An integrated assessment of estrogenic contamination and biological effects in the aquatic environment of The Netherlands.

An extensive study was carried out in the Netherlands on the occurrence of a number of estrogenic compounds in surface water, sediment, biota, wastewater, rainwater and on the associated effects in fish. Compounds investigated included natural and synthetic hormones, phthalates, alkylphenol(ethoxylate)s and bisphenol-A. The results showed that almost all selected (xeno-)estrogens were present at low concentrations in the aquatic environment. Locally, they were found at higher levels. Hormones and nonylphenol(ethoxylate)s were present in concentrations that are reportedly high enough to cause estrogenic effects in fish. Field surveys did not disclose significant estrogenic effects in male flounder (Platichthys flesus) in the open sea and in Dutch estuaries. Minor to moderate estrogenic effects were observed in bream (Abramis brama) in major inland surface waters such as lowland rivers and a harbor area. The prevalence of feminizing effects in male fish is largest in small regional surface waters that are strongly influenced by sources of potential hormone-disrupting compounds. High concentrations of plasma vitellogenin and an increased prevalence of ovotestes occurred in wild male bream in a small river receiving a considerable load of effluent from a large sewage treatment plant. After employing in vitro and in vivo bioassays, both in situ and in the laboratory, we conclude that in this case hormones (especially 17 alpha-ethynylestradiol) and possibly also nonylphenol(ethoxylate)s are primarily responsible for these effects.

Detection of estrogenic potency in wastewater and surface water with three in vitro bioassays.

A study was performed to optimize sample preparation and application of three in vitro assays for measuring estrogenic potency in environmental extracts. The three assays applied were an estrogen receptor (ER)-binding assay and two reporter gene effect assays: a yeast estrogen screen (YES) and the ER-mediated chemically activated luciferase gene expression (ER-CALUX) assay. All assays were able to detect estrogenicity, but the amounts of material needed for the assays differed greatly between the three assays (ER-binding assay >> YES > ER-CALUX). In addition, in the ER-binding assay, both agonists and antagonists give an estrogenic response, resulting in higher estradiol equivalency (EEQ) levels than both the ER-CALUX and the YES assay for the same samples. The EEQs found in wastewater treatment plants (WTPs) with the ER-CALUX assay were in the range of 4 to 440 and 0.11 to 59 pmol/L for influent and effluent, respectively. Water extracts from four large rivers had levels ranging from 0.25 to 1.72 pmol/L. Extracts from suspended matter and sludge contained estrogenic potency of 0.26 to 2.49 and 1.6 to 41 pmol EEQ/g dry weight, respectively. In WTPs, the average reduction of estrogenic potency in effluent compared to influent was 90 to 95% in municipal WTPs and about 50% in industrial WTPs. In influent, 30% of the ER-CALUX activity could not be explained by the calculated potencies based on chemical analysis of a number of known (xeno)estrogens; in effluent the unexplained fraction was 80%. These first results of analyzing estrogenic potency in WTP water and surface water in The Netherlands indicate that further studies are warranted to investigate the actual risks for aquatic systems.

Quantitative risk assessment of FMD virus transmission via water.

Foot-and-mouth disease (FMD) is a viral disease of domesticated and wild cloven-hoofed animals. FMD virus is known to spread by direct contact between infected and susceptible animals, by animal products such as meat and milk, by the airborne route, and mechanical transfer on people, wild animals, birds, and by vehicles. During the outbreak of 2001 in the Netherlands, milk from dairy cattle was illegally discharged into the sewerage as a consequence of transport prohibition. This may lead to contaminated discharges of biologically treated and raw sewage in surface water that is given to cattle to drink. The objective of the present study was to assess the probability of infecting dairy cows that were drinking FMD virus contaminated surface water due to illegal discharges of contaminated milk. So, the following data were collected from literature: FMD virus inactivation in aqueous environments, FMD virus concentrations in milk, dilution in sewage water, virus removal by sewage treatment, dilution in surface water, water consumption of cows, size of a herd in a meadow, and dose-response data for ingested FMD virus by cattle. In the case of 1.6 x 10(2) FMD virus per milliliter in milk and discharge of treated sewage in surface water, the probability of infecting a herd of cows was estimated to be 3.3 x 10(-7) to 8.5 x 10(-5), dependent on dilution in the receiving surface water. In the case of discharge of raw sewage, all probabilities of infection were 100 times higher. In the case of little dilution in small rivers, the high level of 8.5 x 10(-3) is reached. For 10(4) times higher FMD virus concentrations in milk, the probabilities of infecting a herd of cows are high in the case of discharge of treated sewage (3.3 x 10(-3) to 5.7 x 10(-1)) and very high in the case of discharge of raw sewage (0.28-1.0). It can be concluded that illegal and uncontrolled discharges of contaminated milk into the sewerage system may lead to high risks to other cattle farms at 6-50 km distance of the location of discharge within one day. This clearly underlines current measures that prohibit such discharges, and also asks for strict control. This risk assessment clearly demonstrated the potential significance of FMD virus transmission via water, and the results will be useful on an international scale, and could also serve as a basis for other FMD risk-assessment models.