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Freshwater protected areas are designated parts of the inland waters that restrict human activities. They were created as a mechanism to combat the decline of fauna and flora of the world. Some authors have questioned their actual effectiveness in terms of the purpose of protecting endangered fauna and flora. We conducted an experiment in Lipno reservoir in the Czech Republic to evaluate the impact of protection against angling pressure on the fish community. We selected data from two years of gill netting and analyzed the difference between areas of low anthropogenic impact (LAI) and those of high anthropogenic impact (HAI) in terms of abundance, biomass, standard length, and diversity indices. Three groups of fish were found to prefer protected areas with low anthropogenic pressure: 1. YOY (Young-of-the-year) perch (Perca fluviatilis), the dominant of the young-of-the-year fish community. 2. Pike (Esox lucius), wels catfish (Silurus glanis) and rudd (Scardinius erythrophthalmus), which were not found in HAI areas at all. 3. Larger individuals of pikeperch (Stizostedion lucioperca), which survived better in LAI areas. Some factors may affect LAI, such as illegal poaching or setting out food bait to attract the fish outside. Another factor that can be considered is the migration of fish, either to forage or to reproduce, since the LAI areas are open to the reservoir. The areas of LAI act as protective habitats for heavily exploited predatory fish species and increase fish diversity indexes. The example of the protected and low-impact areas of Lipno should be followed in other water bodies with high fishing pressure and anthropogenic impact.

Reservoirs interrupt natural riverine continuity, reduce the overall diversity of the environment, and enhance the spread of non-native fish species through suitable environments. Under favourable conditions, invasive species migrate to tributaries to benefit from local resource supplies. However, the changes in physical conditions in reservoirs that motivate fish species to migrate remain poorly understood. We analysed migration between a reservoir and its tributary in three non-native (asp Leuciscus aspius, ide Leuciscus idus, and bream Abramis brama) and two native (chub Squalius cephalus and pike Esox lucius) species equipped with radio tags. This 5-year study revealed that an increasing day length was the most general predictor of migration into the tributary in all observed species except E. lucius. Only L. aspius responded to the substantially increasing water level in the reservoir, while the migration of L. idus and S. cephalus was attenuated. Abramis brama and S. cephalus occurred more frequently in tributaries with an increase in temperature in the reservoir and vice versa, but if the difference in temperature between the reservoir and its tributary was small, then A. brama did not migrate. Our results showed that migration from the reservoir mainly followed the alterations of daylight, while responses to other parameters were species specific. The interindividual heterogeneity within the species was significant and was not caused by differences in length or sex. Our results contribute to the knowledge of how reservoirs can affect the spread of non-native species that adapt to rapid human-induced environmental changes.

Chemical exposure during the early life stages of development may have long lasting effects on organisms that are rarely studied. The present work intended to evaluate the effect of embryonic exposure to the pesticide carbaryl on adult fish behavior. Zebrafish ( Danio rerio ) embryos were exposed, for 4 days, to sublethal concentrations of carbaryl (0.01, 0.1 and 1.0 mg/L) plus a control and then kept in standard cultivation conditions until adulthood. A battery of behavioral tests was then performed to assess anxiety-like behavior (locomotor activity, thigmotaxis and novel tank diving test), social behavior, and feeding. Developmental exposure of zebrafish to sublethal concentrations of carbaryl produced important behavioral alterations in the adulthood. Main effects included decreased locomotion/hypoactivity (increase in slow movements and decrease of medium and rapid movements), especially in the light periods. Moreover, spatial pattern also changed: while during dark periods control fish increased activity in the outer zone of the tank, this was not observed in exposed fish. Overall, this demonstrated the importance of life stage exposure, clearly demonstrating long lasting effects of a (chemical) stress event at embryonic stages. This data supports the need of considering this scenario in environmental risk evaluations. Further work should focus on the mechanistic effects of developmental disruption responsible for the effects observed.

Treated wastewater ponds (TWPs) serve as recipients and passive tertiary treatment mediators for recycled water. These nutrient-rich habitats are increasingly utilised in aquaculture, nevertheless multiple loads of various contaminants with adverse effects on aquatic fauna, including fish, have been recorded. In the present study, we investigated the effects of fish transfer in response to altered levels of pollution on liver metabolic profiles and tissue-specific oxidative stress biomarkers during short- and long-term exposure. In a field experiment, common carp (Cyprinus carpio) originating in severely polluted TWP were restocked after one year to a reference pond with a background pollutant concentration typical of the regional river. In contrast, fish that originated in the reference pond were restocked to TWP. Fish were sampled 0, 7, 14, 60, and 180 days after restocking and fish liver, kidney, intestine, and gill tissues were subjected to biomarker analysis. Pharmaceutically active compounds (PhACs) and metabolic profiles were determined in fish liver using liquid chromatography high-resolution mass spectrometry (LC-HRMS). Fish transferred from reference to polluted pond increased the antioxidant response and absorbed PhACs into metabolism within seven days. Fish liver metabolic profiles were shifted rapidly, but after 180 days to a lesser extent than profiles in fish already adapted in polluted water. Restocked fish from polluted to reference pond eliminated PhACs during the short phase within 14 days, and the highest antioxidant response accompanied the depuration process. Numerous elevated metabolic compounds persisted in such exposed fish for at least 60 days. The period of two weeks was suggested as sufficient for PhACs depuration, but more than two months after restocking is needed for fish to stabilise their metabolism. This study contributed to determining the safe handling with marketed fish commonly restocked to wastewaters and clarified that water pollution irreversibly altered fish metabolic profile.

Despite the frequent detection of organic ultraviolet-filters (UV-filters) in freshwater sediments, there is a lack of ecotoxicological data undermining a correct risk assessment for these emerging contaminants. The present study assessed the effects of three of the most commonly used UV-filters (benzophenone-3 - BP3; 3-(4-methylbenzylidene)camphor - 4-MBC and octocrylene - OC) on Chironomus riparius life history and biochemical responses. Standard ecotoxicological assays confirmed that all compounds impaired growth of C. riparius larvae and induced developmental effects such as delayed emergence and a reduction of imagoes weight. Concerning the biochemical responses analysed no evidences of oxidative damage in lipids or neurotoxicity (tested assessing acetylcholinesterase activity) were observed for any of the tested compounds. However, 4-MBC exposure induced a decrease in catalase activity and an increase in glutathione-S-transferase activity at 14.13mg/Kg while OC exposure caused an increase in total glutathione levels at 0.23 and 18.23mg/Kg. Exposure to all UV-filters tested, increased energy consumption on C. riparius with significant differences above 1.00mg/Kg for BP3, 0.09mg/Kg for 4-MBC and 2.13mg/Kg for OC. These results suggest that environmental relevant concentrations of UV-filters can cause deleterious effects to aquatic benthic species, such as C. riparius, and call for further research concerning effects of organic UV-filters on natural invertebrate communities and ecosystem functioning.

The insecticide thiamethoxam (TMX) is a systemic neonicotinoid widely used for pest control in several agricultural crops. TMX mimics the action of acetylcholine causing uncontrolled muscular contraction eventually leading to insect death. TMX is being found in freshwater ecosystems at concentrations of up to 225µg/L. Still, chronic toxicity data for freshwater invertebrates is limited. Therefore, the aim of this study was to evaluate the acute and chronic effects (at organismal and biochemical levels) of TMX on the freshwater insect Chironomus riparius. C. riparius life history responses were significantly affected by TMX exposure, namely with a decrease in growth and delay in emergence. Concerning the biochemical responses, after a short exposure (48h) to TMX, our results showed that low concentrations of TMX significantly reduced CAT activity and LPO levels of C. riparius. No effects were observed in AChE, GST and ETS activities. Effects in terms of survival, development rates and biochemical responses of C. riparius exposed to low concentrations of TMX observed in this study suggest potential deleterious effects of this neonicotinoid on aquatic insects inhabiting freshwaters environments near agricultural areas.

Recent advances in video tracking technologies provide the tools for a sensitive and reproducible analysis of invertebrate activity under stressful conditions nurturing the field of behavioural ecotoxicology. This study aimed to evaluate behavioural responses of the freshwater planarian Dugesia subtentaculata exposed to a model compound, chlorantraniliprole (CAP). This compound is an anthranilic diamide insecticide and due to its neurotoxic action can, at low concentrations, impair behaviour of exposed organisms. Behavioural endpoints measured included feeding and locomotor activities. Feeding responses were based on planarian predatory behaviour using Chironomus riparius larvae as prey. Locomotion was measured by the traditional planarian locomotor velocity (pLMV) assay and additionally using an automated video tracking system using a Zebrabox(®) (Viewpoint, France) device. While feeding and pLMV were significantly impaired at 131.7μg/L CAP, the video tracking system showed that total distance covered by planarians was significantly reduced at concentrations as low as 26.2μg/L CAP. Our results show that more advanced automated video recording systems can be used in the development of sensitive bioassays allowing a reliable, time- and cost-effective quantification of behaviour in aquatic invertebrates. Due to their ecological relevance, behavioural responses should not be disregarded in risk assessment strategies and we advocate the suitability of planarians as suitable organisms for behavioural ecotoxicological studies.

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.

Erythromycin (ERY) is one of the most common antibiotics used in human and veterinary practices, leading to ubiquitous environmental distribution and possible toxicity to non-target organisms. The purpose of this study was to determine sub-lethal effects of ERY towards the marine fish Sparus aurata (gilthead seabream). S. aurata were acutely (0.3–323 μg/L, 96 h) and chronically (0.7–8.8 μg/L, 28 d) exposed to ERY. Detoxification [7-ethoxyresorufin O-deethylase (EROD), glutathione S-transferases (GSTs), uridine-diphosphate-glucuronosyltransferase (UGT)], oxidative stress [catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GRed)], lipid peroxidation [thiobarbituric acid reactive substances - (TBARS)], genotoxicity [genetic damage index (GDI) and erythrocytic nuclear abnormalities (ENAs)], neurotransmission [acetylcholinesterase (AChE)] and energy metabolism [lactate dehydrogenase (LDH)] biomarkers were evaluated. Results showed that ERY did not promote significant effects in detoxification biomarkers, but induced slight pro-oxidative effects (decrease of GPx activity in the liver after acute exposure and an increase in gills after chronic exposure; and an increase of hepatic GRed activity following chronic exposure). There was a significant decrease in TBARS after chronic exposure, which contradicts a full scenario of oxidative stress. In terms of genotoxicity, both ERY exposures caused only a significant increase of GDI. Neurotransmission and energy metabolism were not also affected by ERY. Although few toxic effects of ERY have been previously documented (involving different metabolic pathways, as tested in this work), these were mainly observed for freshwater species. These findings suggest low vulnerability of S. aurata to ERY at levels close to the ones found in the wild.

The post-release survival and condition of 1+ year old European grayling Thymallus thymallus reared in a local (Husinec) and two 250–300 km distant, hatcheries (Pardubice and Hynčice) were evaluated by the recapture of tagged fish five months after release into the Blanice River, Czech Republic, the fish population of which was depleted by cormorant predation during several previous winters. The fish were marked using Visible Implant Elastomer tags and released into six sites along the river in May 2014: 100 locally sourced fish and 100 of a strain from a distant source per site. Significantly higher recapture of Husinec (33%) than Pardubice (20%) was found at one site but was offset by results in the remaining two sites into which they were stocked. Significantly higher site fidelity was shown by Husinec (12%) than Hynčice (7%) fish in the three sites. Lower initial weight and condition factor of the Husinec fish was equal to or higher than fish from the distant hatcheries at recapture. Some differences in final weight, length, and condition factor were found among groups regardless of release site. Lower recapture rate, growth, and condition were displayed in fish released into the three downstream sites (Husinec 1 and Hynčice). The results indicated the potential for successful use of non-autochthonous stock for grayling population recovery.

In this study, selected biochemical markers - cytochrome P450 (CYP 450), ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST) and glutathione (GSH) - were measured in chub (Leuciscus cephalus L.) liver samples obtained from three locations on the Vltava river (Podolí, Podbaba and Vraňany) and from a control location nmicals commonly known as their inducers. These include polychlorinated biphenyls, hexachlorobenzene and octachlorostyrene measured in chub muscle and polyaromatic hydrocarbons measured in bear Vodňany. The levels of selected biomarkers should correlate with the concentration of cheottom sediments obtained from the same locations. The highest EROD activity (median 101.37 pmol min-1 mg-1 protein), GST activity (median 42.82 nmol min-1 mg-1 protein), and GSH concentration (median 8.01 nmol mg-1 protein) were found in fish liver from the Podbaba location. There were no significant differences in CYP P450 level or EROD activity among the different locations. In Podbaba, GST activity (P < 0.01) and GSH concentrations (P < 0.001) were significantly higher than in the control location. The results of these analyses were correlated and a significant correlation was found between biochemical markers and their inducers. The results show that the use of biochemical markers in water quality assessment is a convenient method that can supplement classical chemical monitoring.

Celechovska O., Z. svobodova, v. Žlabek, b. Macharackova: distribution of Metals in Tissues of common carp (Cyprinus carpio l.). acta vet. brno 2007, 76: s93-s100. The aim of the present study was to investigate the distribution of selected metals (as, cd, Pb, hg, cr, cu and Zn) in tissues of the common carp (Cyprinus carpio l.) grown in ponds. a total of 125 market-ready carp (aged 3 4 years, weight 2312 ± 583 g) from 10 fishponds in the czech republic were analysed. The tissues analysed included muscle, liver, kidney, spleen, gills, ovaria and testes. Metal concentrations were determined by the aas, mercury was determined using a cold-vapour aas analyse (aMa 254). results showed different affinities of metals to different tissues. The highest concentrations of as and hg were in muscle tissue, cu in the liver, and cd and Zn in the kidneys, compared with levels in other tissues (p < 0.01). The lowest metal concentrations (particularly of cd) were found in the spleen, gills and gonads. lead concentrations in the individual tissues were low and practically identical. chromium concentrations were also balanced with sporadic significant differences (p < 0.05). significantly higher concentrations of cu, cr (p < 0.05) and Zn (p < 0.01) were found in the ovaria compared with the testes. The study showed that meat and gonads of pond carp in the czech republic are safe from the point of view of contamination with the metals investigated. Muscle, parenchymatous tissues, gills, gonads, arsenic, cadmium, lead, mercury, chromium, copper, zinc an increase in metal concentrations in the environment is due primarily to erosion and anthropogenic activities, and because metals are very persistent pollutants, they get accumulated in the soil, water sediments and, last but not least, in the food chain (radike et al. 2002; reinecke et al. 2003; cornelis et al. 2005; swaileh and sansur 2006). recurrent cycles are the raison d’etre for conducting contamination monitoring in both the environment and the links of food chains in order to guarantee food quality and safety (svobodova et al. 2002ab, 2004; andreji et al. 2006a). One of the most closely monitored area is the aquatic ecosystem, and fish as the final link of the food chain in particular (Yilmaz 2006; havelkova et al. 2007). Great attention is paid to hazardous elements such as mercury, lead, cadmium and arsenic (alam et al. 2002; Maffuci et al. 2005). a large majority of metals have a great affinity to bind with amino acids and sh groups of proteins, and therefore act as enzyme inhibitors. an important negative characteristic of metals is their ability to accumulate in organs, especially in the liver, spleen, kidneys and gonads (spurný et al. 2002; Yilmaz 2006; andreji et al. 2006b). To better understand not only physiological effects but also toxicological and hygienic effects of metals, it is necessary to know the distribution patterns of individual metals in the organism. The distribution of metals in tissues of different species of fish has been described by, e.g., staniskiene et al. (2006) (perch, roach, silver bream, chub, smelt, tench and pike) and spurný et al. 2002 (chub). The aim of the present study was to investigate the distribution of selected metals in tissues of the common carp (Cyprinus carpio l.) grown in ponds. like in many other european ACTA VET. BRNO 2007, 76: S93–S100; doi:10.2754/avb200776S8S093 Address for correspondence: doc. Ing. Olga Celechovska, Ph.d. department of biochemistry, chemistry and biophysics Faculty of veterinary hygiene and ecology University of veterinary and Pharmaceutical sciences Palackeho 1-3, 612 42 brno, czech republic Phone +420 541 562 606 e-mail: celechovskao@vfu.cz http://www.vfu.cz/acta-vet/actavet.htm countries, carp is the principal commercially exploited fish in the czech republic. carp production makes up about 87 90% of the total fish production in the czech republic (vacha 1998). Materials and Methods between 2000 and 2006, we examined 125 market-ready carp from 10 uncontaminated fishponds in the czech republic, i.e. bezdrev (17 carp), velký bědný (7 carp), dřemliny (17 carp), horusický (17 carp), Nezmar (7 carp), regent (17 carp), rožmberk (16 carp), spolský (10 carp), Tovarys (17 carp) and vrkoc starý (10 carp)

Progeny from artificial propagation of wild brown trout (Salmo trutta L.) of the Blanice river, Czech Republic, were farmed to maturity and spawned at ages three, four and five during 2002-2004. Reproductive parameters and biological quality of eggs in this farmed population were compared to those of the original wild population. ANCOVA showed no differences between wild and farmed fish in weight of eggs per female, total fecundity, or relative fecundity in any year. Significantly higher egg diameter (4.57 mm, P = 0.001) and weight (69.3 mg, P = 0.0375) were found in the wild population in 2002 and conversely in 2004, the mean egg weight was higher in the farmed population (94.7 mg, P = 0.0021). Differences in egg diameter in this year (4.64 ± 0.06 and 4.82 ± 0.06 in wild and farmed trout, respectively) were close to the level of significance (P = 0.079). Mutual correlations between length or weight and studied reproductive traits were similar in both populations. Fertilization rate, duration of incubation period, egg losses during incubation and mortality of starving hatched fry were monitored in embryos and larvae of farmed population only (FxF), crosses between farmed females and wild males (FxW) and wild population only (WxW). Altogether 6.3%, 5.8% and 5.4% of eggs died during incubation period in FxF, FxW and WxW, respectively. There were also no significant differences in duration of incubation period and mortality of starving fry. It can be concluded that farming conditions did not significantly affect the reproductive parameters and quality of eggs in the first generation of farmed broodstock.

Svobodova Z., V. Ilabek, T., Randak, J. Machova, J. Kolafiova, J. Haj‰lova, P. Suchan, L. Du‰ek, J. Jarkovsk˘ : Profiles of PCBs in Tissues of Marketable Common Carp and in Bottom Sediments of Selected Ponds in South and West Bohemia. Acta Vet. Brno 2004, 73: 133-142. The aim of the study was to evaluate the extent of contamination of six selected ponds in South and West Bohemia by PCBs. For this purpose seven indicator congeners were determined in the tissues of common carp and bottom sediments. The ponds were sampled during autumn periods in the years 1999-2001. Five to seven specimens of three-to four-year-old carp were sampled from every pond. The results demostrated comparable indicative ability of fish tissues and bottom sediments for evaluation of pond load by the monitored PCBs. Considering concentrations of PCBs in fish and bottom sediments, the most contaminated pond was Tovary‰ followed by Buzick˘, Dfiemliny, Regent, Bezdrev and Horusick˘. As regards the distribution of PCBs in fish, the highest values of indicator congeners were found in liver followed by ovaries, muscle and testes. A significant correlation was found between the PCB levels (sum of 7 indicator congeners) and the content of fat in individual tissues of carp (r = 0.83 ‐ 0.94). The pattern of individual indicator congeners in fish was almost the same as that in bottom sediments. The correlation was statistically significant (r = 0.93). This observation is corresponding with slow biotransformation rate of PCBs in fish. The study contributed to the assessment of quality of carp (which is the main representative of marketable fish in the Czech Republic) in terms of its safety as food as well as to evaluation of the pollution of respective aquatic environment.

Biochemical markers, specifically enzymes of the first phase of xenobiotic transformation - cytochrome P450 and ethoxyresorufin-O-deethylase (EROD) - were used to determine the quantities of persistent organic pollutants (POPs) in fish muscle (PCB, HCB, HCH, OCS, DDT). Eight rivers were monitored (Orlice, Chrudimka, Cidlina, Jizera, Vltava, Ohře and Bílina; and the River Blanice was used as a control). The indicator species selected was the chub (Leuciscus cephalus L.). There were no significant differences in cytochrome P450 content between the locations monitored. The highest concentration of cytochrome P450 in fish liver was in the Vltava (0.241 nmol mg-1 protein), and the lowest was in the Orlice (0.120 nmol mg-1 protein). Analysis of EROD activity showed a significant difference between the Blanice and the Vltava (P< 0.05), and also between the Orlice and the Vltava (P< 0.01), the Orlice and the Bílina (P< 0.01), and the Orlice and the Ohře (P< 0.05). The highest EROD activity in fish liver was in the Vltava (576.4 pmol min-1 mg-1 protein), and the lowest was in the Orlice (63.05 pmol min-1 mg-1 protein). In individual locations, results of chemical monitoring and values of biochemical markers were compared. A significant correlation (P< 0.05) was found between biochemical markers and OCS, and PCB. Among the tributaries studied those that contaminated the Elbe most were the Vltava and the Bílina. These tributaries should not be considered the main sources of industrial contamination of the River Elbe, because the most important contamination sources were along the river Elbe itself.

The relative performance of European grayling Thymallus thymallus reared in a hatchery on commercial dry feed or in a pond with natural food and their wild conspecifics, was assessed through recapture of tagged fish 5 months after their release into the Blanice River, Czech Republic. One-year old pond and hatchery reared fish from a resident broodstock were marked using Visible Implant Elastomer tags and released into 3 sections of river in May 2006. Wild one-year-old grayling were also tagged in these sections on the same days. The ratio of hatchery reared, pond reared, and wild fish was 1:1:1 in all sections. The recapture rate (hatchery 14.9%, pond 22.1%, and wild 51.3%) and site fidelity (hatchery 7.8%, pond 13.0%, and wild 35.1%) were significantly different among groups. Wild fish had a higher probability of recapture upstream of their original section than did hatchery or pond reared fish. Pond rearing was superior to conventional hatchery rearing for subsequent stocking of 1 + grayling in running water. Initially different mean condition factors were similar in all groups at recapture, suggesting adaptation of the artificially reared fish that remained in the river sections surveyed.