Background, aim, and scope Chemical oxygen demand (COD) is used as a discharge standard parameter in wastewater treatment plant design, environmental modelling and many other applications. Chloride interference is an important problem of COD measurement for wastewaters containing low organic matter and high chloride concentrations. In case of chloride concentrations up to 2,000 mg/L, mercury sulphate addition at a ratio of 10:1 (HgSO₄:Cl⁻) can adequately mask the interference. When chloride concentration exceeds 2,000 mg/L, this ratio becomes ineffective to hinder the interference. At this point, it is proposed to use a greater and constant ratio of mercury sulphate addition. However, this application sometimes results in extra mercury sulphate addition which is not necessary. Even in some cases, greater addition of mercury sulphate alone is not a solution to erroneous measurement results. The purpose of the study is to determine optimum HgSO₄:Cl⁻ ratios according to the chloride concentrations of the samples and to show the importance of the strength of the digestion solution for the correct determination of the COD parameter. Materials and methods CODs of the synthetic samples containing varying COD and chloride concentrations were measured by closed reflux colorimetric method using three digestion solutions having different strengths. Results It is indicated in this study that a constant ratio of mercury sulphate can only prevent chloride interference up to a specific chloride concentration. Conclusions Achieving high precision results in case of low organic matter and high chloride concentration can only be possible by both decreasing the concentration of oxidant and adding mercury sulphate.

Background In the present study, we report the analytical results of pp′DDT, pp′DDE and pp′DDD determination in lake water, plankton and zooplanktivorous fish of Lake Maggiore (Northern Italy), rather recently polluted by DDT of industrial origin, in order to understand the bottom-up pollution transfer among the abiotic and biotic components of the lake ecosystem. Materials and methods Fourteen water sampling campaigns were carried out from March 2003 to January 2009 in the water column of the deepest point (Ghiffa) of Lake Maggiore. Suspended and dissolved pollutants were determined separately. Three sampling campaigns were carried out from July 2008 to January 2009 for zooplankton and pelagic fish, and DDT residues were analysed by HRGC coupled with ECD and MS. Moreover, food items were detected in fish stomachs. Results and discussion Starting from 2007, the DDT contamination along the water column became rather homogeneous, probably because no flooding or other relevant hydrological events occurred; although steady-state condition should be expected, lipid normalised concentrations of pp′DDE and pp′DDD in zooplankton exceeded the levels of the same compounds in zooplanktivorous fishes. Conclusion As this finding represents a thermodynamic paradox since bioaccumulative pollutant concentrations are expected to increase along the trophic chain, it was supposed that the abiotic and biotic lake components were not in a steady-state condition in Lake Maggiore.

Background, aim, and scope Dissolved humic substances (HSs) are exogenous stressors to aquatic plants and animals which activate a variety of transcriptional and biochemical reactions or block photosynthesis. While there are consistent indications which structures may lead to reduced photosynthetic activity, there is much less clear information available on which HS structures or building blocks act as stressors in animals. Consequently, this work was designed to comparatively study the impact of natural organic matter (NOMs) from different sources on major anti-stress mechanisms in one single animal. We utilized major antioxidant responses and relative expression levels of stress proteins (small HSPs and HSP70) and expected that different HSs provoke different response patterns. Materials and methods We tested the freshwater amphipod Gammarus pulex which was collected from several shallow creeks in Northern Germany. All specimens were maintained in aerated 5-L tanks with filtered water from their natural environment at 10°C with prior acclimation. Animals were fed ad libitum with a commercial preparation once every second day. The exposure water was exchanged with the same frequency. NOMs were isolated from three different sources: two from small brown-water lakes in Northern Germany by reverse osmosis and the third one as an aqueous extract from a black layer of a Brazilian sandbar soil (State of Rio de Janeiro). The rationale was to apply NOMs of contrasting quality. Chemical fingerprint features of the NOMs were taken by high-performance size exclusion chromatography. As stress parameters in the animals, the activities of peroxidase and catalase were recorded quantitatively, and stress proteins, HSP70, as well as small α-crystalline HSPs were analyzed semiquantitatively. Results The three NOMs clearly differed in molecular masses, humic substance contents, the moieties of polysaccharides, and low-molecular-weight substances. With the exception of one short-term response, the peroxidase activity increased after 3 to 12 h exposure, whereas the catalase activity did not show any significant modulation. With one exception, the stress protein expression increased after 30 min exposure in a biphasic pattern, and the sHSPs responded less strongly than HSP70. Discussion Although the quality of the exposed NOMs differs significantly, a rather uniform response pattern appears in the animals. Obviously, the contrasting contents of HSs and polysaccharides did not affect the anti-stress response of the exposed gammarids which is in contrast to previous lifespan studies with Caenorhabditis elegans. Furthermore, all NOM sources led to increased contents of both HSP70 and sHSPs. To the best of our knowledge, this is the first protein study to show that also small HSPs are expressed when the animals are exposed against humic material. Conclusions Since the response patterns of the exposed gammarids, in contrast to the initial hypothesis, are rather uniform and since HSs are parts of life on Earth, we furthermore presume that they may have been a primordial exogenous trigger for the development of anti-stress systems in exposed organisms. Recommendations and perspectives Effect studies of chemical stresses on organisms should consider exposure to both natural triggers and xenobiotic compounds in low concentrations—in order to prospectively differentiate between these triggers and, subsequently, classify them.

INTRODUCTION: Naturally occurring hydroxylated polybrominated diphenyl ethers (OH-PBDEs), their methoxylated counterparts (MeO-PBDEs), and polybrominated dibenzo-p-dioxins (PBDDs), together with their potential precursors polybrominated phenols (PBPs) and polybrominated anisoles (PBAs), were analyzed in blue mussels (Mytilus edulis) gathered along the east coast (bordering the Baltic Sea) and west coast of Sweden (bordering the North Sea). Brown algae (Dictyosiphon foenicolaceus) and cyanobacteria (Nodularia spumigena) from the Baltic Sea, considered to be among the primary producers of these compounds, were also analyzed for comparison. MATERIALS AND METHODS: The samples were liquid–liquid extracted, separated into a phenolic and a neutral fraction, and subsequently analyzed by gas chromatography–mass spectrometry (GS-MS). RESULTS AND DISCUSSION: The levels of OH-PBDEs, MeO-PBDEs and PBDDs were significantly higher in Baltic Sea mussels than in those from the west coast, whereas the levels of PBPs and PBAs displayed the opposite pattern. The blue mussels from the Baltic Sea contained high levels of all analyzed substances, much higher than the levels of, e.g., polybrominated diphenyl ethers. In addition, the GC-MS chromatogram of the phenolic fraction of the west coast samples was dominated by four unknown peak clusters, three of which were tentatively identified as dihydroxy-PBDEs and the other as a hydroxylated-methyl-tetraBDE. CONCLUSIONS: Clearly, all of the compounds analyzed are natural products, both in the Baltic and the North Sea. However, the geographical differences in composition may indicate different origin, e.g., due to differences in the occurrence and/or abundance of various algae species along these two coasts or possibly a more extensive dilution on the west coast.

Background, aim, and scope Glutaraldehyde (GA) often acts as an effective sterilant, disinfectant, and preservative in chemical products. It was found that GA had clearly acute toxicity to aquatic organisms. Furthermore, GA in natural environment could not exist as single species but as complex mixtures. To explore the toxicity interaction between GA and the other environmental pollutant, it is necessary to determine the mixture toxicities of various binary mixtures including GA. Two reference models, concentration addition (CA) and independent action (IA), are often employed to evaluate the mixture toxicity, which can be finished by comparing the concentration-response curves (CRCs) predicted by the reference models with the experimental CRC of the mixture. However, the CRC-based method cannot effectively denote the degree of the deviations from the reference models, especially at very low effect levels. Though the model deviation ratio (MDR) can be used to quantitatively evaluate the deviation of a mixture at EC50 level from the reference model, it is difficult to evaluate the deviations at the lower effect levels. Therefore, the primary aim of this study was to develop a new effect residual ratio (ERR) method to validate the deviations from the reference models at various effect levels. Materials and methods Four chemicals having possible dissimilar mode of actions with GA, acetonitrile (ACN), dodine (DOD), simetryn (SIM), and metham sodium (MET), were selected as another component in the binary mixtures including GA, which constructed four binary mixtures, GA-ACN, GA-DOD, GA-SIM, and GA-MET ones. For each binary mixture, two equipotent mixture rays where the concentration ratios of GA to another mixture component are respectively EC50 and EC5 ones were designed and their toxicities (expressed as a percent inhibition to Photobacterium phosphoreum) were determined by microplate toxicity analysis. The observed concentration-response curve (CRC) of a ray was compared with that predicted by CA or IA model to qualitatively assess the toxicity interaction of the mixture ray. To quantitatively and effectively examine the deviations at various effect levels from the reference models, a new concept, ERR at an effect, was defined, and the ERR was employed to evaluate the deviation at various effects with confidence intervals. Results For three binary mixtures, GA-ACN, GA-DOD, and GA-SIM, the CRCs predicted by IA models were almost located in the 95% confidence intervals of the experimental CRCs for both equipotent mixture rays, which indicated the independent actions between binary mixture components. However, two rays of GA-MET binary mixture displayed a little synergistic action because both CRCs predicted by CA and IA were lower than the experimental CRC. ERR showed the same results as MDR, but ERR results at low effect area were clearer than MDR ones. Discussion In CRC comparison, the deviation of CA (for GA-ACN, GA-DOD, and GA-SIM combinations) or IA (for GA-MET) model from the experimental values could be obviously observed at medium area of the CRC. However, at very low effect levels, both deviations of CA and IA and difference between CA and IA model predictions were not very apparent. Thus, it was difficult to confirm which model, CA or IA, had better predicted power at very low effect levels. MDR in many literatures often refers to a ratio at EC50 level. It was also difficult to reflect not only the deviation fact at the other ECx but also the deviation uncertainty. After we extended the definition of MDR to all ECx and examined the 95% confidence intervals based on observation, the plot of the redefined MDRs at many effect levels could better explain the deviations of CA or IA model from the observation. However, MDRs at very low effect levels did not still reflect the high uncertainty there. The ERRs defined in our paper could explicitly explain the degree of deviation from the reference models and especially reflect the high uncertainty at very low effects. It could be said that the ERR is a better indicator than MDR. Conclusions The new ERR validation method developed in our laboratory could provide us with the information about the toxicity interaction between the mixture components and quantitatively assess the accuracy of the reference models (CA or IA) at whole effect levels. The ERR method conquered the invalidation of the classical CRC comparison method on the deviation decision at low effect levels and also got the advantage over the MDR methods. Recommendations and perspectives It holds promise to become an effective method of hazard and risk assessments of chemical mixtures by well characterizing the uncertainty at very low effect levels.

Background, aim, and scope Polybrominated diphenyl ethers (PBDEs) and their metabolites are toxic to animals, and concentrations of the PBDEs metabolites can exceed those of the parent materials. But no information was available on concentrations of PBDEs metabolites in the lower Yangtze River in the region around Jiangsu Province of China, which is heavily urbanized and industrialized area. The aims of this study were to determine whether PBDEs and their methoxylated PBDEs (MeO-PBDEs) were accumulated in Coilia sp. in this area and to investigate the potential sources for these two kinds of brominated organic pollutants. Materials and methods Samples of four species of anchovy were collected from eight sites in the lower Yangtze River, Taihu Lake, and Hongzehu Lake. Concentrations of 13 PBDEs congeners and eight methoxylated PBDEs were determined by use of organic solvent extraction, followed by gas chromatography and mass spectrometry. Results and discussion The frequencies of detection for PBDEs and MeO-PBDEs were 92% and 53%, respectively. Concentrations of ∑PBDEs ranged from not detected (ND) to 77 ng/g lipids (ND-3.8 ng/g wet weight). Concentrations of ∑MeO-PBDEs in anchovy ranged from ND to 48 ng/g lipids (ND-8.2 ng/g wet weight). The PBDE concentrations in anchovy from the Yangtze River Delta were similar to or less than those reported for other species from other locations around the world, while the concentrations of MeO-PBDEs were comparable to or slightly less than those reported in other studies. This is the first report of MeO-PBDEs in biota of China. Conclusions The results of this study as well as those of other studies suggest that PBDEs in anchovy are primarily of synthetic origin and released by human activities, while MeO-PBDEs in anchovy are primarily from nature as natural products from the sea instead of metabolism of PBDEs in anchovy.

Background, aim and scope Ambient air concentrations of polycyclic aromatic hydrocarbons (PAH) were determined at five elevated mountain sites on the European continent and the Atlantic Ocean. All sites can be considered remote background areas since they are situated above the timberline and they lack local emission sources of these compounds. Results and discussion Average gas phase concentrations of ΣPAH were 165, 1,475, 1,553, 1,822 and 4,443 pg m⁻³ for Tenerife, Pyrenees, Central Norway, Tyrolean Alps and High Tatras, respectively. Particulate phase concentrations were 55, 70, 383, 196 and 708 pg m⁻³, respectively. The PAH profiles of samples from the different sites are very similar, being typical of PAH mixtures after long-range atmospheric transport. Part of the fluctuations in PAH concentrations are explained by the influence of temperature on the particulate/gas phase partitioning. Conclusion The differences in PAH levels between sites, with the lowest concentrations found in Tenerife and the highest in the High Tatras, suggest the geographical influence of regional emissions on the sites, especially in the cold periods and for the sites in the eastern sector of the European continent. This is supported by air mass back-trajectories analysis for the samples on the different sites. The influence of the continent is not detectable in the case of the elevated site of Tenerife where the free troposphere has been sampled. The results in this study are consistent with the PAH levels found in soils and/or high mountain lake sediments from these areas.

Global warming has become a source of awareness regarding the potential deleterious effects of extreme abiotic factors (e.g., temperature, dissolved oxygen (DO) levels) and also their influence on chemicals toxicity. In this work, we studied the combined effects of nickel and temperature (low and high levels) and nickel and low levels of DO to Daphnia magna, and concentration addition and independent action concepts as well as their deviations for synergism/antagonism, dose ratio and dose level dependency, were applied to survival and feeding rate data. Nickel single exposure showed an LC₅₀ value for 48 h of 7.36 mg l⁻¹ and an EC₅₀ value for feeding impairment at 2.41 mg l⁻¹. In the acute exposures to high and low temperatures, 50% of mortality was observed, respectively, at 30.7°C and 4.2°C whereas 50% reduction on the feeding activity was recorded at 22.6°C and 16.0°C. Relatively to low DO levels, a LC₅₀ value for 48 h of 0.5 mg l⁻¹ was obtained; feeding activity EC₅₀ value was 2 mg l⁻¹. On acute combined experiments, antagonism was observed for the combination of nickel and extreme temperatures, whereas a synergistic behaviour was observed in the combined exposure of nickel and low DO levels. At sublethal levels, nickel showed to be the main inducer of toxicity at high and low temperatures but not at low levels of dissolved oxygen. Toxicokinetics and toxicodynamics modelling studies should be made in the future to understand the toxicological pathways involved on complex combinations of stressors and to validate any conclusions.

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.

Purpose The purpose of this paper is to assess fatal cancer risk after external and internal (inhalation and ingestion) exposure from natural radionuclides in soil like ²³⁸U, ²³²Th, ⁴⁰K, and ²²⁶Ra on the territory of Bela Crkva, Serbia. Although receiving doses are low from sources like natural radionuclides in soil, because of stochastic effects of ionizing radiation, risk for developing cancer exists and can be quantified. Methods Concentrations of radionuclides from 80 soil samples are measured using HPGe detector. Fatal cancer risk is assessed from calculated ambient dose rate in the target organs of body due to external and internal exposure. Monte Carlo simulations are used to obtain conversion factors which are required to calculate absorbed dose rate in target organs. Results and discussion Assessed cancer risk for ²³⁸U in the case of both inhalation and ingestion exposure is from 1.11 × 10⁻⁶ to 24 × 10⁻⁶ for minimal and maximal activity in soil samples, from 1.02 × 10⁻⁶ to 23.3 × 10⁻⁶ for exposure to ²²⁶Ra, from 1.89 × 10⁻⁶ to 50.3 × 10⁻⁶ for exposure to ²³²Th, and from 0.265 × 10⁻⁶ to 9.83 × 10⁻⁶ for exposure to ⁴⁰K. Overall risk from ⁴⁰K as external and internal source is from 0.8 × 10⁻⁶ to 31.9 × 10⁻⁶. Calculated cancer risks from both inhalation and ingestion exposure could be related to all tissues that are on the way of distribution of particles within the body but especially to deposition sites in the body. Conclusion Assessed risks for fatal cancer development from inhaled and ingested natural radionuclides originating in soil are not increased.