Graphene oxide induces cytotoxicity and oxidative stress in bluegill sunfish cells.

Graphene oxide (GO) is considered a promising material for biological application due to its unique properties. However, the potential toxicity of GO to aquatic organism particularly bluegill sun fish cells (BF-2) is unexplored or remains poorly understood. GO-induced cytotoxicity and oxidative stress in BF-2 cells were assessed using a battery of biomarkers. Two different biological assays (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red uptake were used to evaluate the cytotoxicity of GO on BF-2 cells. It was found that GO induced dose- and time-dependent cytotoxicity on BF-2 cells. BF-2 cells exposed to lower concentration of GO (40 µg ml-1 ) for 24 induced morphological changes when compared to their respective controls. As evidence for oxidative stress lipid peroxidation, superoxide dismutase, catalase, reactive oxygen species and 8-hydroxy-2'-deoxyguanosine levels were increased and glutathione levels were found to decline in BF-2 cells after treatment with GO. Our findings demonstrate that GO when exposed to BF-2 fish cells cause oxidative stress.

Aluminium oxide nanoparticles induced morphological changes, cytotoxicity and oxidative stress in Chinook salmon (CHSE-214) cells.

Aluminium oxide nanoparticles (Al2 O3 NPs) are increasingly used in diverse applications that has raised concern about their safety. Recent studies suggested that Al2 O3 NPs induced oxidative stress may be the cause of toxicity in algae, Ceriodaphnia dubia, Caenorhabditis elegans and Danio rerio. However, there is paucity on the toxicity of Al2 O3 NPs on fish cell lines. The current study was aimed to investigate Al2 O3 NPs induced cytotoxicity, oxidative stress and morphological abnormality of Chinnok salmon cells (CHSE-214). A dose-dependent decline in cell viability was observed in CHSE-214 cells exposed to Al2 O3 NPs. Oxidative stress induced by Al2 O3 NPs in CHSE-214 cells has resulted in the significant reduction of superoxide dismutase, catalase and glutathione in a dose-dependent manner. However, a significant increase in glutathione sulfo-transferase and lipid peroxidation was observed in CHSE-214 cells exposed to Al2 O3 NPs in a dose-dependent manner. Significant morphological changes in CHSE-214 cells were observed when exposed to Al2 O3 NPs at 6, 12 and 24 h. The cells started to detach and appear spherical at 6 h followed by loss of cellular contents resulting in the shrinking of the cells. At 24 h, the cells started to disintegrate and resulted in cell death. Our data demonstrate that Al2 O3 NPs induce cytotoxicity and oxidative stress in a dose-dependent manner in CHSE-214 cells. Thus, our current work may serve as a base-line study for future evaluation of toxicity studies using CHSE-214 cells.

Metal(oid)s in plastic debris, with distinct features, from Spanish Mediterranean beaches with different anthropogenic pressure: Are these particles potential monitors for metal pollution?

Metal(oid)s concentrations have been quantified in plastic pieces collected from four beaches located in the Mediterranean coast of Spain with different characteristics (i.e. anthropogenic pressure, zone). Metal(oid)s content was also related to selected plastic criteria (i.e. color, degradation status, polymer). The selected elements were quantified with mean concentrations in the sampled plastics with the following order: Fe > Mg > Zn > Mn > Pb > Sr > As > Cu > Cr > Ni > Cd > Co. Moreover, black, brown, PUR, PS, and coastal line plastics concentrated the higher metal(oid)s levels. Local of sampling (influence of mining exploitation) and severe degradation were key factors for uptake of metal(oid)s from water by plastics as modification of surfaces strengths their adsorption capacity. Determined high levels of Fe, Pb and Zn in plastics reflected the pollution degree of the marine areas. Therefore, this study is a contribution for the potential use of plastics as pollution monitors.

Mercury-induced chromosomal damage in wild fish (Dicentrarchus labrax L.) reflecting aquatic contamination in contrasting seasons.

Ria de Aveiro (mainly Laranjo basin, Portugal) has been subjected to mercury contamination from a chlor-alkali plant, currently presenting a well-described mercury gradient. This study aimed to assess mercury genotoxicity in this area by measuring the frequency of erythrocytic nuclear abnormalities (ENA) in the European sea bass (Dicentrarchus labrax), addressing the relation with total mercury concentration in the blood and the modulatory role of seasonal variables. Fish were collected, in warm and cold periods, at three locations differing in their distances to the main mercury source: reference (R), moderately (M), and highly (H) contaminated sites. Genotoxicity was detected in both degrees of contamination (M and H) and in both periods of the year (warm and cold), which is in line with the greater levels of mercury measured in fish blood. No significant seasonal variations were observed for mercury bioaccumulation or ENA frequency. The apparent low imperviousness of ENA frequency to seasonal factors reinforced its consistency as a genotoxicity biomarker, thus enabling a clearer identification of cause-and-effect relationships. Overall, the results reflected a serious environmental risk to native ichthyofauna at Laranjo basin due to mercury contamination, showing a potential of mercury to induce genetic damage in fish blood cells through clastogenic and/or aneugenic actions.

Sources of potentially toxic elements and organic pollutants in an urban area subjected to an industrial impact.

Urban and industrial development has caused a major impact on environmental soil quality. This work assesses the extent and severity of contamination in a small urban area subjected to an industrial impact and identifies the major anthropogenic inputs. Twenty-six soil samples were collected from agricultural and urban sites, and concentrations of potentially toxic elements (As, Cd, Cu, Cr, Fe, Mn, Ni, Pb and Zn), PAHs and PCBs, were determined. In spite of the low median concentrations observed, some sites represent a potential hazard for human health and ecosystems. Concentrations of contaminants were higher than those found in a nearby city, indicating that the study area is affected by the surrounding industry. The use of multivariate statistical analyses allowed for the identification of the main factors controlling the variability of potentially toxic elements and organic pollutants in the soils. The presence of Cr, Fe, Mn and Ni was associated with geogenic inputs, and Cu, Pb, Zn, As, PAHs and PCBs were associated with anthropogenic inputs. Industry and traffic were the most important anthropogenic sources. Soil characteristics were identified as important factors controlling the spatial variability of elements, both from recognised natural and anthropogenic origin. Differences between land uses were observed, which may be attributed to both management practices and proximity to sources.

Evaluation of tertiary treatment by fungi, enzymatic and photo-Fenton oxidation on the removal of phenols from a kraft pulp mill effluent: a comparative study.

Pulp and paper mills generate pollutants associated to their effluents depending upon the type of process, type of the wood materials, process technology applied, management practices, internal recirculation of the effluent for recovery, the amount of water used in the industrial process and type of secondary treatment. This study is the first that reports a simultaneous evaluation of the effects of tertiary treatments by fungi (Rhizopus oryzae and Pleurotus sajor caju), by enzyme (laccase) and by an oxidation process (photo-Fenton) on individual phenols (vanillin, guaiacol, phloroglucinol, vanillic acid and syringic acid) of a Eucalyptus globulus bleached kraft pulp and paper mill final effluent after secondary treatment (BKPME). The tertiary treatments were applied on BKPME samples and in BKPME samples supplemented with extra concentration of each phenol. Tertiary treatments by Rhizopus oryzae and photo-Fenton oxidation were able of complete removal (100%) of phenols on BKPME samples whereas P. sajor caju and laccase were able of 60-85% removal. On BKPME samples with added concentration of each phenol, photo-Fenton was the only treatment capable of total phenols removal (100%), which suggests a great potential for its application.

Mercury organotropism in feral European sea bass (Dicentrarchus labrax).

The knowledge of mercury (Hg) burdens in a wide set of tissues and organs of exposed fish is crucial to understand the internal distribution dynamics and thus predict Hg bioavailability and implications for ecosystem and human health. Total Hg was measured in six tissues of Dicentrarchus labrax captured along an estuarine contamination gradient, revealing the following pattern: liver > kidney > muscle > brain ≈ gills > blood. All of the tissues displayed intersite differences, although brain and muscle seemed to better reflect the extent of contamination. Hg speciation showed that liver presented higher concentrations than muscle for both organic and inorganic forms. Furthermore, liver seemed to exert a protective action in relation to Hg accumulation in the other tissues and organs. This protection seems to be particularly marked in relation to the brain, whereas liver is assisted in that action by kidney and muscle.

Optical fiber based methodology for assessment of thiocyanate in seawater.

A new methodology for the assessment of thiocyanate (SCN(-)) is proposed based on optical fiber (OF) detection coupled to a liquid chromatography system (LC). The developed methodology showed an adequate performance for the analysis of SCN(-) comparable to a high performance liquid chromatography with UV detector (HPLC-UV) methodology: a detection limit of 3 µg L(-1), a linear range from 4 to 400 µg L(-1), and an analytical time of less than 6 min. The OF based methodology was of compact design and easy operation. This simple system has the potential to be used as a sensing approach for SCN(-) in seawater.

Mercury distribution in key tissues of fish (Liza aurata) inhabiting a contaminated estuary-implications for human and ecosystem health risk assessment.

This study brings a new viewpoint based on multiple-tissue analyses to form the basis for a predictive mode of mercury accumulation dynamics in fish body under field conditions. Total mercury (T-Hg) was determined in key tissues of Liza aurata captured along an estuarine contamination gradient, displaying the following hierarchy: kidney > liver > muscle > brain > gills > blood. Brain was the tissue that better reflected the mercury contamination extent, closely followed by liver and muscle. Organic mercury (O-Hg) measured in muscle and liver represented more than 85% and less than 30% of the T-Hg, respectively. The lowest O-Hg percentage was found in the most contaminated area, for both muscle and liver. Mercury distribution and accumulation patterns showed dependence on the specific tissue. The high mercury levels found in organs involved in vital physiological processes point out the risk to autochthonous fish fauna. Human risk associated to the ingestion of fish living in the surveyed areas cannot be excluded.

Source and pathway analysis of lead and polycyclic aromatic hydrocarbons in Lisbon urban soils.

One hundred soil samples were collected from urban spaces, in Lisbon, Portugal, in two surveys that were carried out in consecutive years, to assess the potential adverse human health effects following exposure to potentially toxic elements and organic compounds in the urban soils. The study hereby described follows on from the earlier work of the authors and aims at performing a source-pathway-fate analysis of lead (Pb) and polycyclic aromatic hydrocarbons (PAHs) in the urban soils in order to increase current knowledge on factors influencing exposure of the population. Various techniques were combined to achieve the proposed goal. Geogenic and anthropogenic sources were apportioned by means of Pb isotope mixing models. Isotope data was further coupled with geographic information system mapping to assess local mixed sources of Pb and PAHs. Unleaded vehicle exhaust and cement production show the largest relative contribution to the total soil-Pb, but their respective importance depends on factors such as location and urban landscape. The primary sources of PAHs to the urban soils are probably air and land traffic. Multivariate analysis was used to investigate which soil properties could influence mobility and fate of the contaminants. Whilst principal components analysis indicates carbonates and other calcium phases as probable factors controlling the dispersion of Pb in the urban soils, the linear models obtained from stepwise multiple regression analysis show that soil phosphorous (P) and manganese (Mn) are good predictors of the total soil Pb content. No robust model was obtained for the PAHs, impeding identifying environmental factors most likely to influence their dispersion in the urban soils. The solid-phase distribution study provided critical information to untangle the, at a first glance, contradictory results obtained by the multivariate analysis. Carbonates and other calcium phases, having these a probable anthropogenic origin, are soil components containing major fractions of Pb, P, and Mn.

Changes in zooplankton communities along a mercury contamination gradient in a coastal lagoon (Ria de Aveiro, Portugal).

The main objective of this paper was to evaluate the impact of mercury on the zooplankton communities' structure and functioning and their bioaccumulation patterns along a contamination gradient in a temperate coastal lagoon. Our results demonstrated that total abundance was not negatively affected by Hg contamination, since the most contaminated areas presented the highest values, being the copepod Acartia tonsa the dominant species, which means that it is a very well adapted and tolerant species to mercury. Nevertheless, negative effects were observed in terms of species diversity, since the most contaminated areas presented the lowest values of species richness, evenness and heterogeneity. Moreover, the spatial mercury gradient was reflected on the bioaccumulation patterns of the zooplankton communities. This reinforces the idea that zooplankton can be considered as an important vehicle of mercury transfer through the food pelagic web since it constitutes a primordial food resource for several commercial fish species.

Gas chromatography - optical fiber detector for assessment of fatty acids in urban soils.

Fatty acids have been used as biomarkers of the microbial community composition of soils and they are usually separated and quantified by gas-chromatography coupled to a flame ionization detector (GC-FID). The aim of this study was to develop, validate and apply a methodology based on gas chromatography coupled to optical fiber detection (GC-OF) for screening five fatty acids used as indicators of fungal and bacterial communities in urban soils. The performance of the GC-OF methodology (optical fiber detector at 1,550 nm) was evaluated by comparison with the GC-FID methodology and it was found that they were comparable in terms of linear range, detection limit and analytical errors. Besides these similar analytical characteristics, the GC-OF is much cheaper than the GC-FID methodology. Different concentrations were determined for each fatty acid indicator which in turn varied significantly between the soil samples analyzed from Lisbon ornamental gardens. Additionally, the GC-OF showed a great potential as alternative for determination of eleven or more fatty acids in urban soils.

Assessment of fatty acid as a differentiator of usages of urban soils.

The determination of fatty acids (FA) has been extensively used as a sensitive and reproducible parameter for characterizing the soil microorganism communities and to detect various environmental stresses. The aim of this study was to assess the variability of FA in urban soils, in an attempt to use it as a differentiator of urban soil usage. FA were extracted from soils of five different usages (ornamental gardens, ornamental gardens/roadsides, roadsides, parks and urban agricultural areas) in three Portuguese cities and FA concentration was determined by gas chromatography coupled to mass spectrometry (GC-MS). A total of fifteen FA have been detected and the concentration of each FA, in general, decreased in the following order: Lisbon>Estarreja>Viseu, for all the usages of urban soil. According to soil microbial indicators, the concentration of Gram-negative bacteria was lower than the concentration of Gram-positive bacteria, which together were, in turn, higher than the concentration of fungi for all the usages of urban soil and city. This study assessed the FA profiles of urban soils, which differ as a function of soil usage. The FA profile also is at the source of the inference that stress in soil microorganism communities results from the different urban environment in each city.

Degradation of phenols in olive oil mill wastewater by biological, enzymatic, and photo-Fenton oxidation.

BACKGROUND, AIM, AND SCOPE: Olive oil mill wastewater (OOMW) environmental impacts minimization have been attempted by developing more effective processes, but no chemical or biological treatments were found to be totally effective to mitigate their impact on receiving systems. This work is the first that reports simultaneously the efficiency of three different approaches: biological treatment by two fungal species (Trametes versicolor or Pleurotus sajor caju), enzymatic treatment by laccase, and chemical treatment by photo-Fenton oxidation on phenols removal. MATERIALS AND METHODS: Those treatments were performed on OOMW with or without phenol supplement (p-coumaric, vanillin, guaiacol, vanillic acid, or tyrosol). OOMW samples resulted from treatments were extracted for phenols using liquid-liquid extraction and analyzed by gas chromatography coupled to mass spectrometry. RESULTS: Treatment with T. versicolor or P. sajor caju were able to remove between 22% and 74% and between 8% and 76% of phenols, respectively. Treatment by laccase was able to reduce 4% to 70% of phenols whereas treatment by photo-Fenton oxidation was responsible for 100% phenols reduction. DISCUSSION: Range of phenol degradation was equivalent between T. versicolor, P. sajor caju and laccase for p-coumaric, guaiacol, caffeic acid, and tyrosol in supplemented OOMW, which enhances this enzyme role in the biological treatment promoted by these two species. CONCLUSIONS: Phenols were removed more efficiently by photo-Fenton treatment than by biological or enzymatic treatments. RECOMMENDATIONS AND PERSPECTIVES: Use of fungi, laccase, or photo-Fenton presents great potential for removing phenols from OOMW. This should be further assessed by increasing the application scale and the reactor configurations effect on the performance, besides a toxicity evaluation of treated wastewater in comparison to raw wastewater.

Effects of tertiary treatment by fungi on organic compounds in a kraft pulp mill effluent.

BACKGROUND, AIM AND SCOPE: Pulp and paper mills generate a plethora of pollutants depending upon the type of pulping process. Efforts to mitigate the environmental impact of such effluents have been made by developing more effective biological treatment systems in terms of biochemical oxygen demand, chemical oxygen demand, colour and lignin content. This study is the first that reports an evaluation of the effects of a tertiary treatment by fungi (Pleurotus sajor caju, Trametes versicolor and Phanerochaete chrysosporium and Rhizopus oryzae) on individual organic compounds of a Eucalyptus globulus bleached kraft pulp and paper mill final effluent after secondary treatment (final effluent). MATERIAL AND METHODS: The tertiary treatment with P. sajor caju, T. versicolor and P. chrysosporium and R. oryzae was performed in batch reactors, which were inoculated with separate fungi species and monitored throughout the incubation period. Samples from effluent after secondary and after tertiary treatment with fungi were analysed for both absorbance and organic compounds. The samples were extracted for organic compounds using solid-phase extraction (SPE) and analysed by gas chromatography-mass spectrometry (GC/MS). The efficiencies of the SPE procedure was evaluated by recovery tests. RESULTS: A total of 38 compounds (carboxylic acids, fatty alcohols, phenolic compounds and sterols) were identified and quantified in the E. globulus bleached kraft pulp mill final effluent after secondary treatment. Recoveries from the extraction procedure were between 98.2% and 99.9%. The four fungi species showed an adequate capacity to remove organic compounds and colour. Tertiary treatment with R. oryzae was able to remove 99% of organic compounds and to reduce absorbance on 47% (270 nm) and 74% (465 nm). P. sajor caju, T. versicolor and P. chrysosporium were able to remove 97%, 92% and 99% of organic compounds, respectively, and reduce 18% (270 nm) to 77% (465 nm), 39% (270 nm) to 58% (465 nm) and 31% (270 nm) to 10% (465 nm) of absorbance, respectively. DISCUSSION: The wide variety of organic compounds found in the final effluent must be due to the degradation of E. globulus wood in pulp and paper mill. The concentrations of organic compounds in the final effluent of E. globulus bleached kraft pulp mill were in residual levels maybe due to the secondary treatment. The recovery tests showed the effectiveness of the extraction procedure, and no losses of analyte were suspected in the analytical determinations. Lignin derivatives such as vanilic acid, syringic acid, guaiacol, syringol and phloroglucinol were totally removed by R. oryzae, but the 47% absorbance reduction obtained at 270 nm suggests that these species were not able to complete degradation of lignin macromolecular compounds. CONCLUSIONS: The organic compounds (carboxylic acids, fatty alcohols, phenolic compounds and sterols) were removed more efficiently by tertiary treatment with R. oryzae or P. chrysosporium, followed by P. sajor caju and T. versicolor. Regarding the removal of both colour and organic compounds, the tertiary treatment with R. oryzae was the most efficient. RECOMMENDATIONS AND PERSPECTIVES: In order to reduce the deleterious impacts of paper mill effluents, efforts have been made to develop more effective advanced tertiary treatments. This study may serve as a basis of characterisation, in terms of organic compounds of E. globulus bleached kraft pulp mill final effluent after secondary treatment and as an effort to understand the effects of tertiary treatments with fungi on low concentrations of organic compounds from biological secondary treatment.