Endocrine disruption of water and sediment extracts in a non-radioactive dot blot/RNAse protection-assay using isolated hepatocytes of rainbow trout

Goal, Scope and Background In order to evaluate the estrogenic activity of sediments and XAD water extracts of selected sites of the catchment area of the River Neckar, a river system in Southern Germany, an integrative assessment approach was used to assess the ecological hazard potential of endocrine-disrupting compounds in sediment and water. Methods The approach is based on estrogen receptor-mediated vitellogenin synthesis induced in isolated hepatocytes of rainbow trout and quantified in a non-radioactive dot blot/RNAse protection-assay in parallel to comprehensive chemical analyses of estrogenic substances. Results and Discussion Numerous investigated extracts revealed an estrogen activity comparable to that of the positive control (1 nM 17?-estradiol corresponding to 270 ng/L in the test medium). Based on a concentration factor of 30 in the extracts and a recovery of XAD resins of approximately 80 %, 17?-estradiol equivalent concentrations between 20 and 26.7 ng/L could be calculated downstream of a sewage treatment plant (< 0.1 ng/L for a reference site). A comparison of the bioassay-derived Bio-TEQs (toxicity equivalents) and the Chem-TEQs revealed a high correlation with a Pearson coefficient of 0.85, indicating that the same ranking of the samples could be obtained with respect to the endocrine disrupting potential with both chemical and bioanalytical analysis. However, the TEQ concentrations computed from chemical analyses were significantly lower than the bioassay-derived TEQ concentrations. In fact, in none of the samples, more than 14 % of the vitellogenin-inducing potency could be attributed to the substances (steroids, alkylphenols, bisphenol A, diethylstilbestrol) analyzed. A comparison of the endocrine disrupting potential of sediments extracted by the solvents acetone and methanol revealed lower biological effects for acetone-extracted samples. Possible reasons may be a masking of endocrine effects in acetone extracts by cytotoxicity, a low extraction efficiency of the solvent acetone, or anti-estrogen potencies of some extracted sediment compounds. Using a mass balance approach, the contribution of the compounds analyzed chemically (Chem-TEQs) to the total endocrine activity (Bio-TEQs) was calculated. Based on the very low detection limits, particularly of the steroids with their high TEF factors, results revealed that a calculation of the Chem-TEQs is associated with considerable scale inaccuracy: Whereas only 7-15 % of the biological effectiveness (Bio-TEQs) could be explained by endocrine substances identified above the detection limits, the assumption of concentrations slightly below the given detection limits would result in a significant over estimation (137-197 %) of the Bio-TEQs. Even the interassay variation of the dot blot assay with different fish donors for primary hepatocyte (factor 2 - 2.5) is relatively low, when compared to the large range of the Chem-TEQ concentrations (factor 20) obtained when applying different modes of calculation. Conclusions and Outlook Overall, only a minor portion of the endocrine activity detected by bioassays could be linked to compounds identified by chemical analysis. In vitro assays for assessment of endocrine activities are useful as sensitive integrating methods that provide quantitative estimates of the total activity of particular receptor-mediated responses. Although discrepancies may also result from different bioanalytical approaches, it is overall likely that bioanalytical and not chemical analytical approaches give the correct estimate of endocrine disrupting potencies in environmental samples. As a conclusion, assessment of endocrine disruption based on chemical analysis alone does not appear sufficient and further research into the spectrum of substances with potential endocrine activity as well as into additive or even synergistic effects in complex environmental samples is urgently needed.     

Atmospheric fate of non-volatile and ionizable compounds

A modified version of the Multimedia Activity Model for Ionics MAMI, including two-layered atmosphere, air-water interface partitioning, intermittent rainfall and variable cloud coverage was developed to simulate the atmospheric fate of ten low volatility or ionizable organic chemicals. Probabilistic simulations describing the uncertainty of substance and environmental input properties were run to evaluate the impact of atmospheric parameters, ionization and air-water (or air-ice) interface enrichment.The rate of degradation and the concentration of OH radicals, the duration of dry and wet periods, and the parameters describing air-water partitioning (K_AW and temperature) and ionization (pK_a and pH) are the key parameters determining the potential for long range transport. Wet deposition is an important removal process, but its efficiency is limited, primarily by the duration of the dry period between precipitation events.Given the underlying model assumptions, the presence of clouds contributes to the higher persistence in the troposphere because of the capacity of cloud water to accumulate and transport non-volatile (e.g. 2,4-D) and surface-active chemicals (e.g. PFOA). This limits the efficiency of wet deposition from the troposphere enhancing long-range transport.     

profiling areas of ground water contamination by pesticides in california: phase ii – evaluation and modification of a statistical model

A well sampling study was conducted to evaluate anempirical approach to classifying areasof land in California as vulnerable to ground watercontamination by pesticides (Troiano et al., 1994). Wells were sampled from sections of land that had noprevious detections of pesticideresidues. The sections had been classified into vulnerablesoil clusters or into a not-classified groupusing a procedure based on Principal Components Analysis(PCA). Grape, citrus, and olive growingareas of Fresno and Tulare Counties were targeted, areas wherepre-emergence herbicide residues hadbeen detected in well water. Overall, herbicide residues weredetected in 75 of 176 sampled wells, ahigh frequency of detection in relation to results fromprevious targeted well sampling studies. Sinceresidues were also detected in the not-classified group, theclassification procedure was modified usingan approach based on Canonical Variates Analysis (CVA). Moresections were classified intovulnerable soil clusters with the CVA approach than with thePCA method. Data from two otherexplanatory variables, depth to ground water and amount ofpesticide used per section, were includedto illustrate how additional information can be incorporatedinto this approach of identifying vulnerable areas.     

Concentration Evolution of Gas Species within a Collapsing Bubble in a Liquid Medium

In this study numerical methods are used to investigate the relationship between chemical concentration of gas species within a cavitating bubble, equilibrium radius of the gas bubble and pressure variations in the ambient liquid. For this purpose, governing equations are developed to describe the dynamic equilibrium of a bubble in a flowing fluid and mass transfer between gas and liquid phases, where it was assumed that gases undergo isothermal compression, obey the ideal gas law, Henry law. It is further assumed that the concentration of each phase within the bubble is uniform. The resulting nonlinear equations are solved using implicit Trapezoidal method with Newton iteration. Four gas species are modeled under various initial and ambient pressure variation conditions. These conditions maybe considered to represent typical cavitation events. The numerical results obtained are presented in terms of dimensionless numbers. These results indicate that chemical damage maybe an important component of cavitation surface damage, since high concentration profiles may develop within a collapsing bubble. Proposed formulation and numerical solutions are simple and cost effective to implement. The results presented in this study maybe used to benchmark experimental investigations or other more complex solutions, which are outside the scope of this study.     

Sensitivity analysis of the two dimensional application of the Generic Ecological Model (GEM) to algal bloom prediction in the North Sea

Harmful algae can cause damage to co-existing organisms, tourism and farmers. Accurate predictions of algal future composition and abundance as well as when and where algal blooms may occur could help early warning and mitigating. The Generic Ecological Model is an instrument that can be applied to any water system (fresh, transitional or coastal) to calculate the primary production, chlorophyll-a concentration and phytoplankton species composition. It consists of physical, chemical and ecological model components which are coupled together to build one generic and flexible modelling tool. In this paper the model has been analyzed to assess sensitivity of the simulated chlorophyll-a concentration to a subset of ecologically significant input factors. Only a small number of approaches could be considered as suitable for several reasons including the model complexity, engagement of numerous interacting parameters and relatively long time of a single simulation. Thus, sensitivity analysis has been carried out with the use of the Morris method and later enriched by the computation of the correlation ratios of the selected parameters on the model response at more than a few locations in the modelled area. The obtained results are in agreement with expert knowledge of the ecological processes in the North Sea and correspond well with local characteristics.     

重金属Cr(Ⅵ)、Pb及Cu胁迫对双齿围沙蚕体腔细胞的DNA损伤

为探讨重金属Cr(Ⅵ)、Pb以及Cu对沙蚕体腔细胞DNA的毒性效应,以双齿围沙蚕为受试动物,重金属按不同剂量水平,Cr(Ⅵ):10、100和200 mg· L-1,Pb:5、50和100 mg·L-1,Cu:1、10和20 mg· L-1,分别胁迫沙蚕24 h,以不加任何重金属离子的海水为对照,采用单细胞凝胶电泳技术,检测其体腔细胞DNA损伤程度.结果表明,与空白对照组相比,3种重金属离子的各浓度组都能引起沙蚕体腔细胞DNA损伤,且3种重金属胁迫浓度与细胞DNA损伤程度之间存在显著的剂量-效应关系.双齿围沙蚕可以作为单细胞凝胶电泳的实验材料用于重金属所致环境污染的生物监测指示生物.     

酚类化合物对不同组织细胞DNA损伤的研究

应用单细胞凝胶电泳技术 ,研究了酚类化合物对不同组织细胞DNA的损伤 .试验结果表明 ,酚类化合物均能引起不同组织细胞不同程度的DNA损伤 ,并呈现剂量效应关系 .其高剂量组与对照组相比 ,均有极显著性差异 (P <0 0 1 ) .不同组织细胞对同一种药物呈现出不同的敏感性 .酚类化合物遗传毒性效应与其结构密切相关