Background, aim, and scope Mutagenic nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) have been known to arise in the environment through direct emissions from combustion sources and nitration of PAHs, primarily in the atmosphere. In the marine environment, PAHs are one of the classic anthropogenic organic pollutants, while nitrite (NO ₂ ⁻ ) is produced naturally via various biological processes like imbalance in nitrification/denitrification or eutrophication and subsequent oxygen depletion from an oversupply of nutrients. In this paper, we report the formation of PAH-DNA adducts in fish contaminated with PAHs and exposed to NO ₂ ⁻ in the ambient water. Electrospray ionization tandem mass spectrometric (ESI-MS/MS) analysis of the bile of the euryhaline fish Oreochromis mossambicus exposed simultaneously to field relevant sublethal concentrations of phenanthrene and NO ₂ ⁻ and collision-induced dissociation of selected ions revealed the presence of DNA-PAH adducts. The present study indicates that, although several high sensitivity techniques have been developed for the analysis of PAH derived DNA adducts, MS/MS has emerged as a powerful tool in the detection and structure elucidation of DNA adducts. Materials and methods Juvenile O. mossambicus from a local estuarine fish farm were used with increasing frequency for carcinogenicity testing and comparative cancer research. The fish were exposed to the alkylating agent phenanthrene in the presence of NO ₂ ⁻ . Composite untreated bile samples after dilution with methanol: water (1:1; v/v) were analyzed by ESI-MS. Results Several adducts could be evidenced in the bile by MS/MS. Deoxyadenosine/deoxyguanosine having a mass in the range of 450-650 amu is detected. In addition, a segment of modified dinucleotide with a mass that corresponds to a dimer consisting of a modified guanosine and a normal guanosine has also been identified in the bile. Discussion The formation of certain types of DNA adducts is a crucial step in the induction of cancer and a primary stage in mutagenesis. Phenanthrene injected by i.p. route led to the transformation of phenanthrene to N-formyl amino phenanthrene-N ⁶-deoxyadenosine adduct, whereas the fish co-exposed to phenanthrene and ambient nitrite metabolizes PAH to mono-, di- as well as trinitro derivatives, which then react with DNA leading to the formation of mainly modified guanosine and adenosine adducts. In the present investigation, dinitrophenanthrene diol epoxide (DNPDE) adduct with guanosine (m/z 587) seems to be the dominant adduct in the mixture, and its presence is shown first as a comparatively less stable adduct, which decomposes to give a more stable N² adduct (m/z 567). Conclusions MS/MS has proved to be useful in the rapid determination and discrimination of structurally different phenanthrene/derivatives DNA adducts in a complex mixture of fish bile co-exposed to phenanthrene and nitrite. However, the nature of metabolites formed is likely determined by the route of PAH administration, and there is a need to further define the early biochemical events of carcinogenesis in these species. Recommendations and perspectives DNA adduct analysis in fish bile offers a promising approach to study the risk of potentiation of anthropogenic chemicals into genotoxic compounds in the presence of nitrite in the marine environment. We believe this is the first report on the formation of DNA-phenanthrene adducts on co-exposure of the fish to PAH and nitrite.

Background, aim, and scope The Lower Mississippi River is a major transportation route for commercial goods and petroleum products produced and refined locally. Oil spills caused by vessel accidents and equipment failure at refineries are a serious threat to the drinking water supply of Southern Louisiana, as well as to the many natural, economic, and social resources supported by the river. Providing accurate trajectory modeling to contingency planners is critical to protecting the local environment. The majority of trajectory model results, assuming a uniform shoreline, show 60-70% of spilled oil can be retained. This study examines the impact of detailed shoreline mapping that captures spatial and temporal changes in shoreline type on oil spill trajectory modeling. Materials and methods Detailed shoreline maps based on recent remote sensing imagery were generated to identify spatial changes in shoreline. A hydrodynamic model of the 78 mile reach from Convent, Louisiana to West Pointe a la Hache was developed to obtain the stage levels and velocity fields of four river discharges. Based on river stage level, another layer was added to the shoreline maps, so that shoreline type was accurately represented at each river discharge, a feature not included in previous mapping. An oil spill trajectory model was then used to investigate the effect of implementing different re-floatation half-lives that correlate to the shoreline maps developed for this study at four river discharges. Results Detailed shoreline mapping showed the Lower Mississippi River has four major shoreline types each with different oil re-floatation half-lives: muddy clay, sand, low vegetation, and high vegetation. As flow rate changed, the shoreline spatial variability also changed, from 84% mud/sand and 16% vegetation at low flow rates to 4% mud and 96% vegetation at higher flow rates. At flow rates with large variability in shoreline type, the distribution of oil attached to the shore was significantly different from results of simulations that used a constant shoreline type and re-floatation half-life. Discussion At low flow rates, simulations with the detailed delineation of shoreline type predicted that ~30% of the oil would be beached/retained because the oil was able to travel further down the reach and interact with the shoreline in multiple locations. Simulations at the low flow rates with the existing shoreline mapping predicted approximately 65% of the oil would be retained as did all the simulations at the highest flow rates. At high flow rates, the oil interacted mostly with vegetation and results were very similar to those obtained with a single re-floatation half-life of 1 year. In addition to shoreline type, river geometry and the hydrodynamics were major factors influencing the distribution of oil along the river reach. Conclusions Shoreline re-floatation half-lives have a major impact on simulating the distribution of oil along the shore after a spill, especially in areas with a high variability of shoreline type as in the lower Mississippi River. Assigning the correct re-floatation half-life and retention capacity is only possible when shoreline types have been correctly identified. The maps developed for this study provided an important level of detail and incorporated the change in shoreline type with flow rate, resulting in more detailed trajectory modeling of the study reach. Recommendations and perspectives Shoreline maps should include as much detail about shoreline type as possible. When developing shoreline maps or environmental sensitivity assessments, the focus should include specific characteristics of the study area; using standardized maps or methods of assessment may leave out detail that could negatively impact modeling efforts. Finally, shoreline sensitivity to oiling is an important area of research that will benefit from an improved understanding of oil retention by vegetation.

Background, aim, and scope Impacts on the reproductive health of wild fish are thought to be suitable early-warning tools indicating contamination of surface waters with endocrine-disrupting compounds. Ecotoxicological assessment of these field observations depends on the availability of reliable biomarkers to enable a discrimination of natural variations of reproductive functions from anthropogenic impacts. Materials and methods Roach and perch were caught at eight sampling sites by electrofishing twice a year in summer (July-September) and late autumn/winter (November-December) over a 2-year period. The sites are characterized by different degrees of anthropogenic impact and are situated within the greater Upper Rhine catchment. Age growths, parasitization and gonadal histology of more than 3,000 fish were examined. Results The two dominant fish species in German surface waters perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.) differ considerably regarding their suitability for biomonitoring. Even in pristine habitats, perch show several variants of sex differentiation in terms of (1) the time of first sexual maturation, (2) the course of seasonal gonadal recrudescence, and (3) the occurrence of heterologous germ cells (testes ova). A statistically significant elevated proportion of males were observed in fish obtained from a TBT-contaminated marina and suppression of gonadal ripening was observed in females caught in a sewage-contaminated brook. Both effects appear to be due to chemical contamination. The only “natural” alteration of sex differentiation in roach was related to parasitization with Ligula intestinalis (Eucestoda, Pseudophyllidea). Other deviations from the normal pattern of sex differentiation were (1) suppression of ovarian ripening and (2) asynchronic seasonal gonadal recrudescence. These are strong indicators of an anthropogenically induced impact on reproductive health. Feminization phenomena were not observed at either the individual or the population level. Discussion Interpretation of field monitoring results concerning reproductive health requires large numbers of samples and detailed knowledge of the natural plasticity of sex differentiation in the species under investigation. A better understanding of the mechanisms underlying the plasticity of sex differentiation in perch is indispensable to enable perch to be used as a bioindicator. Conclusions Deviation from the strict and probably endogenous control of sex differentiation in roach is a strong and unequivocal warning signal. Recommendations and perspectives The subject of fish monitoring should be addressed in the context of a broader spectrum of potential risks. Seasonal and ontogenetic integrity of gonadal development and recrudescence are potent biomarkers, provided the natural process is well documented for the species under investigation.

Bitumen is a black, thermoplastic, hydrocarbon material derived from the processing of crude oil. At ambient temperature, bitumen is solid and does not present any health/environmental risks. This is one of the main reasons that bitumen is widely used for road construction all over the world. But during manufacturing/modification according to its application, storage, transportation, and use of bitumen is heated giving off various hydrocarbons emissions. In recent years, there has been increasing interest in investigating the potential of bitumen emissions to cause health effects. This is mainly because of the reason that bitumen has small amount of poly-aromatic hydrocarbons, along with some other volatiles like benzene, toluene, etc., which are known to be carcinogenic in nature. Thus, assessment of the emission characteristics and health hazards of bitumen fumes may have far reaching industrial economic and public health implications. In this review, we will discuss about the emission characteristics from bitumen, asphalts, or road construction, which is mainly contributed by bitumen fumes. Sampling strategies and analytical methods employed are also described briefly.

Background Chlorobenzoic acids (CBA) are intermediate products of the aerobic microbial degradation of PCB and several pesticides. This study explores the feasibility of using basket willows, Salix viminalis, to remove 4-CBA from polluted sites, which also might stimulate PCB degradation. Methods The removal of 4-CBA by willow trees was investigated with intact, septic willow trees growing in hydroponic solution and with sterile cell suspensions at concentrations of 5 mg/L and 50 mg/L 4-CBA. Nutrient solutions with different levels of ammonium and nitrate were prepared to achieve different pH levels. The concentration of 4-CBA was tracked over time and quantified by HPLC. Results and discussion At the low level of 4-CBA (5 mg/L), willows removed 70% (pH 4.2) to 90% (pH 6.8), while 48% (pH 4.2) to 52% (pH 6.8) of the water was transpired. At the high 4-CBA level (50 mg/L), the pH varied between 4.4 and 4.6, and 10% to 30% of 4-CBA was removed, but only 5% to 9% of the water. In sterile cell suspensions, removal of 4-CBA by fresh biomass was much higher than removal by dead biomass. Conclusions The results indicate that 4-CBA is toxic to willow trees at 50 mg/L. The removal of 4-CBA from solution is by both passive processes (uptake with water, sorption to plant tissue) and metabolic processes of the plants. Recommendations and outlook Plants, such as willow trees, might assist in the degradation of PCB and their degradation products CBA.

Background, aim, and scope Many pollutants have received significant attention due to their potential estrogenic effect and are classified as endocrine disrupting compounds (EDCs). Because of possible ecological effects and increased attention for water reuse schemes, it is important to increase our understanding of the EDC removal capacities of various wastewater treatment systems. However, there has so far been little research on the fate and behavior of EDCs in stabilization pond systems for wastewater treatment, which represent an important class of wastewater treatment systems in developing countries because of their cost-effectiveness. The aim of this work is to study the fate and behavior of EDCs in algae and duckweed ponds. Because the synthetic hormone 17α-ethinylestradiol (EE2) and the natural hormones estrone (E1), as well as 17β-estradiol (E2), have been detected in effluents of sewage treatment plants and been suggested as the major compounds responsible for endocrine disruption in domestic sewage; E1, E2, and EE2 were therefore chosen as target chemicals in this current work. Materials and methods Both batch tests and continuous-flow tests were carried out to investigate the sorption and biodegradation of estrogens in algae and duckweed pond systems. The applied duckweed was a Lemna species. The applied algae was a mixture of pure cultures of six different algae genera, i.e., Anabaena cylindrica, Chlorococcus, Spirulina platensis, Chlorella, Scenedesmus quadricauda, and Anaebena var. Synthetic wastewater were used in all tests. The concentrations of estrogens were measured with three different enzyme-linked immunosorbent assay kits specific for E1, E2, or EE2. When the concentrations of estrogens in water samples were below the lowest quantitative analysis range (0.05 µg/l), preconcentration of the water samples were performed by means of solid phase extraction (SPE) with C18 cartridges. Results The 6-day batch tests show that the presence of algae or duckweed accelerated the removal of the three estrogens from the synthetic wastewater. More estrogens were removed in the tests with duckweed than in tests with algae or with wastewater. In the sorption tests, a swift sorption of the three estrogens was observed when the estrogens were contacted with duckweed or algae, while the estrogen concentrations in tap water kept unchanged during the 3-h sorption tests. The mass balances indicated that only about 5% of the estrogens were bound to the algae sediment or duckweed at the end of the 6-day tests. Results of the continuous-flow tests revealed that the algae and duckweed ponds effectively removed E1, E2, and EE2 even at nanograms per liter level. Interconversion of E1 and E2 occurred both in batch and continuous-flow tests. E2 could be readily transformed to E1, especially in the tests with algae. Discussion Different processes like sorption, biodegradation and photolytic degradation might play an important role in the removal of estrogens from the aquatic phase. The 3-h sorption tests support the importance of sorption for estrogen removal, in which a rapid initial sorption was observed over the first 2 min for E1/E2/EE2 to both duckweed and algae. In the 6-day batch tests, estrogens were sorbed by algae or duckweed during the early stage when algae and duckweed were contacted with the synthetic wastewater and the sorbed estrogens were further biodegraded by the microorganisms developed in the wastewater. The persistent estrogen concentrations in tap water, however, implied that no sorption, biodegradation, or photolytic degradation occurred in tap water under the specific experimental conditions. Under aerobic or anoxic conditions, E2 could be first oxidized to E1, which is further oxidized to unknown metabolites and finally to CO₂ and water. Under anaerobic conditions, E1 can also be reduced to E2. However, the interconversion might be much more complex especially in the tests with algae because both aerobic and anaerobic conditions occurred in these tests due to the variation of the dissolved oxygen concentration induced by the light regime. Conclusions This study shows that estrogens, E1, E2, and EE2, can be effectively removed from the continuous-flow algae and duckweed ponds even when their concentrations are at nanograms per liter level. The presence of algae and duckweed accelerate the removal of estrogens from the synthetic wastewater because estrogens can be quickly sorbed on duckweed or algae. The sorbed estrogens are subsequently degraded by microorganisms, algae, or duckweed in the wastewater treatment system. E1 and E2 are interconvertible in both duckweed and algae pond systems. E2 can be readily transformed to E1, especially in the tests with algae. Recommendation and perspectives Based on the tests performed so far, one can conclude that both sorption and biodegradation are important to the estrogens removal from stabilization pond systems for wastewater treatment. Further research using, e.g., radioimmunoassay is needed to investigate the biodegradation pathway of estrogens in algae and duckweed ponds.

Background, aim and scope A new method using sedimentary metals and geographic information system as indicators for assessing temporal and spatial anthropogenic change in estuaries has been applied to a large coastal lake (Lake Macquarie) in New South Wales, Australia. Materials, methods and results Two vintages of data (1975 and 2003) on surficial sediment metal (Cd, Cu, Pb and Zn) concentrations combined with ²¹⁰Pb core profiles were used to determine past changes in sediment quality and to predict possible future relaxation rates for the entire lake area in response to change in anthropogenic pressure. Sediment cores showed distinct vertical profiles; sedimentation rates in the northern part of the lake were consistent (14 mm year⁻¹) over the 55-year period investigated. Discussion and conclusions Surficial metal concentrations were highest in the 1975 sediment than in the 2003 samples, with the northern part of Lake Macquarie having much greater metal concentrations than the rest of the lake. Past and future declining sedimentary metal concentrations in the northern part of the lake were expected due to the closure of a nearby Pb-Zn smelter; however, possible increases in Cu in the south of the lake to the year 2020 were surprising. The new method presented in this study can assist estuary managers by providing data on past, present and future conditions, which are essential in making informed decisions for the improvement of estuarine systems.

Background, aim, and scope For many years, highly concentrated wastewater generated from the pulp and paper industry has become the focus of much concern worldwide. The objectives of this study were to determine the treatment efficiency of the alkaline peroxide mechanical pulp (APMP) process wastewater using a sequencing batch reactor (SBR) and analyze the microbial dynamics of the wastewater treatment system using the random amplified polymorphic DNA (RAPD) method. Materials and methods An SBR was applied to the treatment of APMP pulp and paper process wastewater. The wastewater characteristics and many physicochemical operator indicators in the wastewater treatment process were analyzed and determined according to standard methods. Microbial 16 S rDNA in active sludge was extracted, amplified, and analyzed using the RAPD method for the microbial dynamics of the wastewater treatment system. Results and discussion Ten kinds of natural organic compounds of plants such as monoterpene were detected in the APMP pulp and paper process wastewater. With an influent chemical oxygen demand (COD) that varied in the range of 685.7 to 907.5 mg/L, the corresponding effluent COD was 176.5 to 266.1 mg/L and the removal efficiency was 70.3% to 79.8%. An optimal strain (S308: CAGGGGTGGA) was selected to study the population dynamics and diversity of the bacterial community. The RAPD-polymerase chain reaction (PCR) fingerprints showed very high polymorphism of the genetic bands (78-100%). Four groups of species were clustered using the unweighted pair group method with arithmetic (UPGMA) analysis, and the genetic distance was close between the species within each group. The Shannon-Weaver index was high and varied over time with the COD removal. Conclusions The RAPD-PCR technique can be used to study microbial dynamics, which was shown to vary over time with the removal efficiency of SBR treating APMP pulp and paper process wastewater.

Concept and purpose Virtually all water treatment facilities worldwide generate an enormous amount of water treatment residual (WTR) solids for which environmentally friendly end-use options are continually being sought as opposed to their landfilling. Aluminium-based WTR (Al-WTR) can offer huge benefits particularly for phosphorus (P) removal and biofilm attachment when used as media in engineered wetlands. However, potential environmental risks that may arise from the leaching out of its constituents must be properly evaluated before such reuse can be assured. This paper presents results of an assessment carried out to monitor and examine the leachability and leaching patterns of the constituents of an Al-WTR used as media in laboratory-scale engineered wetland systems. Main features, materials and methods Al-WTR was used as media in four different configurations of laboratory-scale engineered wetland systems treating agricultural wastewater. Selected metal levels were determined in the Al-WTR prior to being used while levels of total and dissolved concentration for the metals were monitored in the influent and effluent samples. The increase or decrease of these metals in the used Al-WTR and their potential for leaching were determined. Leached metal levels in the effluents were compared with relevant environmental quality standards to ascertain if they pose considerable risks. Results Aluminium, arsenic, iron, lead and manganese were leached into the treated effluent, but aluminium exhibited the least leaching potential relative to its initial content in the fresh Al-WTR. Levels of P increased from 0.13 mg-P/g (fresh Al-WTR) to 33.9-40.6 mg-P/g (used Al-WTR). Dissolved levels of lead and arsenic (except on one instance) were below the prescribed limits for discharge. However, total and dissolved levels of aluminium were in most cases above the prescribed limits for discharge, especially at the beginning of the experiments. Conclusions, recommendations and perspectives Overall, the study indicates that leaching is observed when Al-WTR is beneficially reused for enhanced P removal in engineered wetlands. In particular, levels of aluminium in the treated effluent beyond the prescribed limits of 0.2 mg/l were observed. However, since the results obtained indicate that aluminium leached is mostly associated with solids, a post-treatment unit which can further reduce the level of aluminium in the treated effluent by filtering out the solids could serve to mitigate this. In addition, plants used in such wetland systems can uptake metals and this can also be a potential solution to ameliorating such metal releases. Periodic monitoring is thus advised. Notwithstanding, the use of Al-WTR as a media in engineered wetlands can serve to greatly enhance the removal of P from wastewaters and also serve as support material for biofilm attachment.

Background, aim, and scope In response to concerns about chemical substances that can alter the function of endocrine systems and may result in adverse effects on human and ecosystem health, a number of in vitro tests have been developed to identify and assess the endocrine disrupting potential of chemicals and environmental samples. One endpoint that is frequently used in in vitro models for the assessment of chemical effects on the endocrine system is the alteration of aromatase activity (AA). Aromatase is the enzyme responsible for converting androgens to estrogens. Some commonly used aromatase assays, including the human microsomal assay that is a mandatory test in US-EPA's endocrine disruptor screening program (EDSP), detect only direct effects of chemicals on aromatase activity and not indirect effects, including changes in gene expression or transcription factors. This can be a problem for chemical screening initiatives such as the EDSP because chemicals can affect aromatase both indirectly and directly. Here we compare direct, indirect, and combined measurements of AA using the H295R cell line after exposure to seven model chemicals. Furthermore, we compare the predictability of the different types of AA measurements for 17β-estradiol (E2) and testosterone (T) production in vitro. Materials and methods H295R cells were exposed to forskolin, atrazine, letrozole, prochloraz, ketoconazole, aminoglutethimide, and prometon for 48 h. Direct, indirect, and combined effects on aromatase activity were measured using a tritiated water-release assay. Direct effects on aromatase activity were assessed by exposing cells only during the conduct of the tritium-release assay. Indirect effects were measured after exposing cells for 48 h to test chemicals, and then measuring AA without further chemical addition. Combined AA was measured by exposing cells prior and during the conduction of the tritium-release assay. Estradiol and testosterone were measured by ELISA. Results and discussion Exposure to the aromatase inhibitors letrozole, prochloraz, ketoconazole, and aminoglutethimide resulted in greater indirect aromatase activity after a 48-h exposure due to presumed compensatory mechanisms involved in aromatase activity regulation. Forskolin and atrazine caused similar changes in hormone production and enzyme profiles, and both chemicals resulted in a dose-dependent increase in E2, T, and indirect AA. Neither of these two chemicals directly affected AA. For most of the chemicals, direct and combined AA and E2 were good predictors of the mechanism of action of the chemical, with regard to AA. Indirect aromatase activity was a less precise predictor of effects at the hormone level because of presumed feedback loops that made it difficult to predict the chemicals' true effects, mostly seen with the aromatase inhibitors. Further, it was found that direct and indirect AA measurements were not reliable predictors of effects on E2 for general inducers and inhibitors, respectively. Conclusions Differential modulation of AA and hormone production was observed in H295R cells after exposure to seven model chemicals, illustrating the importance of measuring multiple endpoints when describing mechanisms of action in vitro. Recommendations and perspectives For future work with the H295R, it is recommended that a combination of direct and indirect aromatase measurements is used because it was best in predicting the effects of a chemical on E2 production and its mechanism of action. Further, it was shown that direct AA measurements, which are a common way to measure AA, must be used with caution in vitro.