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 Current Australian legislation permits the beneficial application of grease trap waste (GTW) to agricultural soil, viewing it as a beneficial source of organic matter and soil conditioner containing no/low amounts of metals or pathogenic organisms. However, little is known about the influence of GTW on soil bacterial community. A field experiment was established at Menangle in south western Sydney in Australia to quantitatively assess the impacts of different types (GTW CO and GTW CL) and amounts of GTW application on the soil bacterial community and diversity. Furthermore, a municipal solid waste (MSW) compost was simultaneously examined to compare against the other organic wastes. Knowledge about the shifts in microbial community structure and diversity following the applications of organic wastes could help to evaluate the ecological consequences on the soil and thus to develop sound regulatory guidelines for the beneficial reuse of organic wastes in agricultural lands. Materials and methods Soil samples were collected from recycled organics plots treated with different types and quantity of organic wastes. The field experimental treatments included control (CK, without application of any organic wastes), low amount of GTW CO (COL), GTW CL (CLL), and MSW (ML), and high amounts of GTW CO (COH), GTW CL (CLH), and MSW compost (MH). Microbial DNA was extracted from soil samples and the 16S rRNA genes were polymerase chain reaction (PCR)-amplified. The PCR products were analyzed by denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing. The bacterial community structures and diversity were assessed using the DGGE profiles and clone libraries constructed from the excised DGGE bands. Results DGGE-based analyses showed that application of the GTW CO, regardless of the amount applied, had significant negative effects on soil bacterial genotypic diversity and community structure compared with the control, while the applications of other organic wastes including the GTW CL and MSW had no clear effects. The effects of the rate of organic waste application on soil bacterial community characteristics varied with the types of organic wastes applied. Sequence-based analyses of 126 clones indicated that Proteobacteria (53.2%) was the dominant taxa at the experimental site, followed by Actinobacteria (9.5%), Bacteroidetes (7.9%), Firmicutes (7.9%), Gemmatimonadetes (5.6%), Chloroflexi (2.4%), Acidobacteria (1.6%) and the unclassified group (11.9%). In the COH treatment, Acidobacteria, Bacteroidetes, and Gemmatimonadetes were not detected; the percentages of Firmicutes, Proteobacteria, and Actinobacteria in the COH treatment were significantly different from those in CK. There is a significant positive correlation (r = 0.71, p = 0.002) between the C/N ratio of organic wastes and the bacterial genotypic communities. Discussion Both the type and the amount of GTW applied affected soil bacterial genotypic diversity and community structure. The different effects of various types of organic wastes on soil bacterial characteristics may be predicted by the differences in specific properties of organic wastes such as C/N ratio, as evidenced by the strong and significant positive relationship between the bacterial community distance and the environmental distance of C/N ratio. This also indicates that the C/N ratio of GTW applied can be a major driver for the shift in the soil bacterial community. Conclusions Our results revealed that the effects of organic wastes on soil bacterial communities varied with the types of organic wastes, and depending on the rate of application. Application of the GTW CO led to significant shifts in soil bacterial community diversity and structure. The effects of different types of organic wastes on the soil bacterial characteristics can be predicted by the differences of specific properties of organic wastes, such as the C/N ratio. Sequence-based analyses of 126 clones indicated that Proteobacteria was the dominant taxa at the experimental site. Recommendations and perspectives Our results have important implications for developing sound regulatory guidelines for the beneficial reuse of organic wastes, indicating that GTW CO and similar organic waste treatments may not be suitable for application in agricultural soils due to its significant negative effect on soil bacterial community.

Background, aim, and scope Passive air sampling survey of the Central and Eastern Europe was initiated in 2006. This paper presents data on toxic organic compounds such as polychlorinated biphenyls (PCB 28, 52, 101, 118, 153, 138, and 180), hexachlorobenzene (HCB), pentachlorobenzene (PeCB), hexachlorocyclohexane compounds (α-HCH, β-HCH,γ-HCH, δ-HCH), and dichloro-diphenyl-trichloroethane (DDT) compounds (p,p′DDE, p,p′DDD, p,p′DDT, o,p′DDE, o,p′DDD, and o,p′DDT) determined in ambient air and soil samples collected at Estonian monitoring stations. Materials and methods Ambient air and soil samples were collected in five sites in northern Estonia. Passive air samplers were deployed four times over 4-week periods covering the period April-August 2006. Samples were analyzed using gas chromatography-electron capture detector (HP 5890) supplied with a Quadrex fused silica column 5% Ph for organochlorine pesticides (OCPs). Local ground-boundary wind field was modeled for each monitoring station and sampling period on the basis of observed wind data from the nearest meteorological station with a high quality of time series and compared with upper air (at 850- and 500-hPa level) data from Tallinn-Harku aerological station. Results Median levels of PCB at Estonian stations varied between 3 and 9 ng/filter, although the maximum in Kohtla-Järve reached as high as 28 ng/filter. Sampling rates about 3.5 m³/day were determined by empirical measurements, making approximately 100 m³ for a 28-day sampling cycle. In general, OCP levels in soil were at the limit of detection, except Tallinn site and Muuga Port affected mainly by local sources. However, the atmospheric PCB concentrations are in agreement with the soil analyses where highest PCB levels were found in the soil sample for Tallinn (12.0 ng/g dry weight). For HCB, the atmospheric distribution was quite uniform, with the background levels sometimes higher than the urban ones. HCB and PeCB concentrations were very low in May and June when meridional airflow from the southern sector dominated, and concentrations were slightly higher in July and August, most probably due to revolatilization of adsorbed HCB (with PeCB impurities) from former industrial applications during the summer month and possibly enhanced by forest fires in Russia. Also, the highest summary HCH and DDT levels (63.5 and 2.5 ng/filter, respectively) in Estonian monitoring stations were determined at the end of July and beginning of August when the ground-boundary wind direction was from NE with relatively high speed (4-7 m/s). The highest DDT levels in ambient air (3.5 ng/filter) were determined in the spring samples. For DDT and HCH, long-range atmospheric transport clearly dominates persistent OCP, atmospheric input to Estonia as well as for the Scandinavian countries. The DDE/DDT ratio was >1, indicating no fresh input. Discussion The passive air sampling demonstrates uniform distribution of OCPs. In the regional context, there is no indication of increased levels of concentrations of OCPs in the industrial Northeast Estonia where the oil shale processing causes certain pollution impacts. Though the passive sampling does not apply for monitoring of short-term fluxes, the method is capable of reflecting background levels in long-term prospective for potential effect on human health due to long-term exposition of OCPs. Conclusions PCB and its congeners, HCB, PeCB, HCH, and DDT were very low in Estonia. None of the persistent organochlorine pesticides have ever been produced in Estonia, and as of today, all old OCP stocks in the country have been destroyed. Highest concentrations could be expected in March and April when southwestern airflow is still strong and dominant, but air humidity is lower and deposition takes place far from the place of origin of OCPs. In summer, the share of locally formed organic compounds increases and deposition depends strongly on weather conditions. In some cases in Tallinn and Muuga where local anthropogenic impact occurs, HCB and PeCB stem from revolatilization of industrial application. Recommendations and perspectives The passive air sampling could be employed more widely to explore long-term human exposure to OCP deposition and assess potential health risks. The survey based on passive air sampling could be extended from Central and Eastern Europe to other European regions to get methodically adjusted cross-European data coverage. Based on the results of the survey, the Lahemaa reference station is a feasible option to represent background monitoring of persistent organic pollutants.

Background, aim, and scope Photochemical smog, characterized by high concentrations of O₃ and fine particles, is of great concern in the urban areas, in particular megacities and city clusters like the Pearl River Delta. Materials, methods, and results Ambient ozone (O₃) and its precursors were simultaneously measured at two sites in the Pearl River Delta, namely, Wan Qing Sha (WQS) in Guangzhou and Tung Chung (TC) in Hong Kong, from 23 October to 01 December 2007 in order to explore their potential relationship. Eight high O₃ episode days were identified at WQS and two at TC during the sampling campaign, indicating a more serious O₃ pollution in Guangzhou than in Hong Kong. An observation-based model was employed to determine the ozone-precursor relationship. At both sites, O₃ production was found to be volatile organic compound (VOC)-limited, which is consistent with previous observations. Anthropogenic hydrocarbons played a key role in O₃ production, while reducing nitric oxide emissions aided the buildup of O₃ concentrations. Among VOC species, the summed relative incremental reactivity (RIR) of the top 12 compounds accounted for 89% and 85% of the total RIR at WQS and TC, respectively, indicating that local photochemical O₃ formation can be mainly attributed to a small number of VOC species. Discussion and conclusions A large increment in both simulated HO₂ and O₃ concentrations was achieved with additional input of hourly carbonyl data. This suggested that apart from hydrocarbons, carbonyls might significantly contribute to the O₃ production in the Pearl River Delta.

Introduction The effect of oceanic CO₂ sequestration was examined exposing a deep-sea bacterium identified as Vibrio alginolyticus (9NA) to elevated levels of carbon dioxide and monitoring its growth at 2,750 psi (1,846 m depth). Findings The wild-type strain of 9NA could not grow in acidified marine broth below a pH of 5. The pH of marine broth did not drop below this level until at least 20.8 mM of CO₂ was injected into the medium. 9NA did not grow at this CO₂ concentration or higher concentrations (31.2 and 41.6 mM) for at least 72 h. Carbon dioxide at 10.4 mM also inhibited growth, but the bacterium was able to recover and grow. Exposure to CO₂ caused the cell to undergo a morphological change and form a dimple-like structure. The membrane was also damaged but with no protein leakage.

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.

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 Precipitation samples collected from 1995 to 2000 at meteorological station in the eastern outskirts of Herceg Novi (Montenegro) were analysed on Na⁺, K⁺, Mg²⁺, Ca²⁺, Cl⁻, SO₄ ²⁻, NO₃ ⁻ and NH₄ ⁺. Four-day backward trajectory simulations were conducted during the precipitation period to investigate the regional transport of main ions and their deposition in the region of the southeastern Adriatic Sea. The air mass trajectories were classified into six trajectory categories by the origin and direction of their approach to Herceg Novi. Materials and methods A bottle and funnel with a small net between them was used for sampling at a height of 1.5 m above the ground. The concentrations of Cl⁻, NO₃ ⁻, NH₄ ⁺ and SO₄ ²⁻ were determined spectrophotometrically, the concentrations of Na⁺ and K⁺ were determined by the FAES method and the concentrations of Mg²⁺ and Ca²⁺ by the FAAS method. The factor analysis technique (PCA analysis) based on the calculation of the factors was employed to differentiate the contribution of emission sources to the content of the main ions in the precipitation. The obtained data sets were processed using the SPSS 11.5 statistical program. The Hybrid Single-Particle Lagrangian Integrated Trajectory model was used to study the air origin for the city of Herceg Novi (42°27′N, 18°33′E), Montenegro. Discussion The following origins of the air masses were considered: northern Europe (NE), eastern Europe-northeastern Europe (EE-NE); eastern Mediterranean-southeastern Europe (EM-SE); Africa-Central Mediterranean (A-CM); western Mediterranean (WM); western Europe-Central Europe (WE-CE) and undefined. The heights and frequencies of precipitation coming by air masses from northern Europe and eastern-northeastern Europe are the lowest. On the contrary, the heights and frequencies of precipitation coming by air masses from the western Mediterranean (36.6%) and Africa and the Central Mediterranean (30.6%) are the highest. The sea salt components (Na⁺, Cl⁻, Mg²⁺) are significantly correlated, except for air masses originating from the northern and eastern European regions. Significant correlations between SO₄ ²⁻ and NO₃ ⁻ are found in air masses coming from the western Europe and North Africa, over the Mediterranean. Conclusions The highest volume-weighted mean (VWM) of: SO₄ ²⁻ , NH₄ ⁺ and Mg²⁺ are for precipitation from EE-NE while the highest values of VWM of Cl ⁻ are from WM and of K⁺ are from WE-CE. Long-range transport of Sahara dust is confirmed. Recommendations and perspectives For better estimation of origins of water-soluble ions in precipitation expanding list of analysis on anions of organic acids, such as HCOO⁻, CH₃COO⁻, and C₂H₂COO⁻, could be indicative of volatile organic compounds emitted by vegetation but also traffic. The chemical composition of precipitation together with a study of air backward trajectories is the proper tool for tracking the long-range transport of water-soluble ions and estimating transboundary pollution.