Assessment of typical pollutants in waterborne by combining active biomonitoring and integrated biomarkers response

Organic pollutants, heavy metals and pharmaceuticals are continuously dispersed into the environment and have become a relevant environmental emerging concern. In this study, a situ assay to assess ecotoxicity of mixed pollutants was carried out in three typical sites with different priority contaminations in Guangzhou, China. Chemical analysis of organic pollutants, metals and quinolones in three exposure sites were determined by GC-ECD/MS, ICP-AES and HPLC, as well as, a combination of biomarkers including: ethoxyresorufin O-deethylase (EROD); aminopyrine N-demethylase (APND); erythromycin N-demethylase (ERND); glutathione S-transferase (GST); malondialdehyde (MDA); CYP1A; and P-glycoprotein (P-gp) mRNA expressions were evaluated in Mugilogobius abei. Results of chemical analysis in sediment samples revealed that the dominant chemicals were organic pollutants and heavy metals in Huadi River while quinolones in the pond. Bioassays indicated that differences among sites were in relation to some specific biomarkers. EROD and GST activities significantly increased after 72 h in situ exposure, but no difference was observed among the exposure sites. APND, ERND and MDA exhibited dissimilar change patterns for different priority pollutants. CYP1A and P-gp mRNA expressions were significantly induced at all exposure sites, whilst P-gp activity was typical for S2 with the highest levels of quinolones. The molecular biomarkers seemed to be more susceptible than enzyme activities. These assays confirmed the usefulness of applying a large array of various combined biomarkers at different levels, in assessing the toxic effects of mixed pollutants in a natural aquatic environment.     

Differential expression profile of membrane proteins in Aplysia pleural–pedal ganglia under the stress of methyl parathion

This study was aimed to analyze the alteration of membrane protein profiles in Aplysia juliana Quoy & Gaimard (A. juliana) pleural–pedal ganglia under MP exposure. Both the results of GC–MS analysis and the activity assay of acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT) reveal that MP toxicological effects on Aplysia left and right pleural–pedal ganglia are different under 7 and 14 days of exposure. Therefore, Aplysia were subjected for exposure at two concentrations (1 and 2 mg/l) of MP for 7 and 14 days for membrane proteomic study. As a result, 19 and 14 protein spots were differentially expressed in A. juliana left pleural–pedal ganglia under 7 and 14 days treatment, and 20 and 14 protein spots found with differential expressions in their right ganglia under the same treatment, respectively. Several proteins with expression variations were detected from both the left and right pleural–pedal ganglia; however, most proteins have distinctive expressions, indicating different mechanisms might be involved in initiating MP toxicology in left and right ganglia. Among the total differential protein spots obtained, 29 proteins were classed as membrane proteins. These proteins are mainly involved in the metabolism process, cell redox homeostasis, signal transduction, immunology, intracellular transport and catalysis, indicating MP toxicity in mollusks seems to be complex and diverse. Some differentially expressed proteins were further confirmed by Western blotting and quantitative real-time PCR. These results might provide renovated insights to reveal the mechanism of MP-induced neurotoxicity, and the novel candidate biomarkers might have potential application for environmental evaluation of MP pollution level.     

Physiological and molecular responses of the earthworm <Emphasis Type="Italic">Eisenia fetida</Emphasis> to polychlorinated biphenyl contamination in soil

Polychlorinated biphenyls (PCBs) are a class of man-made organic compounds ubiquitously present in the biosphere. In this study, we evaluated the toxic effects of different concentrations of PCBs in two natural soils (i.e. red soil and fluvo-aquic soil) on the earthworm Eisenia fetida. The parameters investigated included anti-oxidative response, genotoxic potential, weight variation and biochemical responses of the earthworm exposed to two different types of soils spiked with PCBs after 7 or 14 days of exposure. Earthworms had significantly lower weights in both soils after PCB exposure. PCBs significantly increased catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (POD) activity in earthworms exposed to either soil type for 7 or 14 days and decreased the malondialdehyde (MDA) content in earthworms exposed to red soil for 14 days. Of the enzymes examined, SOD activity was the most sensitive to PCB stress. In addition, PCB exposure triggered dose-dependent coelomocyte DNA damage, even at the lowest concentration tested. This response was relatively stable between different soils. Three-way analysis of variance (ANOVA) showed that the weight variation, anti-oxidant enzyme activities, and MDA contents were significantly correlated with exposure concentration or exposure duration (P < 0.01). Furthermore, weight variation, CAT activity, and SOD activity were significantly affected by soil type (P < 0.01). Therefore, the soil type and exposure time influence the toxic effects of PCBs, and these factors should be considered when selecting responsive biomarkers.     

Xanthoria parietina as a monitor of ground-level ambient ammonia concentrations

Total nitrogen in transplanted and in situ lichen thalli of Xanthoria parietina were related to ambient ammonia air concentrations measured with passive ALPHA (Adapted Low-cost Passive High Absorption) diffusion samplers in Denmark. Transplants and ALPHA samplers were exposed four months in a transect on heathland close to a pig farm. Monthly mean ammonia concentrations declined exponentially approaching background levels after 300 m. Nitrogen content of the lichen transplants tended to decline with distance, though only a few stations were significantly different from each other. Where ammonia concentrations were high, maximum content of nitrogen was reached after one month of exposure. Conversely, at sites with background concentrations, it took several months to reach a statistically significant uptake. The correlation between ammonia concentration in the air and in situ X. parietina was significant.     

Effects of metal pollution on earthworm communities in a contaminated floodplain area: Linking biomarker, community and functional responses

Effects on earthworms in the contaminated floodplain area the Biesbosch, the Netherlands, were determined at different levels of organization using a combination of field and laboratory tests. The species Lumbricus rubellus, collected from different polluted sites in the Biesbosch, showed reduced values for the biomarker neutral red retention time (NRRT), mainly explained by high metal concentrations in the soil and the resulting high internal copper concentrations in the earthworms. Organic pollutant levels in earthworms were low and did not explain reduced NRRTs. Earthworm abundance and biomass were not correlated with pollutant levels in the soil. Litterbag decomposition and bait-lamina feeding activity, measures of the functional role of earthworms, were not affected by metal pollution and did not show any correlation with metal concentrations in soil or earthworms nor with NRRT. Effects at the biochemical level therefore did not result in a reduced functioning of earthworm communities.     

Toxicological responses of earthworm (Eisenia fetida) exposed to metal-contaminated soils

The aim of this study was to evaluate the toxicological responses of earthworm (Eisenia fetida) induced by field-contaminated, metal-polluted soils. Biochemical responses and DNA damage of earthworm exposed to two multi-metal-contaminated soils in a steel industry park and a natural reference soil in Zijin Mountain for 2, 7, 14, and 28 days were studied. Results showed that three enzyme activities, including superoxide dismutase (SOD), acetylcholinesterase (AChE), and cellulase, in earthworm in metal-contaminated soils were significantly different from those of the reference soil. Cellulase and AChE were more sensitive than SOD to soil contamination. The Olive tail moment of the comet assay after 2-day exposure increased 56.5 and 552.0 % in two contaminated soils, respectively, compared to the reference soil. Our findings show that cellulase and DNA damage levels can be used as potential biomarkers for exposure of earthworm to metal-polluted soils.     

Oxidative stress and lipid peroxidation in the earthworm Eisenia fetida induced by low doses of fomesafen

Formesafen is a diphenyl ether herbicide that has adverse effects on non-target animals. However, knowledge about the effect of fomesafen on the antioxidant defense system in earthworms is vague. Thus, it is essential to investigate the effects of fomesafen on the antioxidant defense system in earthworms as a precautionary method. In the present study, earthworms (Eisenia fetida) were exposed to artificial soil treated with a range of concentrations of fomesafen (0, 10, 100, and 500 μg kg^?1) and were collected on the 3rd, 7th, 14th, 21st, and 28th days of exposure. Subsequently, the antioxidant enzyme activities (superoxide dismutase (SOD); catalase (CAT); and guaiacol peroxidase (POD)), reactive oxygen species (ROS) level, and malondialdehyde (MDA) content due to fomesafen treatment were examined in earthworms. Compared with the control, the SOD activity increased on the third and seventh days but decreased on the 14th day due to treatment with 100 and 500 μg kg^?1 of fomesafen. The activities of CAT and POD increased significantly on the third, seventh, and 14th days of exposure. In addition, the ROS level was significantly enhanced throughout the entire experimental period and showed a statistically dose-dependent relationship on the seventh and 14th days. The MDA content markedly increased on the seventh day of exposure; however, obvious changes were not detected at other exposure period. Low doses of fomesafen (≤500 μg kg^?1) may result in oxidative damage and lipid peroxidation in E. fetida by inducing the generation of ROS at short exposure periods (14 days). However, the adverse effects of fomesafen gradually disappear as the cooperation of antioxidant enzymes and exposure time are prolonged. This result may be helpful for further studies on the toxicological mechanisms of fomesafen to earthworms.     

The effects of tertiary treated municipal wastewater on fish communities of a small river tributary in Southern Ontario, Canada

Fish community changes associated with a tertiary treated municipal wastewater effluent outfall in the Speed River, Ontario, Canada, were evaluated at nine sites over two seasons (2008) using standardized electrofishing. Habitat evaluations were conducted to ensure that the riffle sites selected were physically similar. The fish community was dominated by several species of darters that differed in their response to the effluent outfall. There was a significant decrease in Greenside Darter (Etheostoma blennioides) but an increase in Rainbow Darter (E. caeruleum) abundance directly downstream of the outfall. Stable isotope signatures (δ¹³C and δ¹⁵N), which indicate shifts in energy utilization and flow, increased in Rainbow Darter downstream, but showed no change in Greenside Darter. Rainbow Darter may be exploiting a food source that is not as available at upstream sites giving them a competitive advantage over the Greenside Darter immediately downstream of the outfall.     

A time-course study of immune response in Japanese flounder Paralichthys olivaceus exposed to heavy oil

Purpose

The immunotoxicities of oil and its components on fish immunities have been investigated, but there is little literature on the recovery of the fish from the immune suppression. Therefore, the recovery of Japanese flounder Paralichthys olivaceus from an immunosuppressive effect due to heavy oil (HO) exposure was investigated in this study.

Methods

Fish were exposed to HO at a concentration of 0.385?g/L for 2?days, while control fish received no exposure. Seven fish were sampled at 0, 3, 7, and 14?days post-exposure. The respiratory rate was measured everyday as an indicator of the acute effect of HO exposure. Fish serum was collected and used for antibacterial activity assay against Edwardsiella tarda. Expression changes of respiratory and immune-related genes were evaluated by real-time PCR.

Results and discussion

The respiratory rate was significantly increased in the HO-exposed group until 4?days post-exposure. A respiratory-related gene, ??-hemoglobin, was also significantly downregulated in the spleen both at 0 and 7?days post-exposure and kidney at 3?days post-exposure in HO-exposed fish. Immunotoxicity, including suppression of antibacterial activities and downregulation of the IgM gene, was observed in HO-exposed fish until 3?days post-exposure, but not after that time. From these results, we conclude that the fish likely return to normal status around 1?week.     

Assessing the genotoxicity of urban air pollutants using two in situ plant bioassays

Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone # 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air.     

How to assess the feeding activity in ecotoxicological laboratory tests using enchytraeids?

The risk assessment of pesticides on soil fauna is an issue to protect agroecosystem sustainability. Enchytraeids are recognized as relevant soil bioindicators of chemical stress in agroecosystems. In laboratory, the reproduction test was found to be sensitive to reveal chemical impacts on enchytraeids. However, it does not allow to assess the impacts on ecological functions in which enchytraeids are involved. The objectives of this study were (i) to explore the feasibility of the bait-lamina test with enchytraeids under laboratory conditions and (ii) to compare its sensitivity with the Enchytraeid Reproduction Test. We exposed individuals of Enchytraeus albidus to two commercial formulations of fungicides widely used in Europe. The EC50 reproduction for the Swing^® Gold (50 g L^?1 epoxiconazole and 133 g L^?1 dimoxystrobin) and the Cuprafor micro^® (50% copper oxychloride) were respectively estimated at 1.66?±?0.93 times the recommended dose and >?496 mg kg^?1 of copper. However, no impact was found on the feeding activity of enchytraeids. The bait-lamina test thus appeared less sensitive than the Enchytraeid Reproduction Test to the tested fungicides. Despite that, this test which is achievable under laboratory conditions and allows assessing indirect effects of pesticides is quick, cheap, and easy to perform. It would deserve to be used to explore longer-exposure effects through the repeated addition of bait-lamina sticks.     

Structure-related distribution of PCDD/Fs, PCBs and HCB in a river-sea system

Water concentrations of PCDD/Fs, HCB, and non-ortho, mono-ortho, and non-dioxin-like PCBs were measured four times during 1 year in a coastal area of the Baltic Sea, to investigate background levels and distribution behaviour. Sampling sites included two rivers, an estuary, and the sea. Particulate and apparently dissolved concentrations were determined using active sampling (filters + PUFs), while freely dissolved concentrations were determined using passive sampling (POM-samplers). The distribution between particulate + colloidal and freely dissolved phases, in the form of TOC-normalized distribution ratios (K_TOC), was found to be near or at equilibrium. The observed K_TOC were not significantly different between sampling sites or seasons. For PCDD/Fs, the concentrations were significantly correlated to suspended particulate matter (SPM), while no correlation to organic carbon (TOC) was observed. In the estuary and the sea, PCB concentrations were correlated to TOC. The sorption of various congeners to SPM and TOC appeared to be related to both hydrophobicity and 3D-structure. The PCDD/F concentration in the sea decreased to one third in May, likely connected to the increased vertical flux of particles during the spring bloom.     

Exposure of juvenile Danio rerio to aged TiO2 nanomaterial from sunscreen

The toxicity of dietary exposure to artificially aged TiO₂ nanomaterial (T-Lite?) used in sunscreen cream was studied on Danio rerio. Embryolarval assays were conducted to assess the effects of TiO₂ residues of nanomaterial (RNM) on fish early life stages. Juvenile fishes were exposed by the trophic route in two experiments. During the first experiment, juvenile fishes were exposed to TiO₂ RNM for 14 days by adding RNM to commercial fish food. The second one consisted in producing a trophic food chain. Pseudokirchneriella subcapitata algae, previously contaminated with TiO₂ RNM in growth medium, was used to feed Daphnia magna neonates over a 48-h period. Daphnia were used next to feed juvenile fishes for 7 days. Accumulation of Ti, life traits (survival and growth) and biochemical parameters such as energy reserves, digestive (trypsin, esterase, cellulose and amylase) and antioxidant (superoxide dismutase and catalase) enzyme activity were measured at the end of exposures. As expected in the receiving aquatic system, TiO₂ RNM at low concentrations caused a low impact on juvenile zebrafish. A slight impact on the early life stage of zebrafish with premature hatching was observed, and this effect appeared mainly indirect, due to possible embryo hypoxia. When juvenile fish are exposed to contaminated food, digestive enzyme activity indicated a negative effect of TiO₂ RNM. Digestive physiology was altered after 14 days of exposure and seemed to be an indirect target of TiO₂ RNM when provided by food.     

Variability of Nox and No2 concentrations observed at pedestrian level in the city centre of a medium sized urban area

NO_x and NO₂ concentrations were measured at different locations in a city centre of an urban zone (Population 450 000) in order to study the variation of the outdoor exposure at pedestrian level. These measurements were carried out to understand the influence of traffic emissions at each measured site. The observations were done during four weeks in winter, including several days with high pollution levels. The results at different locations have been used to analyse criteria recommended for locating observation sites in a monitoring network. No large differences in background pollution averaged over several weeks have been found throughout the city centre, even during pollution peaks. Measurements were also carried out inside one street canyon. The contribution of the street traffic to the NO=NO_x−NO₂ concentrations observed at side-walk has been found important, i.e., several times the background level. On the other hand, the majority of observed NO₂ pollution is due to the contribution of background pollution within the street. The pollutant excess at pedestrian level is strongly correlated to the street traffic emission and to the atmospheric turbulence observed at roof level. Application of a box model to the street data demonstrates that such models can be useful to estimate the pollutant accumulation within the street.     

Environmental mutagenicity and toxicity caused by sodium metabisulfite in sea shrimp harvesting in Piauí, Brazil

Sodium metabisulfite is used in marine shrimp harvesting to prevent the occurrence of black spots. Shrimps are soaked in a sodium metabisulfite solution in ice, which is disposed of in sewages that run into marine canals, creating an environmental hazard. This study evaluates the toxicity and mutagenicity caused by sodium metabisulfite in sea waters and sediments collected in a shrimp farm in Cajueiro da Praia (Luis Correia), state of Piauí, Brazil, using the Allium cepa assay. Water and sediment samples were collected in the dry and in the rainy seasons, in three sites: upstream the shrimp farm (Site 1), at the point sodium metabisulfite is discharged (Site 2), and 100 m downstream the farm (Site 3). Three sample dilutions were used (50%, 25% and 10%) for all samples. A negative control (well water) and a positive control (copper sulfate 0.0006 mg mL⁻¹) were used in each experiment. At the end of the 72-h exposure period, onion roots were measured and removed. Mutagenicity analysis included the determination of mitotic index, chromosomal aberrations and the detection of micronuclei; analysis of root size and mitotic index were used as an index of toxicity. The A. cepa assay revealed that the water and sediments samples collected in the Piauí coast contaminated with sodium metabisulfite induce toxicity. The results demonstrate that the assay may be used as a regular tool in the analysis of water parameters in shrimp farms in the coast of Piauí state, and in strategies to preserve the region’s marine ecosystem.     

Can time-weighted average concentrations be used to assess the risks of metsulfuron-methyl to Myriophyllum spicatum under different time-variable exposure regimes?

We tested the effects of the herbicide metsulfuron-methyl on growth of the submerged macrophyte Myriophyllum spicatum under laboratory conditions using different exposure scenarios. The exposures of each scenario were comparable in the concentration × time factor, viz., the same 21-d time-weighted average (TWA) concentrations but variable in peak exposure concentrations (ranging from 0.1 to 21 000 ng ai L⁻¹) and exposure periods (1, 3, 7, 14 or 21 d). To study recovery potential of the exposed M. spicatum plants we continued the observation on shoot and root growth for another 21 d in herbicide-free medium so that the total observation period was 42 d. Non-destructive endpoints, length and number of new shoots and roots, were determined weekly from day 14 onwards. Destructive endpoints, dry-weight (DW) of main shoots, new shoots and new roots, were measured at the end of the experiment (t = 42 d).Metsulfuron-methyl exposure in particular inhibited new tissue formation but was not lethal to main shoots. On days 21 and 42 after start exposure, EC₁₀/EC₅₀ values for new tissues expressed in terms of peak concentration (=measured concentration during exposure periods of different length) showed large differences between exposure scenarios in contrast to EC₁₀/EC₅₀ values for days 21 and 42 expressed in terms of 21-d and 42-d TWA concentrations, respectively. At the end of the experiment (day 42), 42-d TWA EC_x values were remarkably similar between exposure scenarios, while a similar trend could already be observed on day 21 for 21-d TWA EC_x values. For the macrophyte M. spicatum and exposure to the herbicide metsulfuron-methyl the TWA approach seems to be appropriate to use in the risk assessment. However, the data from the toxicity experiment suggest that on day 21 also the absolute height of the pulse exposure played a (minor) role in the exposure - response relationships observed.     

Long-term personal exposure to PM2.5, soot and NOx in children attending schools located near busy roads,a validation study

Several studies have investigated the health of children attending schools located near busy roads. In this study, we have measured personal exposure to traffic-related pollutants in children to validate exposure classification based on school location. Personal exposure to PM_2.5, soot, NO_x and NO₂ was measured during four 48-h periods. The study involved 54 children attending four different schools, two of which were located within 100 m of a major road (one ring road and one freeway) and the other two were located at a background location in the city of Utrecht, The Netherlands. Outdoor monitoring was conducted at all school sites, during the personal measurements. A questionnaire was administered on time activity patterns and indoor sources at home. The outdoor concentration of soot was 74% higher at the freeway school compared to its matched background school. Personal exposure to soot was 30% higher. For NO_x the outdoor concentration was 52% higher at the freeway school compared to its background school. The personal concentration of NO_x was 37% higher for children attending the freeway school. Differences were smaller and insignificant for PM_2.5 and NO₂. No elevated personal exposure to air pollutants was found for the children attending the school near the ring road. We conclude that the school's proximity to a freeway can be used as a valid estimate of exposure in epidemiological studies on the effects of the traffic-related air pollutants soot and NO_x in children.     

Multiparametric approach for assessing environmental quality variations in West African aquatic ecosystems using the black-chinned tilapia (Sarotherodon melanotheron) as a sentinel species

The study highlights the potential of the black-chinned tilapia to be used as a sentinel to assess environmental contaminants based on the use of a set of biomarkers. The usefulness of fish species as sentinels for assessing aquatic environment contamination was tested using a set of biomarkers in Senegalese environments characterized by multi-pollution sources. The black-chinned tilapia (Sarotherodon melanotheron) was selected as a sentinel because of its abundance, wide distribution in all coastal aquatic ecosystems and physiological properties. The potential influence of confounding factors such as salinity on biomarker in the tilapia has been examined. Individuals were sampled during two seasons (dry and wet) in eight sites characterized by various degrees of anthropogenic contamination and different salinities (from 0 to 102?psu). Biomarkers??including growth rate (GR), condition factor (CF), biotransformation enzymes such as 7-ethoxyresorufin-O-deethylase (EROD) and glutathione-S-transferase (GST), lipid peroxidation (TBARS) and acetylcholinesterase (AChE)??were measured. Chemical contaminant [polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs)] levels showed different sources of contamination with relatively high concentrations of PAHs in the Hann Bay and Foundiougne locations. The most sensitive biomarker present in different sites according to the principal component analysis is EROD. Few variations of the AChE activity and TBARS levels were found. No clear relationship was found between biomarker responses and salinity, but GR and CF were lower in hypersaline conditions. Tilapia is responsive to environmental contaminants such as PAHs, OCPs and PCBs. The S. melanotheron multiparametric approach showed a better discrimination of sites.     

Evaluation of the distribution of fecal indicator bacteria in a river system depending on different types of land use in the southern watershed of the Baltic Sea

The aim of the study was to determine the effects of land use management on changes in the fecal contamination of water in the ?yna River, one of the main lowland watercourses in the southern watershed of the Baltic Sea (northern Poland). A total of 120 water samples were collected in different seasons of 2011 and 2012 at 15 sites where the river intersected forest (FA), agricultural (AA), and urbanized (UA) areas. Fecal indicator bacteria (FIB), the counts of Enterobacteriaceae and Escherichia coli, total bacterial counts (TBCs), and domain Bacteria (EUB338) were determined by culture-dependent and culture-independent methods. Temperature, pH, chemical oxygen demand, dissolved oxygen, total dissolved solids, ammonia nitrogen, nitrite nitrogen, nitrate nitrogen, orthophosphate, and total phosphorus were also determined. The lowest bacterial counts were noted in water samples collected in FA, and the highest in samples collected in UA. Statistically significant differences were determined between bacterial populations across the analyzed land use types and in different sampling seasons. Significant correlations were also observed between the populations of FIB and physicochemical parameters. The results indicate that land use type influenced FIB concentrations in river water. The combined use of conventional and molecular methods improves the accuracy of fecal contamination analyses in river ecosystems.