The Urban Wastewater Treatment Directive recent draft issued last October 2022 pays attention to contaminants of emerging concern including organic micropollutants (OMPs) and requires the removal of ...some of them at large urban wastewater treatment plants (WWTPs) calling for their upgrading. Many investigations to date have reported the occurrence of a vast group of OMPs in the influent and many technologies have been tested for their removal at a lab- or pilot-scale. Moreover, it is well-known that hospital wastewater (HWW) contains specific OMPs at high concentration and therefore its management and treatment deserves attention. In this study, a 1-year investigation was carried out at a full-scale membrane bioreactor (MBR) treating mainly HWW. To promote the removal of OMPs, powdered activated carbon (PAC) was added to the bioreactor at 0.1 g/L and 0.2 g/L which resulted in the MBR operating as a hybrid MBR. Its performance was tested for 232 target and 90 non-target OMPs, analyzed by UHPLC-QTOF-MS using a direct injection method. A new methodology was defined to select the key compounds in order to evaluate the performance of the treatments. It was based on their frequency, occurrence, persistence to removal, bioaccumulation and toxicity. Finally, an environmental risk assessment of the OMP residues was conducted by means of the risk quotient approach. The results indicate that PAC addition increased the removal of most of the key OMPs (e.g., sulfamethoxazole, diclofenac, lidocaine) and OMP classes (e.g., antibiotics, psychiatric drugs and stimulants) with the highest loads in the WWTP influent. The hybrid MBR also reduced the risk in the receiving water as the PAC dosage increased mainly for spiramycin, lorazepam, oleandomycin. Finally, uncertainties and issues related to the investigation being carried out at full-scale under real conditions are discussed.
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•A 1-year investigation on micropollutant removal was carried out on a full scale MBR.•232 target and 90 non-target organic micropollutants were monitored.•0.1 g/L and 0.2 g/L of PAC added in the MBR acting as an hybrid MBR•Key organic micropollutants selected by means of three new approaches•Removal significantly enhanced for antibiotics and psychiatric drugs at 0.2 g/L of PAC.
Cardiovascular pharmaceuticals (CVPs) are globally present in inland waters and have also been found in the sediment and plasma of fish from the Sava River, Croatia. Based on the previous research, ...CVPs amiodarone (AMI), ramipril (RAM), simvastatin (SIM), and verapamil (VER) have been selected for this study. Their effect has been investigated, individually and in a mixture, on the development of the zebrafish embryo Danio rerio (Hamilton, 1822) within the first 96 h of development. Upon exposure to environmentally relevant concentrations of tested CVPs (0.1, 1, and 10 μg/L) zebrafish survival and development as apparent from observed morphological abnormalities, heartbeat rates and changes in behavior, hatching success, larval length and oxidative stress level were monitored. The CVP causing the highest mortality and pathological changes was SIM (1 and 10 μg/L), which corresponds well with the observed effects during zebrafish exposure to CVPs' mixtures (4 and 40 μg/L). All pharmaceuticals affected cardiac function and decreased heart rate. SIM (1 μg/L), VER and RAM (10 μg/L) decreased larval length, while induced oxidative stress was recorded in the SIM- and VER-exposed specimens. Behavioral alterations of zebrafish were observed only in AMI-treated group (10 μg/L). Our amino acid sequence comparison and structural and docking analysis showed a highly conserved binding site between human and zebrafish HMG-CoA reductase for SIM and its main metabolite simvastatin acid. Using these ecotoxicological bioassays on a zebrafish model with particular emphasis on sublethal endpoints, the risk of CVPs, especially statins, for fish in inland waters has been identified.
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•Cardiovascular drugs are widespread in the aquatic environment.•Cardiovascular drugs affect cardiac physiology and growth, induced oxidative stress.•Simvastatin caused severe adverse effects on zebrafish embryonal development.•Mixture toxicity analysis highlights interaction of cardiovascular drugs.
Increased contamination of aquatic ecosystems with pharmaceuticals could have been expected due to the COVID-19 pandemic. Surface water from three domains (rural, semirural, urban) of the Mrežnica ...River (Croatia) was screened for 253 pharmaceuticals by SPE-UHPLC-MS/MS. At the beginning of the pandemic, the highest concentration of drugs (excluding veterinary) was detected at urban site (291.4 ng/L), followed by semirural (186.5 ng/L) and rural (141.6 ng/L). With the progression of pandemic, contamination increase was observed at all sites, but it was the most obvious at semirural (approximately 400–700 ng/L). The most pronounced concentration increases were observed for non-opioid analgesics, especially ibuprofen. In September 2021, the first notable occurrence of opioid analgesics was recorded. The most represented group of pharmaceuticals at the start of the pandemic (May 2020) was generally stimulants (caffeine, cotinine). In September 2021, the predominant group was analgesics at all sites (45–84%), whereas stimulants decreased to undetectable levels. The results of this study indicated that the epidemiological measures and medical treatments that were widely imposed/applied caused notable increase of the surface water contamination with drugs of a small river with limited dilution capacity, indirectly pointing to the changes that occurred in the behaviour and habits of the inhabitants of the affected areas.
Quantitative chemical analyses of 428 organic contaminants (OCs) indicated the presence of 313 OCs in the sediment extracts from Sava River, Croatia. Pharmaceuticals were present in higher ...concentrations than pesticides thus confirming their increasing threat to freshwater ecosystems. Toxicity evaluation of the sediment extracts from four locations (Jesenice, Rugvica, Galdovo and Lukavec) using zebrafish embryotoxicity test (ZET) accompanied with semi-quantitative histopathological analyses exhibited correlation with cumulative number and concentrations of OCs at the investigated sites (10.05, 15.22, 1.25, and 9.13 μg/g respectively). Toxicity of sediment extracts and sediment was predicted using toxic unit (TU) approach and persistence, bioaccumulation and toxicity (PBT) ranking. Additionally, influential OCs and genes were identified by graph mining of the prior knowledge informed, site-specific chemical-gene interaction models. Predicted toxicity of sediment extracts (TUext) was similar to the results obtained by ZET and associated histopathology with Rugvica sediment being the most toxic, followed by Jesenice, Lukavec and Galdovo. Sediment TU (TUsed) favoured OCs with low octanol-water partition coefficients like herbicide glyphosate and antibiotics ciprofloxacin and sulfamethazine thus indicating locations containing higher concentrations of these OCs (Galdovo and Rugvica) as the most toxic. Results suggest that comprehensive in silico sediment toxicity predictions advocate providing equal attention to organic contaminants with either very low or very high log Kow.
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•Pharmaceuticals are a major component of river sediment pollution.•ZET and histopathological analysis are good indicators of sediment toxic potential.•In silico sediment toxicity predictions should include OCs with low or high log Kow.
The remnants of historical industrial contamination can be detected in many aquatic ecosystems worldwide even at present time. Mrežnica is a river in Croatia that has been, for more than a hundred ...years, continually exposed to effluents of various industries, which have, in modern time, mostly ceased to operate. Our aim was to establish the level of current contamination and pollution of the Mrežnica river-water and sediments. The study of river contamination at three sites (reference site; site nearby former cotton industry facility in Duga Resa – DRF; industrial zone of Karlovac town - KIZ) in three sampling campaigns (May 2020, April and September 2021) encompassed analyses of physico-chemical water parameters, screening of 369 pesticides, measurement of metal (loid) concentrations in the sediments, and in the dissolved and particulate phases of the river-water. The sediment pollution was assessed through the analyses of total bacteria abundance (by targeting 16S rRNA genes), and their associated metal resistance genes (cnrA, pbrT and czcD) and class 1 integrons (intl1). At the DRF site, industrial organic contaminants that can be traced to textile production were detected (dye and nylon components), as well as increased levels of some metals bound to suspended particulate matter and sediments. At the most downstream KIZ site, occasional high level of industrial herbicide neburon was measured in the river-water, and metal contamination of suspended particulate matter and sediments was evident. Although, based on the comparison with legislation and literature data, the level of contamination was rather mild, the effects on microbial communities were unquestionable, confirmed by increased abundance of the czcD gene at DRF site and the intI1 gene at both industrially impacted sites. Obtained results indicated long-term sediment retention of some industrial contaminants at the places of historical freshwater contamination, and, thus, the necessity for their monitoring even after the termination of contamination sources.
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•The remnants of historical contamination detected in freshwater ecosystem.•Dye and nylon components found in river-water near former cotton industry facility.•High levels of industrial herbicide detected in water near urban industrial zone.•River mildly contaminated with metals at sites of past long-term industrial impact.•Pollution biomarker, microbial gene intI1, higher at industrially impacted sites
This study focused on the short-term whole organism bioassays (WOBs) on fish (Danio rerio) and crustaceans (Gammarus fossarum and Daphnia magna) to assess the negative biological effects of water ...from the major European River Sava and the comparison of the obtained results with in vitro toxicity data (ToxCast database) and Risk Quotient (RQ) methodology. Pollution profiles of five sampling sites along the River Sava were assessed by simultaneous chemical analysis of 562 organic contaminants (OCs) of which 476 were detected. At each sampling site, pharmaceuticals/illicit drugs category was mostly represented by their cumulative concentration, followed by categories industrial chemicals, pesticides and hormones. An exposure-activity ratio (EAR) approach based on ToxCast data highlighted steroidal anti-inflammatory drugs, antibiotics, antiepileptics/neuroleptics, industrial chemicals and hormones as compounds with the highest biological potential. Summed EAR-based prediction of toxicity showed a good correlation with the estimated toxicity of assessed sampling sites using WOBs. WOBs did not exhibit increased mortality but caused various sub-lethal biological responses that were dependant relative to the sampling site pollution intensity as well as species sensitivity. Exposure of G. fossarum and D. magna to river water-induced lower feeding rates increased GST activity and TBARS levels. Zebrafish D. rerio embryo exhibited a significant decrease in heartbeat rate, failure in pigmentation formation, as well as inhibition of ABC transporters. Nuclear receptor activation was indicated as the biological target of greatest concern based on the EAR approach. A combined approach of short-term WOBs, with a special emphasis on sub-lethal endpoints, and chemical characterization of water samples compared against in vitro toxicity data from the ToxCast database and RQs can provide a comprehensive insight into the negative effect of pollutants on aquatic organisms.
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•Chemical analysis detected 476 organic pollutants in surface water.•Pharmaceuticals/Illicit drugs found as the main contaminants in Sava River.•ToxCast indicated steroidal anti-inflammatory drugs as chemicals of concern.•Activation of nuclear receptors as bio-targets of greatest concern.•Potential association of bioassays and ToxCast data for effect-based monitoring.
To better understand the impact of conventional agricultural practices on soil and water resources, two main objectives were established in this study: to measure nitrate (NO3-) and sulphate (SO42-) ...concentrations in lysimeters and drainpipes leachates in response to different nitrogen fertilization levels and soil amendments and to identify organic contaminants that could have originated from the long-term use of agrochemicals in historically intensive agricultural area in Croatia. During the two-year study period leachate samples were collected from lysimeters and drainpipes. Research results indicate NO3- content in both lysimeters and drainpipes increased with rising doses of N fertilizer. The highest concentration of SO42- was recorded in the treatment with added phosphogypsum, while in all other treatments the concentrations were low. Over 40% of the 287 target substances were detected at least once during the two-year study period but only two substances (IPC/propham and carbosulfan) were found to have concentrations above 0.5 μg/L. Additionally, three priority substances were continuously present in the leachate samples: atrazine, simazine and isoproturon.
This study aimed at assessing the toxic effects of industrial effluents using duckweed (Lemna minor L.) plants as a test system. Growth inhibition test according to standardized protocol (ISO 20079) ...was performed. The suitability of the Comet assay (indicates DNA damage) and certain parameters such as peroxidase activity and lipid peroxidation level, as biomarkers for environmental monitoring was evaluated. The water samples were collected monthly over a 3-month period from the stream near the industrial estate of Savski Marof, Croatia. All samples caused inhibition of growth rates based on frond number and biomass as well as decrease of chlorophylls content. In contrast, peroxidase activity, malondialdehyde content and tail extent moment (measure of DNA strand breaks) markedly increased. Obtained data demonstrate the relevance of duckweed as sensitive indicators of water quality as well as the significance of selected biological parameters in the reliable assessment of phyto- and genotoxic potential of complex wastewaters.
The genotoxic and toxic potential of polluted surface water exposed to a fertilizer factory effluent was evaluated using assays with fish (Cyprinus carpio) and plant (Lemna minor) model organisms. ...Beside classical physicochemical parameters, the contents of fluorides, some heavy metals and polycyclic aromatic hydrocarbons were analyzed as well. Surface water caused inhibition of plant growth and decrease of photosynthetic pigment content. Regarding DNA damage and oxidative stress parameters, both fish and plants showed similar response to the surface water. In confirmation to biochemical markers, histopathological analysis of gill and liver tissues revealed a higher incidence of lesions in fish exposed to polluted surface water. Generally, results obtained by biological monitoring were mostly in agreement with chemical analysis of the surface water, although several discrepancies were observed which might be due to difference in sensitivity of model organisms or in experimental conditions (laboratory and field exposure). The results imply that conventional chemical analysis should be extended to genotoxicity/toxicity assays as measured biological effects and the potential health hazard cannot be predicted based on the physicochemical characteristics of water samples alone.
•Chemical analysis and various tests were used in assessment of the surface water.•Micropollutants induced genotoxicity, oxidative stress reactions and tissue damage.•Model organisms showed similar responses to pollutants in surface water.•Interpretation of surface water (geno)toxicity was given.
Chemical analysis of plasma samples of wild fish from the Sava River (Croatia) revealed the presence of 90 different pharmaceuticals/illicit drugs and their metabolites (PhACs/IDrgs). The ...concentrations of these PhACs/IDrgs in plasma were 10 to 1000 times higher than their concentrations in river water. Antibiotics, allergy/cold medications and analgesics were categories with the highest plasma concentrations. Fifty PhACs/IDrgs were identified as chemicals of concern based on the fish plasma model (FPM) effect ratios (ER) and their potential to activate evolutionary conserved biological targets. Chemicals of concern were also prioritized by calculating exposure-activity ratios (EARs) where plasma concentrations of chemicals were compared to their bioactivities in comprehensive ToxCast suite of in vitro assays. Overall, the applied prioritization methods indicated stimulants (nicotine, cotinine) and allergy/cold medications (prednisolone, dexamethasone) as having the highest potential biological impact on fish. The FPM model pointed to psychoactive substances (hallucinogens/stimulants and opioids) and psychotropic substances in the cannabinoids category (i.e. CBD and THC). EAR confirmed above and singled out additional chemicals of concern - anticholesteremic simvastatin and antiepileptic haloperidol. Present study demonstrates how the use of a combination of chemical analyses, and bio-effects based risk predictions with multiple criteria can help identify priority contaminants in freshwaters. The results reveal a widespread exposure of fish to complex mixtures of PhACs/IDrgs, which may target common molecular targets. While many of the prioritized chemicals occurred at low concentrations, their adverse effect on aquatic communities, due to continuous chronic exposure and additive effects, should not be neglected.
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•Ninety pharmaceuticals/illicit drugs and their metabolites were analyzed.•Fish plasma model and ToxCast data were used to identify chemicals of concern.•Nicotine, cotinine, prednisolone and dexamethasone: chemicals of concern.•Fish plasma model pointed psychoactive chemicals as potentially hazardous to fish.
