Effects of sewage effluent remediation on body size, somatic RNA: DNA ratio, and markers of chemical exposure in three-spined sticklebacks.

Body mass, fork length, RNA:DNA ratio, specific growth rate, and hepatic EROD activity and CYP1A expression, were measured in three-spined sticklebacks in the River Ray (south west England) at sites downstream of an urban waste water treatment works (WWTW) prior to, and following, remediation of the effluent with granular activated carbon (GAC) tertiary treatment. During the same two-year period fish were also sampled from a neighbouring reference river (R. Ock). The WWTW effluent elevated water temperatures and nutrient content in the R. Ray and rendered a direct comparison of fish populations in the two rivers untenable. Instead, the stability of population parameters within each river during matched pre- and post-remediation periods was compared. Stickleback populations in both rivers were annual but fish in the R. Ray spawned earlier and were larger than those in the R. Ock. In the R. Ray fish gained mass throughout the winter months whereas in the R. Ock growth was much reduced during this period. In fish from the R. Ray the somatic RNA:DNA ratio remained elevated during May-November after remediation, rather than declining as in the same period pre-remediation and as was the case for fish in the R. Ock during both periods. The specific growth rate of the first post-remediation generation of sticklebacks in the R. Ray was higher than that of the previous pre-remediation generation. Following remediation there was no decline in hepatic EROD activity or in the abundance of hepatic CYP1A transcripts in fish in the R. Ray suggesting that the primary route of exposure to contaminants for these fish was not via the water column, and that the change in performance of the fish post-remediation was not impeded by continued exposure to contaminants. Both EROD activity and CYP1A expression increased in fish in the R. Ock during the later stages of the study suggesting that the fish in this river were exposed to an unidentified contaminant episode. This may have been linked with the poorer performance of fish in the R. Ock during the post-remediation period. The improved performance of fish in the R. Ray suggest that there may be factors in good quality secondary treated sewage effluent which can adversely influence the performance of fish populations, directly or indirectly, and which can be removed by tertiary treatment.

The use of modelling to predict levels of estrogens in a river catchment: how does modelled data compare with chemical analysis and in vitro yeast assay results?

Effluent discharges at Rodbourne sewage treatment works (STWs) were assessed using chemical and in vitro biological analysis as well as modelling predictions. Results showed that Rodbourne STW discharged less estrone (E1) than expected, but similar 17beta-estradiol (E2) and 17alpha-ethinyl estradiol (EE2) to those predicted by a widely cited effluent prediction model. The Exposure Analysis Modelling System (EXAMS) model was set up using measured effluent concentrations as its starting point to predict estrogen concentrations along a 10 km length of the receiving water of the River Ray. The model adequately simulated estrogen concentrations along the river when compared to July 2007 measured data. The model predicted combined estrogen equivalents in reasonable agreement with estrogenicity as measured by passive sampler (POCIS) extracts using the yeast estrogen screen. Using gauged mean flow values for 2007 the model indicated that the most important determinand for estrogen exposure in the Ray was not season, but proximity to the Rodbourne effluent. Thus, fish in the first 3 km downstream of Rodbourne were typically exposed to two or even three times more estrogens than those living 7-10 km further downstream. The modelling indicated that, assuming the effluent estrogen concentrations measured in February 2008 were typical, throughout the year the whole length of the Ray downstream of Rodbourne would be estrogenic, i.e. exceeding the 1 ng/L E2 equivalent threshold for endocrine disruption.

Identification of nonregulated pollutants in North Sea-produced water discharges.

Produced water collected from oil and gas platforms on the United Kingdom Continental Shelf was characterized for nonregulated pollutants through an effects-directed analysis procedure. Produced water samples were characterized for the presence of aryl hydrocarbon receptor (AhR) and estrogen receptor (ER) agonists using the dioxin-responsive, chemical-activated luciferase gene expression assay (DR-CALUX) and yeast estrogen screen (YES) bioassays. The AhR and ER agonists were then isolated by normal-phase, high-performance liquid chromatography and identified using gas chromatography coupled to mass spectrometry in a number of formats. The identified compounds were cross-referenced with those compounds routinely analyzed and regarded by the Oslo and Paris (OSPAR) Commission for the North East Atlantic as priority hazardous substances. The occurrence in produced water of a number of nonregulated compounds with demonstrable potential environmental effects is presented, to our knowledge for the first time. These include persistent organic contaminants, such as hexachlorobenzene, decachlorobiphenyl, and octachlorodibenzofuran.

Bioanalytical characterisation of estrogen and arylhydrocarbon receptor agonists in transplanted blue mussels (Mytilus edulis): proof of concept.

The characterisation of estrogen (ER) and arylhydrocarbon (AhR) receptor agonists was performed in extracts of the tissues of transplanted blue mussels (Mytilus edulis). The yeast estrogen screen (YES) was used to detect the presence of ER agonists, whilst the DR-CALUX assay was used to detect AhR agonists. The concentration of ER agonists in mussel tissue from the Tees estuary, Brancaster and River Crouch (UK) were below the limits of detection for the YES assay (0.87 pg E2 g(-1)). AhR agonists were measured at concentrations of between 1 and 950 pg TCDD g(-1) in mussel tissue. A bioassay-directed fractionation of the sample extracts, followed by advanced broad spectrum gas chromatography-mass spectrometry analysis was then used in an attempt to identify the AhR agonists present. This showed that a complex mixture of AhR agonists occurs in the samples and that further work will be required in order to isolate and identify the individual compounds responsible.