Search Results (17)

As emerging contaminants increasingly detected in aquatic and terrestrial ecosystems, environmental estrogens (EEs) pose significant ecological risks to marine ecosystems, particularly affecting photosynthetic microorganisms occupying fundamental roles in marine food webs. This review summarizes the current knowledge on the toxicological effects of EEs in marine microalgae through a systematic analysis of dose-dependent physiological, biochemical, and molecular responses. Experimental evidence reveals a biphasic response pattern characterized by growth promotion and photosynthetic enhancement in microalgae under low-concentration EE exposure (0.1–10 μg/L), while marked inhibition of both growth and photosynthetic activity was observed at elevated EE concentrations (>50 μg/L). Notably, sustained EE exposure induces metabolic reprogramming, manifested through reduced protein and polysaccharide biosynthesis concurrent with accelerated lipid accumulation. Cellular stress responses include significant ultrastructural alterations such as chloroplast membrane disruption, cell wall thickening, and the formation of multicellular aggregates. The study further elucidates the concentration-dependent modulation of toxin metabolism, with sublethal doses stimulating intracellular microcystin synthesis (1.5–2.3-fold increase), while acute exposure triggers toxin release through membrane permeabilization. At molecular levels, transcriptomic analyses identify the up-regulation of heat shock proteins (HSP70/90) and the differential expression of genes governing cell cycle progression (cyclin-D), apoptotic pathways (caspase-3), photosynthetic electron transport (psbA), and oxidative stress responses (SOD, CAT). These findings demonstrate that EEs exert multilevel impacts on microalgal physiology through interference with fundamental metabolic processes, potentially disrupting marine primary productivity and biogeochemical cycles. The identified response mechanisms provide critical insights for environmental risk assessment and establish a conceptual framework for investigating estrogenic pollutant effects in aquatic ecosystems. Full article

Vitellogenins (Vtgs) are key yolk precursor proteins in fish, serving as critical indicators of gonadal maturation in females and reliable biomarkers for detecting xeno-oestrogenic pollution, particularly through their expression in juveniles or males. The vtg gene family comprises multiple subtypes that are species-specific, necessitating precise characterisation and quantification for effective use as biomarkers in studies on estrogenic endocrine-disrupting chemicals (EEDCs). In this study, we successfully cloned and characterised the full-length cDNAs of three vtg subtypes (vtgAa, vtgAb, and vtgC) from Scatophagus argus. Differential expression analysis revealed that vtgAb exhibited the highest responsiveness to 17α-ethynylestradiol (EE2) exposure, with a 3-fold increase in vivo at 10.0 μg/g EE2 and a 30-fold increase in vitro at 10⁻⁷ mol/L EE2. The expression patterns were dose- and time-dependent, with peak expression observed 72 h post-exposure. While in vivo assays indicated moderate upregulation, in vitro experiments demonstrated significantly higher expression, attributed to direct hepatocyte interaction with EE2. These findings confirm vtgAb as the most responsive subtype to oestrogen exposure in S. argus and highlight the species’ tolerance to EE2, as compared to more sensitive species like Danio rerio. This study shows the evolutionary conservation of vtg transcripts across teleost species and reinforces the importance of subtype-specific characterisation to advance their application as biomarkers for EEDCs, with significant implications for environmental monitoring and pollution regulation. Full article

The male reproductive impairment caused by environmental estrogens (EEs) stands as a pivotal research area in environmental toxicology. Alpha2-macroglobulin (A2M) emerges as a promising molecule capable of counteracting oxidative stress induced by EEs. This study conducted exposure experiments spanning PND1 to PND56 employing ICR mice, aiming to delve into the expression patterns of A2M and its modulated IL-6 in the testicular tissue of mice subsequent to diethylstilbestrol (DES) and benzophenone (BP) exposure, while elucidating the pivotal role of ERs in this intricate process. Our findings revealed that upon DES exposure (10 and 100 nM), there was a pronounced upregulation of A2M (mRNA and in situ protein levels) in mouse testicular tissue. Similarly, exposure to BPs (BP-1, BP-2, and BP-3, each at 10 and 1000 nM) exhibited comparable effects and increasing A2M levels in serum. Notably, BP exposure also caused an elevation in IL-6 levels (which could be directly regulated by A2M) within testicular tissue (mRNA and in situ protein). Remarkably, the specific estrogen receptor antagonist ICI 182780 (0.5 mg/kg/day) was effective in reversing the upregulation of both A2M and IL-6 induced by BP exposure. Significantly, the results of theoretical prediction of the potential ERE site in the A2m gene promoter region and ChIP-qPCR experiment provide essential and strong evidence for the key conclusion that A2m is the target gene of ER. Taken together, our study highlights EEs’ ability to regulate A2M expression in the male reproductive system via the ER signaling pathway. This vital insight deepens our understanding of molecular mechanisms protecting against oxidative stress caused by EEs. Full article

A covalent organic framework (COF) based on imine was synthesized using 2,5-dihexoxyterephthalaldehyde (DHT) and 1,3,5-tris(4-aminophenyl) benzene (TAPB) as starting materials. The TAPB-DHT-COF exhibited satisfactory chemical stability, making it a promising adsorbing material for stir bar sorptive extraction (SBSE) of four estrogens, including estrone (E1), β-estradiol (E2), hexestrol (HES), and mestranol (MeEE2), in ambient water samples. The extracted analytes were subsequently analyzed using a high-performance liquid chromatography-diode array detector (HPLC-DAD). A series of parameters affecting the SBSE process, such as solution pH, ionic strength, extraction time, and desorption solvent, were investigated by the controlled variable method. Under optimal conditions, the limit of detection (LODs) for the four targeted estrogens ranged from 0.06 to 0.15 µg/L, with a linear range from 0.2 to 100 µg/L. The observed enrichment factor (EF) ranged from 39 to 49, while the theoretical EF was estimated to be 50-fold. This methodology can be applied to the identification of estrogens in three environmental water samples. Full article

Three-dimensional (3D) fish hepatocyte cultures are promising alternative models for replicating in vivo data. Few studies have attempted to characterise the structure and function of fish 3D liver models and illustrate their applicability. This study aimed to further characterise a previously established spheroid model obtained from juvenile brown trout (Salmo trutta) primary hepatocytes under estrogenic stimulation. The spheroids were exposed for six days to environmentally relevant concentrations of 17α-ethinylestradiol—EE2 (1–100 ng/L). The mRNA levels of peroxisome (catalase—Cat and urate oxidase—Uox), lipid metabolism (acyl-CoA long chain synthetase 1—Acsl1, apolipoprotein AI—ApoAI, and fatty acid binding protein 1—Fabp1), and estrogen-related (estrogen receptor α—ERα, estrogen receptor β—ERβ, vitellogenin A—VtgA, zona pellucida glycoprotein 2.5—ZP2.5, and zona pellucida glycoprotein 3a.2—ZP3a.2) target genes were evaluated by quantitative real-time polymerase chain reaction. Immunohistochemistry was used to assess Vtg and ZP protein expressions. At the highest EE2 concentration, VtgA and ZP2.5 genes were significantly upregulated. The remaining target genes were not significantly altered by EE2. Vtg and ZP immunostaining was consistently increased in spheroids exposed to 50 and 100 ng/L of EE2, whereas lower EE2 levels resulted in a weaker signal. EE2 did not induce significant changes in the spheroids’ viability and morphological parameters. This study identified EE2 effects at environmentally relevant doses in trout liver spheroids, indicating its usefulness as a proxy for in vivo impacts of xenoestrogens. Full article

Plastic/rubber materials used as fasteners in equipment for analyzing or removing organic pollutants in water treatment technologies form an essential part of the device. Micropollutants in water are typically present at very low concentrations (ng/L to µg/L). Therefore, when designing, for example, units for advanced oxidation processes (AOPs) or planning sample handling, it is necessary to assess whether the material is compatible with the usually hydrophobic nature of the pollutants. As a model example, the possible interactions of estrogens, namely, estrone (E1), 17β-estradiol (E2), estriol (E3) and 17α-ethinylestradiol (EE2) with six commonly used plastic and rubber materials were investigated at environmentally relevant concentrations (100–500 ng/L). In the first phase, we proved that polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF) and ethylene propylene diene monomer (EPDM) materials adsorbed only negligible amounts of estrogens, while significant amounts of E1, E2 and EE2 were adsorbed onto Tygon S3™ material. Another unsuitable material was styrene butadiene rubber (SBR), sorbing a considerable quantity of estrone. A detailed test of EPDM at higher concentrations (300 and 500 ng/L) and prolonged soaking time showed significant sorption of EE2 after 12 h of soaking in both deionized and tap water matrices. Thus, EPDM, PTFE and PVDF are suitable materials for sample handling or producing devices for AOP treatment due to their chemical inertness and mechanical flexibility. The results suggest that plastic materials that come into contact with contaminated water must be carefully selected, especially when working at environmentally relevant concentrations. Full article

Reproductive abnormalities, that could lead to possible effects at the population level, have been observed in wild fish throughout the United States, with high prevalence in largemouth bass (LMB; Micropterus salmoides) and smallmouth bass (Micropterus dolomieu). Estrone (E1) and atrazine (ATR) are common environmental contaminants often associated with agricultural land use. 17alpha-ethinylestradiol (EE2) is a contaminant associated with wastewater treatment effluent, and a representative, well-studied estrogen commonly used for fish toxicity testing. Our objective was to assess whether early gonad recrudescence in adult fish was a period of sensitivity for alterations in reproductive condition and function. Adult male LMB were exposed from post-spawning to early gonad recrudescence to either a mixture of E1 (47.9 ng/L) + ATR (5.4 µg/L), or EE2 (2.4 ng/L) in outdoor experimental ponds. Gonad samples were collected from fish just prior to the start of exposure (July), at the end of the exposure period (December), the following spring just prior to spawning (April), and post spawning (May). Gonadosomatic index (GSI) was significantly reduced in E1 + ATR-exposed and EE2-exposed males compared to control at every post-exposure time point. Reduced sperm count and sperm motility were observed in the mixture treatment (E1 + ATR) compared to the control. Sperm motility was also reduced in the EE2 treatment. These data together indicate that estrogenic endocrine-disrupting compounds can lessen the reproductive condition of adult male LMB, and that effects of exposure during early gonad recrudescence can persist at least through the subsequent spawning cycle. Full article

The negative effects of environmental estrogens on wildlife and human beings are gaining increasing attention. Research on the highly sensitive detection method for Vitellogenin (Vtg), one of the biomarkers of environmental estrogens (EEs), is expected to detect weak estrogens in complex environments. This study aimed to develop a label-free immunosensor with high specificity and sensitivity for testing Vtg. Carbon quantum dots (CQDs) with high fluorescence and excellent stability were synthesized, and antilipovitellin monoclonal antibody (Anti-Lv-mAb) was prepared. Based on the fluorescence resonance energy transfer (FRET) between CQDs-conjugated Anti-Lv-mAb and reduced graphene oxide (RGO), an ultrasensitive fluorescent “ON-OFF” label-free immunosensor for detection of Vtg of marine medaka was established. By modification of RGO with poly dimethyl diallyl ammonium chloride (PDDA), the Zeta potential of RGO was changed and the FRET efficiency was improved. The immunosensor displayed a wide linear response to Vtg of marine medaka from 0.1 to 3000 ng/mL, a low limit of detection (LOD) of 0.04 ng/mL, and excellent sensitivity (28,833.63 CPS/(ng/mL)), selectivity, and reproducibility. The results demonstrated that the fluorescent “ON-OFF” immunosensor is an easy-to-use, relatively fast, ultrasensitive, and accurate detection method for weak estrogenic activity. Full article

The production and use of manufactured chemicals have risen significantly in the last few decades. With interest in preserving and improving the state of the environment, there is also growing interested in new technologies for water purification and wastewater treatment. One frequently discussed technological group is advanced oxidation processes (AOPs). AOPs using sulphur-based radicals appear to reduce the volume of organic contaminants in wastewater significantly. The use of persulfate has excellent potential to successfully eliminate the number of emerging contaminants released into the environment. The main disadvantage of sulphur-based AOPs is the need for activation. We investigated an economically and environmentally friendly solution based on hydrodynamic cavitation, which does not require heating or additional activation of chemical substances. The method was evaluated for emerging contaminant removal research, specifically for the group of steroid estrogens. The mixture of estrone (E1), 17β-estradiol (E2), estriol (E3), and 17α-ethinylestradiol (EE2) was effectively eliminated and completely removed during a treatment that lasted just a few seconds. This novel method can be used in a broad spectrum of water treatment processes or as the intensification of reactions in chemical engineering technologies. Full article

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. Full article

New technologies have been developed around the world to tackle current emergencies such as biowaste recycling, renewable energy production and reduction of environmental pollution. The thermochemical and biological conversions of waste biomass for bioenergy production release solid coproducts and byproducts, namely biochar (BC), hydrochar (HC) and digestate (DG), which can have important environmental and agricultural applications. Due to their physicochemical properties, these carbon-rich materials can behave as biosorbents of contaminants and be used for both wastewater treatment and soil remediation, representing a valid alternative to more expensive products and sophisticated strategies. The alkylphenols bisphenol A, octylphenol and nonylphenol possess estrogenic activity comparable to that of the human steroid hormones estrone, 17β-estradiol (and synthetic analog 17α-ethinyl estradiol) and estriol. Their ubiquitous presence in ecosystems poses a serious threat to wildlife and humans. Conventional wastewater treatment plants often fail to remove environmental estrogens (EEs). This review aims to focus attention on the urgent need to limit the presence of EEs in the environment through a modern and sustainable approach based on the use of recycled biowaste. Materials such as BC, HC and DG, the last being examined here for the first time as a biosorbent, appear appropriate for the removal of EEs both for their negligible cost and continuously improving performance and because their production contributes to solving other emergencies, such as virtuous management of organic waste, carbon sequestration, bioenergy production and implementation of the circular economy. Characterization of biosorbents, qualitative and quantitative aspects of the adsorption/desorption process and data modeling are examined. Full article

Nowadays, there is a growing interest in environmental pollution; however, knowledge about this aspect is growing at an insufficient pace. There are many potential sources of environmental contamination, including sex hormones—especially estrogens. The analyzed literature shows that estrone (E1), estradiol (E2), estriol (E3), and synthetic ethinyloestradiol (EE2) are the most significant in terms of environmental impact. Potential sources of contamination are, among others, livestock farms, slaughterhouses, and large urban agglomerations. Estrogens occurring in the environment can negatively affect the organisms, such as animals, through phenomena such as feminization, dysregulation of natural processes related to reproduction, lowering the physiological condition of the organisms, disturbances in the regulation of both proapoptotic and anti-apoptotic processes, and even the occurrence of neoplastic processes thus drastically decreasing animal welfare. Unfortunately, the amount of research conducted on the negative consequences of their impact on animal organisms is many times smaller than that of humans, despite the great richness and diversity of the fauna. Therefore, there is a need for further research to help fill the gaps in our knowledge. Full article

Autocrine/paracrine factors generated in response to 17β-estradiol (E2) within the fallopian tube (FT) facilitate fertilization and early embryo development for implantation. Since cyclic AMP (cAMP) plays a key role in reproduction, regulation of its synthesis by E2 may be of biological/pathophysiological relevance. Herein, we investigated whether cAMP production in FT cells (FTCs) is regulated by E2 and environmental estrogens (EE’s; xenoestrogens and phytoestrogens). Under basal conditions, low levels of extracellular cAMP were detectable in bovine FTCs (epithelial cells and fibroblasts; 1:1 ratio). Treatment of FTCs with forskolin (AC; adenylyl cyclase activator), isoproterenol (β-adrenoceptor agonist) and IBMX (phosphodiesterase (PDE) inhibitor) dramatically (>10 fold) increased cAMP; whereas LRE1 (sAC; soluble AC inhibitor) and 2’,5’-dideoxyadenosine (DDA; transmembrane AC (tmAC)) inhibitor decreased cAMP. Comparable changes in basal and stimulated intracellular cAMP were also observed. Ro-20-1724 (PDE-IV inhibitor), but not milrinone (PDE-III inhibitor) nor mmIBMX (PDE-I inhibitor), augmented forskolin-stimulated cAMP levels, suggesting that PDE-IV dominates in FTCs. E2 increased cAMP levels and CREB phosphorylation in FTCs, and these effects were mimicked by EE’s (genistein, 4-hydroxy-2’,4’,6’-trichlorobiphenyl, 4-hydroxy-2’,4’,6’-dichlorobiphenyl). Moreover, the effects of E2 and EE were blocked by the tmAC inhibitor DDA, but not by the ERα/β antagonist ICI182780. Moreover, BAPTA-AM (intracellular-Ca²⁺ chelator) abrogated the effects of E2, but not genistein, on cAMP suggesting differential involvement of Ca²⁺. Treatment with non-permeable E2-BSA induced cAMP levels and CREB-phosphorylation; moreover, the stimulatory effects of E2 and EEs on cAMP were blocked by G15, a G protein-coupled estrogen receptor (GPER) antagonist. E2 and IBMX induced cAMP formation was inhibited by LRE1 and DDA suggesting involvement of both tmAC and sAC. Our results provide the first evidence that in FTCs, E2 and EE’s stimulate cAMP synthesis via GPER. Exposure of the FT to EE’s and PDE inhibitors may result in abnormal non-cyclic induction of cAMP levels which may induce deleterious effects on reproduction. Full article

Wastewater treatment plant effluents contain a variety of endocrine disrupting chemicals (EDCs), including chemicals with estrogenic activity such as 17β-estradiol (E2), 17α-ethinyl estradiol (EE2), and nonylphenols. These substances can affect both behavior and physiology in vertebrate animals. To explore the presence and effects of these EDCs in a natural setting, juvenile and adult male fathead minnows, Pimephales promelas, were held in cages upstream and downstream of the effluent site of a wastewater treatment plant for 21 days and subsequently tested for changes in reproductive behaviors and production of vitellogenin. Additionally, estrogenic activity in the stream was measured using a yeast bioassay. Estrogenicity was found to be significantly higher downstream of the wastewater effluent when compared to levels upstream. Vitellogenin levels did not show a correlational pattern with levels of estrogenicity in the water, but two measures of reproductive behaviors occurred significantly less often in downstream males than upstream males. This suggests that a brief (three-week) exposure to stream water containing wastewater treatment plant effluent can bring about changes in reproductive behavior of fish and that behavior may be more sensitive to low levels of environmental endocrine disruptors than vitellogenin production. Full article

In this study, three magnetic ionic liquids (MILs) were investigated for extraction of four estrogens, i.e., estrone (E1), estradiol (E2), estriol (E3), and ethinylestradiol (EE2), from environmental water. The cation trihexyl(tetradecyl)phosphonium ([P₆₆₆₁₄]⁺), selected to confer hydrophobicity to the resulting MIL, was combined with tetrachloroferrate(III), ferricyanide, and dysprosium thiocyanate to yield ([P₆₆₆₁₄][FeCl₄]), ([P₆₆₆₁₄]₃[Fe(CN)₆]), and ([P₆₆₆₁₄]₅[Dy(SCN)₈]), respectively. After evaluation of various strategies to develop a liquid–liquid microextraction technique based on synthesized MILs, we placed the MILs onto a magnetic stir bar and used them as extracting solvents. After extraction, the MIL-enriched phase was dissolved in methanol and injected into an HPLC–UV for qualitative and quantitative analysis. An experimental design was used to simultaneously evaluate the effect of select variables and optimization of extraction conditions to maximize the recovery of the analytes. Under optimum conditions, limits of detection were in the range of 0.2 (for E3 and E2) and 0.5 μg L⁻¹ (for E1), and calibration curves exhibited linearity in the range of 1–1000 μg L⁻¹ with correlation coefficients higher than 0.998. The percent relative standard deviation (RSD) was below 5.0%. Finally, this method was used to determine concentration of estrogens in real lake and sewage water samples. Full article