Analytical and bioanalytical assessments of organic micropollutants in the Bosna River using a combination of passive sampling, bioassays and multi-residue analysis.

Complex mixtures of contaminants from multiple sources, including agriculture, industry or wastewater enter aquatic environments and might pose hazards or risks to humans or wildlife. Targeted analyses of a few priority substances provide limited information about water quality. In this study, a combined chemical and effect screening of water quality in the River Bosna, in Bosnia and Herzegovina was carried out, with focus on occurrence and effects of contaminants of emerging concern. Chemicals in water were sampled at 10 sites along the Bosna River by use of passive sampling. The combination of semipermeable membrane devices (SPMDs) and polar organic chemical integrative samplers (POCIS) enabled sampling of a broad range of contaminants from hydrophobic (PAHs, PCBs, OCPs) to hydrophilic compounds (pesticides, pharmaceuticals and hormones), which were determined by use of GC-MS and LC-MS (MS). In vitro, cell-based bioassays were applied to assess (anti)androgenic, estrogenic and dioxin-like potencies of extracts of the samplers. Of a total of 168 targeted compounds, 107 were detected at least once. Cumulative pollutant concentrations decreased downstream from the city of Sarajevo, which was identified as the major source of organic pollutants in the area. Responses in all bioassays were observed for samples from all sites. In general, estrogenicity could be well explained by analysis of target estrogens, while the drivers of the other observed effects remained largely unknown. Profiling of hazard quotients identified two sites downstream of Sarajevo as hotspots of biological potency. Risk assessment of detected compounds revealed, that 7 compounds (diazinon, diclofenac, 17ß-estradiol, estrone, benzo[k]fluoranthene, fluoranthene and benzo[k]fluoranthene) might pose risks to aquatic biota in the Bosna River. The study brings unique results of a complex water quality assessment in a region with an insufficient water treatment infrastructure.

Evidence for microRNA-mediated regulation of steroidogenesis by hypoxia.

Environmental hypoxia can occur in both natural and occupational environments. Over the recent years, the ability of hypoxia to cause endocrine disruption via perturbations in steroid synthesis (steroidogenesis) has become increasingly clear. To further understand the molecular mechanism underlying hypoxia-induced endocrine disruption, the steroid-producing human cell line H295R was used to identify microRNAs (miRNAs) affecting steroidogenic gene expression under hypoxia. Hypoxic treatment of H295R cells resulted in the downregulation of seven steroidogenic genes and one of these, CYP19A1 (aromatase), was shown to be regulated by the transcription factor hypoxia-inducible factor-1 (HIF-1). Using bioinformatic and luciferase reporter analyses, miR-98 was identified to be a CYP19A1-targeting miRNA from a subset of HIF-1-inducible miRNAs. Gain- and loss-of-function analysis suggested that under hypoxia, the increased expression of miR-98 led to the downregulation of CYP19A1 mRNA and protein expression and that it may have contributed to a reduction in estradiol (E2) production. Intriguingly, luciferase reporter assays using deletion constructs of a proximal 5'-flanking region of miR-98 did not reveal a hypoxia-responsive element (HRE)-containing promoter. Overall, this study provided evidence for the role of miRNAs in regulating steroidogenesis and novel insights into the molecular mechanisms of hypoxia-induced endocrine disruption.

Biological analysis of endocrine-disrupting chemicals in animal meats from the Pearl River Delta, China.

Endocrine disrupting chemicals (EDCs) consist of a diverse group of industrial chemicals and pharmacological agents. The use of instrumental analyses as the first screening tool might not be cost-effective to identify the existence of enormous numbers of chemical contaminants in environments. Also, knowledge of the concentration of individual residues is difficult to use to evaluate biological impacts of contaminants to wildlife and humans. The primary objective of the present study was to develop and to test the feasibility of using a battery of exposure biomarkers for the rapid-screening of various endocrine disrupting activities present in food. The measurement of the EDC-elicited activities involved various (i) receptor-mediated responses, including androgenic, estrogenic, dioxin-like, glucocorticoid-like, progesterone-like, peroxisome proliferator-like and retinoid-like as well as (ii) the non-receptor mediated responses through modulation of cellular reactive oxygen species (ROS) and ATP content. Samples of both local and imported pork, beef and chicken as well as freshwater and seawater fishes were collected. Extracts of different foods exhibited various dioxin-like and "hormonal" activities. Fish and chicken skin were found to be the major source of exogenous "hormonal" and dioxin-like substances in diets. Extracts of beef and pork contained lesser potencies of hormonally-active agents. Our data suggest that the proposed EDC-screening platform may be useful in a risk assessment for the routine monitoring of EDCs in foods. Continuous monitoring and research is warranted to assess the physiological consequences of the consumption.

Characterization of dioxin-like contamination in soil and sediments from the "hot spot" area of petrochemical plant in Pancevo (Serbia).

PURPOSE: Combinatorial bio/chemical approach was applied to investigate dioxin-like contamination of soil and sediment at the petrochemical and organochlorine plant in Pancevo, Serbia, after the destruction of manufacturing facilities that occurred in the spring of 1999 and subsequent remediation actions. MATERIALS AND METHODS: Soil samples were analyzed for indicator polychlorinated biphenyls (PCBs) by gas chromatography/electron capture detection (GC/ECD). Prioritized soil sample and sediment samples from the waste water channel were analyzed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) by high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). Microethoxyresorufin o-deethylase (Micro-EROD) and H4IIE-luciferase bioassays were used for monitoring of dioxin-like compounds (DLC) and for better characterization of dioxin-like activity of soil samples. RESULTS: Bioanalytical results indicated high dioxin-like activity in one localized soil sample, while the chemical analysis confirmed the presence of large quantities of DLC: 3.0 × 10(5) ng/g d.w. of seven-key PCBs, 8.2 ng/g d.w. of PCDD/Fs, and 3.0 × 10(5) ng/g d.w. of planar and mono-ortho PCBs. In the sediment, contaminant concentrations were in the range 2-8 ng/g d.w. of PCDD/Fs and 9-20 ng/g d.w. of PCBs. CONCLUSIONS: This study demonstrates the utility of combined application of bioassays and instrumental analysis, especially for developing and transition country which do not have capacity of the expensive instrumental analysis. The results indicate the high contamination of soil in the area of petrochemical plant, and PCDD/Fs contamination of the sediment from the waste water channel originating from the ethylene dichloride production.

Utilizing the great blue heron (Ardea herodias) in ecological risk assessments of bioaccumulative contaminants.

Selection of an appropriate species is a key element of effective ecological risk assessments (ERA), especially when site-specific field studies are to be employed. Great blue herons (GBH) possess several ideal characteristics of a receptor species for the assessment of bioaccumulative compounds in the environment, such as ease of study, high potential for exposure, widespread distribution, and territorial foraging behavior. Methodologies for assessing exposure and population health are described herein. As outlined, the collection of GBH eggs, GBH nestling blood, and adult GBH blood allows for the determination of contaminant concentrations in various GBH tissues, a top-down assessment, which can be done in conjunction with predicted dietary exposure, a bottom-up assessment, to support a multiple lines of evidence approach. Additionally, population parameters, such as productivity and survival, can also be measured to elucidate if the contaminant exposure may be causing population level effects. Over the course of two years, three GBH rookeries were monitored for productivity and nestling exposure. Nests were monitored from blinds and individually accessed at multiple time points to obtain measures of nestling health, band nestlings, and collect eggs and nestling plasma. Multiple nests could frequently be accessed by climbing one tree, resulting in minimal effort to obtain the necessary sample size. Additionally, 51 adult GBH, captured in their foraging areas, were banded, and provided a blood sample. With these samples, a statistical difference in tissue based exposure was identified between the reference and target area. Statistically significant differences were also identified between the upper and lower reaches of the target area, thereby identifying a range of doses geographically which could be correlated to specific measurement endpoints. The ability to identify a dose response greatly increases the ability of the dataset to determine causation, a key goal of such studies. Overall, the use of the described methods allowed for the collection of a statistically sufficient and ecologically relevant dataset with reasonable effort and minimal impact on GBH.