Consequences of endocrine disrupting chemicals on reproductive endocrine function in birds: establishing reliable end points of exposure

It has been difficult to establish reliable indices of exposure to endocrine disrupting chemicals (EDCs) appropriate for a variety of avian species because of a vast array of reproductive strategies. Data from mammals, reptiles and fish provide insight on likely mechanisms of action for EDCs. However, many of the effects of EDCs are weaker than the actions of the native hormones, making it difficult to assess adverse effects in domestic and wild birds. It is clear that differential sensitivity to EDCs exists across species, due to the timing and mode of exposure, compound toxicity and age of the individual. Our studies on EDCs are conducted in the quail model system, with focus on reproductive endocrine, neuroendocrine and behavioral responses. Studies have included EDC exposure, either by egg injection or via diet. Results from egg injection studies showed the following: (1) estradiol administered by embryonic day 12 demasculinized male sexual behavior, altered hypothalamic neurotransmitters and reduced hen day production and fertility in a dose dependent fashion, (2) methoxychlor (MXC) or vinclozolin impaired male sexual behavior in adult quail and (3) DDE exposure impaired reproductive and immune related end points. Two-generation studies were conducted on Japanese and northern bobwhite quail with dietary methoxychlor (MXC) exposure (0, 5 and 10 ppm) beginning in adults (P1), continuing in their offspring (F1), with F2 offspring raised on control diet. MXC exposure impaired male sexual behavior, hypothalamic catecholamines and plasma steroid hormones. Moreover, MXC exposure had reproductive consequences observable at both the lower and higher doses of MXC in F1 and F2 generations. These data demonstrate that embryonic EDC exposure interferes with sexual differentiation of neural systems that direct reproduction.     

17α-甲基睾酮对稀有鮈鲫体内激素水平的影响

环境内分泌干扰物（EDCs）对动物和人的健康有一定的危害作用,很大一部分环境内分泌干扰物最终都进入到了水环境中,对水生生物的生殖、内分泌等系统造成损伤。以稀有鮈鲫作为毒性试验材料,选择具有代表性的雄激素17α-甲基睾酮（MT）作为研究对象,采用酶联免疫法（ELISA）研究MT处理7 d对稀有鮈鲫体内雌二醇（E2）、睾酮（T）和11-酮基睾酮（11-KT）含量的影响。结果显示,经MT处理7 d后,雌鱼体内的E2、T和11-KT含量均极显著高于对照组（P〈0.01）,而雄鱼体内的E2、T和11-KT含量均极显著低于对照组（P〈0.01）。     

Risk assessment of endocrine-active compounds in feeds

As a key element of the 'farm-to-fork' approach, an assessment of the safety of feed components should consider major long-term toxicological hazards, such as endocrine disrupting chemicals (EDCs). In this review, attention is given to endocrine-modulating trace elements (such as iodine and trivalent chromium) and their potentially sensitive subgroups as nutritional feed additives from the standpoint of farm animal safety and consumer exposure. Feedstuffs can be a major vehicle for persistent EDCs, such as polychlorinated biphenyls, and potential risks include fish farming and ruminants grazing in polluted areas. Limited data still exist on the feed-to-food transfer of other EDCs, such as brominated flame retardants. EDCs of vegetable origin (e.g. zearalenone and isothiocyanates) can have a greater adverse impact on farm animals than environmental xenobiotics but the risk of carry-over to consumers appears low. Topics for further research are suggested, including the need for more refined exposure data, characterization of biomarkers for long-term effects, xenobiotic-nutrient interactions and the search for novel feed ingredients that are less vulnerable to contamination.     

环境内分泌干扰物对睾丸间质细胞功能的影响

近年来,环境内分泌干扰物(environmental endocrine disrupting chemicals,EDCs)对雄性动物的生殖毒性引起了广泛的关注。大量资料显示EDCs可引起雄性生殖发育障碍,如隐睾、尿道下裂、睾丸癌、精液品质不良等。作者就几种典型的EDCs对睾丸间质细胞分泌睾酮的影响机制作一综述。     

Impact of endocrine disrupting chemicals on reproduction in Japanese quail

Many environmental contaminants can interact with the endocrine system, thereby potentially disrupting the reproductive fitness of individuals. In avian species, the egg-yolk is a major route for excretion of lipophilic compounds by the adult female bird and embryos are exposed to contaminants that have been deposited in the eggs. The reproductive and neuroendocrine system of Japanese quail undergoes sexual differentiation during embryo development. The phenotypic sex, including sex-specific adult behavior, is hormonally imprinted already before hatching. The sexual differentiation of the brain in quail is sensitive to estrogens and the presence of estrogen results in a female phenotype. The relatively low concentration of estrogens in male embryos, on the other hand, results in a male behavioral phenotype. The behavior of male quail can be demasculinized by estrogen exposure during the period of sexual differentiation, and estrogen-exposed males are not able to display a male-typical behavior as adults. Also, differentiation of the reproductive organs is sensitive to hormones during embryogenesis, and an excess of estrogens can for instance induce persistent morphological changes in the reproductive organs of females. Our research has focused on effects in adult birds after embryonic estrogen exposure. We have studied sexual behavior and other reproductive variables in adult quail after in ovo injection of known and suspected estrogenic compounds. Synthetic estrogens and insecticides, such as o,p'-DDT altered the development of the neural system and resulted in demasculinization of male quail. In females, o,p'-DDT caused morphological changes of the oviduct and egg laying was reduced. Our studies suggest that the neural system and the female reproductive system of avian embryos are very sensitive to the effects of chemicals with estrogenic activity.     

Is exposure to endocrine disrupting compounds during fetal/post-natal development affecting the reproductive potential of farm animals?

Concerns have been raised about the potential adverse effects on reproductive health and immune status of farm animals following exposure to a range of natural and synthetic environmental compounds that disrupt normal hormonal actions. These compounds range from natural plant oestrogens (e.g. genistein, coumesterol) and mycoestrogens (e.g. Aflatoxins, zearalenone) to growth promoting pharmaceuticals (e.g. trenbolone acetate, melengastrol acetate) to chemicals spread in water, sewage sludge or the atmosphere such as detergents and surfactants (e.g. octylphenol, nonylphenol), plastics (e.g. bisphenol-A, phthalates), pesticides (e.g. methoxychlor, dieldrin, DDT) and industrial chemicals (e.g. PCB, TCDD). These compounds are commonly termed 'endocrine disrupting compounds' (EDCs) or 'endocrine disruptors' due to their ability to act as either hormone agonists or antagonists or the ability to disrupt hormone synthesis, storage or metabolism. A similar group of compounds are called 'immunotoxicants' and are thought to affect the immune system mainly by disrupting B and T cell homeostasis. As more studies are performed it is becoming clear that many compounds can directly or indirectly affect both the endocrine and immune systems. The susceptibility of target tissues is related to the stage of development, the cumulative exposure dose and the immune status of the individual. While some of the effects of the EDCs on the endocrine and immune systems are quite distinct, many are subtle and identifying the causative agent from the vast array of environmental challenges including EDCs, nutrition, temperature, etc. can be problematic. Identifying the causative agent is confounded by the possibility that effects that are observed in the adult may be due to exposure to EDCs during fetal life. This has major implications for the determination of universal end-point measurements to assess exposure to EDCs in farm animals.     

Effects of maternal diet and exposure to bisphenol A on sexually dimorphic responses in conceptuses and offspring

Whereas sexual differentiation is considered as the onset of differentiation of the male or female gonads, mounting evidence indicates that sex differences in developmental programming are established as early as the zygotic stage. Genetic and epigenetic differences between the sexes might govern how each responds to shifts in their early environment, including in the uterus or culture dish, as in the case of in vitro cultured pre-implantational embryos. Even if no differences are evident between the sexes at birth, divergent conceptus responses to surrounding changes, such as maternal diet and exposure to endocrine disrupting compounds (EDC), such as bisphenol A (BPA), might predispose one sex over the other to later adult-onset diseases, otherwise termed developmental origin of health and disease (DOHaD). Overall, males subjected to less than optimal in utero conditions tend to be at greater risk for various diseases, including neurobehavioural disorders. As the placenta is the primary nutrient acquisition and communication organ between the dam and foetus, its ability to adapt rapidly to environmental shifts might buffer the conceptus against environmental insults. The placenta of one sex over the other might possess greater ability to respond to environmental fluctuations. In utero environmental changes, including maternal nutrient excess or reduction or exposure to the EDC, BPA, might govern sex-dependent behavioural alterations. In sum, this review examines the evidence to date that male and female zygotes and conceptuses diverge in their responses to shifting environmental conditions and whether these contrasting sexually dimorphic responses underpin later DOHaD outcomes, namely neurobehavioural changes.     

Morphology of Reproductive Organs,Semen Quality and Sexual Behaviour of the Male Rabbit Exposed to a Soy‐containing Diet and Soy‐derived Isoflavones during Gestation and Lactation

Placental and breastfeeding transfer of soy isoflavones are potential routes for animal and human exposure to phytoestrogens, and reproductive dysfunctions have been linked to early exposure to these compounds. So, the aim of this study was to investigate the effects of perinatal (intrauterine and lactational) exposure to soy‐containing diet and soy‐derived isoflavones on the reproductive parameters of male rabbits. For this purpose, 12 female rabbits were randomly assigned to receive: (1) a soy‐ and alfalfa‐free diet (control diet); (2) a soy‐ and alfalfa‐free diet supplemented with 10 mg/kg body wt/day of soy isoflavones; (3) a soy‐ and alfalfa‐free diet supplemented with 20 mg/kg body wt/day of soy isoflavones; and (4) a diet containing 18% of soy meal, throughout gestation and lactation. Weight and morphology of the reproductive organs of some of the male offspring were evaluated at weaning (between days 29 and 31). The remaining males were placed on the control diet from weaning to adulthood (gestational and lactational exposure only). Sexual behaviour, semen quality and reproductive organs’ morphology were evaluated after puberty. There were no significant differences in litter size and gestation duration between control and treatment groups. Perinatal exposure to soy‐containing diet and soy isoflavones did not alter testis, epididymides, proprostate and prostate weight and gross morphology. After puberty, sexual behaviour and semen parameters did not differ significantly from the control group. These results indicate that intrauterine and lactational exposure to soy‐containing diet and soy‐derived isoflavones may not adversely affect reproductive development and function of male rabbits.     

Are Endocrine Disrupting Compounds a Threat to Farm Animal Health,Welfare and Productivity?

Contents The sources and characteristics of endocrine disrupting compounds (EDCs) are reviewed and discussed with respect to their potential effects on farm animal health, welfare and productivity. The importance of certain properties of these compounds in relation to the expression of their biological effects is addressed together with potential routes of exposure. It is concluded that little is known of factors affecting the tissue concentrations of EDCs in farm animals, the concentrations that are required to perturb physiological function in these species, the effects of prolonged exposure to low doses, the effect of cocktails of EDCs and other pollutants or the responses of specific organs and physiological systems that are affected by EDCs. Much of the available information pertaining to EDCs is derived from epidemiological studies of wildlife species and from laboratory animal studies and while these studies have significant limitations, they are considered to be valuable indicators of potential effects in farm animal species. The results of such studies, together with the small amounts of data from studies of ruminants, indicate that there may be significant effects of exposure to environmental levels of EDCs on farm animal health, even although effects are not generally apparent in practice, at this time.     

植物雌激素对动物生殖及生殖内分泌的影响

植物雌激素是一类存在于豆科植物中的具有雌激素样和抗雌激素样作用的天然活性物质。大量的研究表明 ,植物雌激素具有抗肿瘤和降低心血管疾病、骨质疏松及痴呆症等发病率的作用。然而 ,有些报道认为动物采食富含植物雌激素的日粮可引起生殖功能的紊乱 ,导致繁殖性能降低。但近年来的研究表明 ,某些植物雌激素可作用于性腺轴 ,干扰体内的生殖内分泌功能 ,从而改善动物精液品质、性行为和提高产蛋率等生殖的机能。文章主要综述了植物雌激素对动物生殖及其内分泌的影响 ,为植物雌激素在畜禽业中的应用提供了科学的依据     

Competition Between Different Social Ranked Rams has Similar Effects on Testosterone and Sexual Behaviour Throughout the Year

Dominant rams have preferential access to females, as they frequently interrupt sexual behaviour from subordinated. Testosterone concentrations are directly linked to sexual and aggressive behaviour and have important variations along the year. Therefore, it may be expected that the effects of dominance relationships on reproductive behaviour differ according to testosterone concentrations, and thus to the period of the year. The aim of this study was to compare the effects of dominance relationships on testosterone and sexual behaviour in different moments of the year in rams. Twelve rams were maintained in a single group. Social rank was determined in January (maximum reproductive development), May (regression of the reproductive status) and August (lowest reproductive activity), and the four rams with higher (HR) and the four with lower (LR) success index were used. Testosterone serum concentration was weekly measured four times during each experimental period. Sexual behaviour was evaluated in each period with an oestrous ewe, and with the oestrous ewe and another ram from the other social rank (each HR with each LR ram). Testosterone concentration was greater in HR than LR rams in January (p = 0.03), and all the behaviours were displayed more frequently in non‐competitive than in competitive tests (p < 0.05). Rams modified their sexual strategy in competitive environments decreasing the display of sexual behaviour independently of their social status. This effect was observed consistently throughout the year: high‐ranked rams have greater testosterone concentrations than LR rams only during the pre‐rut, when they naturally compete to join the groups of ewes.     

Endocrine disrupting compounds and farm animals: their properties,actions and routes of exposure

Endocrine disrupting compounds (EDC) comprise a diverse group of compounds of anthropogenic origin, including organochlorine pesticides, alkyl phenols, phthalates, polychlorinated biphenyls (PCB), dioxins and polybrominated diphenyl ethers. EDC are generally present in the environment at low concentrations but they are ubiquitous and persistent and, although environmental concentrations are low, they appear to exert a range of adverse effects on animals of many species, including humans. Their effects include disruption of reproductive function and of the immune system and they can be carcinogenic. Animals may be exposed to relatively high concentrations of EDC because they persist in the environment and when ingested, they may be concentrated in fat tissue and released when the fat is mobilised during pregnancy or lactation, thus exposing, to relatively high concentrations, embryos and neonates. These stages of development are particularly susceptible to EDC effects. Very little is known of EDC body burdens in domestic animal species and particularly in those exposed to unpolluted environments. EDC concentrations in soils and plant material and their rates of ingestion and metabolism have been little studied but it is concluded that there is a potential risk of significant bioaccumulation and of associated effects on the health and reproductive capacity of domestic animals and of humans consuming animal products.     

Exposure to Perchlorate Induces the Formation of Macrophage Aggregates in the Trunk Kidney of Zebrafish and Mosquitofish

Abstract

Environmental contamination of ground and surface waters by perchlorate, derived from ammonium perchlorate (AP) and other perchlorate salts, is of increasing concern. Exposure to perchlorate can impair the thyroid endocrine system, which is thought to modulate renal and immune function in vertebrates. This study with zebrafish Danio rerio and eastern mosquitofish Gambusia holbrooki examined the histological effects of perchlorate on the trunk kidney, which in teleosts serves excretory and hemopoietic functions and therefore may be a target of perchlorate effects. Adult zebrafish of both sexes were exposed in the laboratory to waterborne, AP-derived perchlorate at measured concentrations of 18 mg/L for 8 weeks. Adult male mosquitofish were exposed to waterborne sodium perchlorate at measured perchlorate concentrations of 1–92 mg/L for 8 weeks. Control fish were kept in untreated water. The region of the body cavity containing the trunk kidney was processed from each fish for histological analysis. Macrophage aggregates (MAs), possible markers of contaminant exposure or immunotoxic effect, were present in the hemopoietic region of the kidney in both species exposed to perchlorate. The estimated percent area of kidney sections occupied by MAs was greater in zebrafish exposed to perchlorate at 18 mg/L (P < 0.05) than in controls. In male mosquitofish, the incidence of renal MAs increased proportionally with sodium perchlorate concentration and was significantly different from that of controls at 92 mg/L (P < 0.05). These observations confirm that in fish the kidney is affected by exposure to perchlorate. The concentrations of perchlorate at which the effects were noted are relatively high but within the range reported in some contaminated habitats.     

Effects of Polychlorinated Biphenyls 28, 30 and 118 on Bovine Spermatozoa In Vitro

Decline of semen quality due to endocrine‐disrupting chemicals is of concern globally. Among endocrine disrupting chemicals (EDC), Polychlorinated biphenyls (PCB) are associated with reduced semen quality in various epidemiological studies. In this study, we evaluated the direct effects of selected PCBs (28, 30 and 118) on fresh spermatozoa of Simmetial bulls aged 2–4 years were evaluated in vitro by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and computer assisted semen analysis (CASA) (SCA; Microptics) analysis. IC50 values were found as 8.45, 5.45 and 9.55 ng/ml for PCB 28, 30 and 118, respectively. Total motility, progressive motility and viability decreased dependent on dose and duration of exposure (0, 2, 4 h). Total motility at IC50 doses decreased the most in PCB 28 (72.24%) followed by 30 (60.75%) and 118 (64.77%) at 2nd hour following exposure. Motility results were found to be in accordance with the vitality and morphology data where total abnormalities (especially reacted acrosome rate) were found to have increased.     

An Update on Reproduction in Yak and Mithun

Yak and mithun are two domesticated herbivores of high economic importance to the farming community living in highlands. Improved yak and mithun production could significantly enhance the living standards of these highlanders. Over the years, their dwindling numbers have been a cause of serious concern. In view of the lack of knowledge on the reproductive physiology of these ruminants, studies have been undertaken to investigate their reproductive endocrinology in recent years. This paper attempts to present the latest information on the endocrine changes associated with their various reproductive processes viz. growth and puberty, oestrous cyclicity and oestrous behaviour, ovulation, and, pregnancy and parturition. The paper also provides the recent developments on research done towards the enhancement of yak and mithun reproductive efficiencies through endocrine and embryo biotechniques.     

Effects of perinatal exposure to phthalate/adipate esters on hypothalamic gene expression and sexual behavior in rats

Our previous research has identified the granulin (grn) and p130 genes as sex steroid-regulated genes in the neonatal rat hypothalamus that might be involved in sexual differentiation of the brain. Since phthalate/adipate esters such as di-n-butyl phthalate (DBP), diisononyl phthalate (DINP), and di-2-ethylhexyl adipate (DEHA) are suspected to interfere with the endocrine system as environmental endocrine disruptors having estrogenic or antiandrogenic properties, these chemicals may affect sexual differentiation of the brain. The present study assessed the effects of perinatal exposure to DBP, DINP, and DEHA on grn and p130 mRNA expressions in the hypothalamus on postnatal day (PND) 7 and sexual behaviors after maturation in rats. Maternal rats were given a phytoestrogen-free diet containing different doses of DBP (20, 200, 2,000, and 10,000 ppm), DINP (40, 400, 4,000, and 20,000 ppm) and DEHA (480, 2,400, and 12,000 ppm) from gestational day 15 to the day of weaning (PND 21). DBP and DINP exposure during the perinatal period resulted in an increase in hypothalamic grn and p130 mRNA levels in females and males, respectively, but DEHA exposure decreased expression levels of grn in males and p130 in females, although the effects were not dose-dependent. After maturation, male rats that were exposed to several doses of DBP, DINP, and DEHA displayed decreased copulatory behavior. The lordosis quotient was decreased in females perinatally exposed to DBP, DINP, and DEHA at all the doses used. On the other hand, serum levels of LH and FSH in both sexes and the estrous cycles in females were not affected by the treatments. These results suggest that inappropriate expression of grn and/or p130 genes in the brains of male and female neonatal rats by perinatal exposure to these chemicals may exert permanent effects on the hypothalamus, thereby decreasing sexual behavior after maturation.     

Epigenetic mechanisms in the actions of endocrine-disrupting chemicals: gonadal effects and role in female reproduction

There is a heightened interest and concern among scientists, clinicians and regulatory agencies as well as the general public, regarding the effects of environmental endocrine-disrupting chemicals (EDCs). In this review, we identify the main epigenetic mechanisms and describe key ovarian processes that are vulnerable to the epigenetic actions of EDCs. We also provide an overview of the human epidemiological evidence documenting the detrimental effects of several common environmental EDCs on female reproduction. We then focus on experimental evidence demonstrating the epigenetic effects of these EDCs in the ovary and female reproductive system, with an emphasis on methoxychlor, an organochlorine pesticide. We conclude the review by describing several critical issues in studying epigenetic effects of EDCs in the ovary, including transgenerational epigenetic effects.     

Differential effects of the endocrine-disrupting compounds bisphenol-A and octylphenol on gonadotropin secretion, in prepubertal ewe lambs

This study examined the effects of long-term exposure to the endocrine-disrupting compounds (EDCs) Bisphenol-A (BPA) and Octylphenol (OP) on gonadotrophin secretion in pre-pubertal female sheep. Four-week-old, female lambs were randomly allocated to four groups (n=6), and twice each week treated with i.m. injections of either corn oil (vehicle controls), diethylstilbestrol (DES; 0.175mg/kg), BPA (3.5mg/kg) or OP (3.5mg/kg). After 5 weeks of treatment, animals were ovariectomized (ovx) and ovary weights recorded. Two weeks later, blood samples were collected from lambs every 15min for 6h, for LH pulse analysis. Animals were then euthanased and adrenal and kidney weight recorded. An age-related increase in tonic LH secretion was noted in Control, BPA- and OP-treated lambs, but was absent in DES-treated lambs. Following ovx, LH secretion increased in all except DES-treated lambs; FSH concentrations increased in all groups. BPA and DES significantly suppressed LH pulse frequency (C: 6.7+/-0.3pulses/6h, DES: 1.5+/-0.8pulses/6h, BPA: 2.3+/-0.8pulses/6h) and amplitude (C: 7.1+/-1.0ng/ml, DES: 1.9+/-0.6ng/ml, BPA: 1.6+/-0.4ng/ml). OP had no effect on LH secretion (Frequency: 5.8+/-0.5pulses/6h, amplitude: 8.0+/-2.0ng/ml). Ovary weight was similar among all groups. Results show that chronic in vivo exposure of prepubertal female lambs to BPA, at levels lower than those reported previously, can have significant effects on LH secretion that are comparable to those seen following exposure to the known xenoestrogen, DES. Exposure to an equal dose of the EDC OP, over the equivalent period of time was without effect on gonadotropin secretion in the prepubertal ewe lamb. These results indicate that exposure of prepubertal female lambs to the EDC BPA can induce significant effects on gonadotropin secretion, the potential long-term effects of exposure and the effects of these changes on reproductive performance and efficacy, therefore, merit further study.