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Conventional wastewater treatment methods cannot completely remove the ultraviolet (UV) filters or dissolved organic matter. The transformation characteristics of these substances during chlorination disinfection and the varying species-specific toxicities of their combinations remain unclear. Here, Daphnia magna and zebrafish were exposed to benzophenone-3 (BP-3) and humic acid (HA) before and after chlorination disinfection. The results from chemical indicators showed that chlorination treatment decreased UV254 values and changed the intensity of parallel factors in three-dimensional fluorescence. Based on chemical analysis, the chlorine concentration and chlorination time for the toxicity experiments were set at 5 mg/L and 6 h, respectively. Exposure to HA and BP-3 before and after chlorination decreased the heart rate (by 1.37-28.12 %) in both species. However, species-specific responses, including survival rate, swimming distance, and expression of genes related to neurodevelopment, growth, and oxidative stress, were induced by chlorination. Chlorination reduced the impact of HA exposure but worsened the effects of HA and BP-3 co-exposure on D. magna. However, in zebrafish, the toxic effects intensified in most of the exposure groups after chlorination. Correlation analysis showed that the parallel factors of three-dimensional fluorescence were correlated with toxic effects on zebrafish, whereas UV254 was more significantly correlated with toxic effects on D. magna. This study provides insights into the combined toxicity of UV filters and dissolved organic matter in different aquatic organisms during chlorination, which is useful for risk control and optimization of the chlorination process., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nanoplastics and metal oxide nanoparticles are serious threats that inevitably enter the environment. Their similar particle properties likely lead to interaction and thus cause more unpredictable ecotoxicity to organisms. In this study, it was found that polystyrene nanoplastics (PS NPs) aggravate the toxic effect of iron oxide nanoparticles (Fe 2 O 3 NPs) on Lactuca sativa L. by inducing severe oxidative stress and root deformation, and the expansion of damaged cells from the xylem to the epidermis was observed using confocal laser scanning. Exposure to PS NPs + Fe 2 O 3 NPs correspondingly elevated iron accumulation in the roots and leaves by 1.39 and 1.17 times compared to the amount observed with Fe 2 O 3 NPs individually. Examination of the physicochemical properties, iron ion release, and molecular interactions of the NPs indicated that PS NPs interact with Fe 2 O 3 NPs to form heteroaggregates and facilitate leaching of iron ions, which resulted in aggravating the toxic effect. These were alleviated by the addition of humic acid (HA), which dispersed the heteroaggregates and reduced the release of iron ions. The findings in the present study provide new perspectives for the ecotoxicological risk of binary nano-pollution in the natural environment.

Humic substances, a major component of natural organic matter in surface waters, can induce biotransformation enzyme activities and influence antioxidant defense in fish. The study aimed to provide a molecular basis for the stress responses, the synthesis of biotransformation, and antioxidant enzymes in particular. Adult medaka fish (Hd-rR strain) were exposed to environmentally relevant concentrations of humic acid for 96 h. The actual humic acid concentrations in water were determined photometrically and expressed as organic carbon concentrations. Liquid chromatography with tandem mass spectrometry was used for protein profile analysis of medaka liver samples. The relative amount of isozymes was determined using the label-free quantification approach. Hepatic biotransformation enzyme activities were measured as well. Thus, ethoxyresorufin-O-deethylase activity showed a pronounced induction at the highest tested concentration (9.4 mg C/L). Various biotransformation and antioxidant isozymes responded to humic acid differently, reflecting a balanced interplay of proteins that ensures the metabolism of humic acid in fish liver. Some isozymes were not affected by humic acid. The study provides new insight into the molecular mechanisms of the fish stress response to the humic acid-related challenge., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Aims: This research was performed to investigate the influence of clay and humic acid on Escherichia coli O157 survival in model soils. Additionally, the influence of pH and humic acid on E. coli O157 in liquid culture was investigated., Methods and Results: Artificial soil microcosms were prepared with sand, kaolinite, bentonite and humic acid. Artificial soil microcosms pH was adjusted (6·0-7·0) with aluminium sulphate before E. coli O157 inoculation. After 56 days of incubation at 30°C, significant differences in E. coli O157 log CFU per gram were observed between 0 and 1000 ppm (P < 0·0001) and 0 and 5000 ppm (P < 0·0001) humic acid in 1·5% clay soils, but not in 7·5 or 15% clay soils. Significant differences (P < 0·05) in E. coli O157 log CFU per ml were observed in liquid culture influenced by humic acid concentrations after 8 h at 37°C., Conclusions: The developed model soils support E. coli O157 populations over 28 days, and higher clay soils may aid in E. coli O157 survival., Significance and Impact of the Study: These results provide insights into physicochemical properties of soil that may influence E. coli O157 in the environment and help explain E. coli O157 survival in various soils and geographical regions., (© 2020 The Society for Applied Microbiology.)

Fluoxetine is a widely prescribed antidepressant that has been frequently detected in aquatic environments and is associated with a series of neurological, behavioural and neuroendocrine disruptions in nontarget organisms. However, studies on its effects in fish under realistic environmental conditions are still limited. In this study, we determined the influences of an environmentally relevant concentration of fluoxetine (100 ng/L) on crucian carp (Carassius auratus) in the presence of dissolved organic matter (DOM). Endpoints that were assessed included accumulation of fluoxetine and metabolite formation as well as related biological responses involving neurotransmission and metabolic processes. Fluoxetine was significantly bioconcentrated in the fish brain and liver and largely transformed to the active metabolite norfluoxetine. Brain neurotransmission processes related to serotonin and choline and liver metabolic status were simultaneously altered. DOM added at 1 mg/L had no effect on the accumulation of fluoxetine or its metabolites in different tissues of the fish. However, at 10 mg/L DOM facilitated fluoxetine and norfluoxetine accumulation in the liver, brain, kidney, gill and bile tissues of the fish. The neuroendocrine-disrupting effects on fish caused by fluoxetine were also enhanced by the co-addition of DOM at 10 mg/L. Binding with fluoxetine and the inhibition of metabolic functions caused by DOM may be responsible for this increase in effects. These findings imply that at high concentrations DOM can increase the toxicity of environmentally relevant concentrations of fluoxetine to fish., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Due to the coexistence of organic matter and iron in groundwater, a certain part of the iron is present as iron-organic complexes in the form of colloids and/or dissolved complexes. The study was conducted to evaluate the impact of the type of oxidizing agent: O 2 , Cl 2 , H 2 O 2 , or KMnO 4 , on the efficiency of the oxidation and removal of iron compounds from three groundwaters with significantly different contents and types of organic substances among which humic and fulvic acids occurred. This study shows that after the aeration and the oxidation with Cl 2 and H 2 O 2 , the increasing content of dissolved hydrophilic organic substances containing aromatic rings in the raw water reduced the effectiveness of Fe(II) oxidation and the effectiveness of iron removal during the sedimentation process. This regularity was not found only when KMnO 4 was used as the oxidant. After oxidation with H 2 O 2 , the highest number of organo-iron complexes and an increased concentration of dissolved organic carbon were found. High concentrations of organo-ferrous connections were also found in aerated water samples. The highest KMnO 4 efficiency of removing iron and organic substances and reducing the color intensity and turbidity was due to the catalytic and adsorptive properties of the precipitated MnO 2 , which also improved the sedimentation properties of the resultant oxidation products.

Heavy metals and natural organic matters (NOM) form very stable heavy metal-NOM complexes in aqueous, facilitating the migration of heavy metals and enhancing their potential risks. In this study, non-thermal plasma oxidation was attempted to destroy the heavy metal-NOM complexes, with Cu-humate (Cu-HA) as a model. The decomplexation efficiency reached 86.1 % within 50 min of plasma oxidation at 16 kV. The generated reactive species by the non-thermal plasma, including O 2 - , 1 O 2 , OH, attacked the carboxyl and hydroxyl functional groups of HA, leading to cleavage of the Cu-O bonds, decomplexation of Cu-HA, and release of free Cu(II). Meanwhile, a variety of small molecular intermediates, including phenols, benzoic acids, esters, amines, ketones, acetic acid, formic acid, and oxalic acid, were generated due to attack by the oxidative species on the aromatic moiety and double bonds in Cu-HA. As a consequence of decomplexation, the residual toxicity of Cu-HA to Scenedesmus obliquus was distinctly reduced. This study provides a potential technique to decomplex heavy metal-NOM complexes, and reduces their toxicity to typical Scenedesmus obliquus., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We performed a transcriptome-based bioassay (TSB assay) using human hepatoma HepG2 cells to evaluate the potential toxicity of whole wastewater effluents from two membrane bioreactors (MBRs) and a conventional activated sludge process (AS). The biologically active agent(s) in the wastewater effluents were characterized based on expression of the marker genes (i.e., CYP1A1, AKR1B10, GCLM and GPX2) selected by DNA microarray analysis, after the wastewater effluent samples were concentrated by a reverse osmosis (RO) membrane and further fractionated by various manipulations. The qPCR assay of marker genes demonstrated that the induction of CYP1A1 and GPX2 was mitigated after passing through C18 and chelate columns. In addition, clear induction of CYP1A1 was observed in the smallest size fraction with 1 k Da or smaller organic molecules in all the tested effluents. These results together with the water quality data of the fractionated samples suggested that responsible constituents for potentially adverse and abnormal transcriptomic responses in HepG2 could have hydrophobic nature and act with metal-dissolved organic matter (DOM) complexes in 1 k Da or smaller size fraction. Although DOM is known to play two contradictory roles as a protector and an inducer of toxicants, our present study indicated the DOM in wastewater effluent, particularly humic substances with acidic nature, functioned as a toxicity inducer of residual chemicals in the effluents. This study provided a new insight into the nature of "toxic unknowns" in the wastewater effluents, which should be monitored whole through the reclamation process and prioritized for removal., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Studies have been conducted on the genotoxicity and carcinogenicity of disinfection by-products formed from natural organic matter (NOM) and mitigation effect for mutagens and clastogens by NOM. Whereas, reportedly, synthetic humic acid in high concentration has induced genotoxicity in human cells, and NOM samples have provoked mild oxidative and other physiological responses in aquatic organisms. Our group developed a novel detection method for DNA damage formation, namely enhanced green fluorescent protein (EGFP)-fused mediator of DNA damage checkpoint 1 (MDC1)-expressing human cells as simple and high-sensitive system. By using this method, a significant increase in the foci area was observed after 3 h, but not 24 h for 130 mgC L -1 Suwannee River fulvic acid (SRFA), 38 mgC L -1 humic acid (SRHA), and 19 mgC L -1 NOM (SRNOM). The SRNOM concentration is the original environmental one; therefore, it was suggested that the formation and repair of DNA damage associated with γ-H2AX, a biomarker for DNA double-strand breaks by mild oxidative stress, in Suwannee River (SR) were detected for the first time. The increase in the foci area was not observed for 18 mgC L -1 Lake Biwa fulvic acid (LBFA) and 50 mg L -1 catechin after both 3 h and 24 h. The difference between the SR and Lake Biwa (LB) samples may result from the differences in their electron-accepting capacity. The application of this methodology is expected to elucidate oxidative stress and toxicological effects shortly and in detail for many water samples., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

As emerging contaminants, microplastics (MPs) are predicted to act as vectors for other contaminants and their combined effects are largely unknown. In this study, the combined effects of MPs and natural organic matter (NOM) on the accumulation and toxicity of copper (Cu) in zebrafish (Danio rerio) were investigated. As a result, small-size MPs could absorb more Cu than large-size MPs. The presence of NOM promoted Cu adsorption on MPs in the pH range of 6-8. Our results demonstrate that the combination of MPs and NOM increased Cu accumulation in the livers and guts in a size-depended manner. Correspondingly, the results of biochemical test showed that MPs and NOM could aggravate Cu-toxicity in the livers and guts, which is manifested in the increased levels of malonaldehyde (MDA) and metallothionein (MT) and decreased levels of superoxide dismutase (SOD). Furthermore, the results of transcriptomic analysis suggested that such aggravation of toxicity was mainly attributed to the inhibition of Cu-ion transport and the enhanced oxidative stress. Since the co-existence of MPs and NOM in the environment is inevitable, their enhancement effects on the bioaccumulation and toxicity of other pollutants such as heavy metals deserve more attention., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study evaluated oxidative stress and estrogenic activity induced by hydrophobic (HP), transphilic (TP), and hydrophilic (HL) fractions of effluent organic matter (EfOM) from municipal and industrial wastewater treatment plants. Fluorescence regional integration (FRI) analysis indicated that EfOM showed higher FRI distribution in regions II (aromatic protein-like) and IV (soluble microbial products, SMPs) than Suwannee River natural organic matter (SRNOM), which was primarily in regions III (fulvic acid-like) and V (humic acid-like). The HP and HL fractions of industrial EfOMs significantly increased catalase (CAT) and glutathione S-transferase (GST) activity and lipid peroxidation compared with those of the control (p <  0.05). Estrogenic activity was the highest in industrial EfOM followed by municipal EfOM, and both were significantly higher than that of SRNOM and the control (p <  0.05). FRI distribution in region II was significantly related to CAT, GST, and estrogenic activity (r = 0.7142, 0.7786, and 0.8107, respectively) for the HL fraction of EfOM. For the HP fraction of EfOM, regions II and IV were significantly related to estrogenic activity (r = 0.7221 and 0.6707, respectively). These findings suggest that aromatic protein-like substances and SMPs in EfOM were responsible for the observed oxidative stress and estrogenic activity., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The humic-like substances were the main organic components in most wastewater (e.g. domestic sewage, toilet wastewater and landfill leachate). Two types of actual humic-like substances (fulvic acid (FA) and biologically treated landfill leachate (BTLL)) were selected to describe the changes in the properties of humic-like substances (complexation ability, aromaticity and mobility) during electrochemical oxidation. Meanwhile, the acute cytotoxicity of FA and BTLL was also tested by acute toxicological test of luminescent bacteria. The results showed that the consumption of coordinating groups such as phenolic groups and hydrogen bonds reduced the complexation ability of FA and BTLL. The functional groups were degraded with the removal order of quinone group, phenolic group and aromatic group, and finally realized the molecular saturation and aromaticity decrease for humic-like substances. The mobility of FA and BTLL was decreased because of the enhancement of hydrophobicity during electrolysis process. Furthermore, the available chlorine produced during electrochemical oxidation was the main acute cytotoxicity substance, therefore, it is necessary to remove it before discharge in order to reduce ecological risks. This study provides a basis for understanding and evaluating the electrochemical degradation process of humic-like substances in detail., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The joint toxicity of chemicals mixture in the aquatic environment was still not well clear. To clarify the joint toxicity of the mixtures of metals and organic pollutants, as well as the influence of dissolved organic matter (DOM) in field water-body on their toxic effects, we conducted the toxicity tests with cadmium (Cd) and sodium dodecyl benzene sulfonate (SDBS) on Scenedesmus obliquus (S. obliquus) with or without the presence of fulvic acid (FA), a typical of DOM. Our results showed Cd was more toxic to S. obliquus than SDBS, and the effects of fulvic acid on SDBS were greater than Cd. The joint toxicity of Cd and SDBS expressed a synergistic effect on S. obliquus, which was observed to be increased with the presence of FA. Our results gave an example for the joint toxicity investigations of organics and metals, aiding to understanding the toxicity of pollutant mixtures in field water bodies containing DOM., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The importance of attention to unravel the interaction of nano-plastic particles (NPs) with natural acidic organic polymer (NAOP) in freshwater environment should not be neglected. However, toxicological data available for the interaction between NPs and NAOP remain limited. Here, we investigate the toxicological effects of three model polystyrene (PS) NPs with different functional groups (unmodified, amino- and carboxyl-modified PS NPs) on two freshwater organisms of different trophic levels (Scenedesmus obliquus and Danio rerio) in the absence and presence of two classes of NAOP, namely fulvic acid and humic acid. The NAOP interaction with the NPs is shown to alter oxidative stress and disturb membrane function in S. obliquus cells to a certain extent. Combined oxidative stress responses to the NPs and NAOP in D. rerio as a function of their mixture levels showed inhibition, alleviation, and reinforce. Changes in cellular oxidative stress and membrane function depended on the concentration and types of both NPs and NAOP. Furthermore, the characterization parameters of the NPs were important for the explanation of the ecotoxicological mechanism of the NPs in the presence of NAOP. Our findings emphasized the critical role of NAOP in the fate and toxicity of plastic particles in freshwater environment., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study aimed to assess the effect of encapsulating humic acid inside yeast cell walls (YCW) to detoxify AFB 1 in in vitro gastrointestinal models. Glucan Mannan Lipid Particles (GMLPs) from Saccharomyces cerevisiae cell walls showed the highest AFB 1 adsorption in simulated gastric fluid (SGF) after 10 min, and in simulated intestinal fluid (SIF) after 1 h. GMLPs are hollow 3-4 micron porous microspheres that provide an efficient system for the synthesis and encapsulation of AFB 1 -absorbing nanoparticles (NPs). Humic acid nanoparticles (HA-NPs) were synthesized within the GMLP cavity by complexation with ferric chloride. Encapsulating HA-NPs in GMLPs increased HA-NP stability in SIF. The hybrid GMLP HA-NP formulation synergistically enhanced AFB 1 binding compared to individual GMLP and HA components in SGF and in SIF. Cytotoxicity on a murine macrophage cell line demonstrated that GMLP HA-NP-AFB 1 complexes were stable in both SGF and SIF, detoxified AFB 1 and are suitable for in vivo testing., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

It is now widely accepted that coating on the nano-surface would critically dictate the uptake and cytotoxicity of engineering nanomaterials. However, the influence of natural organic matter (NOM) on the surface is quite limited to humic substances, while the diversity of NOM is neglected. In the present study, we tried to investigate the change of surface in the coexistence of bovine serum albumin (BSA) and humic acid (HA). The isothermal titration calorimetric measurements show that HA can combine with BSA in both freshwater or seawater, however, the patterns are different. In freshwater, HA lowered the adsorption of BSA on PVP-capped AgNPs through complexation with BSA, which prevented the contact of sulfur in BSA with PVP-AgNPs. Then in seawater, BSA retained its sulfur availability to bind with PVP-AgNPs. Furthermore, the toxicity of PVP-AgNPs incubated in the BSA/HA solution was evaluated by measuring the level of reactive oxygen species generated by Escherichia coli. The results indicated that, in seawater, the adsorbed BSA promoted the toxicity of PVP-AgNPs in the presence of Ca 2+ and Mg 2+ , but the presence of HA limited this effect., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The risk assessment of nanomaterials is essential for regulatory purposes and for sustainable nanotechnological development. Although the application of graphene oxide has been widely exploited, its environmental risk is not well understood because several environmental conditions can affect its behavior and toxicity. In the present study, the graphene oxide effect from aquatic ecosystems was assessed considering the interaction with humic acid on 9 organisms: Raphidocelis subcapitata (green algae), Lemna minor (aquatic plant), Lactuca sativa (lettuce), Daphnia magna (planktonic microcrustacean), Artemia salina (brine shrimp), Chironomus sancticaroli (Chironomidae), Hydra attenuata (freshwater polyp), and Caenorhabditis elegans and Panagrolaimus sp. (nematodes). The no-observed-effect concentration (NOEC) was calculated for each organism. The different criteria used to calculate NOEC values were transformed and plotted as a log-logistic function. The hypothetical 5 to 50% hazardous concentration values were, respectively, 0.023 (0.005-0.056) and 0.10 (0.031-0.31) mg L -1 for graphene oxide with and without humic acid, respectively. The safest scenario associated with the predicted no-effect concentration values for graphene oxide in the aquatic compartment were estimated as 20 to 100 μg L -1 (in the absence of humic acid) and 5 to 23 μg L -1 (in the presence of humic acid). Finally, the present approach contributed to the risk assessment of graphene oxide-based nanomaterials and the establishment of nano-regulations. Environ Toxicol Chem 2018;37:1998-2012. © 2018 SETAC., (© 2018 SETAC.)

Ozonation is a promising option to treat reverse osmosis concentrate (ROC). However, a systematic understanding and assessment of ozonation on toxicity reduction is insufficient. In this study, ROC sampled from a typical industrial park wastewater treatment plant of China was fractionated into hydrophobic acid (HOA), hydrophobic base (HOB), hydrophobic neutral (HON), and hydrophilic fraction (HI). Systematic bioassays covering bacteria, algae, fish, and human cell lines were conducted to reveal the role of ozonation in toxicity variation of the four ROC fractions. HOA in the raw ROC exhibited the highest toxicity, followed by HON and HI. Ozonation significantly reduced total organic carbon (TOC) and UV 254 values in HOA, HON, and HI and their toxicity except in HOB. Correlation analysis indicated that chemical data (TOC and UV 254 ) of HOA and HON correlated well with their toxicities; however, poor correlations were observed for HOB and HI, suggesting that a battery of toxicity assays is necessary. This study indicates that TOC reduction during ozonation could not fully reflect the toxicity issue, and toxicity assessment is required in conjunction with the chemical data to evaluate the effectiveness of ozonation., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The present study aimed to investigate the influence of different fractions of dissolved organic carbon (DOC) on the uptake and toxicity of nickel (Ni) in the soil invertebrate Enchytraeus crypticus after different exposure times. The addition of DOC as humic acid or fulvic acid significantly reduced Ni uptake by E. crypticus in the soil-solution test system. Median lethal effect concentrations were calculated based on total dissolved Ni concentrations (LC50[Ni]), free Ni ion activity (LC50{Ni 2+ }), and Ni body concentrations (LC50 Body-Ni ). The LC50[Ni] values increased with increasing DOC levels and decreased with exposure time (4, 7, and 10 d). Humic acid exerted a greater protective effect on Ni toxicity than fulvic acid, but the protective effects decreased with prolonged exposure time. The LC50{Ni 2+ } values also decreased with exposure time but were almost constant with variation in DOC levels, indicating that the protective effect of DOC is mainly through complexation with free Ni ions to reduce Ni bioavailability. The LC50 Body-Ni value was independent of DOC concentration and exposure time, with an estimated overall value of 22.1 µg/g dry weight. The present study shows that body concentration could serve as an effective indicator for predicting Ni toxicity with variations in the exposure environment (e.g., DOC) and exposure time. Environ Toxicol Chem 2017;36:3019-3027. © 2017 SETAC., (© 2017 SETAC.)

Biological process treatment of landfill leachate produces a significant amount of sludge, characterized by high levels of organic matter from which humic acids are known to activate several enzymes of energy metabolism, stimulating plant growth. This study aimed to characterize humic acids extracted from landfill sludge and assess the effects on plants exposed to different concentrations (0.5, 1, 2 and 4 mM C L -1 ) by chemical and biological analysis, to elucidate the influence of such organic material and minimize potential risks of using sludge in natura. Landfill humic acids showed high carbon and nitrogen levels, which may represent an important source of nutrients for plants. Biochemical analysis demonstrated an increase of enzyme activity, especially H + -ATPase in 2 mM C L -1 landfill humic acid. Additionally, cytogenetic alterations were observed in meristematic and F 1 cells, through nuclear abnormalities and micronuclei. Multivariate statistical analysis provided integration of physical, chemical and biological data. Despite all the nutritional benefits of humic acids and their activation of plant antioxidant systems, the observed biological effects showed concerning levels of mutagenicity., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Here we link plant source phylogeny to its chemical characteristics and determine parameters useful for predicting DOM phytotoxicity towards algal monocultures. We found that DOM characterised using UV-visible spectroscopic indices and elemental analysis is useful for distinguishing DOM plant sources. Specifically, combined values of absorbance at 440 nm and coefficients for the spectral slope ratio, were used to distinguish between gymnosperm-leached DOM and that from angiosperms. In our bioassays, DOM leached from 4 g leaf L -1 resulted in over 40% inhibition of photosynthetic yield for the cyanobacterium, Cylindrospermopsis raciborskii, for eight of the nine plants tested. Significant variables for predicting inhibition of yield were DOM exposure time and plant source, or using an alternate model, exposure time and spectroscopic and elemental measures. Our study proposes spectroscopic indices which can estimate a plant source's contribution to aquatic DOM, may provide insights into ecological outcomes, such as phytotoxicity to algae. The cyanobacterium (C. raciborskii) was more sensitive to DOM than a green algae (Monoraphidium spp.), as identified in a subsequent dose-response experiment with five different DOM plant sources. Low level additions of angiosperm derived-DOM (i.e. 0.5 g L -1 ) were slight phytotoxic to Monoraphidium spp. causing 30% inhibition of yield, while C. raciborskii was not affected. Higher DOM additions (i.e. 2 g L -1 ) caused 100% inhibition of yield for C. raciborskii, while Monoraphidium spp. inhibition remained under 30%. The divergence in algal sensitivity to DOM indicates that in aquatic systems, DOM derived from catchment vegetation has the potential to affect algal assemblages., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Humic acid (HA) is a major component of dissolved organic matter, is ubiquitous in the aquatic environment and influences the biological toxicity of organic pollutants. In this study, we investigated the cytochrome P450 1A (CYP 1A) mRNA expression and ethoxyresorufin-O-deethylase (EROD) activity in the gills and liver of zebrafish following exposure to the s-triazine herbicide prometryn with or without HA present. Prometryn induced both CYP 1A mRNA expression and EROD activity. The CYP 1A mRNA expression of zebrafish that were exposed to a combination of prometryn and HA was increased compared to those exposed to prometryn alone. A likely cause for CYP 1A induction is the impact of special components of HA, functioning as aryl hydrocarbon receptor (AHR) agonists. In combination with HA, these increase prometryn levels in tissues. Similar results for EROD activity were evident. In our time course study, CYP 1A mRNA expression reached maximum values during 24h. This revealed CYP 1A mRNA transcription as a comparatively sensitive toxicity index. In a recovery experiment, we found a faster decrease of CYP 1A mRNA expression to control levels (CK) in gills compared to liver tissue. Following exposure to HA, CYP 1A mRNA expression in liver tissue displayed a faster decrease to CK levels. HA induced enhanced metabolic rates for prometryn. In contrast, recovery regularity of CYP 1A expression in gills was independent of the presence of HA. This result indicates different detoxification mechanisms for HA in liver and gills., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Increases in dissolved organic carbon (DOC) in the form of humic substances, causing browning of surface water, have been reported worldwide. Field surveys indicate that higher DOC levels can influence primary production and thus plankton composition. Experimental studies on the direct effects of humic DOC on aquatic organisms have shown varying results depending on concentration and additional environmental factors. Moreover, changes in life-histories and stress responses have usually been tested separately, rather than in combination. We experimentally tested the impact of a sudden increase in humic DOC on two species of the zooplankton cladoceran Daphnia, across several levels of biological organisation, from cellular to population responses. In D. magna, strong impacts on reproduction (delayed maturity and reduced number of offspring) were coupled with overall stress induction (increases in antioxidant capacity and oxidative damage, combined with a reduced amount of available energy). In D. longispina, increased mortality and lowered fecundity were observed. We conclude that a strong input of humic DOC into aquatic systems can have severe negative impacts on zooplankton species, and has the potential to alter zooplankton community structures., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Inhibition effect of humic acid (HA) on anaerobic digestion of cellulose and xylan and the mitigation potential of the inhibition were evaluated in controlled fed batch reactors at 30 °C and a hydraulic retention time (HRT) of 20 days. Reactor performances were evaluated by biogas production and metabolite measurements for 220 days. Microbial population dynamics of the reactors were monitored with next-generation 16S rRNA gene sequencing at nine different sampling times. Our results showed that increasing levels of HA inhibited the hydrolysis efficiency of the digestion by 40% and concomitantly reduced the methane yield. Addition of hydrolytic enzymes helped to reverse the negative effects of HA, whereas calcium addition did not reverse HA inhibition. Microbiological analyses showed that the relative abundance of hydrolytic/fermentative bacterial groups such as Clostridiales, Bacteroidales and Anaerolineales was significantly lowered by the presence of HA. HA also affected the archaeal populations. Mostly hydrogenotrophic methanogens were negatively affected by HA. The relative abundance of Methanobacteriaceae, Methanomicrobiales-WCHA208 and Unassigned Thermoplasmata WCHA1-57 were negatively affected by the presence of HA, whereas Methanosaetacea was not affected., Competing Interests: The authors declare that they have no conflict of interest. Ethical approval This article does not contain any studies with human participants or animals, performed by any of the authors.

In this study, we investigated the influence of heavy metals (HM) on total soil bacterial population and its diversity pattern from 10 km distance of a Zinc smelter in Feng County, Qinling Mountain, China. We characterized and identified the bacterial community in a HM polluted soil using 16S rDNA technology. Out results indicated that the maximum soil HM concentration and the minimum bacterial population were observed in S2 soil, whereas bacterial diversity raised with the sampling distance increased. The bacterial communities were dominated by the phyla Proteobacteria, Acidobacteria and Actinobacteria in cornfield soils, except Fimicutes phylum which dominated in hilly area soil. The soil CEC, humic acid (HA)/fulvic acid (FA) and microbial OTUs increased with the sampling distance increased. Shewanella, Halomonas and Escherichia genera were highly tolerant to HM stress in both cultivated and non-cultivated soil. Finally, we found a consistent correlation of bacterial diversity with total HM and SOM along the sampling distance surrounding the zinc smelter, which could provide a new insight into the bacterial community-assisted and phytoremediation of HM contaminated soils.

Triclosan (TCS) is an ionizable synthetic antimicrobial that has been found to be a persistent environmental contaminant with potential for bioaccumulation. Standard laboratory assays have shown that TCS is toxic to aquatic organisms; however, varied environmental conditions could impact this risk. For example, we would predict that sorption to dissolved organic carbon (DOC) in natural surface waters would reduce the bioavailability and, therefore, toxicity of TCS. To better understand the potential risk that TCS poses to wild fish, we evaluated the toxicity of TCS to zebrafish in the presence of DOC. Zebrafish were exposed to TCS (0-900 μg TCS/L), DOC (0-25 mg/L), or TCS (0-900 μg TCS/L) together with either 10 or 25 mg DOC/L from 8 to 120 h postfertilization through static waterborne exposure. We compared impacts of TCS alone or in conjunction with DOC on mortality, development, and hatching success. Exposure to TCS in the presence of DOC improves survival and hatching success, and reduces the incidence of developmental toxicity. However, since the presence of DOC did not completely prevent sublethal toxicity, our data suggest that given its bioaccumulation potential, developmental toxicity of TCS under environmental conditions still warrants concern.

In a previous study, treatment at higher concentrations of arsenic trioxide or co-exposure to arsenic trioxide and humic acid was found to be inhibited cell growth of cervical cancer cells (SiHa cells) by reactive oxygen species generation. However, treatment at lower concentrations slightly increased cell viability. Here, we investigate the enhancement of progression effects of environmentally relevant concentration of humic acid and arsenic trioxide in SiHa cell lines in vitro and in vivo by measuring cell proliferation, migration, invasion, and the carcinogenesis-related protein (MMP-2, MMP-9, and VEGF-A) expressions. SiHa cells treated with low concentrations of humic acid and arsenic trioxide alone or in co-exposure significantly increased reactive oxygen species, glutathione levels, cell proliferation, scratch wound-healing activities, migration abilities, and MMP-2 expression as compared to the untreated control. In vivo the tumor volume of either single drug (humic acid or arsenic trioxide) or combined drug-treated group was significantly larger than that of the control for an additional 45 days after tumor cell injection on the back of NOD/SCID mice. Levels of MMP-2, MMP-9, and VEGF-A, also significantly increased compared to the control. Histopathologic effects of all tumor cells appeared round in cell shape with high mitosis, focal hyperkeratosis and epidermal hyperplasia in the skin, and some tumor growth in the muscle were observed. Our results may indicate that exposure to low concentrations of arsenic trioxide and humic acid is associated with the progression of cervical cancer. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1121-1132, 2016., (© 2015 Wiley Periodicals, Inc.)

The formation of an arsenic (As)-dissolved organic matter (DOM) complex is important in driving the release of arsenic in groundwater. This study collected groundwater samples from a 20 m deep well throughout 2014 and separated each into three subsamples by ultrafiltration: high molecular weight-DOM (HDOM, 0.45 μm-10 kDa), medium molecular weight-DOM (MDOM, 10-1 kDa), and low molecular weight-DOM (LDOM, <1 kDa) solutions. The fractional DOM was measured with a three-dimensional excitation-emission matrix (EEM) via fluorescence spectroscopy. A fluorescence quenching method was used to calculate the apparent stability constant (Ks) between arsenic and the fractional DOM. Based on the EEM records, three fluorescence indicators were further calculated to characterize the DOM sources, including the fluorescence index (FI), the biological index (BI), and the humification index (HI). The experimental results indicated that arsenic in the groundwater was mainly partitioned into the MDOM and LDOM fractions. All fractional DOMs contained humic acid-like substances and were considered as microbial sources. LDOM had the highest humification degree and aromaticity, followed by MDOM and HDOM. The As and DOM association could be formed by a Fe-bridge, which was demonstrated by the Ks values and fourier transform infrared (FTIR) spectra of the DOM. The formation of AsFe-DOM complex was only significant in the MDOM and LDOM., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Recent studies suggest that direct mortality and physiological effects caused by pollutants are major contributing factors to global amphibian decline. However, even sublethal concentrations of pollutants could be harmful if they combined with other factors to cause high mortality in amphibians. Here we show that sublethal concentrations of pollutants can disrupt the ability of amphibian larvae to recognize predators, hence increasing their risk of predation. This effect is indeed much more important since very low amounts of pollutants are ubiquitous, and environmental efforts are mostly directed towards preventing lethal spills. We analyzed the effects of two common contaminants (humic acid and ammonium nitrate) on the ability of tadpoles of the western spadefoot toad (Pelobates cultripes) to recognize chemical cues from a common predator, nymphs of the dragonfly Anax imperator. We compared the swimming activity of tadpoles in the presence and absence of water-borne chemical cues from dragonflies at different concentrations of humic acid and ammonium nitrate. Tadpoles reduced swimming activity in response to predator cues in the absence of pollutants, whereas they remained unresponsive to these cues when either humic acid or ammonium nitrate was added to the water, even at low concentrations. Moreover, changes in tadpole activity associated with the pollutants themselves were non-significant, indicating no toxic effect. Alteration of the natural chemical environment of aquatic systems by pollutants may be an important contributing cause for declines in amphibian populations, even at sublethal concentrations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The stability and bioavailability of titanium dioxide nanoparticles (TiO2 NPs) suspension could be modified by the physicochemical properties of solution. In the present study, the effect of humic acid (HA) and ionic strength (by adding NaCl) on aggregation and sedimentation of TiO2 NPs suspension were investigated. Accordingly, the sublethal toxicity of TiO2 NPs suspensions with different HA and NaCl concentrations toward zebrafish (Danio rerio) was evaluated by monitoring the changes of superoxide dismutase, catalase, malonaldehyde and glutathione in gill, liver and intestine. The results showed that the aggregations formation and hydrodynamic diameter of TiO2 NPs in suspensions are not essential characteristics to decide toxicity. The varied oxidative stress responses detected in gill, liver and intestine derived from different toxicity mechanisms of TiO2 NPs. Nevertheless, the oxidative stress could be suppressed by the adding of HA and/or the increase of ionic strength, which can decrease the bioavailability of TiO2 NPs in water. The study suggests that the environmental factors, such as HA and ionic strength, are important for the fate (aggregation and sedimentation) and toxicity of nanomaterials in aquatic environment.

Natural organic matter (NOM) can interact with engineered nanoparticles (NPs) in the environment and modify their behavior and toxicity to organisms. In the present study, the phytotoxicity of copper oxide (CuO) NPs to rice seedlings in the presence of humic acid as a model NOM was investigated. The results showed that CuO NPs induced the inhibition of root elongation, aberrations in root morphology and ultrastructure, and losses of cell viability and membrane integrity. The adverse effects partly resulted from the generation of reactive oxygen species caused by CuO NPs, which led to lipid peroxidation, mitochondrial dysfunction, and programmed cell death in rice seedlings. However, all the phytotoxicity was alleviated with the addition of humic acid because humic acid coatings on nanoparticle surfaces enhanced electrostatic and steric repulsion between the CuO NPs and the plant cell wall/membrane, reducing contact between NPs and plant and CuO NP-induced oxidative damage to plant cells. The present study's results shed light on the mechanism underlying NP phytotoxicity and highlight the influence of NOM on the bioavailability and toxicity of NPs., (© 2015 SETAC.)

Engineered nanoparticles (NPs) and natural organic matter (NOM) in the environment may interact with background contaminants such as heavy metals and modify their bioavailability and toxicity. In the present study, the combined influences of 2 common NPs (TiO2 and CeO2 ) and humic acid (HA; as a model NOM) on Cu(II) phytotoxicity to rice were investigated by a 3-d root elongation assay performed on filter paper media. The results showed that the adsorption coefficients of bare TiO2 and CeO2 NPs (100 mg/L) toward Cu(2+) are 2.65 and 4.37, respectively, at an initial concentration of 10 mg/L, suggesting that Cu(II) could be strongly adsorbed by NPs, whereas HA-coated TiO2 and CeO2 NPs further enhanced the adsorption coefficients to 4.37 and 6.85, respectively. In addition, compared with Cu-alone treatment, the addition of bare TiO2 and CeO2 NPs (1000 mg/L) increased the length of rice root by 32.5% and 39.0%, respectively; however, the presence of HA-coated TiO2 and CeO2 NPs increased the root length by 90.2% and 100.1%, respectively, which indicated that the mitigation effect of HA-coated NPs on Cu(II) phytotoxicity was more visible than that of bare NPs. The results demonstrated that coexistence of NPs and HA significantly alleviated Cu(II) phytotoxicity as a result of a decrease in bioavailable soluble Cu(II) concentration, which contributes to an understanding of the potential behavior of NPs in the environment., (© 2015 SETAC.)

Humic acid (HA) is a possible etiological factor associated with for several vascular diseases. It is known that vascular risk factors can directly increase the susceptibility to Alzheimer's disease (AD), which is a neurodegenerative disorder due to accumulation of amyloid β (Aβ) peptide in the brain. However, the role that HA contributes to Aβ-induced cytotoxicity has not been demonstrated. In the present study, we demonstrate that HA exhibits a synergistic effect enhancing Aβ-induced cytotoxicity in cultured human SK-N-MC neuronal cells. Furthermore, this deterioration was mediated through the activation of endoplasmic reticulum (ER) stress by stimulating PERK and eIF2α phosphorylation. We also observed HA and Aβ-induced cytotoxicity is associated with mitochondrial dysfunction caused by down-regulation of the Sirt1/PGC1α pathway, while in contrast, treating the cells with the ER stress inhibitor Salubrinal, or over-expression of Sirt1 significantly reduced loss of cell viability by HA and Aβ. Our findings suggest a new mechanism by which HA can deteriorate Aβ-induced cytotoxicity through modulation of ER stress, which may provide significant insights into the pathogenesis of AD co-occurring with vascular injury.

To incorporate metal mixture toxicity effects into risk-assessment procedures, more information is needed about combined and interactive effects of metal mixtures during chronic exposure. The authors investigated the toxicity of binary Ni-Zn mixtures in 2 independent full-factorial experiments using standard chronic (21-d) Daphnia magna reproduction toxicity tests. Global statistical analysis (i.e., when considering all investigated mixture treatments simultaneously) showed noninteractive effects according to the concentration addition model and significant synergistic effects according to the independent action model. However, treatment-specific statistical analysis revealed that both occurrence and type of interactive effect were dependent on the effect size at which Ni and Zn were combined in the mixture. Only noninteractive or weakly antagonistic effects occurred in mixture treatments in which each of the individual metals produced only weak adverse effects on its own (i.e., ≤20% reduction of reproductive performance). On the other side of the spectrum, synergistic mixture effects occurred in all mixture treatments where both metals already caused a  > 20% (for independent action) and a  > 40% (for concentration addition) effect on reproduction on their own. Because low effect sizes are the most relevant in most regulatory frameworks, the authors' data suggest that the concentration addition and independent action mixture toxicity models can both serve as conservative models for predicting effects of Ni-Zn mixtures. The present study highlights the importance of investigating metal mixture toxicity at low effect sizes and warns against extrapolating conclusions about metal mixture interactions from high to low effect sizes., (© 2015 SETAC.)

The treatment of concentrated landfill leachate from membrane process is a troublesome issue due to high concentrations of biorecalcitrant pollutants. In this study, the effect of Fenton process on dissolved organic matter (DOM) distribution (i.e. humic acid (HA), fulvic acid (FA) and hydrophilic fraction (HyI)), chemical forms of toxic organic compounds and metals, and their biotoxicity were investigated. In the concentrated leachate, toluene, ethylbenzene and chlorobenzene predominated in the HyI fraction, while phthalate esters (PAEs) were mainly absorbed on the HA and FA fractions. PAEs were more readily removed from the HA and FA fractions than that from the HyI fraction in the Fenton process. The complexing abilities of DOM varied with types of metal in the concentrated leachate. The biotoxicities of the DOM fractions to luminescent bacteria (Photobacterium phosphoreum T3 mutation) were HA > FA > - HyI. The biotoxicities of the hydrophobic organic contaminants to luminescent bacteria were not obvious in the concentrated leachate due to their low concentrations. Metals might be the main contributor to the biotoxicity to luminous bacteria in the concentrated leachate. These results indicated that Fenton process could influence the pollutants distribution in DOM and their biotoxicities through the breakdown of HA and FA in the concentrated leachate., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Cultivation medium is one of the first aspects to be considered in zooplankton laboratory cultivation. The use of artificial media does not concern to reproduce natural conditions to the cultivations, which may be achieved by using natural organic compounds like humic substances (HS). This study aimed to evaluate the effects of a concentrate of dissolved organic carbon (DOC) from the Negro River (NR(1)) and an extraction of humic acids (HA) from humus produced by Eisenia andrei on the life history traits of laboratory-based Latonopsis australis SARS (1888). A cohort life table approach was used to provide information about the effectiveness of NR and HA as supplements for the artificial cultivation of L. australis. Additionally, we seek to observe a maximization of L. australis artificial cultivation fitness by expanding the range of HS concentrations. The first experiment demonstrated that the females of L. australis reared under NR10 (mgDOCL(-1)) may have experienced an acceleration of the population life cycle, as the females have proportionally reproduced more and lived shorter than controls. By contrast, the use of the HA did not improve life history traits considered. The expansion of the concentration range (5, 10, 20 and 50 mgDOCL(-1)) corroborated the patterns observed on the first assay. Results for the fitness estimates combined with shorter lifespans than controls demonstrated trade-offs between reproductive output and female longevity reared under NR conditions, with NR20 been suggested as the best L. australis cultivation medium. This response might be associated with hormone-like effects., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Humic acid (HA) has been implicated as a contributory factor for blackfoot disease, which is an endemic peripheral vascular disease. We investigated the effect of HA on the regulation of endothelial nitric oxide (NO) synthase (eNOS) in human umbilical vein endothelial cells (HUVECs) to evaluate the involvement of eNOS and related factors in peripheral vascular impairment with HA exposure. Treatment of HUVECs with HA induced upregulation of eNOS. This result coincides with those of previous studies. Furthermore this is the first study to report that HA induces upregulation of heat shock protein (Hsp)90α, Hsp90β, eNOS phosphorylation at Ser1177, and eNOS phosphorylation at Thr495, as compared to that in the control. In contrast, treatment with BAPTA, an intracellular Ca(2+) chelator, inhibited upregulation of these proteins induced by HA. This study demonstrates that HA treatment leads to increases in both Hsp90α and Hsp90β proteins and indicates that Hsp90α leads to eNOS phosphorylation at Ser1177 and that Hsp90β leads to eNOS phosphorylation at Thr495, respectively. Upregulation of eNOS, Hsp90α, and Hsp90β in HUVECs is regulated by intracellular Ca(2+) accumulation induced by HA. These results suggest that upregulation of eNOS phosphorylation at Ser1177 and eNOS phosphorylation at Thr495 produce NO and superoxide anions, respectively, resulting in generation of peroxynitrite, which causes impairment of vascular endothelial cells., (© 2013 Wiley Periodicals, Inc.)

Humic acid (HA) in well water is associated with Blackfoot disease and various cancers. Previously, we reported that acute humic acid exposure (25-200 µg/mL for 24 hr) induces inflammation in RAW264.7 macrophages. In this study, we observed that prolonged (72 hr) HA exposure (25-200 µg/mL) induces cell-cycle arrest and apoptosis in cultured RAW264.7 cells. We also observed that exposing macrophages to HA arrests cells in the G2 /M phase of the cell cycle by reducing cyclin A/B1 , Cdc2, and Cdc25C levels. Treating macrophages with HA triggers a sequence of events characteristic of apoptotic cell death including loss of cell viability, morphological changes, internucleosomal DNA fragmentation, sub-G1 accumulation. Molecular markers of apoptosis associated with mitochondrial dysfunction were similarly observed, including cytochrome c release, caspase-3 or caspase-9 activation, and Bcl-2/Bax dysregulation. In addition to the mitochondrial pathway, HA-induced apoptosis may also be mediated through the death receptor and ER stress pathways, as evidence by induction of Fas, caspase-8, caspase-4, and caspase-12 activity. HA also upregulates p53 expression and causes DNA damage as assessed by the comet assay. These findings yield new insight into the mechanisms by which HA exposure may trigger atherosclerosis through modulation of the macrophage-mediated immune system., (© 2014 Wiley Periodicals, Inc.)

The species-rich, endemic amphipod fauna of Lake Baikal does not overlap with the common Palearctic fauna; however, the underlying mechanisms for this are poorly understood. Considering that Palearctic lakes have a higher relative input of natural organic compounds with a dominance of humic substances (HSs) than Lake Baikal, we addressed the question whether HSs are candidate factors that affect the different species compositions in these water bodies. We hypothesized that interspecies differences in stress defense might reveal that Baikalian amphipods are inferior to Palearctic amphipods in dealing with HS-mediated stress. In this study, two key mechanisms of general stress response were examined: heat-shock protein 70 (HSP70) and multixenobiotic resistance-associated transporters (ABCB1). The results of quantitative polymerase chain reaction (qPCR) showed that the basal levels (in 3-day acclimated animals) of hsp70 and abcb1 transcripts were lower in Baikalian species (Eulimnogammarus cyaneus, Eulimnogammarus verrucosus, Eulimnogammarus vittatus-the most typical littoral species) than in the Palearctic amphipod (Gammarus lacustris-the only Palearctic species distributed in the Baikalian region). In the amphipods, the stress response was induced using HSs at 10 mg L(-1) dissolved organic carbon, which was higher than in sampling sites of the studied species, but well within the range (3-10 mg L(-1)) in the surrounding water bodies populated by G. lacustris. The results of qPCR and western blotting (n = 5) showed that HS exposure led to increased hsp70/abcb1 transcripts and HSP70 protein levels in G. lacustris, whereas these transcript levels remained constant or decreased in the Baikalian species. The decreased level of stress transcripts is probably not able to confer an effective tolerance to Baikalian species against further environmental stressors in conditions with elevated HS levels. Thus, our results suggest a greater robustness of Palearctic amphipods and a higher sensitivity of Baikalian amphipods to HS challenge, which might prevent most endemic species from migrating to habitats outside Lake Baikal.

Metal oxide nanoparticles (NPs) have been used increasingly and are likely to accumulate in natural water bodies, where they encounter and interact with other environmental chemicals. These interactions result in joint effects on biological systems and the environment. However, compared with the intensive research examining the toxicities of individual NPs, the toxicities of NP mixtures remain relatively unexplored. In this work, we studied the joint effects of NPs and their most likely coexisting chemicals in the environment, including NPs with different compositions, humic substances, and surfactants. Our results indicate that the joint effects of NP mixtures were usually simple addition, which is commonly adopted in real risk assessment. However, the joint effects obtained for mixtures that contained ZnO were exclusively associated with antagonism. In addition, the mixtures of NPs and surfactants resulted in complex joint effects, i.e., antagonistic, additive, and synergistic effects were found for the mixtures with ZnO, NiO, and Fe-oxide, respectively. Our study suggests that the assessments of the ecological risk of NPs, particularly ZnO NPs, should consider the impact of their coexisting chemicals in the environment., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Humic acid (HA), a group of high-molecular weight organic compounds characterized by an ability to bind heavy metals, is normally found in natural water. Although the impairment of vascular endothelial cells in the presence of humic substances has been reported to be involved in some diseases, the mechanisms responsible for this involvement remain unclear. In this study, we examined the cytotoxicity of HA obtained from peatland in Central Kalimantan, Indonesia, to human vascular endothelial cells, as well as the mechanisms behind these effects. It was found that 50 mg/L HA showed cytotoxicity, which we considered to be mediated by apoptosis through the mitochondrial pathway because of an increase in the expression of caspases 6 and 9 in response to HA administration. In addition, this cytotoxicity was enhanced when cells in this experimental system were exposed to oxidative stress, while it was decreased by the addition of vitamin C. Thus, we conclude that the apoptosis induced by HA depends upon oxidative stress. Furthermore, an iron chelator, DFO, showed a tendency to decrease HA-induced cytotoxicity, suggesting that iron may potentially mediate HA-induced oxidative stress. In conclusion, long-term consumption of HA-rich water obtained from our study area may cause damage to endothelial cells and subsequent chronic health problems., (Copyright © 2012 Wiley Periodicals, Inc., a Wiley company.)

Using an ion-exchange-based solid-phase microextraction (SPME) method, the freely dissolved concentrations of C12-benzalkonium were measured in different toxicity assays, including 1) immobilization of Daphnia magna in the presence or absence of dissolved humic acid; 2) mortality of Lumbriculus variegatus in the presence or absence of a suspension of Organisation for Economic Co-Operation and Development (OECD) sediment; 3) photosystem II inhibition of green algae Chlorella vulgaris; and 4) viability of in vitro rainbow trout gill cell line (RTgill-W1) in the presence or absence of serum proteins. Furthermore, the loss from chemical adsorption to the different test vessels used in these tests was also determined. The C12-benzalkonium sorption isotherms to the different sorbent phases were established as well. Our results show that the freely dissolved concentration is a better indicator of the actual exposure concentration than the nominal or total concentration in most test assays. Daphnia was the most sensitive species to C12-benzalkonium. The acute Daphnia and Lumbriculus tests both showed no enhanced toxicity from possible ingestion of sorbed C12-benzalkonium in comparison with water-only exposure, which is in accordance with the equilibrium partitioning theory. Moreover, the present study demonstrates that commonly used sorbent phases can strongly affect bioavailability and observed effect concentrations for C12-benzalkonium. Even stronger effects of decreased actual exposure concentrations resulting from sorption to test vessels, cells, and sorbent phases can be expected for more hydrophobic cationic surfactants., (© 2013 SETAC.)

Humic substances (HS) have been shown to decrease the toxicity of environmental stressors, but knowledge of their ability to influence the toxicity of multiple stressors such as metal mixtures and low pH associated with acid mine drainage (AMD) is still limited. The present study investigated the ability of HS to decrease toxicity of AMD to mayflies (Atalophlebia spp.). The AMD was collected from the Mount Morgan (Mount Morgan, Queensland, Australia) open pit. Mayflies were exposed to concentrations of AMD at 0%, 1%, 2%, 3%, and 4% in the presence of 0 mg/L, 10 mg/L, and 20 mg/L Aldrich humic acid (AHA). A U-shaped response was noted in all AHA treatments, with higher rates of mortality recorded in the 2% and 3% dilutions compared with 4%. This result was linked with increased precipitates in the lower concentrations. A follow-up trial showed significantly higher concentrations of precipitates in the 2% and 3% AMD dilutions in the 0 mg/L AHA treatment and higher precipitates in the 2% AMD, 10 mg/L and 20 mg/L AHA, treatments. Humic substances were shown to significantly increase survival of mayflies exposed to AMD by up to 50% in the 20 mg/L AHA treatment. Humic substances may have led to increased survival after AMD exposure through its ability to influence animal physiology and complex heavy metals. These results are valuable in understanding the ability of HS to influence the toxicity of multiple stressors., (© 2013 SETAC.)

High amounts of humic substances (HS) are commonly found in natural acidic waterways and have been suggested to offer some protection against low pH. This study investigated the ability of HS to decrease respiratory stress in eastern rainbowfish (Melanotaenia splendida splendida) exposed to decreases in pH (range of 7-3.5) in soft and hard water. Repeated measures ANOVA revealed a significant difference in respiration (time taken for ten operculum movements) between pH and HS treatments, with a significant interaction between pH and HS present in 5/6 trials. Respiratory stress was shown to increase with increasing acidity, but significantly decreased in treatments with HS (10 and 20 mg/L) compared to those without. The fish exposed to pH treatments without HS also displayed increased hyperactivity, larger operculum movements and increased mucous production. Increased morbidity was shown in HS treatments at pH 3.5 (soft water) and at pH 4 (hard water) compared to treatment without HS. This indicates that HS is helpful in ameliorating the effects of decreased pH on respiration at sublethal pH levels; however, as pH decreases further, it seems that HS increases the toxicity (morbidity) of the low pH.

Because of large usage as flame retardants and additives, organophosphate esters (OPEs) are widely detected in the environment and regarded as emerging contaminants. However, the sorption of OPEs to organic matter and its effects have scarcely been studied. In the present study, the sorption of 9 commonly used OPEs to 4 representative humic acids--Elliott Soil humic acid, Suwannee River humic acid, Aldrich humic acid, and Acros humic acid--in the range of 0 mg/L to 50 mg/L dissolved organic carbon (DOC), was evaluated with negligible-depletion solid-phase microextraction and verified by its impacts on the toxicity to the aquatic invertebrate Daphnia magna. Whereas OPEs with a high octanol/water partition coefficient (log K(OW)=4.51-6.64) were associated with humic acids mainly by hydrophobic interaction with DOC partition coefficient (K(DOC)) in the range of 10²·²² to 10⁵·³¹, the sorption of low-K(OW) OPEs (log K(OW)=-0.65 to 2.59) to humic acids was not hydrophobic interaction-dominant, with K(DOC) in the range of 10³·⁴⁷ to 10⁴·²⁹. These results were corroborated by the effects of humic acids on the acute toxicity of 3 high-K(OW) OPEs to D. magna. The sorption of OPEs to Suwannee River humic acid was weak and had negligible effects on the toxicity of high-K(OW) OPEs; the presence of terrestrial Acros humic acid (50 mg/L DOC), however, significantly decreased the toxicity by 53% to 60%. The results indicated that the strong sorption between high-K(OW) OPEs and terrestrial humic acid might affect their transportation and bioavailability., (© 2013 SETAC.)

This work investigated the application of a solar driven advanced oxidation process (solar Fenton), for the degradation of the antibiotic ofloxacin (OFX) in various environmental matrices at a pilot-scale. All experiments were carried out in a compound parabolic collector pilot plant in the presence of doses of H2O2 (2.5 mg L(-1)) and at an initial Fe(2+) concentration of 2 mg L(-1). The water matrices used for the solar Fenton experiments were: demineralized water (DW), simulated natural freshwater (SW), simulated effluent from municipal wastewater treatment plant (SWW) and pre-treated real effluent from municipal wastewater treatment plant (RE) to which OFX had been spiked at 10 mg L(-1). Dissolved organic carbon removal was found to be dependent on the chemical composition of the water matrix. OFX mineralization was higher in DW (78.1%) than in SW (58.3%) at 12 mg L(-1) of H2O2 consumption, implying the complexation of iron or the scavenging of hydroxyl radicals by the inorganic ions present in SW. On the other hand, the presence of dissolved organic matter (DOM) in SWW and RE, led to lower mineralization per dose of H2O2 compared to DW and SW. The major transformation products (TPs) formed during the solar Fenton treatment of OFX, were elucidated using liquid chromatography-time of flight-mass spectrometry (LC-ToF-MS). The transformation of OFX proceeded through a defluorination reaction, accompanied by some degree of piperazine and quinolone substituent transformation while a hydroxylation mechanism occurred by attack of the hydroxyl radicals generated during the process leading to the formation of TPs in all the water matrices, seven of which were tentatively identified. The results obtained from the toxicity bioassays indicated that the toxicity originates from the DOM present in RE and its oxidation products formed during the photocatalytic treatment and not from the TPs resulted from the oxidation of OFX., (Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.)

Pesticides are a major stressor for stream ecosystem health. They enter surface waters from diffuse agricultural sources but also from point sources such as municipal wastewater treatment plants (WWTPs). However, to date, no studies have focused on the ecological effects of pesticide-contaminated WWTP effluent on macroinvertebrate communities. On the basis of governmental monitoring data of 328 sites in Hesse, Germany, we identified insecticidal long-term effects on the structure of the macroinvertebrate community up to 3 km downstream of WWTPs. The effects were quantified using the trait-based SPEAR(pesticides) index, which has been shown to be an effective tool for identifying community effects of pesticide contamination. In addition, based on the German Saprobic Index, we revealed that WWTPs are still an important source of oxygen-depleting organic pollution, despite the extensive technological improvements in wastewater management over several centuries. In general, our findings emphasize the need to take municipal WWTPs into consideration in the management of river basins under the EU Water Framework Directive to achieve good ecological and chemical status for European streams and rivers., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

For long, natural organic matter (NOM) composed mainly of humic substances (HS) were regarded as inert in the ecosystems with respect to their possible chemical interaction with exposed organisms. However recently, NOM have been shown to elicit various adverse effects generally attributed to synthetic xenobiotics, including estrogenic effects translating into shifts of the gender ratios in populations. However, the anti/androgenic pathway was not yet evaluated. Here, we applied an anti/androgenic sensitive cell line MDA stably transfected with the firefly luciferase gene under transcriptional control of the androgen responsive element. With five out of twelve tested NOM preparations, particularly with soil and coal isolates, we identified a relatively high, concentration-dependent antiandrogenic effect. This appears to be the first study to show this endocrine disrupting pathway for a ubiquitous biogeochemical matrix, a potential activity which should not be neglected in forthcoming studies on synthetic endocrine disruptors in the environment., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Dissolved organic matter (DOM) has been shown to affect phytoplankton species directly. These interactions largely depend on the origin and molecular size of DOM and are different in prokaryotes and eukaryotes. In a preceding study, however, two humic substance preparations did not adversely affect coccal green algae or cyanobacterial growth even at high concentrations of dissolved organic carbon (DOC). These results contradicted previous findings, showing a clear, negative response of different phototrophs to much lower DOC concentrations. To test whether or not at least defined building blocks of humic substances (HSs) are effective algicidal structures, we enriched two humic preparations with hydroquinone and p-benzoquinone, respectively, and exposed two different green algae, Pseudokirchneriella subcapitata and Monoraphidium braunii, and two cyanobacterial species, Synechocystis sp. and Microcystis aeruginosa, to the unmodified and enriched HSs. As response variables, growth rates in terms of biomass increase, chlorophyll-a content, and photosynthetic yield were measured. The highest concentration (4.17 mM DOC) of the modified HSs clearly inhibited growth; the cyanobacterial species were much more sensitive than the green algal species. However, realistic ecological concentrations did not adversely affect growth. Aerating the exposure solution for 24 h strongly reduced the inhibitory effect of the modified HSs. The algicidal effect was obviously caused by monomers and not by polymerised high molecular weight HSs themselves. Furthermore, the maximum quantum yield (Φ PSII max) was stimulated in the green algal species by low and medium DOC concentrations, but reduced in the cyanobacterial species upon exposure to higher HS concentrations. The quinone- and phenol-enriched HSs only showed algicidal activity at high concentrations of 4.17 mM DOC and lost their effects over time, presumably by oxidation and subsequent polymerisation. This study confirms that the applied humic substances themselves are not effective algicides even if enriched in effective structures., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background, Aim and Scope: There is growing evidence to show that dissolved humic substances, HSs, can directly interact with freshwater organisms, such as phototrophic organisms, cladocerans, amphipods and fish. The responses are-at least in part-transcriptionally controlled. These interactions can lead to stress symptoms in the exposed organisms. In phototrophs, stress symptoms include a reduction in photosynthetic oxygen release and antioxidative stress. Besides the direct effects, HSs also cause indirect effects that provoke different physiological adaptations in the phototrophs., Materials and Methods: The HS-influenced photosynthetic performance and stress response of two different green algae, Pseudokirchneriella subcapitata (Koršikov) Hindák and Monoraphidium braunii (Nägeli in Kützing) Komárková-Legnerová, and two cyanobacterial species, Synechocystis sp. (PCC 6803, Institut Pasteur) and Microcystis aeruginosa (PCC 7806, Institut Pasteur), were tested. Two humic preparations were applied, the synthetic HS1500 and HuminFeed, HF, which had previously been proven effective in bioassays with invertebrates and a water mould., Results and Discussion: When the algae were grown near light saturation, most of the tested species were positively affected by HSs in growth rate or chlorophyll content. Cell sizes decreased with increasing HS concentrations for all eukaryotic phototrophs, except for the cyanobacteria. After 4 to 5 days of cultivation at the highest HS exposure, there was a decrease in total dry weight due to reduced cell sizes in contrast to an increase in cell numbers. With the exception of Synechocystis, the dry weight per cell ratio decreased with increasing HS concentration. The efficiency of utilizing absorbed light quanta increased with increasing HS concentrations; the maximum quantum yield of photosystem II (ΦPSIImax) was higher in all of the tested species, with the exception of M. aeruginosa, after exposure to HS., Conclusion: The applied humic preparations did not interact directly with PSII, but changed the physiological state of the algae, especially the photosynthetic performance. Neither the green algae nor the cyanobacteria were inhibited in growth or negatively affected in their photosynthetic performance. The exposure to lower concentrations of HS stimulated better growth of the phototrophs. The tested humic preparations obviously did not have the potency to act as xenobiotic stressors; furthermore, there was no sign of herbicide potency.