Estradiol Equivalents Research Articles

The use of in vitro bioassays and chemical screening to assess the impact of a minimally processed vegetable facility on wastewater quality.

Fruit- and vegetable-processing facilities may contaminate wastewater via contaminants found in the produce and disinfecting chemicals used. These contaminants may include agrochemicals, pesticides, and disinfectants such as chlorine and quaternary ammonium compounds (QACs). Some compounds may exhibit harmful endocrine-disrupting activity. This study investigated the impact of a minimally processed vegetable facility on wastewater quality via in vitro bioassays and chemical screening. Estrogen activity was assessed via a yeast estrogen screen (YES), and (anti-)androgenic and glucocorticoid activities were evaluated via an MDA-kb2 reporter gene assay. The samples were screened via gas and liquid chromatography-tandem mass spectrometry (GC-MS/MS and LC-MS/MS) to identify target compounds, and GC coupled with time-of-flight mass spectrometry (GC-TOFMS) was used for non-targeted screening. Sample complexity and chemical profiles were assessed using GC-TOFMS. Estrogenic activity was detected in 16 samples (n = 24) with an upper limit of 595 ± 37ng/L estradiol equivalents (EEqs). The final wastewater before discharge had an EEq of 0.23ng/L, which is within the ecological effect-based trigger value range for the estrogenic activity of wastewater (0.2-0.4ng/L EEq). Androgenic activity was detected in one sample with a dihydrotestosterone equivalent (DHTEq) value of 10 ± 2.7ng/L. No antiandrogenic activity was detected. The GC-MS/MS and LC-MS/MS results indicated the presence of multiple pesticides, nonylphenols, triclocarban, and triclosan. Many of these compounds exhibit estrogenic activity, which may explain the positive YES assay findings. These findings showed that wastewater from the facility contained detergents, disinfectants, and pesticides and displayed hormonal activity. Food-processing facilities release large volumes of wastewater, which may affect the quality of the water eventually being discharged into the environment. We recommend expanding conventional water quality monitoring efforts to include additional factors like endocrine activity and disinfectant byproducts.

Human exposure to methyl and butyl parabens and their transformation products in settled dust collected from urban, semi-urban, rural, and tribal settlements in a tropical environment

The present study involved monitoring the distribution of two widely consumed parabens (methyl paraben (MeP) and butyl paraben (BuP)) and their transformation products in indoor dust from different categories of settlement (urban, semi-urban, rural, and tribal homes). The results revealed a prevalent occurrence of parabens in all the settlement categories. A non-normal distribution pattern for MeP and BuP levels across the sampling sites was noted. While comparing the residence time of parabens in dust samples, it was found that the half-lives of the analytes were greater in the dust from urban (MeP t1/2: 47.510 h; BuP t1/2: 22.354 h) and rural (MeP t1/2: 27.725 h and BuP t1/2: 31.500 h) areas. The presence of paraben metabolites, such as hydroxy methylparaben (OH–MeP), para hydroxy benzoic acid (p-HBA), and benzoic acid (BA) in dust samples supports their transformation within indoor spaces. The average daily intake of parabens through dust ingestion and dermal absorption by children was higher than adults. BuP was the prime contributor (>85%) to the total estradiol equivalency quotient (tEEQ) in all the settlement categories.

Exploration of the effect of simultaneous removal of EDCs in the treatment process of different types of wastewater.

The SPE-HPLC-MS/MS method was used to investigate the concentration distribution of nine types of estrogens in 18 locations of pollution source along the Jiuzhou River belonging to river systems in Guangdong province and Guangxi Zhuang autonomous region in China, and the estrogenic activity and potential ecological risks were evaluated by calculating the estradiol equivalency (EEQ). The results showed that the calculated estradiol equivalents (cEEQs) of wastewater treatment plants from 17 locations were all higher than 1 ng/L. To further study the removal effect of the treatment process on the estrogens, the pig breeding wastewater from P4 and the municipal wastewater from P13, as well as the black-odorous water, were sampled and surveyed during the entire process. It turned out that estrogens were effectively removed after nitrification activated sludge treatment. Meanwhile, there was a positive correlation between the removal of NH3-N, total nitrogen (TN) and total phosphorus (TP) and the removal of endocrine disrupting chemicals (EDCs). It is shown in the study the secondary treatment process has achieved a significant effect on the removal of estrogen in both the wet and dry periods and that there has been a positive correlation between the activities of total phosphorus compounds, nitrogen-based compounds and the removal of EDCs.

Hazardous chemicals in non-polar extracts from paper and cardboard food packaging: an effect-based evaluation

BackgroundFood contact articles are used in our everyday life and information regarding the potential health hazards of migrating chemicals for humans is scarce. In this study, an effect-based evaluation of non-polar extracts of food contact articles made of paper and board was conducted with a panel of eight bioassay endpoints. These, health-relevant endpoints, included oxidative stress, inflammation, genotoxicity, xenobiotic metabolism and hormone receptor effects.ResultsIn total, 62 food contact articles were pooled into 19 groups, in which articles intended to be used for similar types of food item(s) were pooled, and extracted with acetone:n-hexane (1:4). These were then tested in the effect-based bioassays. Bioactivities were detected for multiple materials in six out of eight assays, the two assays showing no effects were NFκB and androgen receptor agonistic response. In essence, the detection rates of the tested non-polar extracts were 72% for antagonistic effects on the estrogen receptor, 72% for antagonistic effects on the androgen receptor, 47% for oxidative stress, 28% for agonistic effects on the estrogen receptor and 33% for genotoxicity. The bioequivalent concentrations ranges in extracts of 10 mg food contact article/mL cell culture media were: for oxidative stress from 2.45 to 5.64 µM tBHQ equivalents, estrogen receptor agonistic activity from 1.66 to 6.33 ρM estradiol equivalents, estrogen receptor antagonistic activity from 1.21 × 10–3 to 4.20 × 10–3 μM raloxifene equivalents and androgen antagonistic activity 0.08–0.46 μM hydroxyflutamide equivalents. The extracts that were bioactive in multiple assays were: baking moulds, boxes for popcorn, infant formula/skimmed milk, porridge/flour mixes, pizza, fries’ and hamburgers as well as packages for frozen food.ConclusionNon-polar extracts of food contact articles contain compounds that can activate molecular initiating events in toxicity pathways of high relevance to human health. These events included endocrine-disruptive activities, oxidative stress and genotoxicity. Effect-based methods proved to be a valuable tool for evaluating food package articles, as they can detect potentially hazardous effects of both known and unknown chemicals as well as potential cocktail effects.

Characteristics of Steroid Estrogen Loss, Degradation and Residues during Open-Air Dairy Manure Disposal

Steroid estrogens (SEs) are mainly derived from livestock manure, and composting is the common method of bioresource utilization. In this study, an open-air composting experiment with dairy manure was conducted to observe the degradation and loss of five SEs under the influence of different precipitation intensities and additional mixed fermentation strains. SE determination results for dairy manure after 30 days of composting indicated that the average removal rates of 17α-estradiol (E2α), estrone (E1), 17β-estradiol (E2β), 17α-ethinyl estradiol (EE2), estriol (E3), and estradiol equivalent (EEQ) were 76.67%, 71.07%, 73.88%, 92.02%, 98.77%, and 88.11%, respectively, partly due to SE runoff. The rates of SEs leaching from the open-air composting dairy manure ranged from 0.05% to 4.75% after 10 rounds of 5–60 mm/d simulated rainfall. The total leaching amount of SEs was positively correlated with rainfall, but the leaching concentrations of SEs were just the opposite. As a result of its role as a degradation intermediate of other SEs, E3 was the most prone to run off. By strengthening the action of microorganisms, the total leaching amount of EEQ increased by 5%, E3 increased by five times, and E2β also underwent a transition from a conjugated form to free. However, there were also fewer final SEs remaining in the composted product, as well as the environmental risks of conjugated SEs. These conclusions can provide beneficial suggestions and references for controlling the environmental risks of SEs in the process of composting livestock and poultry manure.

Developing a quantitative framework to track the fate and transport of estrogens on a watershed scale

• The first study to develop an HSPF modeling framework to simulate three estrogens with interconversion. • The modeling framework can be used to characterize the temporal and spatial variation of estrogens in streams. • The modeling framework can help to investigate the factors leading to high estrogen levels, evaluate the potential health risks to aquatic animals, and determine the best management practices for estrogens. The risks associated with estrogens in detectable concentrations in the aquatic environment underscore the need to best manage the release of estrogenic compounds. Modeling work can effectively help to understand the variation of estrogens in the natural water bodies at low costs. This study developed a modeling framework based on the Hydrological Simulation Program – FORTRAN (HSPF) code to simulate the fate and transport of three dominant estrogens, estrone (E1), 17β-estradiol (E2β), and 17α -estradiol (E2α) in surface water by integrating their excretion, transport, interconversion, and attenuation processes. This work focused on estrogens from sewage systems, failing septic systems, grazing animals, and manure land application. The developed modeling framework was applied to the Redwood River Watershed in Minnesota to model the in-stream estradiol equivalents (EEQs). The modeled EEQs were comparable to the values measured at three sampling sites along the Redwood River in 2007. The modeling results indicate that wastewater treatment plant discharges elevate estrogen levels on dry days, and surface runoff caused by storms or snow melting after the manure land application and/or cattle grazing can drastically spike EEQs levels, suggesting the application of buffer stripes and manure storage/composting for estrogen control. The case study shows that the modeling framework can be used to characterize the temporal and spatial variation of estrogens in streams, evaluate their risk to aquatic animals, and determine the best management practices for estrogen control.

Estrogenic Activity and Endocrine Disruptor Compounds Determined in Guanabara Bay (Brazil) by Yeast Estrogen Screen Assays and Chemical Analyses

Studies assessing the presence of endocrine disrupting compounds in marine environments have increased in the last decades. In Brazil, the combination of poor sanitation conditions and low investment in sewage treatment plants leads to significant contamination of receiving waters. The risks of these micropollutants in the aquatic biota include biochemical and histopathological alterations of the liver, gonads, and kidneys, as well as, reproductive process and development modifications, and behavioral changes, among others. The aim of this study was to evaluate the quality of the surface and deep waters of Guanabara Bay, southeastern Brazil, regarding the presence of estrogenic substances. Acute toxicity assays were also conducted employing Vibrio fischeri. The estrogenic activity of the water samples was determined by Yeast Estrogen Screen assay and the quantification of the Bisphenol A, estriol, 17β-estradiol, and 17α-ethinylestradiol by high-performance liquid chromatography, using fluorescence and diode array detectors. Estrogenic activity ranged from 9 to 77 ng L-1 of estradiol equivalents. The highest micropollutants concentrations were detected for bisphenol A (298.5 and 465.5 ng L-1), followed by 17α-ethinylestradiol (248 and 256.9 ng L-1), estriol (70.7 and 179.6 ng L-1), and 17β-estradiol (167 and 174.8 ng L-1) for surface and deep waters, respectively. The findings indicate significant risks for the Guanabara Bay ecosystem. No acute toxicity effects were observed in the V. fisheri assay. These data reflect the current environmental degradation situation of the bay’s waters and highlight the need for the systematic monitoring of this important estuary.

Effect-Directed Analysis Based on the Reduced Human Transcriptome (RHT) to Identify Organic Contaminants in Source and Tap Waters along the Yangtze River.

Since a large number of contaminants are detected in source waters (SWs) and tap waters (TWs), it is important to perform a comprehensive effect evaluation and key contributor identification. A reduced human transcriptome (RHT)-based effect-directed analysis, which consisted of a concentration-dependent RHT to reveal the comprehensive effects and noteworthy pathways and systematic identification of key contributors based on the interactions between compounds and pathway effects, was developed and applied to typical SWs and TWs along the Yangtze River. By RHT, 42% more differentially expressed genes and 33% more pathways were identified in the middle and lower reaches, indicating heavier pollution. Hormone and immune pathways were prioritized based on the detection frequency, sensitivity, and removal efficiency, among which the estrogen receptor pathway was the most noteworthy. Consistent with RHT, estrogenic effects were widespread along the Yangtze River based on in vitro evaluations. Furthermore, 38 of 100 targets, 39 pathway-related suspects, and 16 estrogenic nontargets were systematically identified. Among them, diethylstilbestrol was the dominant contributor, with the estradiol equivalent (EEQ) significantly correlated with EEQwater. In addition, zearalenone and niclosamide explained up to 54% of the EEQwater. The RHT-based EDA method could support the effect evaluation, contributor identification, and risk management of micropolluted waters.

Novel hyphenation of DGT in-situ passive sampling with YES assay to ascertain the potency of emerging endocrine disruptors in water systems in New Zealand

This study represents the first attempt to investigate selected estrogenic compounds that include 17α-ethynylestradiol (EE2), 17β-estradiol (E2) bisphenol A (BPA), and bisphenol AF (BPAF) along the drinkable water, from river-to-tap, and wastewater, from effluent-to-treated wastewater, treatment processes of the Hamilton City Council and the monitoring of the freshwater, from source-to-outfall, of the Waikato River in New Zealand. This was accomplished by the adoption of a novel combination of diffusive gradients in thin films (DGTs) in-situ passive sampling coupled with high-performance liquid chromatography/mass spectrometry analysis (HPLC/MS) and the Yeast Estrogen Screen (YES). Estradiol equivalency quantities, integrated in time, were evaluated theoretically (cEEQ) by DGT-HPLC/MS and experimentally (EEQ) by DGT-YES assay. cEEQ and EEQ highlighted that primary treatments are not suitable for estrogens and bisphenolic plastics removal both at drinkable and wastewater treatment plants in Hamilton where they worsen the water quality in terms of estrogenicity making these pollutants more available in the water phase. All downstream sites monitored along the Waikato River showed higher cEEQ and EEQ, moreover the Waikato River water quality showed a moderate worsening moving from Taupo (source) to Tuakau (outfall). The most polluted sites were downstream of Hamilton city and Huntly township wastewater treatment plants that serve the main conurbations in the area. cEEQ and EEQ generally showed good agreement at low concentrations but differed substantially at more polluted sites where cEEQ consistently underestimated estrogenic potency, possibly due to DGT accumulation of estrogenic compounds not quantified by HPLC/MS.

Determination of bisphenols and parabens in cow urine distillate from India: implication of human exposure and risk assessment.

The exposures of a wide range of endocrine-disrupting chemicals to cows have been linked to diseases and are a major concern. In Indian scripts, cow urine is believed to be a treatment for many diseases. Nonetheless, exposure of bisphenols and parabens to cow urine distillates is unknown. Hence, in this study, we determined the concentration of bisphenols and parabens in commercially available cow urine distillate collected from India. The mean concentration of total bisphenols and parabens ranges from the limit of quantification (LOQ) to 149.3ng/mL and 1479.88 ng/mL, respectively. Predominant bisphenol-F and bisphenol-A were accounted for 88.6% and 6% of total bisphenol concentration, respectively, in marketed cow urine distillate, whereas methyl and propyl parabens were dominant and accounted for 33% and 65%, respectively. The estimated mean daily intake (EDI) of bisphenol for males and females were 45.94 ng/kg-bw/day and 54.29 ng/kg-bw/day, respectively, while for parabens EDI was 455.35ng/kg-bw/day and 538.14 ng/kg-bw/day for males and females, respectively. Hazard quotient, to evaluate the potential risk of exposure, showed no risk in the studied samples. Even though the EDI results from the Monte-Carlo risk assessment analysis did not exceed the acceptable daily intake, their estrogenic actions cannot be ignored in general populations. The estrogenic activities contributed by parabens and bisphenol A measured by estradiol equivalency quotient (EEQ) ranged from 0.00033-42 pg/mL and 2.3 pg/mL, respectively. Our results revealed higher concentrations of bisphenols and parabens in cow urine distillates; hence, special attention should be given to the quality and safety of cow urine distillates. Moreover, strict guidelines should be enforced for the quality of cow urine distillates.

Monitoring estrogen and androgen residues from livestock farms in Phayao Lake, Thailand.

The aim of the present study was to investigate steroid hormone residues from livestock farms and assess their risks to the surface water of Phayao Lake. These steroid hormones are endocrine-disrupting compounds (EDCs), which can be found in natural and synthetic forms. This research focused on examining the residues of seven steroid hormones (five estrogens and two androgens-estrone (E1), 17α-estradiol (αE2), 17β-estradiol (βE2), estriol (E3), 17α-ethinyl estradiol (EE2), testosterone (T), and 17α-methyltestosterone (MT)) from four types of livestock farms around Phayao Lake, Thailand. The samples collected from the livestock farms included feces, soil, and wastewater and were extracted by the solid phase extraction (SPE) technique and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The risks from the residual steroid hormones were also characterized by estradiol equivalents (EEQs), testosterone equivalents (TEQs), and risk quotients (RQs). The results indicated that most hormone contamination from the farms' livestock was due to the estrogen hormones E1 (1.38-97.10ng/g), βE2 (10.08-1366ng/g), and EE2 (1.50-99.92ng/g), which originate from the natural excretion and admixture of steroids in feedstock or medicines. Steroid hormones were not detected in the wastewater from cleaning processes on farms with wastewater treatment plants, whereas farms without wastewater treatment plants showed high values of estrogen hormone contamination, with EEQs of 128.8-472.9ng/L and RQs of 208.3-294.3. However, the analysis of steroid hormone residues in Phayao Lake demonstrated that the residues did not severely affect aquatic organisms (with RQs of 0.002-144.5), and no estrogen or androgen residues were observed in the water treatment plant or tap water.

Occurrence, distribution, and risk assessment of bisphenol analogues in Luoma Lake and its inflow rivers in Jiangsu Province, China.

Bisphenol analogs (BPs) are widely used in industrial and commercial products and have been detected in surface water, sediment, sewage, and sludge. The presence of BPs in the natural environment poses threats to the aquatic ecosystem and human health. The concentration, distribution, seasonal variation, and risk assessment of BPA and BPA structural analogs including BPB, BPF, BPS, BPZ, BPAF, and BPAP in surface water and sediment during dry season and flood season in Luoma Lake and its inflow rivers in Jiangsu Province, China, were investigated in this study. The detection frequency of BPA and BPF was 100%. Although the use of BPA is restricted, BPA is still the dominant BPs in surface water and sediment. The concentration of BPs in surface water during flood season was higher than that in dry season. The concentrations of BPs in Fangting River, Zhongyun River, and Bulao River were higher than those in Luoma Lake. The average concentrations of BPs in surface water were in the order of BPA > BPF> BPS> BPB > BPZ > BPAF> BPAP. Compared with other studies, the concentration of BPs in Luoma Lake was moderate. There is no significant spatial distribution and difference in seasonal variation of BPs concentration in sediment (p > 0.05). Compared with other studies, the contamination of BPs in sediment of Luoma Lake was relatively low. Risk quotient (RQ) was used to evaluate the ecological risk of BPs in water environment, and the 17β estradiol equivalent (EEQ) method was used to estimate the estrogenic activity of BPs. The risk assessment showed no high ecological risk (RQ < 1.0) and estrogenic risk (EEQ < 1.0 ng/L) in dry season and flood season. The estimated RQ and EEQt indicated that the ecological and human health impacts were negligible in the short term.

Lethal and long-term effects of landfill leachate on Eisenia andrei earthworms: Behavior, reproduction and risk assessment

Leachate is difficult to biodegrade, and presents variable physical, chemical and biological characteristics, as well as high toxicological potential for soil, groundwater and water bodies. In this context, untreated leachate toxicity was evaluated through acute and chronic exposures in Eisenia andrei earthworms. Physico-chemical leachate characterizations indicate a complex composition, with high organic matter (COD – 10,634 mg L−1) and ammoniacal nitrogen (2388 mg L−1) concentrations. Metals with carcinogenic potential, such as Cr, As and Pb, were present at 0.60, 0.14 and 0.01 μg L−1, respectively and endocrine disrupting compounds were detected in estradiol equivalents of 660 ± 50 ng L−1. Acute tests with Eisenia andrei indicated an LC50 (72 h) of 1.3 ± 0.1 μL cm−2 in a filter paper contact test and 53.9 ± 1.3 mL kg−1 in natural soil (14 days). The EC50 in a behavioral test was estimated as 31.6 ± 6.8 mL kg−1, indicating an escape effect for concentrations ranging from 35.0 to 70.0 mL kg−1 and habitat loss from 87.5 mL kg−1 of leachate exposure. Chronic exposure (56 days) led to reproduction effects, resulting in a 4-fold decreased cocoon production and 7-fold juvenile decrease. This effect was mainly attributed to the possible presence of endocrine disrupting compounds. An estimated NOAEL of 1.7 mL L−1 and LOAEL of 3.5 mL L−1 were estimated for earthworms exposed to the assessed effluent. Extremely high-risk quotients (RQ ≥ 1) were estimated based on leachate application in irrigation. Thus, adequate municipal solid waste management is paramount, especially with regard to generated by-products, which can result in high toxicological risks for terrestrial organisms.

Occurrence and distribution of estrogenic substances in the northern South China Sea

Estrogenic substances are today among the contaminants of emerging concern. Besides naturally occurring estrogens, other natural and synthetic substances can mimic a hormonal action due to their structural resemblance to hormones, possibly affecting the endocrine system of living organisms. Estrogens have been detected in inland water bodies such as influents and effluents of waste water treatment plants as well as in rivers, but data on their distribution and variability in the marine ecosystem are still limited. Surface water samples obtained during two research cruises on the northern shelf of the South China Sea (SCS) near the Pearl River Estuary, in September 2018 and in August 2019, were investigated for estrogenic substances, namely estrone (E1), 17β-estradiol (E2), 17α-ethinylestradiol (EE2), genistein (GEN), daidzein (DAI) and zearalenone (ZEN). Among the target analytes, the natural hormones E1 and E2, as well as the synthetic EE2, were the most abundant with maximum concentrations of 1.1 ng L−1, 0.7 ng L−1 and 0.6 ng L−1, respectively. Of substances produced by plants and fungi, GEN, DAI and ZEN, only GEN was detected (1.2 ng L−1). High concentrations occurred predominantly close to the coast, which was also reflected in the calculated estradiol equivalent quotients (up to 1.4 ng L−1). In general, the distribution of estrogenic substances observed in both years shows a regional and inter-annual variability consistent with the modeled surface current data for the SCS. Regarding single estrogenic compounds and estradiol equivalents, marine organisms in the northern SCS might be exposed to high potential risk.

Surveillance of plasticizers, bisphenol A, steroids and caffeine in surface water of River Ganga and Sundarban wetland along the Bay of Bengal: occurrence, sources, estrogenicity screening and ecotoxicological risk assessment

The transboundary River Ganga serves as a conduit for meltwater from the Himalayas and is a major freshwater source for two thirds of Indian population before emptying into the Sundarban Delta, the largest estuary in the Bay of Bengal. Endocrine disrupting compounds (EDCs) such as phthalic acid esters (PAEs) and bisphenol A (BPA) used as organic plastic additives can pollute the aquatic environment receiving plastic litter. Hence, we have investigated these EDCs in water samples from Ganga and Sundarban wetland of India. Since these compounds exhibit estrogenic potential, we have further measured steroids and evaluated the estrogenic activity (estradiol equivalents, BioE2Eqs) using an in-vitro bioassay (E-Screen). Further BioE2Eqs were compared with the sum of predicted estradiol equivalents based on the chemical concentrations of PAEs and BPA by E-Screen (ChemE2Eq) and YES factors (ChemYES). Caffeine was measured as a marker for anthropogenic wastewater discharge. Results showed that the highest BioE2Eq (below the lowest observable effect of E2 on fish) was associated with sites having sewer outfalls in the middle stretch of the river, and concomitantly coinciding with the elevated concentrations of caffeine. Neither ChemE2Eq nor ChemYES correlated with measured BioE2Eqs. River concentrations of BPA (0.04–4.46 µg/L) and ∑7plasticizers (0.43–7.63 µg/L) were higher than BPA (0.21–2.82 µg/L) and ∑7plasticizers (0.85–2 µg/L) in the Sundarban wetland. The only steroids detected were androgens, found at four sites in Ganga (0.007 µg/L± 0.003, mean ± S.D.). The highest estimated ecotoxicological risk to aquatic insect and fish stemmed from BPA. A secondary effect, and a potential impact on human health could be reflected via fish consumption from the productive fisheries region along the lower stretch of River Ganga. Identification of areas of elevated estrogenicity, plasticizer and steroid concentrations in River Ganga can be used to design and implement interventions for the remediation of such emerging contaminants.

Bacterial community composition and function succession under aerobic and anaerobic conditions impacts the biodegradation of 17β-estradiol and its environmental risk

The widespread detection of 17β-estradiol (E2) in the environment has become an emerging concern worldwide due to its endocrine disrupting effects. This work focuses on the aerobic and anaerobic biodegradations of E2 in various sedimentary environments with different availabilities of electron acceptors, including O2, NO3−, Fe3+, SO42−, or HCO3−. The highest removal efficiency (98.9%) and shortest degradation half-life of E2 (t1/2 = 5.0 d) were achieved under aerobic condition, followed by nitrate-reducing, ferric-reducing, sulfate-reducing and methanogenic conditions. We propose four different degradation pathways of E2 based on the metabolites identified under various redox conditions. Although most of E2 was effectively removed under aerobic condition, the potential environmental risk still needs to be considered due to the residual estrogenic activity induced by estrone (E1) formation. The endocrine-disrupting activities, as indicated by estradiol equivalent (EEQ) values, were related to E2 degradation rate and metabolite formation. We further analyzed the succession of bacterial community compositions and functions using Illumina HiSeq sequencing and Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). The findings herein evidenced that bacterial community compositions and metabolic functions associated with different redox conditions impact the biodegradation of E2 and its endocrine-disrupting activity. This knowledge will be useful in predicting the environmental fates of estrogenic hormones in various sedimentary environments and aid in establishing appropriate strategies for eliminating potential environmental risks.

Nonylphenol ethoxylates biodegradation increases estrogenicity of textile wastewater in biological treatment systems

The formation of estrogenic intermediates, i.e. nonylphenol diethoxylate (NP2EO), nonylphenol monoethoxylate (NP1EO), and nonylphenol (NP), following nonylphenol ethoxylates (NPEOs) biodegradation in textile wastewater raises concerns about its endocrine disruptive activity, but the estrogenicity changes of textile wastewater throughout biological treatment processes remain unknown. In the present study, the estrogenicity of textile wastewater sampled from 10 wastewater treatment plants (WWTPs) were investigated using the reporter gene-based T47D-KBluc bioassay. Results showed that the estrogenicity of the textile wastewater significantly increased after either anaerobic or aerobic treatment in all WWTPs, with an average fold change of 3.21, although traditional pollutants were effectively removed. The estradiol equivalents of the effluent (ranging from 1.50 to 4.12 ng-E2/L) were generally higher than published effect based trigger values, indicating an increased risk for the receiving waters. Removal efficiency was high (84.46%) for NPEOs, but was low for NP2EO and NP1EO in the biological treatment processes. Nevertheless, NP had increased concentrations after the treatment. Bioanalytical equivalent concentration of the textile wastewater and that of NP2EO, NP1EO, and NP showed a good linear correlation, of which NP alone contributed more than 70% to the observed estrogenicity. Extending hydraulic retention time was found effective in reducing the estrogenicity as it allows relatively complete degradation of NP, which was further confirmed by running lab-scale A/O reactors fed with NP10EO. The results may extend our knowledge regarding the estrogenicity of textile wastewater and its reduction technologies used in WWTPs.

Estrogenic activity of food contact materials\u2014evaluation of 20 chemicals using a yeast estrogen screen on HPTLC or 96-well plates

Food contact materials (FCM) may contain complex mixtures of estrogenic chemicals. A yeast estrogen screen performed on high performance thin-layer chromatography plates (planar-YES, P-YES) is promising for analysis of such mixtures, as it could allow for better elucidation of effects compared with established methods in microtiter plates. However, the P-YES has not been directly compared with established methods. We compared the performance of a microtiter plate YES (lyticase-YES, L-YES) to P-YES on silica gel HPTLC plates using 17β-estradiol (E2), 20 chemicals representative of migrants from plastic FCM, and three migrates of coated metal food cans. Effective doses (ED10, ED50) and estradiol equivalencies were calculated for each chemical. Thirteen chemicals had calculable EDs in the L-YES or P-YES, with average EDs 13-fold (range 0.63–36) more potent in P-YES than in the L-YES. Normalized to E2, the median estrogenicity was within 1.5-fold (0.43–8.8) between the assays. Therefore, P-YES was as or more sensitive than L-YES but potencies relative to E2 were comparable between assays. With chromatography, the P-YES detected estrogenicity in coated metal cans, effects that were unmeasurable in L-YES. With the sample preparation methods used in this study, both YES assays are sufficiently sensitive to detect bisphenol A below the specific migration limit for plastic packaging (0.05 mg/kg food). This study demonstrates that P-YES outperforms L-YES because it is more sensitive, provides comparable estradiol equivalents, and circumvents confounding mixture effects. The P-YES will be useful for routine monitoring of FCM and toxicant identification in problematic materials.Graphical abstract

Endocrine disrupting compounds in the middle and lower reaches of the Lhasa River Basin: Occurrence, distribution, and risk assessment

The contamination of endocrine disrupting compounds (EDCs) in frigid alpine areas is poorly understood compared with that in industrialized regions. In this study, the occurrence, distribution, and risk assessment of EDCs were investigated in the middle and lower reaches of the Lhasa River Basin in the Tibetan Plateau. Eight EDCs were analyzed in samples of surface water, sediment, natural soils, and farmland soils during the dry season and rainy season. Bisphenol A (BPA) and estriol were the most frequently detected compounds. The EDCs concentrations in the sediment and surface water varied with the rainy and dry seasons. The BPA concentration in the surface water of the lower reaches in the dry season was higher than that in the rainy season, which could be attributed to the low dilution by low water flow in the dry season and high degree of urbanization and industrialization in the lower reaches. The estradiol equivalents of EDCs in the surface water were below the predicted no-effect concentration (2 ng/L) of 17β-estradiol in the two seasons. Compared with other research results in the world, EDCs contamination in the Lhasa River Basin is at relatively low levels, and the risk in the middle and lower reaches of the Lhasa River Basin is generally low.

Efficiency of selected wastewater treatment processes in removing estrogen compounds and reducing estrogenic activity using the T47D-KBLUC reporter gene assay

The occurrence of endocrine-disrupting compounds (EDCs) consisting of natural and synthetic estrogens, namely estrone (E1), 17β-estradiol (E2), estriol (E3) and 17α-ethinylestradiol (EE2) was quantified in wastewater samples. The aim of this study was to assess the removal efficiency for the selected estrogens (E1, E2, E3 and EE2) and reduction of estrogenic activity in wastewater samples from wastewater treatment plants (WWTPs) using different processes. Solid-phase extraction (SPE) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were used to quantify the selected estrogens in wastewater samples. Estrogenic activity was assessed using the T47D-KBluc gene reporter assay. Results revealed a decrease in estrogen concentrations observed in the effluents of all the WWTPs, except for E2 at Daspoort where no removal was noted. In general, the highest removal for total estrogens was observed at Phola (84%) combining three processes (AP, BF and wetland). The AS at Daspoort had a highest removal of 75% for E3; while at Zeekoegat the highest removal reached 61% for EE2. The PST at Daspoort had no removal recorded for all the compounds, except for the EE2 (33%). The AP and BF systems at Phola contributed to a higher removal of selected compounds. Downstream of the wetland at Phola no removal was recorded for E3; while the highest removal reached 61% for E1. The best performance in terms of the overall influent-to-effluent removal efficiency was observed at Phola WWTP, where E1 removal of 85% was recorded.The highest estrogenic activity in the effluent was reported at Phola, with an average estradiol equivalent (EEQ) value of 6.3 ± 6.7 ng/L. However, no anti-estrogenic activity was detected in any of the samples. The daily mass load discharged from the effluent of the three WWTPs was higher for E1 recorded at Zeekoegat (8002.3 ± 6416.3 mg/d), followed by Daspoort (3509.8 ± 849.0 mg/d) and finally Phola (176.1 ± 34.9).