Purpose Dissolved humic acids abiotically reduced inorganic arsenic to varying degrees, depending on solution pH, ionic strength, and type of humate used. The functionalities of dissolved organic matter responsible for these redox reactions remained in question, but quinoid moieties undoubtedly played an important role. It is not fully understood whether the quinoids are solely responsible for arsenate reduction, and what the kinetics of the relevant processes are. Methods Electron spin resonance (ESR) spectroscopy was used to monitor the radical content of the humates, both as bulk material and as size fractions. Information on the redox status of the humates was obtained from fluorescence excitation-emission matrices and correlated with the observed spin count. Size data were obtained from fractionation and UV-Vis spectrometry. Arsenic speciation was carried out by ion chromatography. Results ESR spectroscopy showed a free radical content of 3.4 × 1,017-20 × 1,017 spins/g for bulk and fractionated aqueous humic acids. The number of electrons corresponding to these counts could not account for the entire charge transferred to arsenate during abiotic reduction. The rate constants of the reactions were found to be independent of the humic concentration. Leonardite humic acid separated on a XAD-8 resin yielded fractions that on the short time frame (0-5 h) had rate constants of 0.035 h⁻¹ for the hydrophobic fraction compared to 0.0052 h⁻¹ for the hydrophilic fraction. The rate constants for the hydrophobic and hydrophilic fractions over the longer time frame (100-200 h) were similar—7.3 × 10⁻⁴ and 7.2 × 10⁻⁴ h⁻¹, respectively. Fluorescence excitation-emission matrices of humates involved in arsenate reduction exhibited shifts typical of quinoid components undergoing redox transformations. These gradual shifts took place during the first 24 h of reduction process, after which the spectra no longer changed. The reduction of arsenate, however, continued after this period, indicating that species other than quinoids were involved. This was consistent with the fact that the rate constants for the later processes were smaller. Conclusions The existence of different redox pools within the humate was confirmed, with the quinoid-centered redox entities showing the fastest kinetics. The results pertained to all size and polarity fractions.

PURPOSE: Organophosphate pesticides (OPs) are among the most used insecticides in agriculture, causing the inhibition of esterases like acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and carboxylesterase (CbE). Pesticides can reach the aquatic environment, posing risks to non-target organisms, including tadpoles. METHODS: In this work, we characterized the activities of AChE, BChE and CbE in tadpoles of the snouted treefrog Scinax fuscovarius, and verified their in vitro sensibility to different inhibitors [phenylmethane sulfonyl fluoride (PMSF), tetra-isopropylpyrophosphamide (iso-OMPA) and the OP diazinon]. In vivo effects of diazinon and esterase recovery after 2-pyridine-aldoxime (2-PAM) treatment of the protein extract were also studied in tadpoles with distinct stages of development exposed to 1 and 3 mg/l for 2 and 7 days. RESULTS: Optimal conditions were established for AChE and CbE; BChE activity was negligible. PMSF affected esterase activities and is not recommended for homogenization buffers. Iso-OMPA treatment caused no changes in AChE and CbE activities, but diazinon inhibited these enzymes in a dose-responsive manner. In vivo, CbE activity was insensitive to diazinon in younger tadpoles, but inhibited after 2 days of exposure in more developed tadpoles. AChE activity was inhibited after 2 and 7 days of exposure, in a dose-responsive manner. Esterase reactivation by 2-PAM was obtained both in vitro and in vivo. CONCLUSIONS: (1) Tadpoles can be adequate sentinel organisms in biomonitoring studies of OP contamination; (2) AChE was more sensitive than CbE to diazinon; (3) tadpoles from earlier developmental stages seems to be less responsive to OPs; (4) AChE activity was sensitive to diazinon in both development stages, being a better OP biomarker.

Background, aim and scope Polychlorinated biphenyls (PCB) and dichlorodiphenyltrichloroethane (DDT) concentrations were determined in the liver of two deep sea fish species, Mediterranean slimehead and blackfin sorcerer, from the Adriatic Sea (southeastern Mediterranean Sea). Results and discussion The examination of congener profiles showed that hexachlorinated molecules were dominant (hexa-CBs, 55.3-56.2%), followed by penta-CBs (21.5-21.8%) and hepta-PCB 180 (14.9-16.0%). PCB 138, 153 and 180 were the prominent congeners accounting for 69.3% of the total PCBs. Among the compounds of DDT, p,p′-DDE was the most dominant molecule (Mediterranean slimehead, 86.6%; blackfin sorcerer, 92.8%), demonstrating the old age of these compounds in the environment. In both species, PCB contents were higher than those of DDTs. Contaminant load was higher in Mediterranean slimehead (PCBs, 1,086 ng g⁻¹ lipid weight; DDTs, 799 ng g⁻¹ lipid weight) than in blackfin sorcerer (PCBs, 561 ng g⁻¹ lipid weight; DDTs, 224 ng g⁻¹ lipid weight). Conclusion The high ratios ΣPCBs/ΣDDTs indicated predominantly industrial versus agrarian activities in the area. Dioxins toxic equivalent (TEQ) concentrations (8.1-18.7 pg TEQ per gram wet weight) reached those encountered in marine organisms at higher levels in the trophic chain, revealing the onerous status of contamination by PCBs in Mediterranean deep sea biota.

Introduction Within the last decade, numerous studies have investigated the role of environmental history on tolerance to stress of many organisms. This study aims to assess if Manila clams Ruditapes philippinarum may react differently to cadmium exposure and trematode parasite infection (Himasthla elongata) depending on their origin and environmental history in Arcachon Bay (France). Materials and methods Clams were exposed to Cd (15 µg L⁻¹) and parasites (25 cercariae per clam), alone or in combination, at 15°C under controlled laboratory conditions for 7 days. Metal accumulation and success of parasite infestation were examined, also physiological parameters such as metallothionein response and hemocyte counts and activities (phagocytosis, oxidative burst, viability, and adhesion). Results and Discussion Sensitivity of Manila clams to both stressors differed from one site to another, suggesting local adaptation of populations. Clams from the more parasitized site presented better resistance to trematodes than the others in terms of first line defense, i.e., avoidance of infection. On the other hand, clams that adapted to chronic Cd contamination showed better detoxification mechanisms, both in a faster transfer of metal from gills to visceral mass and in a higher metallothionein baseline, than clams which had never experienced Cd contamination. Finally, hemocyte concentration and viability differed between clam origin site, highlighting the fact that populations living in different environments may adapt their physiological and biochemical responses to environmental stressors. Conclusion It is therefore important to be cautious when extrapolating results from field studies of one species and one site, if the life history of the organisms is not taken into account.

Introduction and background Primary producers play critical structural and functional roles in aquatic ecosystems; therefore, it is imperative that the potential risks of toxicants to aquatic plants are adequately assessed in the risk assessment of chemicals. The standard required macrophyte test species is the floating (non-sediment-rooted) duckweed Lemna spp. This macrophyte species might not be representative of all floating, rooted, emergent, and submerged macrophyte species because of differences in the duration and mode of exposure; sensitivity to the specific toxic mode of action of the chemical; and species-specific traits (e.g., duckweed's very short generation time). Discussion and perspectives These topics were addressed during the workshop entitled “Aquatic Macrophyte Risk Assessment for Pesticides” (AMRAP) where a risk assessment scheme for aquatic macrophytes was proposed. Four working groups evolved from this workshop and were charged with the task of developing Tier 1 and higher-tier aquatic macrophyte risk assessment procedures. Subsequently, a SETAC Advisory Group, the Macrophyte Ecotoxicology Group (AMEG) was formed as an umbrella organization for various macrophyte working groups. The purpose of AMEG is to provide scientifically based guidance in all aspects of aquatic macrophyte testing in the laboratory and field, including prospective as well as retrospective risk assessments for chemicals. As AMEG expands, it will begin to address new topics including bioremediation and sustainable management of aquatic macrophytes in the context of ecosystem services.

Background, aim, and scope Lungs are permanently and simultaneously challenged by airborne microorganisms and airborne pollutants. Temporal increase of airborne particulate matter (APM), a potential carrier for extractable organic matter (EOM), degrades the situation of pulmonary patients. The Ah receptor (AhR) has been described as an important factor influencing the immunological challenge by viral infections. Molecular mechanisms underlying epidemiological observations are not well understood. Cytokine secretion (IL-6, IL-8, and TGF-β) from human bronchial epithelial cells (Beas2B) was determined as an indicator for immune responses upon co-stimulation with an artificial analog of viral dsRNA [polyinosinic/polycytidylic acid, (PIC)] and EOM of Standard Reference Material 1649a (SRM). Since polycyclic aromatic hydrocarbons are major components of APM usually acting via the AhR, particular focus was on AhR involvement. Materials and methods Cytokine secretion was demonstrated by enzyme-linked immunosorbent assay. To mimic the activation of organic matter during contact of particles with the human lung, Soxhlet extraction of SRM was performed. In some experiments, the AhR was blocked by α-naphthoflavone. Results Microbial stimulation (PIC) induced Beas2B cytokine release, whereas isolated exposure to EOM of APM did not. Co-stimulation with EOM and PIC increased IL-8 secretion, whereas neither IL-6 nor TGF-β was affected. Blocking of the AhR suppressed the release of IL-8. Discussion Organic compounds adsorbed on airborne particulate matter influence the cytokine secretion of lung epithelial cells induced by pathogen-associated molecular patterns. Recommendations and perspectives Further investigation of these observations is required to understand the molecular mechanisms underlying adverse health effects of APM reported in epidemiological studies.

Introduction From the metallurgic industry zone of Dambovita County, we harvested and analyzed seven herbaceous plants species (Lolium perenne, Festuca pratensis, Stipa capillata, Agrostis alba, Cynodon dactylon, Luzula campestris, and Agrostis tenuis) to establish the heavy metal accumulation levels in these species. Materials and method The heavy metal contents (for Cr, Mn, Zn, Sr, Cu, Ba, and Sn) were determined by analyzing the dry matter with an inductively coupled plasma-atomic emission spectrometer. This method has detection limits of 0.4-0.6 mg/kg for the analyzed metals. The heavy metal concentrations in plants harvested from the industrial area are in milligram per kilogram of dry matter and ranged from 10.03 to 191.98 mg/kg of dry matter for Cr, 165.89 to 1,103.92 mg/kg of dry matter for Mn, 62.09 to 921.67 mg/kg of dry matter for Zn, 29.21 to 50.12 mg/kg of dry matter for Sr, 0.99 to 113.83 mg/kg of dry matter for Cu, 58.66 to 133.51 mg/kg of dry matter for Ba, and 8.38 to 276.44 mg/kg of dry matter for Sn. The heavy metal accumulation levels in the studied species of plants were calculated by the rapport between the concentration level of the metal in plant samples and the level of the same metal in the soil, near the radicular system for each species of plants. Results The highest accumulation levels were found in A. alba for Cr (267.69%); in L. perenne for Mn (51.45%), Sr (114.35%), and Ba (60.81%); and in C. dactylon for Zn (136.62%), Cu (97.65%), and F. pratensis for Sn (704.00%).

Background, aim and scope Chlormequat (Cq) is a plant growth regulator used throughout the world. Despite indications of possible effects of Cq on mammalian health and fertility, little is known about its fate and transport in subsurface environments. The aim of this study was to determine the fate of Cq in three Danish subsurface environments, in particular with respect to retardation of Cq in the A and B horizons and the risk of leaching to the aquatic environment. The study combines laboratory fate studies of Cq sorption and dissipation with field scale monitoring of the concentration of Cq in the subsurface environment, including artificial drains. Materials and methods For the laboratory studies, soil was sampled from the A and B horizons at three Danish field research stations—two clayey till sites and one coarse sandy site. Adsorption and desorption were described by means of the distribution coefficient (K d) and the Freundlich adsorption coefficient (K F,ads). The dissipation rate was estimated using soil sampled from the A horizon at the three sites. Half life (DT₅₀) was calculated by approximation to first-order kinetics. A total of 282 water samples were collected at the sites under the field monitoring study— groundwater from shallow monitoring screens located 1.5-4.5 m b.g.s. at all three sites as well as drainage water from the two clayey sites and porewater from suction cups at the sandy site, in both cases from 1 m b.g.s. The samples were analysed using LC-MS/MS. The field monitoring study was supported by hydrological modelling, which provided an overall water balance and a description of soil water dynamics in the vadose zone. Results The DT₅₀ of Cq from the A horizon ranged from 21 to 61 days. The Cq concentration-dependant distribution coefficient (K d) ranged from 2 to 566 cm³/g (median 18 cm³/g), and was lowest in the sandy soil (both the A and B horizons). The K F,ads ranged from 3 to 23 (µg¹ ⁻ ¹/n (cm³)¹/n g⁻¹) with the exponent (1/n) ranging from 0.44 to 0.87, and was lowest in the soil from the sandy site. Desorption of Cq was very low for the soil types investigated (<10%w). Cq in concentrations exceeding the detection limit (0.01 µg/L) was only found in two of the 282 water samples, the highest concentration being 0.017 µg/L. Discussion That sorption was highest in the clayey till soils is attributable to the composition of the soil, the soil clay and iron content being the main determinant of Cq sorption in both the A and B horizons of the subsurface environment. Cq was not detected in concentrations exceeding the detection limit in either the groundwater or the porewater at the sandy site. The only two samples in which Cq was detected were drainage water samples from the two clayey till sites. The presence of Cq here was probably attributable to the hydrogeological setting as water flow at the two clayey till sites is dominated by macropore flow and less by the flow in the low permeability matrix. In contrast, water flow at the sandy site is dominated by matrix flow in the high permeability matrix, with negligible macropore flow. Given the characteristics of these field sites, Cq adsorption and desorption can be expected to be controlled by the clay composition and content and the iron content. Combining these observations with the findings of the sorption and dissipation studies indicates that the key determinant of Cq retardation and fate in the soil is sorption characteristics and bioavailability. Conclusions The leaching risk of Cq was negligible at the clayey till and sandy sites investigated. The adsorption and desorption experiments indicated that absorption of Cq was high at all three sites, in particular at the clayey till sites, and that desorption was generally very limited. The study indicates that leaching of Cq to the groundwater is hindered by sorption and dissipation. The detection of Cq in drainage water at the clayey till sites and the evidence for rapid transport through macropores indicate that heavy precipitation events may cause pulses of Cq. Recommendations and perspectives The present study is the first to indicate that the risk of Cq leaching to the groundwater and surface water is low. Prior to any generalisation of the present results, the fate of Cq needs to be studied in other soil types, application regimes and climatic conditions to determine the Cq retardation capacity of the soils. The study identifies bioavailability and heavy precipitation events as important factors when assessing the risk of Cq contamination of the aquatic environment. The possible effects of future climate change need to be considered when assessing whether or not Cq poses an environmental risk.

Introduction In the present work, we explored the kinetics of dichlorvos (2,2-dichlorvinyl dimethyl phosphate, DDVP) decay through UV-A light-induced TiO₂ photocatalysis at pH 4 and 9, and the formation of degradation intermediates and final products under specific experimental conditions. Experimental observations and theoretical considerations allowed us to suggest the degradation mechanism of DDVP by the UV/TiO₂ process in aqueous solution. Methods The irradiation experiments were carried out in a photoreactor using a 228-W medium-pressure Hg vapor lamp. The concentration of DDVP, phosphate ion and formaldehyde as reaction intermediate, are determined spectrophotometrically. Chloride ion concentration was measured potentiometrically. Results The photocatalytic degradation rate of dichlorvos (DDVP) under UV irradiation (360-380 nm) was optimized with respect to the flow rate of O₂ gas sparged into the solution and photocatalyst concentration for a constant dichlorvos concentration (1.66×10⁻⁴ M) at pH 4. Kinetic data were obtained at pH 4 and pH 9 for dichlorvos and the inorganic species released through its photo-induced degradation. The proposed mechanism which assumes the formation of some toxic intermediates resistant to mineralization is supported by theoretical calculations and the observed inorganic mass balances. Conclusions The calculated pseudo-first-order rate constants were dependent on the dissolved oxygen level at low O₂ flow rate, but somewhat independent on the initial pH. The decrease of pH during the irradiation suggests the formation of organic acids. The presence of organic intermediates was confirmed also by TOC measurements. A plausible reaction mechanism of DDVP degradation through the UV-A/TiO₂ process was proposed.

Background, aim and scope The paper presents concentrations of trace elements in blood of homebred animals (cows and sheep) from Southern Serbia (Bujanovac) and the contents of natural and anthropogenic radionuclides and some heavy metals in feed. The region of Southern Serbia was exposed to contamination by depleted uranium ammunition during NATO attacks in 1999 and therefore, is of great concern to environmental pollution and human and animal health. Materials and methods Conventional instrumental and epithermal neutron activation analyses are used to measure trace elements in cow and sheep blood samples collected randomly at six locations in the region of Bujanovac (village of Borovac) in the spring of 2005. Samples of feed (grass and crops: corn, wheat and oats), collected on the same locations (households), are analysed for the contents of radionuclides on an HPGe detector (Ortec, relative efficiency 23%) by standard gamma spectrometry. The content of Hg, Pb and Cd in feed is determined by standard atomic absorption spectrometry on the VarianSpectra220/ThermoSolar GFS97 spectrometer. Results Concentrations of 29 elements (Na, Al (P), Cl, K, Sc, Cr, Mn, Ni, Fe, Co, Zn, Se, As, Br, Sr, Rb, Sb, In, I, Ba, Cs, La, Nd, Eu, Sm, Tb, Hf, Ta and Th) are determined in blood of the examined animals. In feeds, natural ⁴⁰K is found in all of the samples, cosmogenic ⁷Be and fission product ¹³⁷Cs are detected only in the grass samples, while heavy metals Hg, Cd and Pb are found in the range of 0.01-0.02, 0.84-1.15 and 0.74-7.34 mg/kg, respectively. Calculated soil-to-blood transfer factors are in a wide range of 8·10⁻⁶ to 64, as a result of varying significance of the elements in animal metabolism and feeding habits. Discussion The results of trace elements' concentrations in animal blood are in good agreement with available data for K, Ni, Zn, Se and Rb. Higher Br concentrations in animal blood are most probably caused by large biomass burning events during blood sampling. Very low concentration of Fe in cows and sheep confirms the results of previous biochemical studies on animal anaemia in the region. High concentration of As correlates with geochemical peculiarities of the Balkans and is also likely influenced by the use of pesticides in the agricultural production. For some of the elements (La, Nd, Eu, Sm, Tb, Sb, Hf, Ta, Th, In, Ba, Sr, Sc and Cs), there are few or no literature data. Therefore, some of the presented data are significant not only for the country and the region, but on a wider scale. Activities of natural radionuclides in feeds are within the average values reported for the region, while the activities of ²¹⁰Pb and ²³⁵/²³⁸U are below the limit of detection. This is in accordance with previous investigations showing no widespread contamination by depleted uranium in the area. Contents of Hg and Pb in feeds are below the nationally permissible levels, unlike the content of Cd which exceeds it, probably caused by the use of phosphate fertilisers and fossil fuel combustion in the area. Conclusions In general, the concentrations of trace elements in blood of homebred cows and sheep are in good agreement with reference materials, available literature data and the results of previous studies in the area. The exceptions are Fe, As and Br. The contents of natural and anthropogenic radionuclides in feeds are within the expected levels, and there are no signs of contamination by depleted uranium or other fission products. Apart from Cd, there are no signs of pollution by heavy metals in feeds. The highly sensitive method of instrumental neutron activation analysis provides data on the concentration of some elements in animal blood not previously reported for the region and elsewhere. Recommendations and perspectives The presented study is a part of the long term ongoing project on the health risk assessment on animals and humans in the region. The collected data is intended to provide a base for the animal and human risk assessment as well as an estimate of the general pollution status of the environment in the region. Since some of the investigated elements are classified as important trace elements for livestock, the results could also be used to balance and improve the animal diet and thus, improve the growth and reproduction rate.