Biomonitoring of estrogenic exposure and identification of responsible compounds in bream from Dutch surface waters.

The exposure to and effects of estrogenic compounds in male breams from Dutch freshwater locations were investigated. Ovotestis was observed infrequently (maximum frequency 16%). However, plasma vitellogenin (VTG) concentration was elevated highly at some locations. Estrogenic activities in male bream plasma, liver, and in gastrointestinal content were measured in the estrogen-responsive chemical-activated luciferase gene expression (ER-CALUX) assay. Plasma concentrations of vitellogenin correlated very well with the estrogenic activities in gastrointestinal content. The ER-CALUX activity in gastrointestinal content thus could provide a biomarker for recent exposure to estrogenic compounds, and the gastrointestinal content was chosen as investigative matrix for the toxicity identification and evaluation ([TIE]; bioassay-directed fractionation) of estrogenic compounds in bream. The approach consisted of a reversed-phase high-performance liquid chromatography fractionation of gastrointestinal content extract, directed by ER-CALUX and followed by gas chromatography analysis. The estrogenic hormones 17beta-estradiol and its metabolite estrone were identified as major contributors to the activity at all locations (except the reference location), independent of the presence or absence of a known source of estrogenic activity, such as a sewage treatment plant. Chemical screening showed the presence of other pollutants, such as a lower chlorinated dioxin and the disinfectants clorophene and triclosan. However, these compounds did not have high estrogenic potencies and their concentrations were not high enough to contribute significantly to the observed estrogenic activity.

Effects of copper and temperature on aquatic bacterial communities.

The present study aimed to characterise effects of copper and temperature on bacterial communities in photosynthetic biofilms using a suit of supplementary methods: pollution-induced community tolerance (PICT), DNA profiles with denaturing gradient gel electrophoresis (DGGE) and physiological profiles with community-level physiological profiling (CLPP). Biofilms of algae and bacteria were grown in a ditch of a Dutch polder and exposed in the laboratory to copper (3 microM and a reference) at three different temperatures (10, 14 and 20 degrees C). Bacterial communities sampled from the field showed heterogeneity in their physiological profiles, however the heterogeneity decreased during laboratory incubation. After 3 days laboratory incubation, the copper treated biofilms were different from the reference biofilms, as revealed by DGGE and CLPP analyses. Effects of temperature were not observed in the CLPPs, or in the DGGE profiles. PICT was observed for the bacterial communities at all temperatures. The copper-tolerance at 10 and 14 degrees C increased about 3 times, whereas copper-tolerance at 20 degrees C increased about 6 times. Temperature had an effect on the community tolerance, but not on the structure or on the physiological profile, suggesting that temperature was not a major factor causing successional changes under these laboratory conditions. In contrast, temperature had an effect on tolerance development indicating that the exposure to copper was enhanced at higher temperature.

Copper-induced modifications of the trophic relations in riverine algal-bacterial biofilms.

The effects of copper (Cu) on photosynthetic riverine biofilms were studied in artificial stream channels. Direct effects on the composition and functioning of the biofilms were investigated using plant pigments, community-level physiological profiles (CLPP), and pulse-amplitude-modulated (PAM) fluorescence. Copper caused a significant reduction of microalgal biomass and induced a shift in the population from diatoms to cyanobacteria. However, a decrease in biomass indicated that the replacement of species was not totally effective to counteract the toxic effects of Cu. A direct effect of Cu could also be shown in the bacterial community, and, furthermore, changes in the CLPP could be related to the Cu treatment. Copper-exposed biofilms lost the capacity to use between 11 and 15% of the substrates, but many of the remaining capacities became more robust, indicating an increased Cu tolerance due to the exposure. The change in the biofilm microbial composition points to the indirect effects of Cu on biofilms due to the close interdependence between biofilm autotrophic and heterotrophic compartments. Grazing by snails, which appeared to be an important factor structuring biofilms without any Cu addition, had a very minor effect on Cu-exposed biofilms. Although grazing changed the bacterial composition, its effects were not detected either on the algal community or on the biofilm community tolerance to Cu.