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.     

Impact of Endocrine‐Disrupting Chemicals on Reproductive Function in Zebrafish (Danio rerio)

The prevalence of endocrine‐disrupting chemicals (EDCs) in the aquatic environment has been associated with the wide detection of alterations in the development and physiology of vertebrates. Zebrafish, as a model species, has been extensively used in toxicological research. In this review, we focus on recent published evidence of the harmful effects of EDCs on reproductive function in zebrafish, including skewed sex ratio, immature gonads, diminished sexual behaviour, decreased sperm count, reduced spawning and fertilization. These impairments mostly result from disruption to sex‐steroid hormones induced by endocrine disruptors. We also discuss other effects of exposure to EDCs. In EDC exposure research, despite incomplete assessments of altered gonad histopathology and sexual behaviour, these present potential effective biomarkers or pathways for evaluating the reproductive function in zebrafish on EDC exposure. To date, the pernicious effects of some EDCs on the reproductive performance in laboratory zebrafish are well understood; however, similar alterations remain for further determination in wild‐type fish and more kinds of EDCs. More studies should be performed under established scientific regulatory criteria to investigate the impact of EDCs on reproduction in zebrafish. Moreover, further research is required to explain the definite mechanism of sexual differentiation, which helps in understanding the shift of sexual phenotype with EDC exposure.     

Effects of methoxychlor exposure during perinatal period on reproductive function after maturation in rats

Methoxychlor (MXC) is a non-steroidal pesticide that is known to possess estrogenic activity, and therefore may be potentially hazardous to the development and/or reproduction. The present study assessed the effects of perinatal exposure to MXC on reproductive function after maturation in both male and female rats. Pregnant rats were fed a phytoestrogen-free diet containing different doses of MXC (24, 240, and 1200 ppm) from day 15 of gestation to day 10 after parturition, and reproductive functions of offspring were examined after maturation. In males, MXC exposure during the perinatal period decreased serum LH and FSH, but not testosterone levels, but it did not affect copulatory behavior. In females, MXC exposure prolonged the days exhibiting cornified vaginal smears during the estrous cycle. In addition, both the lordosis reflex and preovulatory LH surge on the presumptive proestrous evening were suppressed in MXC-exposed females. These results suggest that perinatal exposure to MXC exerts permanent effects on several aspects of the reproductive function in both male and female rats.     

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.     

Effect of coccidiosis on reproductive maturation of male Japanese quail

The effects of coccidiosis on reproductive development of male Japanese quail were examined. Male Japanese quail were exposed to high (5 x 10(5) sporulated oocysts/quail) or low (5 x 10(3) sporulated oocysts/quail) doses of Eimeria uzura at 16 or 30 days of age and sampled at 37 days. Quail given high doses of coccidia had reduced testes weight and lowered circulating concentrations of androgen compared with control males. Low doses of coccidia did not affect testes weight but did result in elevated plasma androgen levels. There were no differences in average testes weights by 51 days; however, plasma androgen was still reduced in most groups. To study the effects of coccidiosis on egg production, males exposed to high doses of coccidia at 16 (16H) or 30 (30H) days of age were mated with control females, and control males were mated with control or 16H females. The onset of laying was delayed for 5 days in the control male: 16H female group. During the first week of production, eggs from females bred to 30H males had lower fertility and hatchability than those bred to control or 16H males. By the third week of production, levels of fertility were similar. Apparently, exposure of quails to coccidiosis before sexual maturation might result in long-term effects on later reproductive capability.     

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）。     

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。     

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.     

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.     

Behavioral, ovarian and endocrine relationships in the pubertal bitch

Behavioral, ovarian and endocrine correlates were determined in seven bitches experiencing first estrus, and the results compared with previous data obtained on multiparous females. One or more atypical reproductive patterns were observed in certain bitches at the pubertal estrus. The ability of pubertal dogs to display normal reproductive relationships appeared to be related to age, because animals that exhibited normal sexual behavior and endocrine profiles tended to be older than females that produced aberrant patterns. For the latter group, atypical observations included lack of sexual receptivity and reduced or inconsistent patterns of circulating concentrations of estradiol-17 beta, luteinizing hormone (LH) or progesterone. Ovulation was confirmed in all four pubertal dogs subjected to laparoscopy, including two bitches that failed to demonstrate estrous behavior. In individual animals, it appeared that inadequate or sporadic elevations in titers of estradiol-17 beta during the preovulatory phase of the cycle were associated with reduced or abbreviated LH surge concentrations, and the latter were subsequently related to an absent or only minor preovulatory rise in serum progesterone. Reduced preovulatory estradiol-17 beta concentrations and a delayed increase in circulating progesterone were also associated with a lack of sexual receptivity. These results (1) suggest that hormonal insufficiencies or irregularities exist in certain bitches at the pubertal estrus and that such atypical activity appears more characteristic of the first than later adult estrous periods, and (2) are consistent with the theory that estrogen preconditioning and a decrease in the endogenous estrogen to progesterone ratio are important in the initiation of sexual receptivity in this species.     

Effects of 3-methyl-4-nitrophenol in diesel exhaust particles on the regulation of reproductive function in immature female Japanese quail (Coturnix japonica)

In a previous study, we found that 3-methyl-4-nitrophenol (PNMC), a component of diesel exhaust particles and also a degradation product of the insecticide fenitrothion, exhibits reproductive toxicity in the adult male Japanese quail. The present study investigated the toxicity of PNMC in the female Japanese quail and its ability to influence reproduction in immature females. The quail (21-day-old) were injected intramuscularly (im) with PNMC at doses 0.1, 1 or 10 mg/kg body weight daily for 3 days. There was no significant difference in body growth between the PNMC-administered and control birds. However, the weights of the oviducts were significantly lower in the PNMC-treated birds at all doses. Furthermore, the plasma concentrations of luteinizing hormone (LH) and estradiol-17 beta were significantly decreased with 1 and 10 mg/kg of PNMC. These findings suggest that PNMC might influence the hypothalamo-pituitary-gonadal axis with decreasing in secretion of GnRH, LH and ovarian steroid hormones and subsequently disturb growth of the reproductive organs of immature female quail. This study indicates that PNMC induces reproductive toxicity at the central level and disrupts reproductive function in the immature female quail.     

Effect of organic selenium in quail diet on its accumulation in tissues and transfer to the progeny

1. The aim of the present study was to investigate the effects on the eggs and hatchlings (up to 2 weeks post-hatch) of feeding a relatively large amount of so-called organic selenium to breeder quail. 2. Two groups of quail (3 families in each group consisting of 4 females and 1 male) were formed at the beginning of their reproductive period. The quail were fed on a commercial maize-based diet containing 0.096 mg/kg feed-derived selenium (Se), supplemented with 0.2 mg/kg selenite (control group) or 0.5 mg/kg organic selenium in the form of Sel-Plex (Alltech Ltd, USA) for 6 months. Eggs were collected at 6 months of age and Se in the egg yolk, egg white and shell was analysed. Five quail at 1, 7 and 14 d post-hatch were killed to provide samples of liver, brain, breast and leg muscles for Se analysis. After egg collection for analysis and incubation, adult quail were killed and liver, kidney, lung, brain, breast and leg muscles were collected for Se analyses. 3. Inclusion of high doses (0.5 mg/kg) of organic Se in the quail diet was associated with a significant increase in Se concentration in all tissues studied of adult quail as well as in egg yolk, egg albumin and eggshell. 4. Increased Se concentration in the quail egg was associated with increased Se concentration in the liver, breast and leg muscles and brain of newly hatched quail. This difference was shown to be significant for 2 weeks post-hatch. Therefore, it has been suggested that the maternal effect of dietary selenium can be seen beyond the hatching time and more emphasis should be given to this effect in future. 5. It was shown that it is possible to produce Se-enriched quail meat and eggs by adding organic selenium to the diet.     

Ivermectin impairs sexual behavior in sexually naïve, but not sexually experienced male rats

Ivermectin (IVM) is an antiparasitic drug, widely used in domestic animals. In mammals, IVM act as a GABA agonist. This neurotransmitter has an important role in the regulation of sexual behavior. Thus, this study sought to investigate the effects of various medically relevant doses IVM on the sexual behavior of male rats. In particular, we also wished to examine if previous sexual experience modulated responses to IVM. In the first experiment, the sexual behavior of inexperienced male rats was analyzed after they received 0.2, 0.6, 1.0 or 2.0 mg/kg IVM, 15 min prior to behavioral testing. In the second experiment, the effects of four previous sexual experiences on IVM treated rats (1.0 or 2.0 mg/kg, 15 min prior to the 5th session) were assessed. The standard therapeutic dose (0.2 mg/kg) did not impair the sexual behavior of inexperienced male rats. At a more concentrated dose (0.6 mg/kg), which is still within the therapeutic range, the appetitive phase of sexual behavior of inexperienced male rats was impaired. Likewise, 1.0 mg/kg impaired the appetitive phase. Previous sexual experience blocked almost entirely this sexual impairment, suggesting that previous sexual experience exerts a positive effect in attenuating the sexual impairment produced by IVM treatment. Therefore, the standard therapeutic dose of IVM can be used without producing side effects on sexual behavior. Use of more concentrated therapeutic doses is not recommended during reproductive periods, unless the animals have had previous sexual experience.     

Ontogeny of copulatory behaviour in male Japanese quail classified by their T-maze performance as hatchlings

1. This study examines the time courses of male mating behaviours using quail that were categorised at 2 d of age as high performance (HP) or low performance (LP) individuals in a T-maze. 2. Individually caged males from each T-maze category were observed for 5 min after a female of the same category (HP or LP) was introduced into their home cages as a sexual partner at 4, 5, 6, 8, 10 and 12 weeks of age. The number of grabs, mounts and cloacal contacts performed by each male and his copulatory efficiency (number of cloacal contacts/number of grabs; CE) were determined at each time interval. 3. No differences were observed in the proportion of HP and LP males performing grabs at any of the ages evaluated. However, a greater proportion of HP males showed mounting and cloacal contact behaviour, coupled with a higher CE than did their LP counterparts at 5 and 6, 6 and 8, and 8 weeks of age, respectively. At those ages, a higher mean number of mounts and cloacal contacts were also observed. No significant sexual behaviour differences between HP and LP quail were observed at 10 and 12 weeks of age. 4. The results suggest that rapid negotiation of the T-maze in hatchlings is associated with accelerated puberty in male quail. The differential reproductive behaviour of HP males during the development of puberty does not remain extant in fully developed quail. 5. The finding of accelerated male developmental sexual activity responses in HP quail indicates that selection for T-maze HP behaviour may allow males to be successfully bred at an earlier age.     

Endocrine changes in late bovine pregnancy with special emphasis on fetal well-being

During late bovine pregnancy, several hormones are involved to maintain and develop a successful result with a live calf. These hormones are e.g., progesterone, high levels during the whole pregnancy period, originating from the corpus luteum, maternal adrenals and placenta. Oestrone sulphate, oestrone in its conjugated form, shows elevated levels from about mid-pregnancy until the third stage of parturition (expelling of the fetal membranes). For the onset of normal parturition and the parturition process as such, a change from progesterone to oestrone synthesis is crucial. The increasing levels of oestrone are time-related to an increased synthesis of prostaglandin F(2alpha) (reflected as elevated levels of 15-ketodihydro-PGF(2alpha)) causing prepartal luteolysis and several hormones are then involved in the labour process such as prostaglandin F(2alpha), cortisol and oxytocin. Cortisol might also be an indicator of stressful events for the dam. Levels of pregnancy associated glycoproteins (PAGs), originating from the trophoblastic binucleate cells, are increasing during the last 10 days prior to parturition. All the mentioned hormones have certain functions during pregnancy, more or less understood. However, could deviations from the expected profiles during late bovine pregnancy indicate impaired fetal well-being or be of importance for reproductive performance during the postpartum period? Abortions, stillbirths or dystocia are situations where endocrine profiles might predict the status of the calf. There are two possible approaches to study the endocrine changes in late pregnancy-to follow spontaneous cases of normal or impaired pregnancies or to experimentally disturb the gestation or induce parturition. We have in one study followed pregnant animals to depict reproductive disturbances, both animals with expected normal parturitions and animals where the sire of the calf has given rise to a high incidence of stillborn calves. The number of stillborn calves or dystocia has been small and so far it has not been possible to obtain a clear picture of the usefulness of endocrine parameters to follow fetal well being, but some of the hormonal parameters show a deviating profile. In a small group of animals with induced parturition (PGF(2alpha)), two out of three had parturition problems and one of these animals had a stillborn calf. All three animals had retained fetal membranes. It was possible to demonstrate a deviating endocrine profile in the cow having the stillborn calf in the sense of higher levels of progesterone, cortisol and 15-ketodihydro-PGF(2alpha) at the time of parturition. In both animals with dystocia the levels of oestrone sulphate after parturition were more sustained. Increasing and high levels of PAGs were only demonstrated in the animal with a normal parturition. These studies are ongoing, aiming at finding changes in endocrine profiles related to impaired pregnancies.     

Effects of poppy seed meal on egg production and hatching results in quail (Coturnix coturnix japonica)

This study was carried out to determine the effects of poppy seed meal (PSM) on the egg production and hatching of quail. Two experiments were undertaken. In the first experiment, 240 quail, reared under the same conditions, were divided into four groups of 60 birds each (which were, in turn, subdivided into four subgroups of 15 each for experimental replicates). The first, second, and third experimental groups were fed ratios containing 5%, 15%, and 25% PSM, respectively, over 12 weeks. The control group received no PSM. Feed and water were supplied ad libitum and artificial light was provided for 16 h a day. During the first experiment, egg production, egg weight, and feed conversion ratio (FCR) were measured. In the second experiment, 240 female and 48 male quail were divided into four groups, as in the preceding experiment, and the birds were housed in cages at a ratio of one male to five females. The diets and their presentation to the quails were the same as in the preceding experiment. The results of the two experiments showed that the addition of PSM to the feed ratios increased egg production, feed consumption, and feed conversion per kg of eggs and per dozen eggs. However, PSM addition at all percentages significantly reduced (P<0.001) hatchability, mainly because of increased infertility and embryonic deaths.     

Ram mating behavior after long-term selection for reproductive rate in Rambouillet ewes

Mating behavior is known to be heritable in several species, but it is not known if selection schemes for ewe reproductive traits affect mating behavior of rams. Therefore, our objectives were to determine if divergent (high vs low index) selection of a female reproductive trait (lambs born divided by age of ewe minus one) affects mating behavior patterns of male offspring during training to service an artificial vagina (AV) in a less than natural setting (Exp. 1) or during serving capacity tests (SCT) in simulated natural mating conditions (Exp. 2). The method for evaluating male mating behavior was a series of serving capacity tests to estimate sexual performance. For Exp. 1, five, 30-min observations were conducted to assess the mating behavior of 29 rams (22 mo old) being trained to service an AV. In Exp. 1, percentages of rams mounting (73 vs 36%) and ejaculating (67 vs 29%) differed (P < 0.05) between rams from high and low Rambouillet ewe selection lines, respectively. In Exp. 2, ram classification consisted of exposing each 22- to 24-mo-old ram (n = 48) to three unrestrained ewes in estrus for 18, 30-min tests. Any ram that had not mounted or ejaculated during SCT was evaluated for sexual orientation in a 30-min preference test in which each ram had access to both restrained estrual ewes and restrained rams. In Exp. 2, there was no difference (P > 0.2) in percentages of rams mounting (92 vs 78%) and ejaculating (88 vs 74%) between rams from high and low Rambouillet ewe selection lines, respectively. There was no difference (P > 0.33) between high and low ewe selection line rams for mounts or ejaculations for 18 SCT. Only one male-oriented ram was identified, which was from the high line. In Exp. 2, six SCT were necessary to obtain 95% reliability in sexual performance scores. Results of these studies indicate that long-term selection of ewes for litter size did not alter ram sexual performance after SCT under simulated natural mating conditions or affect incidence of male-oriented rams. Mating behavior observed during training to service an AV was lower for rams from ewes of the low selection line than for rams from ewes of the high selection line. We concluded that long-term selection for reproductive rate in ewes did not alter mating behavior patterns of male offspring; however, test conditions, such as restrained and unrestrained ewes and number of serving capacity tests, may affect conclusions in studies that evaluate sexual performance of rams.     

Kisspeptin-10对蛋鹌鹑卵巢发育与内分泌的影响

为研究Kisspeptin-10(Kp-10)对禽类生殖内分泌的影响,将75羽22日龄雌鹌鹑随机分为3组,分别腹腔注射300μL生理盐水(对照组),注射0.1 nmol Kp-10(低剂量组)和1 nmol Kp-10(高剂量组),每日注射1次,连续注射3周。记录鹌鹑开产情况,并于60日龄时采集血液和卵泡组织,测定生殖激素与受体的mRNA表达。结果:与对照组相比,注射Kp-10组鹌鹑产蛋率显著提高,且大卵泡数显著增加,血液中雌激素水平显著升高;低剂量组输卵管重显著增加,但体重、肝脏重和小卵泡重无显著变化。Real-time PCR结果显示,Kp-10处理显著下调小卵泡及F1级卵泡上促黄体激素受体(LHR)、促卵泡激素受体(FSHR)和促性腺激素释放激素Ⅰ(GnRH-Ⅰ)的基因表达。