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European grayling, Thymallus thymallus (Linnaeus, 1758) (Actinopterygii: Salmoniformes: Salmonidae), represents one of the highly attractive riverine fishing species in Europe. Its populations have declined in the Czech Republic due to various adverse factors. Current approaches for strengthening these populations based on restocking with artificially reared 1+ or 2+ old individuals have proven to be ineffective. This study focuses on the possibilities of supporting or restoring these populations by reintroducing two-month-old, fast-growing fry reared in ponds. In June 2021, 5400 advanced fry marked with Alizarin Red (ARS) were introduced into three free-flowing South Bohemian streams. The recapture rates and biometric data (length and weight) of stocked grayling were assessed at the release sites as well as further downstream. The first electrofishing monitoring was conducted at the end of the growing season (October) and after their first winter (March). Water temperature and flow rate at the stocking sites were monitored throughout the growing season. The substantial number of recaptured individuals across all monitored sites suggests that the fry successfully adapted to their new habitat and significantly contributed to the composition of local fish communities. Downstream movement of stocked fry correlated with lower water temperatures and higher flows. Our findings demonstrate that some introduced individuals successfully overwintered at all three reintroduction sites. Marking with ARS has proven to be a very effective non-invasive method of group marking juvenile fish and is suitable for monitoring stocking programs. The production and stocking of fast-growing advanced fry of European grayling is thus a promising strategy for revitalizing and strengthening the populations of this threatened fish in running waters.

Fry of rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), was subjected to one-hour and four hours immersion in Alizarin red S (ARS) bath 150 mg · L–1. The experiment involved seven age groups (40, 50, 60, 70, 80, 90, and 100 days) and was conducted to estimate the minimal age of salmonids for mass marking with ARS bath enabling subsequent effective, simple field nonlethal identification, based on fin rays checking. The fish were examined at the ages of 200 and 300 days. The results showed a high retention level of ARS traces in caudal fin rays ensured satisfactory visibility and quick detection. A success rate of marking detection was >90% at 200 and 300 days of age and the fish were immersed in ARS solution from 60 days of age (685°D). This treatment provided better results in fish bathed for four hours. Recognition of marks using a laser pointer and protective glasses was successful even in fish, with a 3.5–5.0 times length increase compared to the marking time. The results indicate a high potential for ARS marking and its field identification for juvenile salmonids, which can significantly expand the possibilities of field experiments.

Pharmaceutically active compounds are common and increasing in the aquatic environment. Evidence suggests they have adverse effects on non-target organisms, and they are classified as emerging pollutants for a variety of aquatic organisms. To determine the effects of environmentally relevant levels of psychoactive compounds on non-target organisms, we analyzed cardiac and locomotory activity in early developmental stages of marbled crayfish Procambarus virginalis. Responses to sertraline, methamphetamine, and a mixture of citalopram, oxazepam, sertraline, tramadol, venlafaxine, and methamphetamine at a concentration of 1 µg L-1 of each compound were assessed. On day four of exposure, cardiac activity was recorded for 5 min, and on day eight, locomotory activity was recorded for 15 min. There was a significant increase (p 0.05) in exposed and control animals. These findings revealed that low concentrations of chemicals and their mixtures can modify the physiological state of aquatic animals without outward manifestations (activity, distance moved, and velocity). Aquatic animals can be impacted earlier than is visible, but effects can potentially lead to substantial changes in populations and in ecosystem processes. Additional research to investigate chemical combinations, exposure systems, and organism physiological and molecular responses may provide evidence of broad impact of environmental pharmaceuticals.

The response of parasite communities to aquatic contamination has been shown to vary with both type of pollutant and parasite lifestyle. In this semi-experimental study, we examined uptake of pharmaceutical compounds in common carp (Cyprinus carpio L.) restocked from a control pond to a treatment pond fed with organic pollution from a sewage treatment plant and assessed changes in parasite community composition and fish biometric parameters. The parasite community of restocked fish changed over the six-month exposure period, and the composition of pharmaceutical compounds in the liver and brain was almost the same as that in fish living in the treatment pond their whole life. While fish size and weight were significantly higher in both treatment groups compared to the control, condition indices, including condition factor, hepatosomatic index, and splenosomatic index, were significantly higher in control fish. Parasite diversity and species richness decreased at the polluted site, alongside a significant increase in the abundance of a single parasite species, Gyrodactylus sprostonae. Oviparous monogeneans of the Dactylogyridae and Diplozoidae families and parasitic crustaceans responded to pollution with a significant decrease in abundance, the reduction in numbers most likely related to the sensitivity of their free-living stages to pollution.

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.

Background The 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. Methods The 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. Results The 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.

The aim of the present study was to investigate the concentrations of total mercury and methylmercury in muscle of 55 chub (Leuciscus cephalus) from seven main tributaries (Orlice, Chrudimka, Cidlina, Jizera, Vltava, Ohře, Bílina) of the Elbe River and to evaluate the health risks of eating fish from the tributaries monitored. Mercury was determined by means of cold vapour atomic absorption spectrometry using AMA-254, methylmercury in the form of CH3HgCl by gas chromatography. The highest mean concentrations of total mercury and methylmercury were found in the Jizera (0.27 ± 0.19 mg/kg and 0.23 ± 0.15 mg/kg, respectively) and the lowest mean concentrations of total mercury and methylmercury were found in the chub from the Cidlina (0.07 ± 0.05 mg/kg and 0.06 ± 0.04 mg/kg, respectively). The average methylmercury-to-total mercury ratio was 83 ± 15%. The fish intake hazard indexes calculated for the individual tributaries monitored were between 0.01 and 0.03. The results of this study show that the Elbe River is not significantly affected by mercury contamination from its main tributaries

Polar organic compound integrative samplers (POCIS) in combination with instrumental techniques such as LC-MS–MS were previously used to monitor environmental pollutants but the performance of alternative immunochemical methods such as ELISA (enzyme-linked immunosorbent assay) has been explored less. In the present study, POCIS technology was applied to surface water sampling in the Czech Republic, and ELISA was used as a detection technique for the herbicide atrazine. In the first study, 28 samples from streams around small municipal waste water treatment plants (WWTPs) were collected using two different devices (POCISpest and POCISpharm) over the course of 21 days. Elevated atrazine concentrations (up to 25 ng per POCIS) were found in samples down-stream of WWTPs. This observation was also confirmed in another two year study (4 sampling periods) investigating 7 river sites around a major city of Brno as well as the inlet and outlet of the city’s WWTP. High atrazine levels were systematically determined at the outlet from the WWTPs (120–605 ng per POCIS). A decreasing trend in the atrazine concentrations in rivers around the city of Brno has been observed, with the highest levels observed within the first sampling period in spring 2007 (100–600 ng per POCIS, with an extreme value of 2760 ng per POCIS). Results of the atrazine ELISA were closely correlated with LC-MS/MS, which confirmed good applicability of ELISA as a cost-effective screening tool. [ABSTRACT FROM AUTHOR]