Concentrations Of EE2 Research Articles

Estrogen induces shift in abundances of specific groups of the coral microbiome

Synthetic estrogens such as ethinylestradiol (EE2) are persistent micropollutants that are not effectively removed from wastewater by conventional treatments. These contaminants are released into waterbodies, where they disrupt endocrine systems of organisms and cause harmful effects such as feminization, infertility, reproduction problems and genital malformations. The consequences of this pollution for key marine ecosystems such as coral reefs and their associated microbiomes are underexplored. We evaluated the effects of EE2 concentrations of 100 ng L−1 and 100 µg L−1 on the coral metaorganism Mussismilia harttii. The results indicated no effects on visible bleaching or Fv/Fm ratios in the corals during a 17-day microcosm experiment. However, next-generation sequencing of 16S rDNA revealed a statistically significant effect of high EE2 concentrations on OTU richness, and shifts in specific microbial groups after treatments with or without EE2. These groups might be bioindicators of early shifts in the metaorganism composition caused by EE2 contamination.

Ethinylestradiol removal of membrane bioreactor effluent by reverse osmosis and UV/H2O2: A technical and economic assessment

Synthetic hormone 17α-ethinylestradiol (EE2) is not completely removed by conventional wastewater treatment plants and therefore is often detected in surface and groundwater, sludge and sediments. Due to its persistence in the environment and its estrogenic potential, a high removal of EE2 from wastewaters before its disposal has become a concern from an environmental point of view, particularly when considering urban reuse applications. This work investigated the application of advanced processes to treat synthetic municipal wastewater containing EE2 after treatment in a membrane bioreactor (MBR). Two advanced processes were assessed: the first is advanced oxidation process (AOP), using hydrogen peroxide (H2O2) and ultraviolet (UV) light (route MBR-AOP) and the second, reverse osmosis (RO), in this case using UV/H2O2 to treat the retentate from RO (route MBR-RO). EE2 concentration in final effluent was one order of magnitude lower in route MBR-AOP than in route MBR-RO. Implications for disposal or water reuse were discussed considering the importance of other water quality parameters as well. Economic estimates for CAPEX, OPEX and total cost were made. The introduction of the oxidative step (UV/H2O2) after MBR caused an increase in the total cost of US$ 0.39/m3. In turn, route MBR-RO increased the total process cost by US$ 0.86/m3, showing that reduction of volume to be treated by UV/H2O2 in this route did not offset the cost associated with the acquisition and operation of RO. The total cost was estimated at US$ 2.47/m3 for MBR-AOP and US$ 2.94/m3 for MBR-RO for a design flow of 10 m³/h.

Ethinylestradiol (EE2) residues from birth control pills impair nervous system development and swimming behavior of zebrafish larvae

The ubiquitous use of ethinylestradiol (EE2), an active constituent of birth control preparations, results in continuous release of this synthetic estrogen to surface waters. Many studies document the untoward effects of EE2 on the endocrine system of aquatic organisms. Effects of environmental EE2 on the nervous system are still poorly documented. We studied effects of pico- to nanomolar concentrations of EE2 on early nervous system development of zebrafish larvae. EE2 disrupted axonal nerve regeneration and hair cell regeneration up to 50%. Gene expression in larval brain tissues showed significantly upregulated expression of target genes, such as estrogen and progesterone receptors, and aromatase B. In contrast, downregulation of the tyrosine hydroxylase, involved in the synthesis of neurotransmitters, occurred concomitant with diminution of proliferating cells. Overall, the size of exposed fish larvae decreased by 25% and their swimming behavior was modified compared to non-treated larvae. EE2 interferes with nervous system development, both centrally and peripherally, with negative effects on regeneration and swimming behavior. Survival of fish and other aquatic species may be at risk in chronically EE2-contaminated environments.

Assessment of 17α-ethinylestradiol effects in Daphnia magna: life-history traits, biochemical and genotoxic parameters.

The occurrence of pharmaceuticals in aquatic ecosystems and the need to study them have increased over the years since they enter continuously the environment. Besides, these compounds are not intended for applications with environmental purposes, and therefore, little is known about their ecological effects, particularly in non-target organisms, as invertebrate species. Inside these substances, endocrine disrupting compounds (EDCs) have recently come into the limelight, due to environmental concentrations and consequently their detrimental effects on different organisms. 17α-ethinylestradiol (EE2) has been detected in the aquatic environment in various locations around the globe since it is the main synthetic hormone used as a female oral contraceptive and is also applied in veterinary medicine and animal production. The present study was intended to assess the chronic effects of EE2, in the non-target organism as Daphnia magna. Thus, to analyze the individual and subindividual impact, this aquatic organism was chronically exposed (21days) to 0.00 (control group), 0.10, 1.00, 10.0, and 100μg/L of EE2. Results here obtained demonstrated that D. magna exposed to the EE2 concentrations had significant effects in individual (life-history) and sub-individual (biochemical levels) parameters. Alterations as anticipation in the age at first reproduction, a decrease of the growth rate, oxidative stress, and lipid peroxidation were detected, as well as genotoxic damage. Therefore, it was possible to infer that EE2 can disrupt several metabolic pathways and physiological functions of D. magna, since EE2 demonstrated ecotoxicity, at environmentally relevant concentrations. This work reinforces the importance of examining the effects of more relevant exposures (more prolonged and with ecologically pertinent concentrations) of potential endocrine disruptors like EE2, to the freshwater organisms and ecosystem.

Analytical procedures for the determination of estrogen compounds in a surface water reservoir in Southeast Brazil

In this study, an analytical methodology was validated to determine and quantify four estrogen hormones using high-performance liquid chromatography (HPLC) with detections by diode array detector (DAD) and by fluorescence detector (FLD). For validation of the method, the following parameters were evaluated: linearity, selectivity, precision, accuracy, limit of detection (LOD), limit of quantification (LOQ) and robustness. Environmental samples were preconcentrated using solid phase extractions and for that, an experimental design was planned to determine the best recovery conditions by varying cartridge types, flow of eluent, pH of the samples, and eluting solvent. Five surface water sampling campaigns were carried out in five different sites of Furnas Reservoir over the months of December 2015 and May 2016. Sample point 1 was located near the sewage treatment plant of the city of Alfenas - MG, while sample point 5 was the most distant from this location. All estrogens, except for E1, were found in all water samples of at least one of the sampling sites. The concentrations of E3, E2 and EE2 ranged from 11-366, 63-422 and 75-9998 ng L-1, respectively. These results are consistent with several studies published in the scientific literature.

Detecting Traces of 17α-Ethinylestradiol in Complex Water Matrices

Hormones have a harmful impact on the environment and their detection in water bodies is an urgent matter. In this work, we present and analyze a sensor device able to detect traces of the synthetic hormone 17α-ethinylestradiol (EE2) below 10−9 M in media of different complexities, namely, ultrapure, mineral and tap waters. This device consists of solid supports with interdigitated electrodes without and with a polyethylenimine (PEI) and poly (sodium 4-styrenesulfonate) (PSS) layer-by-layer film deposited on it. Device response was evaluated through capacitance, loss tangent and electric modulus spectra and the data were analyzed by principal component analysis method. While the three types of spectra were demonstrated to be able to clearly discriminate the different media, loss tangent spectra allow for the detection of EE2 concentration, with a sensitivity of −0.072 ± 0.009 and −0.44 ± 0.03 per decade of concentration, for mineral and tap water, respectively. Detection limits values were found to be lower than the ones present in the literature and presenting values of 8.6 fM (2.6 pg/L) and of 7.5 fM (22.2 pg/L) for tap and mineral waters, respectively. Moreover, the obtained response values follow the same behavior with EE2 concentration in any medium, meaning that loss tangent spectra allow the quantification of EE2 concentration in aqueous complex matrices.

17\u03b1-ethynylestradiol prevents the natural male-to-female sex change in gilthead seabream (Sparus aurata L.)

Exposure to 17α-ethynylestradiol (EE2, 5 μg/g food) impairs some reproductive events in the protandrous gilthead seabream and a short recovery period does not allow full recovery. In this study, spermiating seabream males in the second reproductive cycle (RC) were fed a diet containing 5 or 2.5 μg EE2/g food for 28 days and then a commercial diet without EE2 for the remaining RC. Individuals were sampled at the end of the EE2 treatment and then at the end of the RC and at the beginning of the third RC, 146 and 333 days after the cessation of treatment, respectively. Increased hepatic transcript levels of the gene coding for vitellogenin (vtg) and plasma levels of Vtg indicated both concentrations of EE2 caused endocrine disruption. Modifications in the histological organization of the testis, germ cell proliferation, plasma levels of the sex steroids and pituitary expression levels of the genes coding for the gonadotropin β-subunits, fshβ and lhβ were detected. The plasma levels of Vtg and most of the reproductive parameters were restored 146 days after treatments. However, although 50% of the control fish underwent sex reversal as expected at the third RC, male-to female sex change was prevented by both EE2 concentrations.

Study on the Release Potential of BPA and Steroid Estrogens in the Sediments of Erhai Lake, a Typical Plateau Lake of China.

On-site sampling analysis and laboratory-scale experiments were conducted to study the pollution status and release potential of EDCs in Erhai Lake. We found that nitrogen and phosphorus pollution in Erhai Lake sediment were both at a high level, as well as EDCs pollution. The concentrations of BPA, E2α, E1, E2β, EE2, and E3 were 36.84ng/g(DW), 13.04ng/g(DW), 128.97ng/g(DW), 52.57ng/g(DW), 18.48ng/g(DW) and 5.36ng/g(DW), respectively. The concentrations of E2α, E1, E2β and EE2 in the bottom water were higher than the surface water due to the impact of sediment release. The results of the 20days release test indicated that BPA release from the sediment had a greater correlation with the original concentration and the particle size of sediment, while the steroid EDCs had no obvious correlation with these two factors, probably due to the difference in hydrophobicity between them. Under hydraulic disturbance and aerobic conditions, the release process of EDCs was accompanied by a large amount of microbial degradation, and degradation amount > released amount. BPA was released quickly, 9.56% was released in 20days, but only 3.37% of steroid EDCs released. In comparison, the release process of steroids was longer and posed a greater threat to aquatic ecology.

Augmenting the biodegradation of recalcitrant ethinylestradiol using Rhodopseudomonas palustris in a hybrid photo-assisted microbial fuel cell with enhanced bio-hydrogen production

This study presents the biodegradation potential of ethinylestradiol (EE2) in anaerobic environments using exoelectrogenic activity of Rhodopseudomonas palustris. EE2, a basic ingredient in oral contraceptives, is a significant estrogenic micropollutant in various wastewaters and is considered highly recalcitrant. This recalcitrance of EE2 has caused anoxic areas to become repositories for these pollutants. Thus, it is essential to find the microorganisms and suitable methods to degrade this compound. An initial EE2 concentration of 1 mg/L, used in an anaerobic photobioreactor, resulted in 70% EE2 degradation over a period of 16 days with an increase of 63% in hydrogen production when EE2 was used with glycerol as the main carbon source in the culture medium. Furthermore, in the novel setup of hybrid photo-assisted microbial fuel cell (h-PMFC) employed here, EE2 degradation enhanced to 89.82% with a maximum power density of 0.633 ± 0.04 mW/m2. The hybrid MFC employed here could metabolize EE2 and sustained the bio-hydrogen production for 14 days to run the hydrogen fuel cell which otherwise could not be sustained with glycerol only and thus increased the overall power output. The current work highlights the use of R. palustris and the significance of co-metabolism in bioremediation of pollutants and bioenergy generation.

17α-ethinylestradiol and 4-tert-octylphenol concurrently disrupt the immune response of common carp

The aquatic environment is massively polluted with endocrine-disrupting compounds (EDCs) including synthetic estrogens (e.g. 17α-ethinylestradiol, EE2) and alkylphenols (e.g. 4-tert-octylphenol, 4t-OP). A major mechanism of action for estrogenic EDCs is their interaction with estrogen receptors and consequently their modulation of the action of enzymes involved in steroid conversion e.g. aromatase CYP19.We now studied the effects of EE2 and 4t-OP on the anti-bacterial immune response of common carp. We investigated effects on the number/composition of inflammatory leukocytes and on the gene expression of mediators that regulate inflammation and EDC binding. In vitro we found that high concentrations of both EE2 and 4t-OP down-regulated IFN-γ2 and IFN-γ-dependent immune responses in LPS-stimulated monocytes/macrophages. Similarly, during bacterial infection in fish, in vivo treated with EE2 and 4t-OP, decreased gene expression of il-12p35 and of ifn-γ2 was found in the focus of inflammation. Moreover, during A. salmonicida-induced infection in EE2-treated carp, but not in fish fed with 4t-OP-treated food, we found an enhanced inflammatory reaction manifested by high number of inflammatory peritoneal leukocytes, including phagocytes and higher expression of pro-inflammatory mediators (inos, il-1β, cxcl8_l2). Furthermore, in the liver, EE2 down-regulated the expression of acute phase proteins: CRPs and C3. Importantly, both in vitro and in vivo, EDCs altered the expression of estrogen receptors: nuclear (erα and erβ) and membrane (gpr30). EDCs also induced up-regulation of the cyp19b gene.Our findings reveal that contamination of the aquatic milieu with estrogenic EDCs, may considerably violate the subtle and particular allostatic interactions between the immune response and endogenous estrogens and this may have negative consequences for fish health.

Effects of Whole-Lake Additions of Ethynylestradiol on Leech Populations.

Leeches are widespread, found in many freshwater habitats, and have diverse dietary habits. Despite their close phylogenetic relationships to Mollusca, a phylum with species affected by exogenous estrogens, it is unclear whether Hirudinea may also be impacted. A whole-lake experiment was done at the Experimental Lakes Area in Ontario, Canada, to assess whether 17α-ethynylestradiol (EE2) affected fishes and other species. Herein, we examined whether EE2 impacted leech community composition, species abundance, growth rates, gonad size, and cocoon production, when compared with reference lakes using a before-after-control-impact design. Each month baited leech traps were set overnight in the littoral zone at 10 sites around experimental Lake 260 and 2 reference lakes, and individuals were identified, weighed, and measured. Male and female organs of common species Haemopis marmorata were measured. Across all lakes, 9 species representing 3 families were collected. There were no apparent effects of EE2 on numbers, species richness, or community composition; however, condition in 1 of 5 species increased significantly after EE2 exposures. Total gonadosomatic index (GSI) and the GSI for all male or all female organs combined for H. marmorata were not affected by EE2 additions. However, some individual reproductive organs including relative sperm sac length (+), relative epididymis weight (-), relative vaginal bulb length (+), and relative ovisac + albumen gland length (+) changed after EE2 additions. Finally, whereas overall cocoon production was similar, it occurred earlier in the EE2-amended lake. In summary, few individual through community measures of leeches responded to low ng/L concentrations of EE2, suggesting that Hirudinea are less sensitive to this endocrine disruptor than other invertebrates and vertebrates. Environ Toxicol Chem 2020;39:1608-1619. © 2020 SETAC.

Anticancer Activity of Solvent Extracts of Hexogonia glabra against Cervical Cancer Cell Lines.

Objective:In this study, we aimed to harness some solvent extracts of one wild mushroom Hexagonia glabra and test their anti-cancer activity against cervical human cell lines, namelyHeLa, SiHa, and CaSki. Methods:It includes cell morphological study by microscope, nuclear morphology by DAPI staining under fluorescence microscopy, apoptosis assay by fluorescence technique, anti-proliferation by MTT assay and expression of apoptotic and anti-apoptotic genes by Western blotting and cell cycle analysis was done. Results:The selected cervical cancer cells were treated separately with 150 µg/mL of three extracts, namely of ethanolic (EE), ethyl acetate (EAE), and water extract (WE) and exhibited features like round, shrink and dead. All extracts caused apoptosis in cell lines and EE had the highest effect in this regard. The percentages of apoptotic cells in HeLa, SiHa and CaSki, at the same concentration of EE were 79.23, 75.42, and 76.36% respectively. Cytotoxicity assay showed that all three extracts (50 – 250 μg/mL) were potent for inhibition of cell growth of three cell lines and again EE had the highest effect. The percentages of cell growth inhibition in HeLa, SiHa, and CaSki cells treated with EE at 24 h at 50 µg/mL were 45.79±4.11, 41.66±4.03, and 36.72±2.67, while they were 74.23±7.45, 62.31±5.97, and 54.23±5.04 at 150 µg/mL concentration. At 250 µg/mL concentration, the percentages of cell growth inhibition were 94.25 ±8.11, 90.02 ±8.67, and 85.43±6.21, respectively. The expression of apoptotic gene (Caspase 3, 9) and tumor guard gene (p53), as their proteins in Western blotting increased . However, anti-apoptotic BcL2 gene of all cell lines was decreased following treatment with extracts. In addition, the cell cycle analysis (CaSki cell) showed that treatment (EE) arrested at G2/M check point cell cycle. Conclusion:All extracts of this mushroom were active in arresting growth of three cell lines and EE had the highest effect, indicating that this mushroom can be a valuable source of anticancer agents.

Transcriptome analysis of testis reveals the effects of developmental exposure to bisphenol a or 17α-ethinylestradiol in medaka (Oryzias latipes)

Endocrine disrupting chemicals (EDCs) can induce abnormalities in organisms via alteration of molecular pathways and subsequent disruption of endocrine functions. Bisphenol A (BPA) and 17α-ethinylestradiol (EE2) are ubiquitous EDCs in the environment. Many aquatic organisms, including fish, are often exposed to varying concentrations of BPA and EE2 throughout their lifespan. Both BPA and EE2 can activate estrogenic signaling pathways and cause adverse effects on reproduction via alteration of pathways associated with steroidogenesis. However, transcriptional pathways that are affected by chronic exposure to these two ubiquitous environmental estrogens during embryonic, larval, and juvenile stages are not clearly understood. In the present study, we examined transcriptional alterations in the testis of medaka fish (Oryzias latipes) chronically exposed to a low concentration of BPA or EE2. Medaka were exposed to BPA (10 μg/L) or EE2 (0.01 μg/L) from 8 h post-fertilization (as embryos) to adulthood 50 days post fertilization (dpf), and transcriptional alterations in the testis were examined by RNA sequencing (RNA-seq). Transcriptomic profiling revealed 651 differentially expressed genes (DEGs) between BPA-exposed and control testes, while 1475 DEGs were found between EE2-exposed and control testes. Gene ontology (GO) analysis showed a significant enrichment of “intracellular receptor signaling pathway”, “response to steroid hormone” and “hormone-mediated signaling pathway” in the BPA-induced DEGs, and of “cilium organization”, “microtubule-based process” and “organelle assembly” in the EE2-induced DEGs. Pathway analysis showed significant enrichment of “integrin signaling pathway” in both treatment groups, and of “cadherin signaling pathway”, “Alzheimer disease-presenilin pathway” in EE2-induced DEGs. Single nucleotide polymorphism (SNP) and insertion-deletion (Indel) analysis found no significant differences in mutation rates with either BPA or EE2 treatments. Taken together, global gene expression differences in testes of medaka during early stages of gametogenesis were responsive to chronic BPA and EE2 exposure.

Long-term exposure to low 17α-ethinylestradiol (EE2) concentrations disrupts both the reproductive and the immune system of juvenile rainbow trout, Oncorhynchus mykiss

Estrogenic endocrine disrupting compounds (EEDCs), such as ethinylestradiol (EE2), are well studied for their impact on the reproductive system of fish. EEDCs may also impact the immune system and, as a consequence, the disease susceptibility of fish. It is currently not yet known whether the low concentrations of EEDCs that are able to disrupt the reproductive system of trout are effective in disrupting the immune system and the fish host resistance towards pathogens, too, or whether such immunodisruptive effects would occur only at higher EEDC concentrations. Therefore, in the present study we compare the effect thresholds of low 17α-ethinylestradiol concentrations (1.5 and 5.5 EE2 ng/L) on the reproductive system, the immune system, the energy expenditures and the resistance of juvenile rainbow trout (Oncorhynchus mykiss) against the parasite Tetracapsuloides bryosalmonae – the etiological agent of proliferative kidney disease (PKD) of salmonids. The parasite infection was conducted without injection and under low pathogen exposure concentrations. The disease development was followed over 130 days post infection – in the presence or absence of EE2 exposure. The results show that the long-term EE2 exposure affected, at both concentrations, reproductive parameters like the mRNA levels of hepatic vitellogenin and estrogen receptors. At the same concentrations, EE2 exposure modulated the immune parameters: mRNA levels of several immune genes were altered and the parasite intensity as well as the disease severity (histopathology) were significantly reduced in EE2-exposed fish compared to infected control fish. The combination of EE2 exposure and parasite infection was energetically costly, as indicated by the decreased values of the swim tunnel respirometry. Although further substantiation is needed, our findings suggest that EE2 exerts endocrine disruptive and immunomodulating activities at comparable effect thresholds, since reproductive and immune parameters were affected by the same, low EE2 concentrations.

Encapsulation of biogenic manganese oxide and Pseudomonas putida MnB1 for removing 17 α-ethinylestradiol from aquatic environments

In this study, we fabricated BMO-MnB1 beads by encapsulating biogenic manganese oxide (BMO) and Pseudomonas putida MnB1 into a calcium alginate hydrogel matrix and evaluated its possible use as a novel material for removing 17α-ethinylestradiol (EE2) from aqueous environments. A response surface methodology (RSM) was used to investigate the individual and interaction effects of the four independent encapsulation conditions (i.e., sodium alginate concentration, CaCl2 concentration, curing time, and bead size) on the removal efficiency of EE2 to obtain optimal encapsulation products. The effects of different experimental conditions (i.e., pH, temperature, initial EE2 concentration, and BMO-MnB1 bead concentration) on the removal of EE2 were also examined. The reusability of BMO-MnB1 beads was also studied in comparison with free BMO and P. putida MnB1 and purified BMO. The results suggested that the optimal conditions for the preparation of BMO-MnB1 beads were the sodium alginate concentration of 3%, the CaCl2 concentration of 0.2 M, the curing time of 3 h, and the bead size of 2.5 mm. Successful fabrication of BMO-MnB1 beads was confirmed via optical microscopy and field emission scanning electron microscopy analysis. Initial pH, initial EE2 concentration, bead concentration, and temperature markedly affected the EE2 removal rates by the BMO-MnB1 beads. The BMO-MnB1 beads exhibited good separation and satisfactory reusability; they could be recycled up to 14 times without obvious changes in the bead structure. This study demonstrates that encapsulation of BMO and P. putida MnB1 into alginate beads can be an excellent strategy with great efficiency and reusability, and high potential for industrial applications for EE2 removal.

Quantification of estrogen concentration in a creek receiving wastewater treatment plant effluent.

Estrogen in streams threatens aquatic animals, especially where wastewater treatment plant (WWTP) effluent contributes to baseflow. We investigated total estrogen (E1+E2+E3) as estradiol equivalent (E2) and ethynylestradiol (EE2) concentration in Cibolo Creek (Cibolo), a groundwater-fed stream near San Antonio, TX, receiving effluent via two WWTP. We collected water samples bi-monthly from late spring to early fall 2018 in Cibolo and WWTP effluent, and used ELISA analysis and discharge measurements to determine concentrations and loads of estrogens. We measured several environmental variables to investigate what factors influenced estrogen concentrations in Cibolo downstream from WWTP inputs. Mean concentrations of WWTP effluent (E2, 41.43 ± 15.48; EE2, 11.40 ± 2.07ngL-1) were higher compared with concentrations in Cibolo, both downstream (E2, 30.09 ± 25.85; EE2, 6.33 ± 1.92ngL-1) and upstream (E2, 12.91 ± 11.12; EE2, 4.5 ± 1.38ngL-1) of WWTP inputs. Both E2 and EE2 concentrations decreased downstream from WWTP inputs, a section of stream without large quantities of fine sediments for sorption, indicating potential dilution or chemical and biological degradation. Effluent into Cibolo via the first, and older, WWTP contributed the most estrogen load in Cibolo. Median concentrations of E2 and EE2 were 19 and 5ngL-1, respectively, downstream of WWTP inputs, concentrations known to affect reproductive processes of aquatic biota and impair human health. Results suggest estrogens may pose a risk to aquatic ecosystems wherever WWTP effluent comprises a majority of baseflow, though further studies are required in this stream to verify biological impacts.

Occurrence and distribution of estrogenic chemicals in river waters of Malaysia

This study investigated the occurrence of eight endocrine-disrupting compounds (EDCs) in river water across five major regions in Malaysia, which are recognized as emerging micropollutants in the environment. The water samples were collected from surface water of Klang River, Pahang River, Kuantan River, Kelantan River and Terengganu River from Malaysia. The samples were extracted by liquid–liquid extraction using hexane and ethyl acetate and loaded on silica column for column chromatography. Gas chromatography–flame ionization detector was used to determine the concentration of the target compounds. The maximum concentration of E1, E2, E3, EE2, 1,2,4-triazole, MP, 3,4,4-trichlorocarbanilide and BPA detected is 26.5 ng/L, 15.5 ng/L, 3.6 μg/L, 68.8 ng/L, 6.5 ng/L, 5.75 ng/L, 392.5 ng/L and 37 μg/L, respectively. The high discharge of various emerging organic pollutants in the Klang River estuary ecosystem was caused by heavy industrial, plantation, residential and commercial activities from urbanized and populated cities. Overall, the occurrences show a higher concentration of EDCs when compared to developed countries. Other rivers such as Kuantan, Terengganu, Pahang and Kelantan are known as the major sources of water supply for domestic, industrial and agricultural purposes. Eight endocrine-disrupting chemicals were detected and estimated in five rivers of Malaysia. These target compounds are widely present in various rivers in the range of ND to 37 μg/L. The difference of occurrence of EDCs in these selected sampling sites reflects the distinct usage patterns of various chemicals, pesticide, cosmetic products, drugs all over Malaysia. The presence of BPA, E1 and E3 was observed to be a major contributor of endocrine disruptors.

No evidence that vitellogenin protein expression is induced in marine mussels after exposure to an estrogenic chemical

A wide variety of endocrine disrupting chemicals reach the marine environment and can cause harmful effects in different marine organisms. Vitellogenin (Vtg), the egg-yolk precursor, is a commonly used endocrine disruption biomarker in fish and more recently in marine invertebrates under the assumption of high expected similarities in the endocrine system of vertebrates and invertebrates. However, this assumption has been recently questioned. The results from previous studies focused on bivalve molluscs showed that Vtg induction could be misleading because of the use of either non-robust or indirect techniques to measure Vtg. In this study, mussels (Mytilus galloprovincialis) were exposed to either 10 or 100 ng/L of the synthetic hormone 17α-ethinylestradiol (EE2) at different exposure times (4 and 24 days) and under different feeding regimes (representing different energy balances), and Vtg levels in both male and female mussel gonads were quantified by label free shotgun LC-MS/MS proteomic analysis. Vtg protein was not detected in male gonads. In female gonads, Vtg levels were not significantly affected by EE2 at any exposure time or EE2 concentration tested, whereas a significant correlation was found between the degree of maturation of the gonad and Vtg levels in females. Results obtained in the present study critically question the use of Vtg as a biomarker of endocrine disruption in marine mussels, and show that the degree of maturation of the gonad can be an important confounding factor in the attempts to evaluate estrogenic effects through Vtg measurement in mussel gonads.

Biodiesel Co-products Modified the Rumen Parameters of Feedlot Lambs but did not Change Methane Production In Vitro*

Background: Intensification of livestock is a strategy that increases productivity, but the diets used to increase animal production efficiency are composed mainly of corn and soybean, thereby increasing competition between animals and humans for the same food crops. This study evaluated nutrient intake, apparent digestibility of dry matter (DM) and nutrients, kinetics of gas production, and concentration of volatile fatty acids on diets with or without inclusion of biodiesel co-products formulated for feedlot lambs. So, the hypothesis is that replace of traditional ingredients by biodiesel co-products changes rumen parameters and methane emissions.Material, Methods & Results: The experiment was developed in São Paulo State University (Unesp), Jaboticabal/SP, Brazil, in Sheep Production Laboratory, which is owned to Animal Science Department. All trials developed in this study used a feedlot system, where animals were kept in individual pen. Forty Ile de France lambs male non-castrated were used in in vivo trial. To obtain rumen fluid, that was used in in vitro trial, four Santa Inês lambs with rumen cannula were used. The treatments evaluated were four diets: Control diet: roughage + concentrate; PM20: roughage + concentrate with peanut meal (PM) at 20% of DM; CG25: roughage + concentrate with crude glycerin (CG) at 25% of DM; and PMCG: roughage + concentrate with PM at 10% of DM and CG at 12.5% of DM. The roughage:concentrate ratio was 40:60 for all these diets. The parameters of the in vitro and in vivo experiments used were completely randomized with four treatments. When significant, the means between treatments were compared using Tukey test (P < 0.05). There was no effect of co-product inclusion on intake, except ether extract and neutral detergent fiber, which were higher for PM20 compared with CG25 diet. Apparent digestibility of dry matter (79.87%) and some nutrients (organic matter, crude protein, and neutral detergent fiber) was higher (P < 0.05) with CG25 diet. In vitro cumulative gas production was greater in CG25 and PMCG compared to the other diets, at early measurement points (2, 4, 6, and 10 h). The concentrations of methane, volatile fatty acids, and acetate:propionate ratio in vitro did not differ (P > 0.05) among diets.Discussion: Probably the high quality of the glycerin used (83.9% glycerol, 12.01% humidity, 3.79% salts, and 0.28% organic matter, no fat, as described by the manufacturer) may explain the low EE concentration observed in the diet using only crude glycerin and the observed lack of DMI effects in all diets. About apparent digestibility, the greater values measured for crude protein can be explained by superior synchronism during fermentation of the proteins and carbohydrates in the diet. The data showed that treatment CG25 obtained higher initial gas production, followed by treatment PMCG which contained 12.5% crude glycerin. Probably these results were caused by the greater apparent digestibility of DM in treatments that included crude glycerin. Our results of volatile fatty acid concentration are different from the decrease in molar proportion of acetic acid and increase of propionic acid described by several authors, with the inclusion of glycerin in the diet. However, the absence of effect by co-product inclusion on the molar proportion of acetic, propionic, and butyric acids measured in this trial reinforce the report from other authors that affirmed the same situation. Hence, the inclusion of PM at 20% of DM and CG at 25% of DM could successfully replace the traditional diets of feedlot lambs such as soybean and corn, respectively, without damages to intake and ruminal parameters in vitro.

Kinetic Model of the Action of 17α-Ethynylestradiol on the Capacitation of Mouse Sperm, Monitored by HPLC-MS/MS

17α-Ethynylestradiol (EE2), a synthetic estrogen used in contraceptive pills, is resistant to hepatic degradation and is excreted in the urine. It is chemically stable and has a negative impact on the endocrine system. The aim of this work was to mathematically describe the possible interaction of EE2 (200, 20, and 2 μg/L) with sperm estrogen receptors during sperm maturation, which is called capacitation. The concentrations of the unbound EE2 remaining in capacitating medium during 180 min of sperm capacitation were determined at 30 min intervals by high performance liquid chromatography with tandem mass spectrometric detection (HPLC-MS/MS) and the data obtained (relative concentrations Bt) were subjected to kinetic analysis. The suggested kinetic schema was described by the system of differential equations with the optimization of rate constants used to calculate the theoretical Bt values. Optimal parameters (overall rate constants K1–K5 and molar ratio n) were determined by searching the minimum of absolute values of the difference between theoretical and experimental Bt values. These values were used for the design of the theoretical B(t) curves which fit to experimental points. The proposed kinetic model assumes the formation of an unstable adduct between EE2 and the receptor in cytoplasm, which acts as an autocatalytic agent and gradually decomposes.