E2 Equivalent Concentration Research Articles

Potential risk of estrogenic compounds produced by water blooms to aquatic environment

Some freshwater phytoplankton species have been suggested to produce estrogenic compounds in concentrations which could cause adverse effects to aquatic biota, while other studies showed no estrogenic effects after exposure to phytoplankton extracts or pointed out possible sources of the overestimation of the estrogenic activity. This study aimed to clarify these research inconsistencies by investigating estrogenicity of biomass extracts from both environmental freshwater blooms and laboratory cyanobacterial and algae cultures by in vitro reporter bioassay. Biomasses of 8 cyanobacterial and 3 algal species from 7 taxonomic orders were extracted and tested. Next to this, samples of environmental water blooms collected from 8 independent water bodies dominated by phytoplankton species previously assessed as laboratory cultures were tested. The results showed undetectable or low estrogenicity of both freshwater blooms and laboratory cultures with E2 equivalent concentration (EEQ) in a range from LOQ up to 4.5 ng EEQ/g of dry mass. Moreover, the co-exposure of biomass extracts with environmentally relevant concentration of model estrogen (steroid hormone 17β-estradiol; E2), commonly occurring in surface waters, showed simple additive interaction. However, some of the biomass extracts elicited partially anti-estrogenic effects in co-exposure with higher E2 concentration. In conclusion, our study documents undetectable or relatively low estrogenic potential of biomass extracts from both environmental freshwater blooms and studied laboratory cultured cyanobacterial and algae species. Nevertheless, in case of very high-density water blooms, even this low estrogenicity (detected for two cyanobacterial species) could lead to EEQ content in biomass reaching effect-based trigger values indicating potential risk, if recalculated per water volume at field sites. However, these levels would not occur in water under realistic environmental scenarios and the potential estrogenic effects would be most probably minor compared to other toxic effects caused by massive freshwater blooms of such high densities.

Oestrogenic Activity in Girls with Signs of Precocious Puberty as Exposure Biomarker to Endocrine Disrupting Chemicals: A Pilot Study.

The relationship between endocrine disrupting chemical (EDC) exposure and Precocious Puberty (PP) was investigated in this pilot study, involving girls with signs of PP (P) and pre-pubertal girls (C). Risk factors for PP were assessed through questionnaires, while 17β-oestradiol (E2) levels and oestrogenic activity were quantified on sera. The oestrogenic activity, expressed as E2 equivalent concentration (EEQ), was applied as EDC exposure biomarker. Questionnaires showed a low EDC knowledge, a high EDC exposure, and a potential relationship between some habits at risk for EDC exposure and PP. EEQs were similar between C and P; however, they were significantly higher in girls living in an urban environment than in girls living in a rural environment, suggesting a potential higher EDC exposure in cities. The results of this pilot study highlighted the need to raise awareness on EDCs and can be considered a starting point to clarify the relationship between EDC exposure and PP.

Estrogens in municipal wastewater and receiving waters in the Beijing-Tianjin-Hebei region, China: Occurrence and risk assessment of mixtures

The potentially high release of estrogens to surface waters due to high population density and local livestock production in the Beijing-Tianjin-Hebei region may pose adverse effects on reproductive systems of aquatic organisms. This study found that total measured concentrations of estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinylestradiol (EE2) and diethylstilbestrol (DES) were 468 ± 27 ng/L in treated wastewater and 219 ± 23 ng/L in river waters in this region. E2, E3 and EE2 were the predominant estrogens in river waters. The restriction of DES for human use should have been enforced, however concentrations of DES were relatively high compared to other studies. Haihe and Yongdingxin Rivers delivered approximately 1.8 tonnes of estrogens to the Bohai Bay annually. Concentrations of individual estrogens were significantly higher in river waters in the dry season, however, mass loadings were significantly higher in the wet season. The average E2-equivalent concentrations reached 1.2 ± 0.2 and 0.64 ± 0.08 μg-E2/L following long-term and short-term exposure estimates, respectively, in river waters with an EE2 contribution of over 90 %. This could give rise to high risks to fish. Estrogens in river waters largely derived from human excretion. Field studies on estrogenic effects on fish reproductive systems are required locally considering high estrogen contamination levels.

Triple functional small-molecule-protein conjugate mediated optical biosensor for quantification of estrogenic activities in water samples

Establishing biosensors to map a comprehensive picture of potential estrogen-active chemicals remains challenging and must be addressed. Herein, we describe an estrogen receptor (ER)-based evanescent wave fluorescent biosensor by using a triple functional small-molecule–protein conjugate as a signal probe for the determination of estrogenic activities in water samples. The signal probe, consisting of a Cy5.5-labelled streptavidin (STV) moiety and a 17β-estradiol (E2) moiety, acts simultaneously as signal conversion, signal recognition and signal report elements. When xenoestrogens compete with the E2 moiety of conjugate in binding to the ER, the unbound conjugates are released, and their STV moiety binds with desthiobiotin (DTB) modified on the optical fiber via the STV–DTB affinity interactions. Signal probe detection is accomplished by fluorescence emission induced by an evanescent field, which positively relates with the estrogenic activities in samples. Quantification of estrogenic activity expressed as E2 equivalent concentration (EEQ) can be achieved with a detection limit of 1.05 μg/L EEQ by using three times standard deviation of the mean blank values and a linear calibration range from 20.8 to 476.7 μg/L EEQ. The optical fiber system is robust enough for hundreds of sensing cycles. The biosensor-based determination of estrogenic activities in wastewater samples obtained from a full-scale wastewater treatment plant is consistent with that measured by the two-hybrid recombinant yeast bioassay.

Modeling the exposure of wild fish to endocrine active chemicals: Potential linkages of total estrogenicity to field-observed intersex

Decades of studies on endocrine disruption have suggested the need to manage the release of key estrogens from municipal wastewater treatment plants (WWTP). However, the proposed thresholds are below the detection limits of most routine chemical analysis, thereby restricting the ability of watershed managers to assess the environmental exposure appropriately. In this study, we demonstrated the utility of a mechanistic model to address the data gaps on estrogen exposure. Concentrations of the prominent estrogenic contaminants in wastewaters (estrone, estradiol, and ethinylestradiol) were simulated in the Grand River in southern Ontario (Canada) for nine years, including a period when major WWTP upgrades occurred. The predicted concentrations expressed as total estrogenicity (E2 equivalent concentrations) were contrasted to a key estrogenic response (i.e., intersex) in rainbow darter (Etheostoma caeruleum), a wild sentinel fish species. A predicted total estrogenicity in the river of ≥10 ng/L E2 equivalents was associated with high intersex incidence and severity, whereas concentrations <0.1 ng/L E2 equivalents were associated with minimal intersex expression. Exposure to a predicted river concentration of 0.4 ng/L E2 equivalents, the environmental quality standard (EQS) proposed by the European Union for estradiol, was associated with 34% (95% CI:30–38) intersex incidence and a very low severity score of 0.6 (95% CI:0.5–0.7). This exposure is not predicted to cause adverse effects in rainbow darter. The analyses completed in this study were only based on the predicted presence of three major estrogens (E1, E2, EE2), so caution must be exercised when interpreting the results. Nevertheless, this study illustrates the use of models for exposure assessment, especially when measured data are not available.

Potential bioactivity and association of 17β-estradiol with the dissolved and colloidal fractions of manure and soil

The dissolved (DF) and colloidal fractions (CF) of soil and manure play an important role in the environmental fate and transport of steroidal estrogens. The first objective of this study was to quantify the association of 17β-estradiol (E2) with the DF and CF isolated from (i) liquid swine manure (LSM), (ii) a soil:water mixture (soil), and (iii) a LSM:soil:water mixture (Soil+LSM). The appropriate CF and DF size fractions of the Soil, Soil+LSM, and LSM media were obtained by first filtering through a 0.45μm filter, which provided the combined DF and CF (DF/CF). The DF/CF from the three media was spiked with carbon-14 ([14C]) radiolabeled E2 ([14C]-E2), and then ultrafiltered to isolate the CF (<0.45μm and >1kDa) from the DF (<1kDa). The average recoveries of the [14C] associated with the DF were 67%–72%, 67%–79%, and 76%–78% for the Soil, Soil+LSM and LSM, respectively. For the CF that was retained on the 1kDa filter, organic carbon and [14C]-E2 were dislodged with subsequent water rinses the Soil+LSM and LSM, but not the Soil. The second objective was to evaluate whether the E2 associated with the various fractions of the different media could still bind the estrogen receptor using an E2 receptor (17β-ER) competitor assay, which allowed E2 equivalent concentrations to be determined. The estrogen receptor assay results indicated that E2 present in the DF of the Soil and Soil+LSM solutions could still bind the estrogen receptor. Results from this study indicated that E2 preferentially associated with the DF of soil and manure, which may enhance its dissolved advective transport in surface and subsurface water. Furthermore, this study indicated that E2 associated with DF solutions in the environment could potentially induce endocrine responses through its interactions with estrogen receptor.

Relative Contribution of Endocrine-disrupting Chemicals to the Estrogenic Potency of Marine Sediments of Osaka Bay, Japan

Although concentrations of endocrine-disrupting chemicals (EDCs) in seawaters of Japan are relatively low, vitellogenin and ovotestis inductions are still being observed in some males of mullet and flounder collected in coastal areas. These fish species are benthic and could be affected by EDCs in marine sediments. Therefore, the concentrations of EDCs in marine sediments of Osaka Bay were determined by LC-MS/MS. In addition, the estrogen receptor binding potencies as estrogenic potencies of these sediments were assessed by the medaka estrogen receptor-α binding assay. Results show that estrogenic potencies were higher in sediments of the inner part of the bay especially at station 13 (off Sakai City) where quite strong estrogenic potency was detected. Through calculation of total E2 equivalent concentration (EEQ) in sediments, it was established that approximately 50 % of estrogenic potency was due to nonylphenol (NP), estrone and 17β-estradiol, suggesting that these compounds play important roles as endocrine disruptors in coastal environments of Osaka Bay.

Removal of Endocrine-Disrupting Chemicals during Ozonation of Municipal Sewage with Brominated Byproducts Control

The decomposition of endocrine-disrupting chemicals (EDCs) including estrone (E1), 17beta-estradiol (E2), estriol (E3), nonylphenol (NP), and bisphenol A (BPA) during ozonation of municipal sewage grabbed from the outlets of primary sedimentation tanks was studied through laboratory-scale experiments. A newly developed in vitro bioassay called nuclear receptor-ligand assay and GC-MS were both utilized to respectively determine the estrogenicity and individual EDCs in the wastewater samples. The original estrogenicity, expressed as the E2 equivalent concentration (EEQC), in the primary effluents was 315-1018 ng/L. Results indicate that the EEQC can be reduced rapidly to below 10 ng/L after ozonation. The appearance of 0.1 mg/L dissolved ozone (DO3), which corresponds to a consumed ozone amount of 0.4 mg per initial TOC (total organic carbon) of wastewater samples, was an appropriate operational parameter to simultaneously achieve efficient EDC removal and control of BrO3- and total organic bromine (TOBr). The presence of suspended solids in the range of 38-67 mg/L exhibited no obvious impact on the removal of nonsorbed estrogenicity. A complete decomposition of E2, E3 and BPA was achieved once 0.1 mg/L DO3 appeared in the primary effluent. The oxidative decomposition of NP was relatively less efficient with a residual concentration of 100 ng/L. This work investigates the feasibility of EDC removal and brominated byproduct control during ozonation of original municipal sewage prior to biological treatment.

The use of in vitro bioassays to quantify endocrine disrupting chemicals in municipal wastewater treatment plant effluents

In vitro bioassays are widely used to detect and quantify endocrine disrupting chemicals (EDCs) in the influents and effluents of municipal wastewater treatment plants (WWTP). These assays have sometimes led to false positive or negative results, partly due to the low EDC concentrations in the samples. The objectives of the present study were: (a) to compare the estrogen screen (E-Screen) and the yeast estrogen screen (YES) bioassays using the 17beta-estradiol (E2) or its equivalence and (b) to investigate if a combination of the E-Screen and YES assays can be used to improve the accuracy of EDC detection and quantification. The E-Screen bioassay was conducted with the MCF-7 (BOS) human breast cancer cell line while the YES bioassay employed two different types of recombinant yeast. The influent and effluent samples collected from the five WWTPs operated by the Greater Vancouver Regional District (GVRD) were analyzed by both the E-Screen and the YES bioassays. Since the results of the E-Screen and YES bioassays varied by up to 4-fold on the same split sample of a nominal E2 concentration, the mean value of the E-screen and YES bioassays was used to represent the EDC activity of a given WWTP sample. Results of these studies showed that the E2 equivalent concentration in each WWTP sample was consistently higher than 1 ng/L, a concentration that may potentially cause endocrine disruption in different aquatic species. The composition of selected EDCs in a subset of effluent samples was examined using a gas chromatograph-high resolution mass spectrometer (GC-HRMS). EDC composition in 10 WWTP samples correlated with the mean endocrine disrupting activities of the E-Screen and YES bioassays. Results also indicated that secondary treatment plants are comparable to the primary treatment plants in removing EDCs from the final effluents.

Ozonation parameter for removal of oestrogenicity from secondary effluent without by-products

The purposes of this study are to investigate ozonation characteristics, to evaluate oestrogenicity and to confirm behaviours of by-products during ozonation of secondary effluent. For the ozonation of secondary effluent, TOC was decreased only 10% when 4 (mgO3/mgC) of ozone consumption per initial TOC. However, UV254 and SUVA was decreased approximately 65% until ozone consumption of 2 (mgO3/mgC). Ozonation was also shown to be very effective for decrease of oestrogenicity in secondary effluent. Bromate ion started to form obviously when the ozone consumption per initial TOC exceeded 2 (mgO3/mgC) and increased while the time of ozonation became longer. From these results, ozone consumption per initial TOC was shown to be an appropriate operation parameter to reduce SUVA effectively and E2 equivalent concentration to less than 0.1 nM without significant formation of bromate ion, and its value was determined to be 2 (mgO3/mgC) in ozonation of secondary effluent.

Evaluation of Estrogenicity for 17 β-Estradiol Decomposition During Ozonation

ABSTRACT To evaluate the decomposition and formation characteristics of estrogenic compounds during ozonation, semibatch ozonation experiments were conducted using the prototypical estrogen, 17β-estradiol (E2). Ozonation was shown to be very effective for decomposition of E2 and decrease of estrogenicity. An E2 equivalent concentration index was introduced based on the estrogenicity intensity measured with Estrogen Competition Binding Assay (ECB Assay). The importance of evaluation of estrogenicity by ECB Assay was shown by the difference in removal efficiencies between E2 equivalent concentration and the E2 itself. Through this research, ozonation was shown to be a promising method to remove E2 without any formation of estrogenic by-products from secondary-treated sewage, which contained E2 of ngL level and total organic carbon (TOC) of several mgL.

Evaluation of Wetland and Tertiary Wastewater Treatments for Estrogenicity Using In Vivo and In Vitro Assays

The effects of wetland wastewater treatment on estrogenic activity and estrogenic activity of water after tertiary treatment were evaluated using in vivo (rainbow trout vitellogenin [VTG] expression) and in vitro (yeast estrogen screening) assays. Juvenile rainbow trout exposed to tertiary-treated wastewater from the Green Acres Treatment Plant in Orange County Water District had increased plasma VTG levels compared with control fish. When trout were exposed to wastewater-dominated water before it entered into Prado Wetland (Riverside County, CA), VTG concentrations were increased above those of controls and were not significantly different from fish exposed to water exiting the wetland. VTG E2-equivalent concentrations (EEQs) of the water samples from the Green Acres Plant were 16.92 +/- 16.49 ng/L. Activity of water entering Prado Wetland was 29.80 +/- 28.41 ng/L EEQ, and water exiting was 24.34 +/- 23.17 ng/L EEQ. In vitro assays estimated that estrogenic activity of water from the Green Acres Plant was <1 ng/L EEQ, whereas water entering and exiting Prado Wetland had yeast estrogen screening EEQs of 2.57 and <1 ng/L, respectively. Our results suggest that environmental estrogens that are not potent estrogen-receptor ligands exist in wastewaters from the Green Acres Plant as well as water entering and exiting Prado Wetland. Wetland treatment did not remove environmental estrogens in the water. Our results also suggest that in vitro assays may underestimate estrogenic activity of sampled water.

Radiation-induced decomposition of trace amounts of 17 β-estradiol in water

The radiation-induced decomposition of trace amounts of 17 β-estradiol (E2) in water was studied as a function of the dose of 60Co γ-rays. The rate constant of the reaction of the OH radicals with E2 was estimated to be 1.6×10 10 mol dm −3 s −1 by a comparison with the known rate constant for the reaction with phenol. Both E2 and E2-equivalent concentrations were estimated by LC-MS and ELISA, and decreased with an increase in γ-rays dose. E2 (1.8 nmol dm −3) in water was degraded almost completely by irradiations up to 10 Gy. The estrogen activity of the same sample solution still remained at a dose of 10 Gy, but decreased at 30 Gy to the lower than the threshold level of contamination to induce some estrogenic effects on the environmental ecology.