Comparison of two methods of measuring the urinary protein concentration for the determination of the urinary protein to creatinine ratio in various animal species.

Determination of the urinary protein-to-creatinine ratio (UPC) is an important tool in the quantification of proteinuria in animals. However, the result may be affected by the different methods of determining the urinary protein concentration. The aim of this study was to compare the turbidimetric method using benzethonium chloride and the colorimetric method using pyrogallol red in the measurement of the urinary protein concentration in dogs, cats, guinea pigs and horses. A total of 464, 192, 216 and 119 urine samples from dogs, cats, guinea pigs and horses were examined in the study, respectively. The group consisted of animals of both sexes and different ages, and, in the dogs and cats, it included both healthy animals and those with various health problems. In the group of horses and guinea pigs, only clinically healthy animals were included. A total of 347, 185, 103 and 100 samples from the dogs, cats, guinea pigs and horses were used in the statistical analysis; the other values were excluded as they were below the detection limit. According to the Passing-Bablok analysis, there was a significant constant and proportional difference in the horses. In the dogs, cats and guinea pigs, there was a significant constant difference, but no proportional difference. The Bland-Altman method showed significant bias between the two methods in the horses and cats, but not in the dogs and guinea pigs. In the dogs and cats, the agreement between the two methods was tested and expressed as Cohen's kappa (κ). In the cats, it was almost perfect for the proteinuric samples (κ = 0.823 3) and significant for the non-proteinuric samples (κ = 0.804 9). In the dogs, the agreement was significant for the non-proteinuric samples (κ = 0.621 5) and only moderate for the proteinuric samples (κ = 0.527 5). The influence of the method used to determine the urinary protein concentration should be taken into account when evaluating the UPC. Repeated examinations in one patient should be performed with the same method.

Pharmacokinetics of praziquantel and its metabolites in grass carp (Ctenopharyngodon idella) following the oral administration of a single bolus.

The study aimed to evaluate and compare the routes and rates of the depletion of the antiparasitic praziquantel (PZQ), a derivative of pyrazinoisoquinoline, following its oral administration in grass carp (Ctenopharyngodon idella). We focused on the depletion of PZQ and its major metabolites - cis-hydroxy praziquantel (CPZQ) and trans-hydroxy praziquantel (TPZQ), in water, the plasma, hepatopancreas, kidney, muscle, and skin, following a single oral administration of PZQ in a concentration of 50 mg/kg. Fish were sampled before the drug administration and then eight times in the course of the 30-day-long experiment. Our results indicate the rapid absorption and elimination of PZQ and its metabolites in all the analysed matrices. The most PZQ-burdened tissue was the hepatopancreas, the gill and the skin. In all the samples, the concentration of the drug and its metabolites consistently declined over time. The residue of the parent compound was detected for the longest time in all the tissues. During the study, a significant (P < 0.01) correlation was found within the concentration of PZQ, CPZQ, and TPZQ in the water and all the biological matrices. It was also found that the PZQ residue was not detected below the maximum residue levels (i.e., 20 µg/kg) until 16 days after exposure in the muscle and skin.

Long-term exposure to polycyclic musk tonalide - A potential threat to juvenile zebrafish (Danio rerio)?

Polycyclic musk compounds are commonly used in personal care products to replace expensive natural fragrances. Due to their huge consumption, they have become a part of the aquatic environment. In the present study, a two-month exposure effect of tonalide on juvenile zebrafish (Danio rerio) was investigated. We determined the vitellogenin concentration to define the potential endocrine-disrupting effect of tonalide and also analysed selected indices to evaluate the induction of oxidative stress. The environmentally relevant concentration of tonalide (i.e., 500 ng/l) caused a significant decrease in the catalase activity (P < 0.05) and a significant increase (P < 0.05) in the lipid peroxidation. An increasing lipid peroxidation was also recorded for the highest concentration group tested (i.e., 50 000 ng/l). On the other hand, no significant changes were recorded in vitellogenin in all the exposed groups. Thus, based on these results, we have not demonstrated the endocrine-disrupting activity of tonalide in zebrafish. The results of the oxidative stress indices showed a significant impairment of the antioxidant defence after the two-month tonalide exposure, which could indicate part of the adaptive response to the tonalide toxicity.

The effect of sulfamethoxazole on oxidative stress indices in zebrafish (Danio rerio).

The aim of this study was to assess the impact of sulfamethoxazole (SMX) on oxidative stress indices in zebrafish (Danio rerio). The test was completed after 14 days. The tested concentrations were 50, 100 and 500 µg/L of SMX. Glutathione peroxidase, glutathione reductase, glutathione S-transferase and lipid peroxidation were investigated to determine the effects of SMX on oxidative stress in zebrafish. Lipid peroxidation gradually increased slightly (but non-significantly) at all tested concentrations during the test as compared to the control. The evaluation of oxidative stress biomarkers showed no significant changes in the activity of antioxidant enzymes in any experimental group exposed to SMX as compared to the control. The gradual increase in lipid peroxidation after 3 and 14 days in the SMX treated groups as compared to the control group indicates increasing cell membrane damage.

Functional rate-code models of the auditory brainstem for predicting lateralization and discrimination data of human binaural perception.

This paper presents a rate-code model of binaural interaction inspired by recent neurophysiological findings. The model consists of a peripheral part and a binaural part. The binaural part is composed of models of the medial superior olive (MSO) and the lateral superior olive (LSO), which are parts of the auditory brainstem. The MSO and LSO model outputs are preprocessed in the interaural time difference (ITD) and interaural level difference (ILD) central stages, respectively, which give absolute values of the predicted lateralization at their outputs, allowing a direct comparison with psychophysical data. The predictions obtained with the MSO and LSO models are compared with subjective data on the lateralization of pure tones and narrowband noises, discrimination of the ITD and ILD, and discrimination of the phase warp. The lateralization and discrimination experiments show good agreement with the subjective data. In the case of the phase-warp experiment, the models agree qualitatively with the subjective data. The results demonstrate that rate-code models of MSO and LSO can be used to explain psychophysical data considering lateralization and discrimination based on binaural cues.

Ergodicity and parameter estimates in auditory neural circuits.

This paper discusses ergodic properties and circular statistical characteristics in neuronal spike trains. Ergodicity means that the average taken over a long time period and over smaller population should equal the average in less time and larger population. The objectives are to show simple examples of design and validation of a neuronal model, where the ergodicity assumption helps find correspondence between variables and parameters. The methods used are analytical and numerical computations, numerical models of phenomenological spiking neurons and neuronal circuits. Results obtained using these methods are the following. They are: a formula to calculate vector strength of neural spike timing dependent on the spike train parameters, description of parameters of spike train variability and model of output spiking density based on assumption of the computation realized by sound localization neural circuit. Theoretical results are illustrated by references to experimental data. Examples of neurons where spike trains have and do not have the ergodic property are then discussed.

Multibiomarker Responses of Juvenile Stages of Zebrafish (Danio rerio) to Subchronic Exposure to Polycyclic Musk Tonalide.

Synthetic polycyclic musks, widely used as additives in personal care products, are present in both biotic and abiotic matrices of the aquatic environment at concentrations of ng/l to µg/l. Although they are determined at comparatively low concentrations, these levels are biologically relevant and pose a significant growing risk as stressors to aquatic organisms. The purpose of our study was to evaluate the effects of 28-day-long exposure to polycyclic musk tonalide in zebrafish juvenile stages (Danio rerio) using selected biomarkers. Environmentally relevant concentrations of tonalide caused significant changes in selected enzyme activities in the experimental groups exposed to the highest concentrations. The activity of glutathione S-transferase and lipid peroxidation increased significantly (p < 0.05) after exposure to the highest concentration (50,000 ng/l) compared with the control. A similar trend was observed in catalase activity; there was a significant increase (p < 0.05) after exposure to two highest concentrations of tonalide (5000 and 50,000 ng/l). In addition, a statistically significant decrease (p < 0.05) in glutathione reductase activity was found in the lowest test concentration of tonalide (50 ng/l). None of the tested concentrations resulted in histopathological changes in liver, kidney, skin, or gill. Furthermore, no effects on body weight, body length, specific growth rate, and behavior were observed. Our results showed that tonalide exposure induced profound changes in the activities of antioxidant and detoxifying enzymes, such changes representing an adaptive response of the fish organism to tonalide toxicity.

The Effects of Salicylic Acid on Juvenile Zebrafish Danio rerio Under Flow-Through Conditions.

The aquatic environment is becoming increasingly contaminated with pharmaceuticals. Salicylic acid (SA), which can be used individually or appear as a degradation product of the widely used acetylsalicylic acid was chosen for testing. Juvenile zebrafish Danio rerio were subjected to OECD test No. 215 (fish, juvenile growth test) with salicylic acid concentrations of 0.004; 0.04; 0.4; 4 and 40 mg/L. Specific growth rate (SGR), histological changes, and parameters of oxidative stress were evaluated. SA had no effects on histological changes, SGR, glutathione reductase, and lipid peroxidation. Increased catalytic activity of GPx was found at 0.04 mg/L compared to control, increased catalytic activity of catalase was found at 0.04 and 4 mg/L compared to control, and increased catalytic activity of glutathione-S-transferase was found at 0.004 and 0.04 mg/L compared to control (P < 0.05). Juvenile zebrafish turned out to be relatively insensitive to both environmentally relevant (0.004 mg/L) and higher concentrations of salicylic acid.

Simultaneous determination of ten anticoagulant rodenticides in tissues by column-switching UHPLC-ESI-MS/MS.

This paper describes the development of a method for the simultaneous determination of ten anticoagulant rodenticides (coumafuryl, warfarin, pindone, coumatetralyl, coumachlor, difenacoum, bromadiolone, brodifacoum, chlorophacinone and flocoumafen) in the liver and kidney based on column-switching liquid chromatography coupled with heated electrospray ionization tandem mass spectrometry. The simple sample preparation includes extraction with methanol. A C18 trapping column was used for online solid-phase extraction before analytical separation with the mobile phase comprising a mixture of 0.1% formic acid in water, methanol and acetonitrile. Chromatographic separation was achieved using a Thermo Hypersil ultra high-performance liquid chromatography (UHPLC) C18 column with the mobile phase consisting of 5 mM ammonium formate buffer (pH = 9) and methanol. The column-switching procedure ensured no matrix effects during electrospray ionization (ESI). Extraction recoveries ranged between 91 and 100% for liver and between 89 and 97% for kidney. The method showed good linearity up to 750 ng g(-1). The limit of detection ranged between 0.001 and 0.022 ng g(-1) for liver and between 0.001 and 0.028 ng g(-1) for kidney. The developed method was successfully used in several animal poisoning cases.

Degradation rate of praziquantel and fenbendazole in rainbow trout following oral administration.

OBJECTIVES: The aim of this study was to evaluate and compare the rate of degradation and elimination of praziquantel and fenbendazole antiparasitics following oral administration to salmonids. In addition, we determine whether the length of the legal withdrawal period is sufficient for complete elimination of antiparasitic residue from the body. The use of these drugs in fish is currently considered off-label and data on degradation are not available for rainbow trout. METHODS: The model species for this experiment was the rainbow trout (Oncorhynchus mykiss) and praziquantel and fenbendazole were chosen for experimental therapy. Both drugs were administered into the gastrointestinal tract using a stomach tube. Concentrations of fenbendazole and praziquantel were established through high performance liquid chromatography-tandem mass spectrometry. RESULTS: Our results show that concentrations of praziquantel and fenbendazole reach their maximum in the body within 24 hours of administration, with concentrations dropping sharply over the following 24 hours. With one exception, when trace amounts of both substances were found in blood plasma, the drugs were completely degraded and eliminated from the body by the end of the experiment (corresponding to 497.6 degree days). CONCLUSIONS: Praziquantel and fenbendazole both show a high rate of degradation and elimination from fish. As both substances were eliminated from the body within the required withdrawal period (i.e. within 500 degree days) they can be safely used based on current knowledge of their therapeutic effect for treating helminth infections.

Oxidative stress indices and histopathological effects of the nonsteroidal antiinflammatory drug naproxen in adult zebrafish (Danio rerio).

OBJECTIVES: The aim of this study was to investigate the effects of the nonsteroidal antiinflammatory drug naproxen on adult zebrafish (Danio rerio). METHODS: Three months old zebrafish (Danio rerio) were exposed to naproxen at concentrations of 0.001, 0.1 and 5 mg.L(-1). We focused on the changes in oxidative stress indices during and at the end of the experiment and histopathological examination of tissues after a two week long exposure period. RESULTS: We found that a 3 day long exposure to naproxen causes mild oxidative stress and affects detoxification in zebrafish, which is demonstrated by the increased activity of glutathione peroxidase and glutathione S-transferase at 0.001 and 0.1 mg.L(-1) of naproxen, respectively. After a 7 day long exposure to 0.1 and 5 mg.L(-1), more potent effects on enzymes occur. However, these effects are only short lasting. At the end of the experiment, the activities of the target enzymes recover back to homeostatic baseline levels. Except catalase, which is induced only after a two week long exposure to the environmental concentration of naproxen. Despite the fact that naproxen causes mild oxidative stress in zebrafish, exposure to this drug does not result in lipid peroxidation. Histopathological examination revealed obvious changes to the gills and liver even at exposure to the environmental concentration of naproxen. CONCLUSION: This study demonstrates that the environmental concentration of naproxen can slightly influence both the antioxidant defense system and histopathology of non-target fish.

The effect of the fluoroquinolone norfloxacin on somatic indices and oxidative stress parameters in early stages of common carp (Cyprinus carpio L.).

OBJECTIVES: The aim of this study was to assess the effect of the subchronic exposure of early stages of common carp (Cyprinus carpio L.) to norfloxacin using morphometric data and oxidative stress parameters. METHODS: A subchronic toxicity test was performed on fertilized embryos of common carp according to the OECD Guidelines No. 210. Embryos were exposed to norfloxacin concentrations of 0.0001 (environmental), 0.1, 1.0, 5.0, and 10.0 mg.L(-1) for 34 days. RESULTS: At the end of the test (day 34), significant (p<0.05) stimulation of development was observed in all experimental groups, in contrast to the control. Significantly greater (p<0.01) total body length was also observed in the group exposed to 10.0 mg.L(-1) of norfloxacin compared to the control. A significant increase in the activity of glutathione S-transferase in all carp exposed to norfloxacin concentrations of 0.1 and 1.0 mg.L(-1) (p<0.01), and 5.0 mg.L(-1) (p<0.05) compared to control group was revealed. The activity of glutathione peroxidase was significantly lower (p<0.01) in experimental carp exposed to a norfloxacin concentration of 10.0 mg.L(-1). In experimental carp exposed to a norfloxacin concentration of 0.0001 mg.L(-1), a significant increase (p<0.05) in glutathione reductase activity was found. Significant (p<0.01) decreases in the content of thiobarbituric acid reactive substances in the groups exposed to norfloxacin concentrations of 1.0, 5.0, and 10.0 mg.L(-1) were revealed. CONCLUSION: From the results, we can conclude that norfloxacin has a negative impact on selected biochemical processes related to the production of reactive oxygen species in early-life stages of common carp.

Persistent organic pollutants in muscle of fish collected from the Nové Mlýny reservoir in Southern Moravia, Czech Republic.

The aim of the present study was to investigate the content of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in muscle tissues of fish, i.e. carp (Cyprinus carpio, L.), bream (Abramis brama, L.) and pike perch (Stizostedion lucioperca, L.) from the middle Nové Mlýny reservoir and compare our results with previous corresponding studies. Samples were analysed by gas chromatography with ion trap tandem mass spectrometry. The highest contents of all pollutants were determined in muscle tissue of bream. The analysis of HCHs showed that ß-HCH was the most abundant. PCB congener 28 was evaluated as the predominant PCB congener. The metabolite p,p'-DDE exhibited the highest concentration of all the monitored metabolites. The monitoring confirmed significantly (P < 0.01) higher concentrations of OCP and PCB in muscle of bream compared to carp and pike perch. The contents of pollutants in our study were found to be lower in comparison to the findings of some previous studies.

The effects of subchronic exposure to ciprofloxacin on zebrafish (Danio rerio).

OBJECTIVES: The aim of this study was to assess the impact of ciprofloxacin in Danio rerio on growth, on the development of histopathological changes in selected organs (gills, kidney, liver), and on the activity of some oxidative stress markers during a 28-day toxicity test. METHODS: Juvenile growth tests were performed on D. rerio according to OECD guideline No. 215. Fish at the age of 30 days were exposed for 28 days to a range of concentrations of ciprofloxacin (0.7 µg x L(-1)--environmental concentration, 100, 650, 1100 and 3 000 µg x L(-1)). RESULTS: There were no significant differences between the specific growth rates of fish from the test groups and from the control group. Histopathological examination revealed no pathological changes in organs of fish exposed to any ciprofloxacin concentration. The activity of glutathione S-transferase increased at the ciprofloxacin concentrations of 0.7 and 100 µg x L(-1) compared to the control. A significant decrease in glutathione reductase activity was obtained in fish exposed to ciprofloxacin at 1100 and 3000 µg x L(-1); a significant decrease in glutathione peroxidase activity was also found, but at all tested concentrations except for 100 µg x L(-1). A decrease in the concentration of thiobarbituric acid reactive substances was noted only at 100 µg x L(-1) compared to the control. CONCLUSIONS: According to our results, all tested concentrations of ciprofloxacin, including the environmental concentration, had an influence on oxidative stress markers and detoxifying enzymes in exposed fish, but did not affect fish growth or cause the development of histopathological changes in the fish organism.

Effects of subchronic exposure of diclofenac on growth, histopathological changes, and oxidative stress in zebrafish (Danio rerio).

The aim of this study was to investigate effects of subchronic exposure to sublethal levels of diclofenac on growth, oxidative stress, and histopathological changes in Danio rerio. The juvenile growth tests were performed on Danio rerio according to OECD method number 215. Fish at the age of 20 days were exposed to the diclofenac environmental concentration commonly detected in the Czech rivers (0.02 mg L(-1)) and the range of sublethal concentrations of diclofenac (5, 15, 30, and 60 mg L(-1)) for 28 days. A significant decrease (P < 0.01) in the fish growth caused by diclofenac was observed in the concentrations of 30 and 60 mg L(-1). The identified value of LOEC (lowest observed effect concentration) was 15 mg L(-1) of diclofenac and NOEC (no observed effect concentration) value was 5 mg L(-1) of diclofenac. We did not find histopathological changes and changes of selected parameters of oxidative stress (glutathione S-transferase, glutathione reductase) in tested fish. The environmental concentration of diclofenac in Czech rivers did not have any effect on growth, selected oxidative stress parameters (glutathione S-transferase, glutathione reductase), or histopathological changes in Danio rerio but it could have an influence on lipid peroxidation.

New insights into the mechanisms and prevention of central nervous system oxygen toxicity: A prospective review.

Hyperbaric oxygen therapy (HBOT) elevates the pressure of life-sustaining oxygen (pO2), thereby saving lives. However, HBOT can also cause toxic effects like lung and retinal damage (peripheral oxygen toxicity) and violent myoclonic seizures (central nervous system (CNS) toxicity). The mechanisms behind these effects are not fully understood, hindering the development of effective therapies and preventive strategies. Herein, we critically reviewed the literature to understand CNS oxygen toxicity associated with HBOT to elucidate their mechanism, treatment, and prevention. We provide evidence that (1) increased pO2 increases reactive oxygen species (ROS) concentration in tissues, which irreversibly alters cell receptors, causing peripheral oxygen toxicity and contributing to CNS oxygen toxicity. Furthermore, (2) increased ROS concentration in the brain lowers the activity of glutamic decarboxylase (GD), which lowers concentrations of inhibitory neurotransmitter γ-aminobutyric acid (GABA), thereby contributing to the onset of HBOT-derived seizures. We provide long-overlooked evidence that (3) elevated ambient pressure directly inhibits GABAA, glycine and other receptors, leading to the rapid onset of seizures. Additionally, (4) acidosis facilitates the onset of seizures by an unknown mechanism. Only a combination of these mechanisms explains most phenomena seen in peripheral and CNS oxygen toxicity. Based on these proposed intertwined mechanisms, we suggest administering antioxidants (lowering ROS concentrations), pyridoxine (restoring GD activity), low doses of sedatives/anesthetics (reversing inhibitory effects of pressure on GABAA and glycine receptors), and treatment of acidemia before routine HBOT to prevent peripheral and CNS oxygen toxicity. Theoretically, similar preventive strategies can be applied before deep-sea diving to prevent life-threatening convulsions.

Long-term dietary exposure to the non-steroidal anti-inflammatory drugs diclofenac and ibuprofen can affect the physiology of common carp (Cyprinus carpio) on multiple levels, even at "environmentally relevant" concentrations.

The aim of the study was to evaluate the effects of emerging environmental contaminants, the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP), on physiological functions in juvenile common carp (Cyprinus carpio). Fish were exposed for 6 weeks, and for the first time, NSAIDs were administered through diet. Either substance was tested at two concentrations, 20 or 2000 µg/kg, resulting in four different treatments (DCF 20, DCF 2000, IBP 20, IBP 2000). The effects on haematological and biochemical profiles, the biomarkers of oxidative stress, and endocrine disruption were studied, and changes in RNA transcription were also monitored to obtain a comprehensive picture of toxicity. Fish exposure to high concentrations of NSAIDs (DCF 2000, IBP 2000) elicited numerous statistically significant changes (p < 0.05) in the endpoints investigated, with DCF being almost always more efficient than IBP. Compared to control fish, a decrease in total leukocyte count attributed to relative lymphopenia was observed. Plasma concentrations of total proteins, ammonia, and thyroxine, and enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase, and alkaline phosphatase (ALP) were significantly elevated in either group, as were the activities of certain hepatic antioxidant enzymes (superoxide dismutase, glutathione-S-transferase) in the DCF 2000 group. The transcriptomic profile of selected genes in the tissues of exposed fish was affected as well. Significant changes in plasma total proteins, ammonia, ALT, and ALP, as well as in the transcription of genes related to thyroid function and the antioxidant defense of the organism, were found even in fish exposed to the lower DCF concentration (DCF 20). As it was chosen to match DCF concentrations commonly detected in aquatic invertebrates (i.e., the potential feed source of fish), it can be considered "environmentally relevant". Future research is necessary to shed more light on the dietary NSAID toxicity to fish.

The effects of mycotoxin deoxynivalenol (DON) on haematological and biochemical parameters and selected parameters of oxidative stress in piglets.

OBJECTIVES: Deoxynivalenol (DON) - trichothecene mycotoxin, is frequently detected in high concentrations in cereals in the temperate region of Europe. The aim of this study was to determine the effect of DON in feed on haematological and biochemical parameters and on oxidative stress in piglets. METHODS: Two concentrations of DON in feedstuff for pigs were chosen: 0.6 mg/kg (group C) and 2.0 mg/kg (group M). Twelve weaned pigs were used in each group. Pigs were fed with naturally contaminated feed for 4 weeks. On days 14, 21 and at the end of the experiment (day 28) samples of blood were taken to determine haematological parameters, plasma biochemical parameters, ceruloplasmin activity and FRAP (ferric reducing ability of plasma). RESULTS: The haematological variables did not show changes in response to contaminated diet with exception of the mean corpuscular volume, which was significantly decreased at the end of the experiment in the group M. A significant increase of alkaline phosphatase activity (140%, p<0.01) was found in the group M compared to the group C at the end of the experiment. A significant decrease was found on the day 21 in FRAP (85%, p<0.001) and on the day 28 in ceruloplasmin (75%, p<0.01) in the group M compared to the group C. CONCLUSIONS: The decrease of FRAP and ceruloplasmin indicate a lowered ability of organism to scavenge reactive oxygen species. The higher concentration of DON in feedstuffs had a negative influence on the antioxidant ability of piglet's plasma.

Evaluation of ibuprofen toxicity for zebrafish (Danio rerio) targeting on selected biomarkers of oxidative stress.

OBJECTIVES: The aim of the study was to investigate the effects of subchronic exposure of zebrafish to ibuprofen, using selected oxidative stress parameters as a target. DESIGN: Toxicity tests were performed on Danio rerio according to OECD No. 203 and No. 215. In the growth test, fish were exposed to subletal concentrations of ibuprofen (0.0001, 0.05, 1, 8, and 25 mg.L-1) for 28 days. For the assessment of free radical defense in fish, the catalytic activities of glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPx), and catalase (CAT), as well as the concentration of malondialdehyde (MDA) were measured. RESULTS: Ibuprofen did not affect the activity of glutathione reductase and catalase. A significant (p<0.01) increase in the activity of glutathione peroxidase was found, which was proved dose-dependent (10.58 nmol NADPH per min per mg protein in the control and 20.53, 26.36, 26.89, and 45.87 nmol NADPH per min per mg protein in the ibuprofen concentrations of 0.5, 1, 8, and 25 mg.L-1. An increased (p<0.05) activity of glutathione S-transferase in the highest concentration was found compared to control. Malondialdehyde levels were found significantly (p<0.01) decreased from control in the concentrations of 0.0001 and 8 mg.L-1, but no dose-dependence was found. CONCLUSION: The results suggest that ibuprofen causes the increase in the activity of some antioxidative and biotransformation enzymes in zebrafish (GPx and GST). We also found a significant decrease in lipid peroxidation in the concentrations of 0.0001 and 8 mg.L-1 compared to control.

Biochemical markers for the assessment of pollution of selected small streams in the Czech Republic.

OBJECTIVES: The objective of the presented study was to investigate levels of the most important biochemical markers in fish which were caught upstream and downstream from sites near waste water treatment plants situated at 3 selected small streams of the Czech Republic. Organic pollutants and mercury were determined in muscle tissue of fish for complex assessment. DESIGN: Levels of biochemical markers (cytochrome P450 (CYP450), ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), vitellogenin (VTG)) were measured in selected tissues (liver, plasma, brain, gill and kidney) samples of brown trout. The concentrations of persistent organic pollutants (POPs) and mercury were determined in muscle samples. RESULTS: Significantly higher levels of biochemical markers (EROD, GST, VTG) were measured downstream from waste water treatment plant (WWTP) in Vlachovo Brezí (the small Libotýnský brook). Values of biochemical markers at the other localities Cerný brook and Moravice River were measured downstream from WWTP consistently or slightly lower with values of upstream. The values of POPs and mercury were in all monitored sites higher upstream compared with downstream from WWTP. The highest values of POPs and mercury were found on Cerný brook in Bruntál (more industrial and agricultural activity). CONCLUSION: The highest occurrence of selected POPs and mercury was generally upstream suggesting that small WWTP are able to partially remove these substances in those locations. But finding higher values of biochemical markers in downstream shows that there are many other substances endocrine disrupting chemicals (EDCs) that WWTP are unable to remove.