Assessment of a potential preventive ability of amygdalin in mycotoxin-induced ovarian toxicity

The possible effects of a natural substance amygdalin and its combination with the mycotoxin deoxynivalenol (DON) on the steroid hormone secretion (progesterone and 17-β-estradiol) by porcine ovarian granulosa cells (GCs) were examined in this in vitro study. Ovarian GCs were incubated without (control group) and with amygdalin (1, 10, 100, 1,000 and 10,000 μg mL¹), or its combination with DON (1 μg mL¹) for 24 h. The release of steroid hormones was determined by ELISA. The progesterone secretion by porcine ovarian GCs was not affected by amygdalin in comparison to the control. However, the highest amygdalin dose (10,000 μg mL¹) caused a significant stimulation of the 17-β-estradiol release. A combination of amygdalin with DON significantly (P < 0.05) increased the progesterone release at all concentrations. Similarly, a stimulatory effect of amygdalin co-administered with DON was detected with respect to the 17-β-estradiol secretion at the highest dose (10,000 μg mL¹) of amygdalin and 1 μg mL¹ of DON. Noticeable differences between the effects of amygdalin alone and its combination with DON on the progesterone release were detected. In contrast, no differences between the stimulatory effects of amygdalin and its combination with DON on the 17-β-estradiol synthesis by porcine GCs were observed. Findings from this in vitro study did not confirm the expected protective effect of amygdalin on mycotoxin induced reprotoxicity. Our results indicate that the stimulatory effect of amygdalin combined with DON on the progesterone release was clearly caused by the DON addition, not by the presence amygdalin per se. On the other hand, the stimulation of 17-β-estradiol production was solely caused by the presence of amygdalin addition. These findings suggest a possible involvement of both natural substances into the processes of steroidogenesis and appear to be endocrine modulators of porcine ovaries.     

Endocrine modulation, inhibition of ovarian development and hepatic alterations in rainbow trout exposed to polluted river water

Under laboratory conditions, female rainbow trout were exposed to graded concentrations of water from the River Lambro, a polluted tributary of the River Po, and to the effluent of a large wastewater treatment plant which flows into the River Lambro. In field exposures, trout were held in cages in the River Po upstream and downstream from the confluence of the River Lambro. After 10-day (laboratory) and 30-day (laboratory and field) exposures, trout were examined for several chemical, biochemical and histological endpoints. The results indicated that exposure to complex mixtures of chemicals, including estrogen receptor agonists, aryl-hydrocarbon receptor agonists, and probably antiandrogens, had occurred. Exposure altered the plasma levels of 17β-estradiol and testosterone, and some treatments also enhanced the activity of hepatic ethoxyresorufin O-deethylase. Gonadal histology showed varying levels of degenerative processes characterised by oocyte atresia, haemorrhages, melano-macrophage centres (MMCs), and oogonia proliferation. Liver histology showed less severe effects.     

Statewide Survey of Hormones and Antibiotics in Surface Waters of Delaware

Water‐quality surveys have confirmed the presence of hormones and antibiotics in surface waters of the United States, which may be of concern to aquatic life. We investigated the concentrations of hormones and antibiotics in surface waters of the state of Delaware to determine – how they compared against environmental thresholds, how they varied across the state, and if they were correlated with land use type. Fifty surface water locations were sampled during early spring and late summer. Water samples were initially screened with ELISA followed by analysis with LC/MS/MS. The measured ranges of hormone concentrations were: 0‐3.71 ng/l for estrone, 0‐4.65 ng/l for estrone‐3‐sulfate, and 0‐6.27 ng/l for 17β‐estradiol. The measured ranges of antibiotics were: 0‐3.30 ng/l for sulfamerazine, 0‐10.74 ng/l for sulfamethoxazole, and 0‐2.29 ng/l for tetracycline. The predicted no‐effect concentration (PNEC) for estrone was exceeded for three samples and the PNEC for 17β‐estradiol was exceeded for 11 samples. In general, concentrations and detection frequencies were lower in the summer than the spring. The highest concentrations of hormones and antibiotics were spatially distributed in agricultural and urban areas; however, the correlations between land use type and the concentrations were weak. This study was the first statewide survey of hormones and antibiotics for Delaware and provided important baseline data on these emerging contaminants.     

The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

天然类固醇雌激素源解析、环境行为及其污染控制

天然类固醇雌激素是一种在生物体内具有极强雌激素效应的内分泌干扰物,其在环境中的广泛存在严重影响野生动物尤其是鱼类的正常生长和发育,对环境生态系统所产生的破坏也日趋严重,然而其危害依然没有引起足够的重视。文章研究在阐述天然类固醇雌激素基本理化性质和环境存在形态的基础上,解析其在人类和畜禽的环境来源,分析其在地表水和地下水的污染状况以及环境迁移和生物积累等环境行为,并综合探讨了现今较为常见的如活性污泥法、光降解法以及微滤、纳滤和反渗透等与天然类固醇雌激素相关的污染控制技术,以期为天然类固醇雌激素的毒理性质、污染现状及相关控制方法的深入研究提供支撑。在此基础上,提出深入天然类固醇雌激素微量检测技术研究、加强国家行政管理层面上的污染控制等已十分急迫。     

环境激素对水生动物干扰效应及机制研究进展

环境激素是环境中的激素类似物,它干扰体内正常分泌的激素,引起种种负面的生物学效应,威胁水生动物和人类的健康。文章介绍了环境激素的概念、种类及其对水生动物的影响和作用机制,提出了环境激素污染的防治措施和研究前景。     

Anti-implantation and anti-fertility potentials of Anethum graveolens L. extracts in rats

Ovarian hormones affected some enzymes in reproductive tissues. Previous studies revealed Anethum graveolens (dill) seed extract elevated ovarian hormones. In this study, we compared the effects of dill seed and leaf extracts on some enzymes activities and infertility potential. Female Wistar rats were divided into three groups of control, dill seed and dill leaf aqueous extracts treated groups. Experimental groups received 1 mL of 0.5 g kg^?1 of each extract orally for 25 days (five estrous cycles). Estradiol, progesterone, luteinizing hormone, and follicle stimulating hormone concentrations were determined. Uterus and ovaries were removed and their glucose-6-phosphate dehydrogenase, lactate dehydrogenase, and alkaline phosphatase activities were measured. Female rats of three groups were mated with untreated males. Fetuses were extracted in the 20th gestational day and their weights and crown rump lengths were measured. Anti-fertility and anti-implantation activities and fertility indices were calculated. Glucose-6-phosphate activity of dill seed treated group increased 1.6 times in the endometrium and 1.7 times in the myometrium. Progesterone concentration and lactate dehydrogenase activity of serum increased in the dill seed treated group compared to the control and dill leaf treated group. A significant decrease was observed in weights and crown rump lengths of fetuses, weights of placenta, and fertility index of the dill seed treated group compared to the control and dill leaf treated group, but its anti-implantation and anti-fertility indices increased. We suggested that high level of glucose-6-phosphate could entail increased oxidative stress in uterus and ovary. High lactate dehydrogenase activity in serum may affect cell membranes. Higher secretion of progesterone caused hormonal imbalance and produced estrous cycle irregularity. It was concluded that dill seed but not dill leaf extract could induce infertility in an animal model.     

The influence of deoxynivalenol and zearalenone on steroid hormone production by porcine ovarian granulosa cells in vitro

Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) are frequently occurring in feed of pigs together. The aim of this study was to evaluate the possible in vitro effects of DON and ZEA, alone or their combination on steroid secretion of porcine ovarian granulosa cells (GCs). A species-specific model with porcine ovarian GCs was used to study the potential endocrine disrupting effects of DON and ZEA alone and in co-exposure. Progesterone (P4) and estradiol (E2) were determined by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). The results of this study demonstrate that DON alone at the higher concentrations may act to stimulate P4 (at 1,000, 2,000, 3,000 and 5,000 ng mL^?1 but not 10 and 100 ng mL^?1) and E2 (at 2,000, 3,000 and 5,000 ng mL^?1 but not 10, 100 and 1000 ng mL^?1) secretion. The effects of ZEA on P4 and E2 secretion were not confirmed. DON in combination with the other fusariotoxin ZEA may impair steroidogenesis. Results aslo demonstrate different toxicological effects of fusariotoxins on follicle stimulating hormone-induced secretion of P4 and E2. All these results taken together suggest that fusariotoxin and their interactions can impact ovarian steroidogenesis, thereby demonstrating their potential reproductive effects in pigs.     

中国天然雌激素排放清单和风险评价

环境雌激素由于其较强的内分泌干扰特性,越来越受到普遍关注。环境雌激素中,人和动物排放的天然类固醇雌激素（E1、E2、EE2、E3）干扰性最强。建立天然雌激素排放清单,对于了解中国环境雌激素的排放状况,以及进一步的污染风险分析与控制,具有重要意义。本研究将天然雌激素的主要排放源按人、猪、牛和家禽分为10种不同类型,根据各...