Selected endocrine disrupting compounds (Vinclozolin,Flutamide, Ketoconazole and Dicofol): Effects on survival,occurrence of males,growth, molting and reproduction of <Emphasis Type="Italic">Daphnia magna</Emphasis>

BACKGROUND, AIM AND SCOPE: Pollution-induced endocrine disruption in vertebrates and invertebrates is a worldwide environmental problem, but relatively little is known about effects of endocrine disrupting compounds (EDCs) in planktonic crustaceans (including Daphnia magna). Aims of the present study were to investigate acute 48 h toxicity and sub-chronic (4-6 days) and chronic (21 days) effects of selected EDCs in D. magna. We have investigated both traditional endpoints as well as other parameters such as sex determination, maturation, molting or embryogenesis in order to evaluate the sensitivity and possible use of these endpoints in ecological risk assessment. MATERIALS AND METHODS: We have studied effects of four model EDCs (vinclozolin, flutamide, ketoconazole and dicofol) on D. magna using (i) an acute 48 h immobilization assay, (ii) a sub-chronic, 4-6 day assay evaluating development and the sex ratio of neonates, and (iii) a chronic, 21 day assay studying number of neonates, sex of neonates, molting frequency, day of maturation and the growth of maternal organisms. RESULTS: Acute EC50 values in the 48 h immobilization test were as follows (mg/L): dicofol 0.2, ketoconazole 1.5, flutamide 2.7, vinclozolin >3. Short-term, 4-6 day assays with sublethal concentrations showed that the sex ratio in Daphnia was modulated by vinclozolin (decreased number of neonate males at 1 mg/L) and dicofol (increase in males at 0.1 mg/L). Flutamide (up to 1 mg/L) had no effect on the sex of neonates, but inhibited embryonic development at certain stages during chronic assay, resulting in abortions. Ketoconazole had no significant effects on the studied processes up to 1 mg/L. DISCUSSION: Sex ratio modulations by some chemicals (vinclozolin and dicofol) corresponded to the known action of these compounds in vertebrates (i.e. anti-androgenicity and anti-oestrogenicity, respectively). Our study revealed that some chemicals known to affect steroid-regulated processes in vertebrates can also affect sublethal endpoints (e.g. embryonic sex determination and/or reproduction) in invertebrates such as D. magna. CONCLUSIONS: A series of model vertebrate endocrine disrupters affected various sub-chronic and chronic parameters in D. magna including several endpoints that have not been previously studied in detail (such as sex determination in neonates, embryogenesis, molting and maturation). Evaluations of traditional reproduction parameters (obtained from the 21 day chronic assay). as well as the results from a rapid, 4-6 day, sub-chronic assay provide complementary information on non-lethal effects of suspected organic endocrine disrupters. RECOMMENDATIONS AND PERSPECTIVES: It seems that there are analogies between vertebrates and invertebrates in toxicity mechanisms and in vivo effects of endocrine disruptors. However, general physiological status of organisms may also indirectly affect endpoints that are traditionally considered 'hormone regulated' (especially at higher effective concentrations as observed in this study) and these factors should be carefully considered. Further research of D. magna physiology and comparative studies with various EDCs will help to understand mechanisms of action as well as ecological risks of EDCs in the environment.     

Europe-wide survey of estrogenicity in wastewater treatment plant effluents: the need for the effect-based monitoring

A pan-European monitoring campaign of the wastewater treatment plant (WWTP) effluents was conducted to obtain a concise picture on a broad range of pollutants including estrogenic compounds. Snapshot samples from 75 WWTP effluents were collected and analysed for concentrations of 150 polar organic and 20 inorganic compounds as well as estrogenicity using the MVLN reporter gene assay. The effect-based assessment determined estrogenicity in 27 of 75 samples tested with the concentrations ranging from 0.53 to 17.9 ng/L of 17-beta-estradiol equivalents (EEQ). Approximately one third of municipal WWTP effluents contained EEQ greater than 0.5 ng/L EEQ, which confirmed the importance of cities as the major contamination source. Beside municipal WWTPs, some treated industrial wastewaters also exhibited detectable EEQ, indicating the importance to investigate phytoestrogens released from plant processing factories. No steroid estrogens were detected in any of the samples by instrumental methods above their limits of quantification of 10 ng/L, and none of the other analysed classes of chemicals showed correlation with detected EEQs. The study demonstrates the need of effect-based monitoring to assess certain classes of contaminants such as estrogens, which are known to occur at low concentrations being of serious toxicological concern for aquatic biota.     

Multisource emission retrieval within a biogas plant based on inverse dispersion calculations—a real-life example

Open digestate storage tanks were identified as one of the main methane (CH₄) emitters of a biogas plant. The main purpose of this paper is to determine these emission rates using an inverse dispersion technique in conjunction with open-path tunable diode laser spectroscopy (OP-TDLS) concentration measurements for multisource reconstruction. Since the condition number, a measure of “ill-conditioned” matrices, strongly influences the accuracy of source reconstruction, it is used as a diagnostic of error sensitivity. The investigations demonstrate that the condition number for a given source-sensor configuration in the highly disturbed flow field within the plant significantly depends on the meteorological conditions (e.g., wind speed, stratification, wind direction, etc.). The CH₄ emissions are retrieved by removing unrepresentative periods with high condition numbers, which indicate uncertainty in recovering the individual sources. In a final step, the CH₄ emissions are compared with the maximum biological methane potential (BMP) in the digestate analyzed under laboratory conditions. The retrieved methane emission rates represent an average of 50 % of the maximum BMP of the stored digestate in the winter months, while they comprised an average of 85 % during the measurement campaigns in the summer months. The results indicate that the open tanks have the potential to represent a substantial emission source even during colder periods.     

Validation of the species sensitivity distribution in retrospective risk assessment of herbicides at the river basin scale—the Scheldt river basin case study

Species sensitivity distribution (SSD) is commonly used in prospective risk assessment to derive predicted no-effect concentrations, toxicity exposure ratios, and environmental quality standards for individual chemicals such as pesticides. The application of SSD in the retrospective risk assessment of chemical mixtures at the river basin scale (i.e., the estimation of “multiple substance potentially affected fractions” [msPAFs]) has been suggested, but detailed critical assessment of such an application is missing. The present study investigated the impact of different data validation approaches in a retrospective model case study focused on seven herbicides monitored at the Scheldt river basin (Belgium) between 1998 and 2009. The study demonstrated the successful application of the SSD approach. Relatively high impacts of herbicides on aquatic primary producers were predicted. Often, up to 40 % of the primary producer communities were affected, as predicted by chronic msPAF, and in some cases, the predicted impacts were even more pronounced. The risks posed by the studied herbicides decreased during the 1998–2009 period, along with decreasing concentrations of highly toxic pesticides such as simazine or isoproturon. Various data validation approaches (the removal of duplicate values and outliers, the testing of different exposure durations and purities of studied herbicides, etc.) substantially affected SSD at the level of individual studied compounds. However, the time-consuming validation procedures had only a minor impact on the outcomes of the retrospective risk assessment of herbicide mixtures at the river basin scale. Selection of the appropriate taxonomic group for SSD calculation and selection of the species-specific endpoint (i.e., the most sensitive or average value per species) were the most critical steps affecting the final risk values predicted. The present validation study provides a methodological basis for the practical use of SSD in the retrospective risk assessment of chemical mixtures.     

Utilization of the solid sorbent media in monitoring of airborne cyclophosphamide concentrations and the implications for occupational hygiene

The main objective of the study was to evaluate the applicability of two solid sorbent media (Anasorb 708 and Strata X), the impinger filled with distilled water and PTFE filters for determination of airborne cyclophosphamide (CP) in the hospital working environment. For this purpose, air contamination of Masaryk Memorial Cancer Institute (Czech Republic) was monitored using the sampling apparatus containing the samplers described above. In addition, the surface contamination was also determined using the wipe sampling technique. During the monitoring, contamination of three different workplaces (storage room, preparation room and outpatient clinic) was studied. Using Strata X solid sorbent tubes, airborne CP was determined in all (n = 5) samples collected at the outpatient clinic over a 5 day monitoring period (concentration range: 0.3-4.3 ng m(-3)). Other samplers (including PTFE filters) did not collect any detectable amount of CP (the limit of detection, LOD ≤ 0.1 ng m(-3)). Negative results detected at filter samples indicate that CP determined at Strata X samples was most probably of gaseous origin. Surface contamination ranged from <2 to 19, <8 to 418 and 133-15,500 pg cm(-2) at the storage room, preparation room and outpatient clinic, respectively. The study showed that evaporation of antineoplastic drugs should not be neglected, albeit the concentrations determined in our study are relatively low. Therefore, proper monitoring of airborne contamination should involve simultaneous sampling of both particle-bonded and gaseous phases. In this way, Strata X sorbent tubes seem to be an effective tool for the sampling of gaseous CP in the indoor air.     

Determination of anthropogenic boundary depth in industrially polluted soil and semi-quantification of heavy metal loads using magnetic susceptibility

This study focuses on magnetic susceptibility processing and analysis towards fast and cost-efficient discrimination and semi-quantification of anthropogenic heavy metal loads in soil. Spatial variability of magnetic susceptibility was investigated on sets of soil cores from both “polluted” and “less polluted” forest soil close to a steel mill near Leoben, Austria. Test sites of ∼10 m² represent “site scale” dimensions. Statistical analysis of magnetic data provides a boundary depth indicating the transition from the “polluted” to the deeper, “unpolluted” zone in contaminated natural soil. Introduction of a block master curve simplifies the complex variations of individual curves, and represents magnetic susceptibility at “site scale”. For linking the block master curve to heavy metals we only require magnetic susceptibility data from one soil core and heavy metal data from two sub-samples from the same core. Our optimized magnetic susceptibility data processing scheme provides an applicable tool to semi-quantify anthropogenic heavy metal loads in soil.     

Removal of Microcystins by Phototrophic Biofilms. A Microcosm Study (6 pp)

Background, Aims and Scope Microcystins (MCs) are a family of natural toxins produced by cyanobacteria (blue-green algae). As a result of eutrophication, massive cyanobacterial blooms occur more frequently and MCs represent important contaminants of freshwater ecosystems. Bacterial biodegradation is considered a main mechanism for MC breakdown in environmental conditions. While existing studies were mostly focused on MC biodegradation by planktonic bacteria, our experiments examined the fate and kinetics of MC degradation in river-originated phototrophic biofilms and investigated factors influencing the rate of MC removal. Methods The fate of dissolved MCs was studied in laboratory microcosms with different composition (containing water only, water with phytoplankton and/or phototrophic biofilms). Biofilms originated from river ecosystem were pre-incubated under various conditions (with/without presence of cyanobacterial biomass or model organic substrates: glucose and protein - casein). Changes in MC concentration (0-14 days) in water columns were measured by HPLC DAD after external additions of purified MCs (160 μg L-1, MC-LR and MC YR), and halftimes (t1/2) of MC removal were estimated. Results and Discussion The slow degradation of MC was revealed in tap water (t1/2 ~ 14 days) and river water without cyanobacteria (t1/2 ~ 8 days). Enhanced removal occurred in the presence of natural planktonic cyanobacteria (t1/2 ~ 44 h), most probably due to microorganisms associated with the biomass of cyanobacterial bloom. More rapid MC elimination occurred in the variants containing phototrophic biofilms, and was particularly pronounced at those biofilms pre-cultivated in the presence of cyanobacterial blooms (t1/2 ~ 20 h). Much slower removal was observed in the variants simulating possible substrate-dependent induction of microorganism metabolism (biofilms pre-incubated with glucose: t1/2 ~ 35 h, and casein: t1/2 ~ 80 h). After termination of experiments, total amounts of MCs accumulated in the biofilms were below 5% of the initial toxin level revealing significant biodegradation processes. Conclusion The microcosm studies contributed to understanding of the environmental fate of MCs and revealed a rapid biodegradation by phototrophic biofilms. The rate of MC elimination depends on history of biofilm community, previous contact with cyanobacteria seems to be a selective factor improving the biodegradation potential. Recommendation and Outlook Our results experimentally showed a positive role of biofilms in MC elimination during water treatment processes such as bank filtration or slow sand filtration, and could eventually serve for further research of biofilm-based technological applications for MCs removal in small-scale drinking water treatment facilities.