Combined exposure to engineered nanoparticles (ENPs) and anthropogenic contaminants can lead to changes in bioavailability, uptake and thus effects of both groups of contaminants. In this study we investigated effects of single and combined exposures of silver (Ag) nanoparticles (AgNPs) and the synthetic hormone 17a-ethinylestradiol (EE2) on tissue uptake of both contaminants in juvenile turbot (Scophthalmus maximus). Silver uptake and tissue distribution (gills, liver, kidney, stomach, muscle and bile) were analyzed following a 14-day, 2-h daily pulsed exposure to AgNPs (2 mg L 1 and 200 mg L 1), Agþ (50 mg L 1), EE2 (50 ng L 1) and AgNP þ EE2 (2 or 200 mg L 1þ50 ng L 1). Effects of the exposures on plasma vitellogenin (Vtg) levels, EE2 and steroid hormone concentrations were investigated. The AgNP and AgNP þ EE2 exposures resulted in similar Ag concentrations in the tissues, indicating that combined exposure did not influence Ag uptake in tissues. The highest Ag concentrations were found in gills. For the Agþ exposed fish, the highest Ag concentrations were measured in the liver. Our results show dissolution processes of AgNPs in seawater, indicating that the tissue concentrations of Ag may partly originate from ionic release. Plasma EE2 concentrations and Vtg induction were similar in fish exposed to the single contaminants and the mixed contaminants, indicating that the presence of AgNPs did not significantly alter EE2 uptake. Similarly, concentrations of most steroid hormones were not significantly altered due to exposures to the combined contaminants versus the single compound exposures. However, high concentrations of AgNPs in combination with EE2 caused a drop of estrone (E1) (female fish) and androstenedione (AN) (male and female fish) levels in plasma below quantification limits. Our results indicate that the interactive effects between AgNPs and EE2 are limited, with only high concentrations of AgNPs triggering synergistic effects on plasma steroid hormone concentrations in juvenile turbots. | acceptedVersion

The present study aimed to evaluate biochemical and cellular responses of the freshwater mussel, Hyriopsis bialata, to the herbicide atrazine (ATZ). The mussels were exposed to environmentally-relevant concentrations of ATZ (0, 0.02 and 0.2 mg/L) and a high concentration (2 mg/L) for 0, 7, 14, 21 and 28 days. Tissues comprising male and female gonads, digestive glands and gills were collected and assessed for ethoxyresorufin-O-deethylase (EROD) activity, glutathione S-transferase (GST) activity, multixenobiotic resistance mechanism (MXR), histopathological responses, DNA fragmentation and bioaccumulation of ATZ and its transformation derivatives, desethylatrazine (DEA) and desisopropylatrazine (DIA). Additionally, circulating estradiol levels were determined. It appeared that ATZ did not cause significant changes in activities of EROD, GST and MXR. There were no apparent ATZ-mediated histopathological effects in the tissues, with the exception of the male gonads exhibiting aberrant aggregation of germ cells in the ATZ-treated mussels. Contrarily, ATZ caused significant DNA fragmentation in all tissues of the treated animals in dose- and time-dependent manners. In general, the circulating estradiol levels were higher in the females than in the males. However, ATZ-treated animals did not show significant alterations in the hormonal levels, as compared with those of the untreated animals. Herein, we showed for the first time differentially spatiotemporal distribution patterns of bioaccumulation of ATZ, DEA and DIA, with ATZ and DEA detectable in the gonads of both sexes, DEA and DIA in the digestive glands and only DEA in the gills. The differential distribution patterns of bioaccumulation of ATZ and its derivatives among the tissues point to different pathways and tissue capacity in transforming ATZ into its transformation products. Taken together, the freshwater mussel H. bialata was resistant to ATZ likely due to their effective detoxification. However, using DNA damage as a potential biomarker, H. bialata is a promising candidate for biomonitoring aquatic toxicity.

Accumulating evidences indicated that heavy metals may disrupt human sex hormones. However, the combined effects of heavy metals on sex hormones remain to be clarified. To explore the independent and combined associations between heavy metal exposure and serum sex hormones among adults, data of 2728 adults from the National Health and Nutrition Examination Survey (NHANES) was applied. We examined independent and combined associations of fourteen urinary heavy metals and three serum sex steroid hormones (total testosterone (TT), estradiol (E2) and sex hormone-binding globulin (SHBG)). Multivariate linear regression was used to evaluate the independent associations between metal exposure and sex hormone alterations. Principle component analysis -weighted quantile sum regression (PCA-WQSR) model was performed to estimate the combined associations in our individuals. In the co-exposure model, we determined that weighted quantile sum (WQS) index of industrial pollutants was negatively associated with E2 in females (WQS Percent change₈₋ₘₑₜₐₗ = -20.6%; 95% CI: -30.1%, -9.96%), while in males WQS index of water pollutants was negatively related to SHBG (WQS Percent change₈₋ₘₑₜₐₗ = -5.35%; 95% CI: -9.88%, -0.598%). Cadmium (Cd), tin (Sn) and lead (Pb) were the dominating metals of female E2-negative association while Ba was the leading contributor related to male SHBG reduction, which was consistent with the results of multivariate linear regression. Additionally, in postmenopausal women, the associations of E2 decrease with heavy metal co-exposure remained significant while Cd and monomethylarsonic acid (MMA) were identified as hazardous metals in the mixture. We concluded that the exposure to heavy metals was associated with human sex hormone alterations in independent or combined manners. Considering the design of NHANES study, further studies from other national-representative surveys are necessary.

Endocrine-disrupting chemicals (EDCs) are compounds that interfere with the expression, synthesis, and activity of hormones in organisms. They are released into the environment from flame retardants and products containing plasticizers. Persistent pesticides, such as dichlorodiphenyltrichloroethane (DDT) and hexachlorobenzene, also disrupt the endocrine system through interaction with hormone receptors. Endogenous hormones, such as 17β-estradiol (E2), are released in the urine and feces of farm animals and seep into terrestrial and aquatic ecosystems through sewage. Pigs are widely used as animal models to determine the effects of EDCs because they are physiologically, biochemically, and histologically similar to humans. EDCs primarily disrupt the reproductive and nervous systems of pigs. Moreover, embryonic development during the prenatal and early postnatal periods is particularly sensitive to EDCs. Mycotoxins, such as zearalenone, are food contaminants that alter hormonal activities in pigs. Mycotoxins also alter the innate immune system in pigs, making them vulnerable to diseases. It has been reported that farm animals are exposed to various types of EDCs, which accumulate in tissues, such as those of gonads, livers, and intestines. There is a lack of an integrated understanding of the impact of EDCs on porcine reproduction and development. Thus, this article aims to provide a comprehensive review of literature regarding the effects of EDCs in pigs.

In this study, a multidisciplinary approach investigated the enzymatic degradation of micropollutants in real, not modified, municipal wastewaters of a plant located in Italy. Stir Bar Sorptive Extraction combined to Gas Chromatography-Mass Spectrometric detection (SBSE-GC-MS) was applied to profile targeted pollutants in wastewaters collected after the primary sedimentation (W1) and the final effluent (W2). Fifteen compounds were detected at ng/L - μg/L, including pesticides, personal care products (PCPs) and drugs. The most abundant micropollutants were bis(2-ethylhexyl) phthalate, diethyl phthalate and ketoprofen. Laccases of Trametes pubescens MUT 2400 were very active against all the target micropollutants: except few cases, their concentration was reduced more than 60%. Chemical analysis and environmental risk do not always come together. To verify whether the treated wastewaters can represent a stressor for the aquatic ecosystem, toxicity was also evaluated. Raphidocelis subcapitata and Lepidium sativum tests showed a clear ecotoxicity reduction, even though they did not evenly respond. Two in vitro tests (E-screen test and MELN assay) were used to evaluate the estrogenic activity. Treatments already operating in the plant (e.g. activated sludge) partially reduced the estradiol equivalent concentration, and it was almost negligible after the laccases treatment. The results of this study suggest that laccases of T. pubescens are promising biocatalysts for the micropollutants transformation in wastewaters and surface waters.

Perfluorooctane sulfonate (PFOS) has been reported to be widely distributed in the environment and wildlife with persistence. PFOS has various biological toxicity, especially disturbing the endocrine system. However, few studies have systematically evaluated its effect on sexual behaviors alteration and reproduction-related genes. This study was performed to assess the effect of PFOS exposure on sexual behaviors and genes in hypothalamic–pituitary–gonadal–liver (HPGL) axis in adult zebrafish.Male adult zebrafish were exposed to PFOS (0, 2, 20, and 200 μg/L) and 5 μg/L estradiol (E₂) continuously for 21 days. Sexual behaviors were analyzed by zebrafish behavior tracking system and the mRNA levels of HPGL-related genes was detected by RT-qPCR.Body weight of the fish was increased in 2, 200 μg/L PFOS and E₂ groups, and body length was increased with exposure to 2 μg/L PFOS and E₂. The hepatic-somatic index was decreased significantly after 2 and 20 μg/L PFOS treatments. Highest PFOS (200 μg/L) and E₂ exposure impaired standard zebrafish sexual behaviors significantly such as chasing, nose-tail and tail-touching. In brains, the genes gonadotropin-releasing hormone (GnRH), gonadotropin-releasing hormone receptor (GnRHr) were down-regulated with exposure to PFOS with linear trend and E₂ exposure, and follicle-stimulating hormone and luteinizing hormone were also down-regulated with exposure to 20 and 200 μg/L PFOS. In livers, the genes vitellogenin 1 and 3 were upregulated with some concentrations of PFOS and E₂, but estrogenic receptor α, β2 were upregulated in any concentration of PFOS and E₂. In testes, the expressions of follicle-stimulating hormone receptor, luteinizing hormone receptor, and androgen receptor genes were all significantly down-regulated with any exposure concentration of PFOS and E₂.PFOS may alter the zebrafish reproductive system by disrupting endocrine activity and impairing sexual behaviors.

Developmental exposures to estrogenic chemicals possibly cause structural and functional abnormalities of reproductive organs in vertebrates. Bisphenol AF (BPAF), a bisphenol A (BPA) analogue, has been shown to have higher estrogenic activity than BPA, but little is known about the effects of BPAF on gonadal development, particularly gonadal differentiation. We aimed to determine whether low concentrations of BPAF could disrupt gonadal differentiation and subsequent development using Xenopus laevis, a model species for studying feminizing effects of estrogenic chemicals. X. laevis tadpoles were exposed to BPAF (1, 10, 100 nM) or 17β-estradiol (E2, positive control) from stages 45/46 to 53 and 66 in a semi-static exposure system, with a prolonged treatment with the highest concentration to the eighth week post-metamorphosis (WPM8). Gonadal morphology and histology as well as sexually dimorphic gene expression were examined to evaluate the effects of BPAF. All concentrations of BPAF caused changes in testicular morphology at different developmental stages compared with controls. Specifically, at stage 53, BPAF like E2 resulted in decreases in both the size and the number of gonadal metameres (gonomeres) in testes, looking like ovaries. Some of BPAF-treated testes remained segmented and even became discontinuous and fragmented at subsequent stages. Histological abnormalities were also observed in BPAF-treated testes, such as ovarian cavity at stages 53 and 66 and poorly developed seminiferous tubules on WPM8. At the molecular level, BPAF inhibited expression of male highly expressed genes in testes at stage 53. Correspondingly, BPAF, like E2, inhibited cell proliferation in testes at stage 50. All results show that low concentrations of BPAF inhibited testicular differentiation and subsequent development in X. laevis, along with feminizing effects to some degree. Our finding implies a risk of BPAF to the male reproductive system of vertebrates including humans.

The present endocrine disrupting chemicals (EDCs) in wastewater effluents due to incomplete removal during the treatment processes may cause potential ecological and human health risks. This study evaluated the removal and uptake of seven EDCs spiked in two types of wastewater effluent (i.e., ultrafiltration and ozonation) and effluent cultivated with the freshwater green alga Nannochloris sp. In ultrafiltration effluent cultivated with Nannochloris sp. for 7 days, the removal rate of 17β-estradiol (E2), 17α-ethinylestradiol (EE2), and salicylic acid (SAL) was 60%; but Nannochloris sp. did not promote the removal of other EDCs studied. The algal-mediated removal of E2, EE2, and SAL was attributed to photodegradation and biodegradation. Triclosan (TCS) underwent rapid photodegradation regardless of adding algae in the effluent with 63%–100% removal within 7 days. Triclosan was also found associated with algal cells immediately after adding algae, and thus the primary mechanisms involved were photodegradation and bioremoval (i.e., bioadsorption and bioaccumulation). After algal cultivation, TCS still has a bioaccumulation potential to pose high risks within the food web and the endocrine disrupting properties of the residual estrogens in the effluent are not eliminated. Algal cultivation can be exploited to treat wastewater effluents but the removal efficiencies of EDCs highly depend on chemical types.

The information acquired by high resolution quadrupole-time of flight mass spectrometry (QTOF-MS) allows target analysis as well as retrospective screening for the presence of suspect or unknown emerging pollutants which were not included in the target analysis. Targeted quantification of eight azole antifungal drugs in wastewater effluent as well as new and relatively simple retrospective suspect and non-target screening strategy for emerging pollutants using UHPLC-QTOF-MS is described in this work. More than 300 (parent compounds and transformation products) and 150 accurate masses were included in the retrospective suspect and non-target screening, respectively. Tentative identification of suspects and unknowns was based on accurate masses, peak intensity, blank subtraction, isotopic pattern (mSigma value), compound annotation using data bases such as KEGG and CHEBI, and fragmentation pattern interpretation. In the targeted analysis, clotrimazole, fluconazole, itraconazole, ketoconazole and posaconazole were detected in the effluent wastewater sample, fluconazole being with highest average concentration (302.38 ng L⁻¹). The retrospective screening resulted in the detection of 27 compounds that had not been included in the target analysis. The suspect compounds tentatively identified included atazanavir, citalopram, climbazole, bezafibrate estradiol, desmethylvenlafaxine, losartan carboxylic acid and cetirizine, of which citalopram, estradiol and cetirizine were confirmed using a standard. Carbamazepine, atrazine, efavirenz, lopinavir, fexofenadine and 5-methylbenzotriazole were among the compounds detected following the non-targeted screening approach, of which carbamazepine was confirmed using a standard. Given the detection of the target antifungals in the effluent, the findings are a call for a wide assessment of their occurrence in aquatic environments and their role in ecotoxicology as well as in selection of drug resistant fungi. The findings of this work further highlights the practical benefits obtained for the identification of a broader range of emerging pollutants in the environment when retrospective screening is applied to high resolution and high accuracy mass spectrometric data.

The ubiquity of microplastics in the world's ocean has aroused great concern. However, the ecological effects of microplastics at environmentally realistic concentrations are unclear. Here we showed that exposure of marine medaka (Oryzias melastigma) to environmentally relevant concentrations of 10 μm polystyrene microplastics for 60 days not only led to microplastic accumulation in the gill, intestine, and liver, but also caused oxidative stress and histological changes. Moreover, 2, 20, and 200 μg/L microplastics delayed gonad maturation and decreased the fecundity of female fish. Alterations of the hypothalamus-pituitary-gonadal (HPG) axis were investigated to reveal the underlying mechanisms, and gene transcription analysis showed that microplastic exposure had significantly negative regulatory effects in female HPG axis. Transcription of genes involved in the steroidogenesis pathway in females were also downregulated. This disruption resulted in decreased concentrations of 17β-estradiol (E₂) and testosterone (T) in female plasma. Furthermore, parental exposure to 20 μg/L microplastics postponed the incubation time and decreased the hatching rate, heart rate, and body length of the offspring. Overall, the present study demonstrated for the first time that environmentally relevant concentrations of microplastics had adverse effects on the reproduction of marine medaka and might pose a potential threat to marine fish populations.