between both groups. Locomotor activity during tramadol treatment differed from that during depuration, in general showing less time spent in locomotion and lower distance moved. The tramadol exposed crayfish exhibited higher velocity during depuration than during the exposure period. Results may suggest a potential shift in prey-predator relationships.

Interspecific relationships frequently determine the effect a pollutant can have on an organism, and this is especially true in closely interacting species such as hosts and parasites. The high spatial and temporal variability of contaminant concentrations combined with the movement of aquatic biota can further influence the consequences that are associated with contamination. We used a full factorial design for the exposed and unexposed partners of the relationship between the parasitic larvae (glochidia) of the European freshwater mussel (Anodonta anatina) and its host fish (Squalius cephalus) to identify the sources of variation in the sublethal endpoints of species interaction (the intensity of parasite attachment, the spatial position of glochidia on the host body, and encapsulation success). We used the water-borne human pharmaceutical compounds methamphetamine (a central nervous system stimulant) and tramadol (an opioid) at environmentally relevant concentrations (˜ 6.7 and 3.8 nmol L−1 of methamphetamine and tramadol, respectively) as a proxy for contaminant exposure because these compounds are emerging aquatic stressors that are known for high spatial and temporal variability in their detected concentration levels. The relationship between the bivalve and the fish species was influenced by the preceding contact with both methamphetamine and tramadol, but this effect was highly asymmetric. Our experimental design enabled us to identify the specific changes in the relationship outcome that are elicited by the exposure of individual partners, such as the significant increase in glochidia infection success rate from 59.6 ± 3.9% to 78.7 ± 2.8% (means ± s.e.) that was associated with host exposure to methamphetamine. Additionally, the significant interaction effect of the exposure was demonstrated by the lowered proportion of glochidia attached to gills after the coexposure of both partners to tramadol. The impact of pharmaceuticals on wild aquatic host–parasite relationships provides an example of the risks that are associated with the unintentional discharge of biologically active compounds into freshwater habitats. Given the increasing evidence showing the ecological impact of waste pharmaceuticals, the use of multitrophic interaction endpoints after joint and unilateral exposures provides an important step towards the realistic risk assessment of these compounds.

Reservoirs are known to alter temperature and flow regimes, shift nutrient cycles, reduce downstream species diversity and enable a predominantly upstream spread of non-native species. However, information about the seasonal dynamics of the spread of non-natives from a reservoir to its tributaries and the further consequences regarding the spatial distribution of native species is rare. We observed the occurrence of fish in the Vltava River and its tributaries (Elbe catchment area, central Europe) upstream of the Lipno Reservoir for five consecutive years. We radio-tagged two non-native and four native species. To detect assemblage spatial variability, we sampled sites in the study area by electrofishing twice per year (spring and autumn). We expected seasonal trends in non-native species appearance in upstream reservoir tributaries and, conversely, low motivation of native fishes to descend to the reservoir. By analysing nearly 3000 individuals of 21 species from the longitudinal profile of the study area, we observed an effect of reservoir distance on the native species ratio in the upper Vltava catchment area, i.e., an increase in distance increased the native species proportion, and the opposite was observed for non-native species. Analyses of 3798 tracking positions of 193 tagged individuals showed massive spring dispersal of non-native species from the reservoir to the main tributary, the Vltava River, and their return to the reservoir for wintering. Their upstream movement positively correlated with an increase in flow rate. Native Salmo trutta showed a specific shift from the Vltava River to smaller streams during the summer, when the presence of non-native species in the Vltava River was most significant. These findings indicate that non-native species repeatedly spread from the reservoir to the upstream river stretch and its tributaries and potentially compete with native species for resources.

With increasing demand for aquaculture products, water reuse is likely to increase for aquaculture operations around the world. Herein, wastewater stabilization ponds (WSP) represents low cost and sustainable treatment technologies to reduce nutrients and various contaminants of emerging concern from effluent. In the present study, we examined bioaccumulation of selected pharmaceuticals from several therapeutic classes by two important fish species in aquaculture with different feeding preferences (Cyprinus carpio and Sander lucioperca) and their common prey to test whether species specific accumulation occurs. Forty and nineteen from 66 selected pharmaceuticals and their metabolites were positively found in water and sediment samples, respectively from the representative WSP. After a six-month study, which corresponds to aquaculture operations, fourteen pharmaceuticals and their metabolites were detected (at a frequency of higher than 50% of samples) in at least one fish tissue collected from the WSP. We observed striking differences for species and organ specific BAFs among study compounds. Though muscle tissues consistently accumulated lower levels of the target analytes, several substances were elevated in brain, liver and kidney tissues (e.g., sertraline) of both species. Low residual concentrations of these target analytes in aquaculture products (fish fillets) suggest WSPs are promising to support the water-food nexus in aquaculture.

The aim of this study was to characterize biomarker responses, haematological profiles, structural changes and uptake in juvenile rainbow trout exposed to clotrimazole (CLO) at three concentrations (0.01 - [lowest environmentally relevant concentration], 1.0 [highest environmentally relevant concentration] and 10 μg L(-1)) in a semi-static system over a period of 42 days. Antioxidant defence enzymes, which responded to CLO exposure, changed the oxidative stress status of cells, but no differences were observed in lipid peroxidation. Clotrimazole triggered a biphasic response of CYP3A-like activity in liver microsomes, which may indicate a detoxification process in the liver. Histopathological alterations were most pronounced in kidneys and testes in the group exposed to 10 μg L(-1). Structural changes in the kidney included tubulonephrosis and hyaline droplet degeneration in the tubular epithelial cells. The relative proportions of germ cells in testes were changed: The number of spermatozoa was reduced, and the spermatogonia and spermatocytes were increased. The highest CLO concentration was detected in fish liver (3710 ng per gram wet tissue) and kidney (4280 ng per gram wet tissue). Depuration half-life was estimated to be 72, 159, and 682 h in liver, muscle, and kidney, respectively. Taken together, these results provide valuable toxicological data on the effects of CLO on aquatic non-target organisms, which could be useful for further understanding of the potential risks in the real aquatic environment.

RATIONALE Diltiazem, a calcium channel blocker drug, is widespread in the environment because of its incomplete elimination during water treatment. It can cause negative effects on aquatic organisms; thus, a rapid and sensitive liquid chromatography/mass spectrometry (LC/MS) method to detect its presence was developed. Our approach is based on accurate mass measurements using a hybrid quadrupole-orbital trap mass spectrometer that was used to measure diltiazem and its metabolites in fish tissue. METHODS Blood plasma, muscle, liver, and kidney tissues of rainbow trout (Oncorhynchus mykiss), exposed for 42 days to 30 μg L(-1) diltiazem, were used for the method development. No metabolite standards were required to identify the diltiazem biotransformation products in the fish tissue. RESULTS Overall, 17 phase I diltiazem metabolites (including isomeric forms) were detected and tentatively identified using the MassFrontier spectral interpretation software. A semi-quantitative approach was used for organ-dependent comparison of the metabolite concentrations. CONCLUSIONS These data increase our understanding about diltiazem and its metabolites in aquatic organisms, such as fish. These encompass desmethylation, desacetylation and hydroxylation as well as their combinations. This study represents the first report of the complex diltiazem phase I metabolic pathways in fish.

Stream ecosystems face ever-increasing pressures by the presence of emergent contaminants, such as, personal care products. N, N-diethyl-3-methylbenzamide (DEET) is a synthetic insect repellent that is being found in surface waters environments in concentrations up to 33.4 μg/L. Information concerning DEET's toxicity in the aquatic environment is still limited and focused only on its acute effects on model species. Our main objective was to assess the effects of DEET exposure to a caddisfly non-target species using sub-lethal endpoints. For that, we chose Sericostoma vittatum, an important shredder in Portuguese freshwaters that has been already used in different ecotoxicological assays. Besides acute tests, S. vittatum were exposed during 6 days to a gradient of DEET concentrations (8, 18 and 40.5 mg/L) to assess effects on feeding behaviour and biochemical responses, such as, lipid peroxidation levels (LPO), catalase and acetylcholinesterase (AChE) activities, and also assess effects on energy reserves and consumption. Acute tests revealed a 48 h-LC50 of 80.12 mg/L and DEET exposure caused feeding inhibition with a LOEC of 36.80 mg/L. Concerning the biochemical responses, DEET caused no effects in LPO nor on catalase activity. A non-significant decrease in AChE activity was observed. Regarding energetic reserves, exposure to DEET caused a significant reduction in S. vittatum carbohydrates levels. These results add important information for the risk assessment of insect repellents in the aquatic environment and suggest that reported environmental concentrations of DEET are not toxic to non-target freshwater insects.

The concentrations of seven perfluoroalkyl substances (PFASs) were investigated in 36 European chub (Squalius cephalus) individuals from six localities in the Czech Republic. Chub muscle and liver tissue were analysed at all sampling sites. In addition, analyses of 16 target PFASs were performed in Polar Organic Chemical Integrative Samplers (POCISs) deployed in the water at the same sampling sites. We evaluated the possibility of using passive samplers as a standardized method for monitoring PFAS contamination in aquatic environments and the mutual relationships between determined concentrations. Only perfluorooctane sulphonate was above the LOQ in fish muscle samples and 52% of the analysed fish individuals exceeded the Environmental Quality Standard for water biota. Fish muscle concentration is also particularly important for risk assessment of fish consumers. The comparison of fish tissue results with published data showed the similarity of the Czech results with those found in Germany and France. However, fish liver analysis and the passive sampling approach resulted in different fish exposure scenarios. The total concentration of PFASs in fish liver tissue was strongly correlated with POCIS data, but pollutant patterns differed between these two matrices. The differences could be attributed to the metabolic activity of the living organism. In addition to providing a different view regarding the real PFAS cocktail to which the fish are exposed, POCISs fulfil the Three Rs strategy (replacement, reduction, and refinement) in animal testing.

Methamphetamine, mainly consumed as an illicit drug, is a potent addictive psychostimulant that has been detected in surface water at concentrations ranging from nanograms to micrograms per litre, especially in Middle and East Europe. The aim of this study was to expose brown trout (Salmo trutta fario) to environmental (1 μg L−1) and higher (50 μg L−1) concentrations of methamphetamine for 35 days with a four-day depuration phase to assess the possible negative effects on fish health. Degenerative liver and heart alterations, similar to those described in mammals, were observed at both concentrations, although at different intensities. Apoptotic changes in hepatocytes, revealed by activated caspase-3, were found in exposed fish. The parent compound and a metabolite (amphetamine) were detected in fish tissues in both concentration groups, in the order of kidney > liver > brain > muscle > plasma. Bioconcentration factors ranged from 0.13 to 80. A therapeutic plasma concentration was reached for both compounds in the high-concentration treatment. This study indicates that chronic environmental concentrations of methamphetamine can lead to health issues in aquatic organisms.

Environmental monitoring and surveillance studies of pharmaceuticals routinely examine occurrence of substances without current information on human consumption patterns. We selected 10 streams with diverse annual flows and differentially influenced by population densities to examine surface water occurrence and fish accumulation of select psychoactive medicines, for which consumption is increasing in the Czech Republic. We then tested whether passive sampling can provide a useful surrogate for exposure to these substances through grab sampling, body burdens of young of year fish, and tissue specific accumulation of these psychoactive contaminants. We identified a statistically significant (p 0.05) relationship between ambient grab samples and passive samplers in these streams when psychoactive contaminants were commonly quantitated by targeted liquid chromatography with tandem mass spectrometry, though we did not observe relationships between passive samplers and tissue specific pharmaceutical accumulation. We further observed smaller lotic systems with elevated contamination when municipal effluent discharges from more highly populated cities contributed a greater extent of instream flows. These findings identify the importance of understanding age and species specific differences in fish uptake, internal disposition, metabolism and elimination of psychoactive drugs across surface water quality gradients.

Although pharmaceuticals are recognized as a major threat to aquatic ecosystems worldwide, little is known about their ecological effect on aquatic biota and ecosystems. Drug-induced behaviour changes could have a substantial impact on consumer-resource interactions influencing stability of the community and ecosystem. We combined laboratory experiments and functional response modelling to investigate effects of real wastewater treatment plant (WWTP) effluent, as well as environmentally relevant concentrations of the antidepressants citalopram and opioid pain medication tramadol, on trophic interactions. Our biological system consisted of dragonfly Aeshna cyanea larvae as predator of common carp Cyprinus carpio fry. Exposure to WWTP effluent significantly increased A. cyanea maximum feeding rate, while those parameters in tramadol and citalopram-exposed larvae were significantly lower from unexposed control group. This suggested the potential of all tested pollutants to have an effect on consumer-resource equilibrium in aquatic ecosystems. While WWTP effluent strengthened interaction strength (IS) of consumer-resource interaction dynamics making the food web more vulnerable to fluctuation and destabilization, tramadol and citalopram could inhibit the potential oscillations of the consumer-resource system by weakening the IS. Similar studies to reveal the potential of pervasive pharmaceuticals to change of consumer-resource interactions dynamics are needed, especially when real WWTP effluent consisting of mixture of various pharmaceuticals displayed very different effect from single compounds tested.

A simple, robust and effective extraction procedure for the determination of 74 pharmaceuticals in different fish tissues by ultrasensitive high performance liquid chromatography with electrospray high resolution product scan (HPLC-ESI-HRPS) was developed and validated. Different extraction solvent mixtures were tested to achieve the highest recoveries of the selected analytes, to minimize the influence of a complex matrix and to reduce the total analysis time as well as cost of analysis. A mixture of acetonitrile + isopropanol (3:1 v/v) acidified with 0.1% formic acid was the best extraction solvent among the five solvents tested for most of the tissues with the exception of plasma samples, where only acidified acetonitrile exhibited the best performance. The developed method was validated at three concentration levels (5, 20 and 50 ng g-1) in five different fish tissues (liver, kidney, brain, muscle and plasma). Most of the target analytes were extracted with a recovery between 60 and 130%. Very low limits of quantification (LOQs) were obtained for the majority of the pharmaceuticals in all of the studied matrices. The developed analytical method was successfully applied for the analysis of common carp (Cyprinus carpio) originating from the waste water effluent-dominated pond Cezarka (Czech Republic). The results confirmed the importance of multi-tissue analysis to obtain complex information on the distribution of pharmaceuticals in fish.

Environmental pollution by pharmaceutically active compounds, used in quantities similar to those of pesticides and other organic micropollutants, is increasingly recognized as a major threat to the aquatic environment. These compounds are only partly removed from wastewaters and, despite their low concentrations, directly and indirectly affect behaviour of freshwater organisms in natural habitats. The aim of this study was to behaviourally assess the effects of an opioid painkiller (tramadol) and antidepressant drug (citalopram) on behaviour patterns of a clonal model species, marbled crayfish. Animals exposed to environmentally relevant concentrations of both tested compounds (∼1 μg l−1) exhibited significantly lower velocity and shorter distance moved than controls. Crayfish exposed to tramadol spent more time in shelters. Results were obtained by a simple and rapid method recommended as suitable for assessment of behaviour in aquatic organisms exposed to single pollutants and combinations.

Aquatic organisms can be affected not only via polluted water but also via their food. In the present study, we examined bioaccumulation of seventy pharmaceuticals in two benthic organisms, Hydropsyche sp. and Erpobdella octoculata in a small stream affected by the effluent from a sewage treatment plant (STP) in Prachatice (South Bohemia region, Czech Republic). Furthermore, water samples from similar locations were analyzed for all seventy pharmaceuticals. In water samples from a control locality situated upstream of the STP, ten of the seventy pharmaceuticals were found with average total concentrations of 200 ng L−1. In water samples collected at STP-affected sites (downstream the STP′s effluent), twenty-nine, twenty-seven and twenty-nine pharmaceuticals were determined at average total concentrations of 2000, 2100 and 1700 ng L−1, respectively. Six of the seventy pharmaceuticals (azithromycin, citalopram, clarithromycin, clotrimazole, sertraline, and verapamil) were found in Hydropsyche. Four pharmaceuticals (clotrimazole, diclofenac, sertraline, and valsartan) were detected in Erpobdella. Using evaluation criterion bioconcentration factor (BCF) is higher than 2000 we can assign azithromycin and sertraline as bioaccumulative pharmaceuticals. Even pharmaceuticals present at low levels in water were found in benthic organisms at relatively high concentrations (up to 85 ng g−1 w.w. for azithromycin). Consequently, the uptake of pharmaceuticals via the food web could be an important exposure pathway for the wild fish population.

Juvenile wild and hatchery-reared European grayling Thymallus thymallus were tagged with radio-transmitters and tracked in the Blanice River, River Elbe catchment, Czech Republic, to study their behavioural response to stocking and environmental variation. Both wild and hatchery-reared T. thymallus increased their diel movements and home range with increasing light intensity, flow, temperature and turbidity, but the characteristics of their responses differed. Environmental variables influenced the movement of wild T. thymallus up to a specific threshold, whereas no such threshold was observed in hatchery-reared T. thymallus. Hatchery-reared fish displayed greater total migration distance over the study period (total migration) than did wild fish, which was caused mainly by their dispersal in the downstream direction.

The treated effluent from sewage treatment plants (STP) is a major source of active pharmaceutical ingredients (APIs) that enter the aquatic environment. Bioaccumulation of 11 selected psychoactive pharmaceuticals (citalopram, clomipramine, haloperidol, hydroxyzine, levomepromazine, mianserin, mirtazapine, paroxetine, sertraline, tramadol and venlafaxine) was examined in Zivny Stream (tributary of the Blanice River, the Czech Republic), which is a small stream highly affected by effluent from the Prachatice STP. Six of the 11 pharmaceuticals were detected in grab water samples and in passive samplers. All pharmaceuticals were found in fish exposed to the stream for a defined time. The organs with highest presence of the selected pharmaceuticals were the liver and kidney; whereas only one pharmaceutical (sertraline) was detected in the brain of exposed fish. Fish plasma and muscle samples were not adequate in revealing exposure because the number of hits was much lower than that in the liver or kidney. Using the criterion of a bioaccumulation factor (BAF) ≥ 500, citalopram, mianserin, mirtazapine and sertraline could be classified as potential bioaccumulative compounds. In combination, data from integrative passive samplers and fish liver or kidney tissue samples were complimentary in detection of target compounds and simultaneously helped to distinguish between bioconcentration and bioaccumulation.

Pharmaceuticals as environmental contaminants have received a lot of interest over the past decade but, for several pharmaceuticals, relatively little is known about their occurrence in European surface waters. Benzodiazepines, a class of pharmaceuticals with anxiolytic properties, have received interest due to their behavioral modifying effect on exposed biota. In this study, our results show the presence of one or more benzodiazepine(s) in 86% of the analyzed surface water samples (n = 138) from 30 rivers, representing seven larger European catchments. Of the 13 benzodiazepines included in the study, we detected 9, which together showed median and mean concentrations (of the results above limit of quantification) of 5.4 and 9.6 ng L−1, respectively. Four benzodiazepines (oxazepam, temazepam, clobazam, and bromazepam) were the most commonly detected. In particular, oxazepam had the highest frequency of detection (85%) and a maximum concentration of 61 ng L−1. Temazepam and clobazam were found in 26% (maximum concentration of 39 ng L−1) and 14% (maximum concentration of 11 ng L−1) of the samples analyzed, respectively. Finally, bromazepam was found only in Germany and in 16 out of total 138 samples (12%), with a maximum concentration of 320 ng L−1. This study clearly shows that benzodiazepines are common micro-contaminants of the largest European river systems at ng L−1 levels. Although these concentrations are more than a magnitude lower than those reported to have effective effects on exposed biota, environmental effects cannot be excluded considering the possibility of additive and sub-lethal effects.

Seasonal changes in the concentration of 21 pharmaceuticals in a wastewater treatment plant (WWTP) in Ceske Budějovice were investigated over 12 months. The target compounds were 10 antibiotics, 4 antidepressants, 3 psychiatric drugs, 2 antihistamines and 2 lipid regulators. 272 Wastewater samples (136 influents and 136 effluents) were collected from March 2011 to February 2012 and analyzed using two-dimensional liquid chromatography coupled with tandem mass spectrometry. All studied pharmaceuticals were frequently detected in both the influent and the effluent wastewater samples, except for meclozine, which was only found in the influent. The mean concentration of pharmaceuticals varied from 0.006 μg L −1 to 1.48 μg L −1 in the influent and from 0.003 μg L −1 to 0.93 μg L −1 in the effluent. The concentration of most pharmaceuticals was higher during winter.

Tissue-specific bioconcentration of selected antidepressants was studied in rainbow trout (Oncorhynchus mykiss) exposed to undiluted effluent from a Swedish municipal sewage treatment plant for 13 days. Citalopram, sertraline and venlafaxine were found in the brains and livers of most fish, but not in blood plasma or muscle. Venlafaxine was the only drug found in plasma (3/20 fish). Fluoxetine was not detected in any fish tissue, in accordance with a low concentration in the effluent and a comparably high limit of quantification in tissues. Concentrations of citalopram, sertraline and venlafaxine in fish brain were up to 1/12, 1/8 and 1/26, respectively, of the lowest concentrations found in the brains of mammals treated with therapeutic doses. Thus, given co-exposure to several antidepressants and an assumed similar potency in fish, the margin of safety for target-related effects in fish residing in effluent-dominated streams is relatively low. Furthermore, the non-detectable levels of these drugs in blood plasma suggest that analyses of concentrations in target tissues (brain) would be more informative in field studies and other studies with environmentally realistic exposure concentrations.

UV filters belong to a group of compounds that are used by humans and are present in municipal waste-waters, effluents from sewage treatment plants and surface waters. Current information regarding UV filters and their effects on fish is limited. In this study, the occurrence of three commonly used UV filters - 2-phenylbenzimidazole-5-sulfonic acid (PBSA), 2-hydroxy-4-methoxybenzophenone (benzophenone-3, BP-3) and 5-benzoyl-4-hydroxy-2-methoxy-benzenesulfonic acid (benzophenone-4, BP-4) - in South Bohemia (Czech Republic) surface waters is presented. PBSA concentrations (up to 13μgL(-1)) were significantly greater than BP-3 or BP-4 concentrations (up to 620 and 390ngL(-1), respectively). On the basis of these results, PBSA was selected for use in a toxicity test utilizing the common model organism rainbow trout (Oncorhynchus mykiss). Fish were exposed to three concentrations of PBSA (1, 10 and 1000µgL(-1)) for 21 and 42 days. The PBSA concentrations in the fish plasma, liver and kidneys were elevated after 21 and 42 days of exposure. PBSA increased activity of certain P450 cytochromes. Exposure to PBSA also changed various biochemical parameters and enzyme activities in the fish plasma. However, no pathological changes were obvious in the liver or gonads.

RATIONALE Analysis of drugs in wastewater is gaining more interest, as new approaches to estimate drug consumption from the amount of drug residues in wastewater have been proposed. The aim of this ...

An experiment on the effects of hormonal preparations (Gonazon™ and Supergestran containing gonadotropin-releasing hormone) on the course of reproduction of farmed grayling (Thymallus thymallus L.) broodstock and their post-stripping mortality was performed at the hatchery in spring 2009. Four-year-old marked grayling females (n = 80) were randomly divided into four groups. The fish were intramuscularly injected with Supergestran at a dose of 30 μg·kg-1 body weight (Group 1), with GonazonTM at a dose of 30 μg·kg-1 (Group 2) and with saline 0.9% NaCl (Group 3), while fish of Group 4 were left without treatment (control group). No effect of treatment on the total number of ovulated females (70–80%) was found at the end of the stripping period. Slightly higher (non-significant) percentage of ovulated fish in the first stripping time (3 days post injection) was observed in fish treated with Gonazon. The 30-days post-ovulatory mortality remained unaffected by hormone treatments in all groups. Significant differences (P < 0.001) were found in the fertilization rate of egg samples from the first stripping time. The highest fertilization rate (98.6%) was found in fish treated with Supergestran, lower fertilization rates (61% and 65%) in fish treated with saline and control, respectively, and the lowest fertilization rate (39%) in fish treated with Gonazon. Other reproductive indicators remained unaffected by treatment in all groups. Based on our results, Supergestran is the most suitable preparation for the enhancing of artificial reproduction efficiency of farmed European grayling in fishery practice. The study brings important information about artificial reproduction of grayling broodstocks reared in controlled conditions.

Muscle tissue and pectoral fins of two important indicator fish species, frequently used in biomonitoring programs, were sampled and analysed for total mercury content (THg) at six localities within the Czech Republic. The relationship between mercury concentration in muscle and in fin-clips was described. Mean values of THg fin-clip concentration correlate significantly (p

Toxic metals (Hg, Cd, Pb) and fifteen perfluoroalkyl substances (PFASs) were determined in different fish samples at two locations on the Elbe River in the Czech Republic. The muscle tissue of the two adult fish species most commonly used as bioindicators in central Europe and whole body homogenates of various species of young-of-the-year (YOY) fish were used. The purpose of this study was to evaluate the potential to replace adult fish muscle tissue with YOY fish for contamination monitoring. All of the toxic metals and five of the fifteen PFASs were found in the YOY fish samples while only mercury and PFOS were detected in the muscle tissue of adults. The concentration of total mercury (THg) in the YOY fish homogenates ranged between 0.014 and 0.062 μg g−1. Of the spectrum of analysed pollutants, only the THg concentrations were lower in YOY fish homogenates than in adult muscle tissue. The cadmium concentration varied from 0.004 to 0.024 μg g−1 and the lead concentration varied from 0.032 to 0.396 μg g−1 in YOY fish homogenates, while in most of the adult samples, Cd and Pb were below the detection limit of the analytical methods employed. The PFOS concentrations in YOY fish homogenates were comparable to the concentrations frequently found in adult liver tissue. These results show that mixed shoals of YOY fish can be successfully used for aquatic bio-monitoring. Interspecific variability in the concentrations of the target pollutants in YOY fish whole body homogenates is usually lower than the intraspecific variability of the concentrations of the pollutants in adult fish muscle. YOY fish were found to be a suitable bioindicator and have several advantages compared to adult fish.

Diltiazem is a pharmaceutical belonging to a group of calcium channel blockers (CCB) that is widely used in the treatment of angina pectoris and hypertension. The objective of the present study was to assess the effect of diltiazem on rainbow trout (Oncorhynchus mykiss). Juvenile trout were exposed for 21 and 42 days to three nominal concentrations of diltiazem: 0.03 μg L(-1) (environmentally relevant concentration), 3 μg L(-1), and 30 μg L(-1) (sub-lethal concentrations). The number of mature neutrophilic granulocytes was significantly increased by 450 and 400% in fish exposed to 3 μg L(-1) and 30 μg L(-1) diltiazem compared to the control, respectively. Antioxidant enzyme activity was affected in liver and gills of fish exposed to all tested concentrations of diltiazem but the changes were mostly transient and not concentration dependent. Creatine kinase activity was markedly increased (ranging from 520 to 845%) at all tested diltiazem concentrations at the end of the exposure indicating muscle and/or kidney damage. The highest concentration was associated with histological changes in heart, liver, and kidney. These alterations can be attributed to the effects of diltiazem on the cardiovascular system, similar to those observed in the human body, as well as to its metabolism. At the environmentally relevant concentration, diltiazem was found to induce some alterations in the blood, gills, and liver of fish, indicating its potential for adverse effects on non-target organisms in the aquatic environment.