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.

Synthetic Biomimetic Polymethacrylates: Promising Platform for the Design of Anti-Cyanobacterial and Anti-Algal Agents.

Extensive, uncontrolled growth of algae and cyanobacteria is an environmental, public health, economic, and technical issue in managing natural and engineered water systems. Synthetic biomimetic polymers have been almost exclusively considered antimicrobial alternatives to conventional antibiotics to treat human bacterial infections. Very little is known about their applicability in an aquatic environment. Here, we introduce synthetic biomimetic polymethacrylates (SBPs) as a cost-effective and chemically facile, flexible platform for designing a new type of agent suitable for controlling and mitigating photosynthetic microorganisms. Since SBPs are cationic and membranolytic in heterotrophic bacteria, we hypothesized they could also interact with negatively charged cyanobacterial or algal cell walls and membranes. We demonstrated that SBPs inhibited the growth of aquatic photosynthetic organisms of concern, i.e., cyanobacteria (Microcystis aeruginosa and Synechococcus elongatus) and green algae (Chlamydomonas reinhardtii and Desmodesmus quadricauda), with 50% effective growth-inhibiting concentrations ranging between 95 nM and 6.5 µM. Additionally, SBPs exhibited algicidal effects on C. reinhardtii and cyanocidal effects on picocyanobacterium S. elongatus and microcystin-producing cyanobacterium M. aeruginosa. SBP copolymers, particularly those with moderate hydrophobic content, induced more potent cyanostatic and cyanocidal effects than homopolymers. Thus, biomimetic polymers are a promising platform for the design of anti-cyanobacterial and anti-algal agents for water treatment.

Spatial and Temporal Trends in Contamination of the Czech Part of the Elbe River by Mercury Between 1991 and 2016.

We summarized available data concerning contamination of the Czech part of the Elbe River by mercury and assessed the potential risks to humans connected with the consumption of contaminated fish. Total mercury concentrations were evaluated in dorsal muscle of chub (Squalius cephalus) and bream (Abramis brama) that were collected at 11 sampling sites in the period from 1991 to 2016. Significant differences among sites were found for both species. The highest contamination was observed near the non-ferrous Kovohute metal works at Celakovice and also downstream from the Spolana chemical plant at Neratovice. A significant decline in contamination with time (concentrations decreasing from 1.65 to 0.22 mg kg-1; p = 0.004) was revealed in chub downstream from the Spolana plant. Calculated hazard indices (HIs) were below 1, therefore, adverse health effects on the general Czech population connected with the consumption of fish from the Elbe River were not expected.

The effect of the antidepressant venlafaxine on gene expression of biotransformation enzymes in zebrafish (Danio rerio) embryos.

The effect of venlafaxine, a pharmaceutical commonly found in aquatic environment, was analyzed on non-target organism, Danio rerio (Hamilton, 1822). D. rerio embryos were treated by two different concentrations of venlafaxine: either concentration relevant in aquatic environment (0.3 µg/L) or concentration that was two orders of magnitude higher (30 µg/L) for the evaluation of dose-dependent effect. Time-dependent effect was rated at 24, 96, and 144 h post-fertilization (hpf). For gene expression, genes representing one of the phases of xenobiotic biotransformation (0 to III) were selected. The results of this study showed that the effect of venlafaxine on the zebrafish embryos is the most evident at hatching (96 hpf). At this time, the results showed a downregulation of gene expression in each phase of biotransformation and in both tested concentrations. In contrast, an upregulation of most of the genes was observed 144 hpf for both tested venlafaxine concentrations. The study shows that venlafaxine can affect the gene expression of biotransformation enzymes in D. rerio embryos even in the environmentally relevant concentration and thus disrupt the process of biotransformation. Moreover, the pxr regulation of genes seems to be disrupted after venlafaxine exposure in dose- and time-dependent manner.

Resazurin assay for assessment of antimicrobial properties of electrospun nanofiber filtration membranes.

We developed a simple and fast microplate assay for evaluation of the antimicrobial activity of electrospun nanofiber filtration membranes or similar porous materials for water treatment technologies. Resazurin (alamarBlue®) was used as an indicator of the amount of viable experimental microorganisms Gram-negative Escherichia coli, Gram-positive Enterococcus faecalis, and natural wastewater treatment plant effluent bacteria. A bacterial inoculum of concentration 1-3 × 105 CFU mL-1 was pipetted onto the surface of assessed both functionalized and respective control membranes and incubated in 12-well plates for 4 h at 37 °C. Kinetics of resazurin metabolization, i.e. its reduction to fluorescent resorufin, was evaluated fluorimetrically (λex520/λem590 nm). A number of viable bacteria on the membranes expressed as CFU mL-1 was calculated from the kinetic curves by using calibration curves that were constructed for both experimental bacterial species. Antimicrobial activities of the membranes were evaluated by either resazurin assay or modified ISO 20743 plate count assay. Results of both assays showed the significant antimicrobial activity of membranes functionalized with silver nanoparticles for both bacterial species and wastewater treatment plant effluent bacteria as well (log CFU reduction compared to control membrane > 4), while membranes containing specific quaternary ammonium salts were inefficient (log CFU reduction < 1). The suitability of resazurin microplate assay for testing nanofiber filtration membranes and analogous matrices has proven to be a faster and less demanding alternative to the traditionally used approach providing comparable results.

Branched Poly(ethylene imine)s as Anti-algal and Anti-cyanobacterial Agents with Selective Flocculation Behavior to Cyanobacteria over Algae.

Poly(ethylene imine)s (PEIs) have been widely studied for biomedical applications, including antimicrobial agents against potential human pathogens. The interactions of branched PEIs (B-PEIs) with environmentally relevant microorganisms whose uncontrolled growth in natural or engineered environments causes health, economic, and technical issues in many sectors of water management are studied. B-PEIs are shown to be potent antimicrobials effective in controlling the growth of environmentally relevant algae and cyanobacteria with dual-functionality and selectivity. Not only did they effectively inhibit growth of both algae and cyanobacteria, mostly without causing cell death (static activity), but they also selectively flocculated cyanobacteria over algae. Thus, unmodified B-PEIs provide a cost-effective and chemically facile framework for the further development of effective and selective antimicrobial agents useful for control of growth and separation of algae and cyanobacteria in natural or engineered environments.

Effects of selected tricyclic antidepressants on early-life stages of common carp (Cyprinus carpio).

Despite the fact that pharmaceuticals occur in surface water in low concentrations, they can still have adverse effect on aquatic biota, because these substances are designed to have a specific mode of action even at low concentrations. To our knowledge, only little is known about the long-term effect of tricyclic antidepressant residues on non-target animals, especially fish. Hence, the aim of this study was to assess the effect of subchronic exposure to selected tricyclic antidepressants - namely, amitriptyline, nortriptyline, and clomipramine and their mixture - on early-life stages of common carp (Cyprinus carpio). Two embryo-larval toxicity tests with common carp were performed, each taking 30 days. For the experiment, three tricyclic antidepressants as well as their mixtures were tested at three different concentrations - 10, 100 and 500 µg/L. Hatching and mortality were recorded twice a day. In addition, samples were taken at regular intervals in order to record developmental stage, morphometric and condition characteristics and morphological anomalies. At the end of the test, additional samples were taken for histopathological examination and also for the determination of antioxidant and biotransformation enzyme activity and lipid peroxidation and protein carbonylation. Long-term exposure resulted in a significant increase in mortality, developmental retardation, morphological anomalies, and pathological changes in brain, heart, and cranial and caudal kidney. In addition, changes in antioxidant enzyme activity as well as increased lipid peroxidation were observed, even at the lowest tested concentrations. Hence, environmentally relevant concentrations of TCAs have the potential to cause harmful effects on early-life stages of fish.

Evaluation of antibacterial properties of novel phthalocyanines against Escherichia coli--comparison of analytical methods.

We analyzed antibacterial effects of several novel phthalocyanines against Escherichia coli and evaluated the suitability of flow cytometry for the detection of antibacterial effects of phthalocyanines in comparison with routinely used cultivation. After 3h of exposure under cool white light eight cationic phthalocyanines showed very high antibacterial activity in the concentration of 2.00 mg L(-1) and four of them were even efficient in the concentration of 0.20 mg L(-1). Antibacterial activity of neutral and anionic compounds was considerably lower or even negligible. No antibacterial effect was detected when bacteria were exposed without illumination. Binding affinity to bacterial cells was found to represent an important parameter influencing phthalocyanine antibacterial activity that can be modified by total charge of peripheral substituents and by the presence of suitable functional groups inside them. Agglomeration of cells observed in suspensions treated with a higher concentration of certain cationic phthalocyanines (the strongest binders to bacterial membrane) affected cytometric measurements of total cell counts, thus without appropriate pretreatment of the sample before analysis this parameter seems not to be fully valid in the evaluation of phthalocyanine antibacterial activity. Cytometric measurement of cell membrane integrity appears to be a suitable and even more sensitive parameter than cultivation.

A novel electrospun polyurethane nanofibre membrane--production parameters and suitability for wastewater (WW) treatment.

The aim of this study was to investigate the suitability of a novel electrospun polyurethane nanofibre material for water-treatment purposes. Bacterial removal efficiency was tested in the laboratory by filtering artificial water spiked with Escherichia coli through a 0.25 µm nanofibre membrane. The results were compared with those obtained using a commercial microfiltration material (MV020T) with a similar pore size (0.20 µm). Alongside the laboratory experiments, we also determined filtration efficiency with semi-pilot scale experiments using actual wastewater from the secondary sedimentation tank of a wastewater treatment plant. The laboratory experiments indicated very high log10 removal efficiency, ranging from 5.8 to 6.8 CFU (colony-forming units)/ml. These results were better than those of the commercial membrane (3.8-4.6 CFU/ml). The semi-pilot scale experiment confirmed the membrane's suitability for microbial filtration, with both E. coli and total culturable microorganisms (cultured at both 22 and 36 °C) showing a significant decline compared to the non-filtered control (wastewater from the secondary outlet).

Effects of subchronic metribuzin exposure on common carp (Cyprinus carpio).

OBJECTIVES: Effects of metribuzin on biochemical and haematological indices, induction of specific biomarkers and impacts on biometric parameters of Cyprinus carpio were investigated for subchronic conditions. METHODS: Juvenile fish were exposed to 0.175 mg.L-1 or 1.75 mg.L-1 metribuzin for 28 days. Haematological indices were assessed using unified methods of haematological examination in fish. Biochemical indices were determined by biochemical analyzer, hepatic vitellogenin content was estimated by direct sandwich ELISA. Cytochrome P450 concentration and ethoxyresorufin O-deethylase activity were measured in liver spectrophotometrically and spectrofluorimetrically, respectively. RESULTS: Increased haematocrit and RBC count were found in fish exposed to 1.75 mg.L-1 metribuzin compared to control fish (p<0.05). Other indices contents and biomarkers levels were not significantly changed by either concentration of metribuzin. CONCLUSION: Sublethal metribuzin pollution may have adverse impacts on haematological parameters in common carp.

Effects of subchronic exposure to LASSO MTX (alachlor 42% W/V) on hematological indices and histology of the common Carp, Cyprinus carpio L.

The aim of the study was to evaluate subchronic toxic effects of the preparation LASSO MTX (alachlor 42% W/V) on hematological indices and histology of the common carp (Cyprinus carpio L.). In carp exposed for 28 days to LASSO MTX in the concentration of 2,400 microg L(-1), significant differences (p < 0.05), were detected in all indices of the erythrocyte profiles tested except hematocrit (PCV) when compared to controls. At a lower exposure (240 microg L(-1)) concentration of blood hemoglobin and mean corpuscular hemoglobin were significantly reduced. In contrast, no influence of the preparation on leucopoiesis was demonstrated, and only slight changes were observed in histopathological indices.

Evaluation of xenoestrogenic potential of propylparaben in zebrafish (Danio rerio).

OBJECTIVES: Xenoestrogenic potential of propylparaben (PP), one of the most commonly used preservatives in drugs, cosmetics and food, was investigated in vivo using zebrafish (Danio rerio). METHODS: Juvenile zebrafish (20 days post hatching) were exposed to three different concentrations of propylparaben (PP) dissolved in ethanol and added into the water. After 20 days of exposure the fish were euthanized and vitellogenin concentrations in their whole body homogenates were measured by the method of direct sandwich ELISA. Simultaneously, vitellogenin concentrations in either fish from the control group (exposed to solvent without the substance tested) and in fish from the positive control group (exposed to natural estrogen 17beta-estradiol) were measured. RESULTS: Vitellogenin concentration in whole body homogenates of control fish was 400 (396-540) ng/ml(-1) (geometric mean (95% CI)). Zebrafish exposure to propylparaben at the concentrations of 0.1; 0.4 and 0.9 mg/l(-1) elicited statistically significant decline (P<0.001) of vitellogenin production, i.e. geometric means of vitellogenin concentrations in whole body homogenates were 240 (186-311); 218 (175-270) and 270 (234-311) ng/ml(-1), respectively. Conversely, the geometric mean of vitellogenin concentration in whole body homogenates of zebrafish exposed to 100 ng/ml(-1) of 17beta-estradiol (positive control) was significantly higher (P<0.001) than values in all other groups, i.e. 35,553 (16,860-74,968) ng/ml(-1). CONCLUSIONS: Our results suggest an antiestrogenic potential of propylparaben tested in vivo in juvenile zebrafish (Danio rerio). The estrogenic effect of 17beta-estradiol was confirmed.