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      Oestrogenic activity in drinking waters from a rural area in the Waterberg District, Limpopo Province, South Africa

      research-article
      , ,
      Water SA
      Water Research Commission (WRC)
      recombinant yeast screen, oestrogenic activity, water, rural, Limpopo, South Africa

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          Abstract

          In South Africa, limited data are available regarding possible oestrogenic activity in the aquatic systems and especially drinking water. Water in the rural areas is often contaminated with a complex mixture of toxic compounds originating from nearby industries, agriculture and households. In these rural areas the only access to drinking water is boreholes, natural springs and rivers. Thus human exposure to environmental contaminants in drinking water is potentially high. Two rural communities near Mokopane in the Waterberg district of the Limpopo Province were selected in order to screen for oestrogenic activity in drinking water sources in a rural area. Eleven 1 ℓ water samples (Molekane n= 4; Sekuruwe n= 7), were collected in prepared glass bottles and extracted on a SPE C18 cartridge and reconstituted into ethanol. The recombinant yeast oestrogen screen was used to determine the oestrogenic activity in the extracts. 17β-estradiol (E2) was used as a positive control and the results were expressed as estradiol equivalents (EEq). The EEq of the water from both the communities ranged between 0.63 - 2.48 x10-9 g/ℓ. These concentrations are similar to other studies conducted in Korean river waters in rural and city areas and Flemish surface waters. The recombinant yeast screen confirmed oestrogenic activity in the drinking water samples; further investigation is necessary to determine the source of the contamination and association with impaired growth.

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          Most cited references70

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          The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants.

          Estrogens are defined by their ability to induce the proliferation of cells of the female genital tract. The wide chemical diversity of estrogenic compounds precludes an accurate prediction of estrogenic activity on the basis of chemical structure. Rodent bioassays are not suited for the large-scale screening of chemicals before their release into the environment because of their cost, complexity, and ethical concerns. The E-SCREEN assay was developed to assess the estrogenicity of environmental chemicals using the proliferative effect of estrogens on their target cells as an end point. This quantitative assay compares the cell number achieved by similar inocula of MCF-7 cells in the absence of estrogens (negative control) and in the presence of 17 beta-estradiol (positive control) and a range of concentrations of chemicals suspected to be estrogenic. Among the compounds tested, several "new" estrogens were found; alkylphenols, phthalates, some PCB congeners and hydroxylated PCBs, and the insecticides dieldrin, endosulfan, and toxaphene were estrogenic by the E-SCREEN assay. In addition, these compounds competed with estradiol for binding to the estrogen receptor and increased the levels of progesterone receptor and pS2 in MCF-7 cells, as expected from estrogen mimics. Recombinant human growth factors (bFGF, EGF, IGF-1) and insulin did not increase in cell yields. The aims of the work summarized in this paper were a) to validate the E-SCREEN assay; b) to screen a variety of chemicals present in the environment to identify those that may be causing reproductive effects in wildlife and humans; c) to assess whether environmental estrogens may act cumulatively; and finally d) to discuss the reliability of this and other assays to screen chemicals for their estrogenicity before they are released into the environment.
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            Bisphenol-A: an estrogenic substance is released from polycarbonate flasks during autoclaving.

            In studies to determine whether Saccharomyces cerevisiae produced estrogens, the organism was grown in culture media prepared using distilled water autoclaved in polycarbonate flasks. The yeast-conditioned media showed the presence of a substance that competed with [3H]estradiol for binding to estrogen receptors (ER) from rat uterus. However, it soon became clear that the estrogenic substance in the conditioned media was not a product of the yeast grown in culture, but was leached out of the polycarbonate flasks during the autoclaving procedure. [3H]Estradiol displacement activity was monitored by ER RRA, and the active substance was purified from autoclaved medium using a series of HPLC steps. The final purified product was identified as bisphenol-A (BPA) by nuclear magnetic resonance spectroscopy and mass spectrometry. BPA could also be identified in distilled water autoclaved in polycarbonate flasks without the requirement of either the organism or the constituents of the culture medium. Authentic BPA was active in competitive RRAs, demonstrating an affinity approximately 1:2000 that of estradiol for ER. In functional assays, BPA (10-25 nM) induced progesterone receptors in cultured human mammary cancer cells (MCF-7) at a potency of approximately 1:5000 compared to that of estradiol. The BPA effect on PR induction was blocked by tamoxifen. In addition, BPA (25 nM) increased the rate of proliferation of MCF-7 cells assessed by [3H]thymidine incorporation. Thus, BPA exhibited estrogenic activity by both RRA and two functional bioresponse assays. Finally, MCF-7 cells grown in media prepared with water autoclaved in polycarbonate exhibited higher progesterone receptor levels than cells.grown in media prepared with water autoclaved in glass, suggesting an estrogenic effect of the water autoclaved in polycarbonate. Our findings raise the possibility that unsuspected estrogenic activity in the form of BPA may have an impact on experiments employing media autoclaved in polycarbonate flasks. It remains to be determined whether BPA derived from consumer products manufactured from polycarbonate could significantly contribute to the pool of estrogenic substances in the environment.
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              Developmental effects of endocrine-disrupting chemicals in wildlife and humans.

              Large numbers and large quantities of endocrine-disrupting chemicals have been released into the environment since World War II. Many of these chemicals can disturb development of the endocrine system and of the organs that respond to endocrine signals in organisms indirectly exposed during prenatal and/or early postnatal life; effects of exposure during development are permanent and irreversible. The risk to the developing organism can also stem from direct exposure of the offspring after birth or hatching. In addition, transgenerational exposure can result from the exposure of the mother to a chemical at any time throughout her life before producing offspring due to persistence of endocrine-disrupting chemicals in body fat, which is mobilized during egg laying or pregnancy and lactation. Mechanisms underlying the disruption of the development of vital systems, such as the endocrine, reproductive, and immune systems, are discussed with reference to wildlife, laboratory animals, and humans.
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                Author and article information

                Journal
                wsa
                Water SA
                Water SA
                Water Research Commission (WRC) (Pretoria, Gauteng, South Africa )
                0378-4738
                1816-7950
                April 2009
                : 35
                : 3
                : 245-251
                Affiliations
                [01] Pretoria orgnameUniversity of Pretoria orgdiv1Department of Urology South Africa
                [02] Pretoria orgnameUniversity of Pretoria orgdiv1School of Health Systems and Public Health South Africa
                Article
                S1816-79502009000300002 S1816-7950(09)03500302
                2baa0af8-9453-4af5-9677-9128e4128d86

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 04 February 2009
                : 27 February 2007
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 51, Pages: 7
                Product

                SciELO South Africa


                recombinant yeast screen,South Africa,Limpopo,rural,water,oestrogenic activity

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