Plasma Estradiol-17β Concentrations Research Articles

Application of blood parameters for the early diagnosis of natural ascending placentitis in pregnant mares

Placental infection is an important cause of late-term pregnancy loss and neonatal diseases in horses. Detection of changes in blood parameters especially during early placentitis could improve the diagnostic accuracy, treatment decision, and potential outcomes. The objectives of this 2-part study were to identify differences in circulating immunological, inflammatory, and hormonal parameters between mares with natural ascending placentitis and control mares; evaluate each and combination of parameters as predictors of placentitis; and determine how these parameters indicate severity of placentitis. Reproductive examination and blood sampling were prospectively performed on pregnant mares in a natural setting. Study 1 enrolled mares diagnosed with early stage of ascending placentitis based on ultrasonographic findings (n = 12), and gestationally age-matched mares with healthy pregnancies as controls (n = 12). Blood samples were classified as pre-onset (before) and early onset (at the time of ultrasonographic changes) of placentitis. There were no detected statistically significant differences between groups and timepoints in immunological and inflammatory parameters, including peripheral lymphocyte subpopulations, cytokine, and serum amyloid A concentrations. The placentitis group showed a reduced (P = 0.01) DHP/20α-DHP ratio when compared to the control group at the early onset timepoint. Plasma estradiol-17β concentration <359 pg/mL predicted ascending placentitis with acceptable accuracy (area under the curve, AUC = 0.71). Combined albumin <3.7 g/dL, estradiol-17β < 499 ng/mL, and DHP <33 ng/mL predicted 100 % of cases of ascending placentitis. In study 2, samples were classified according to the presence and severity of the abnormal ultrasonographic findings as mild (n = 11) and moderate-severe (n = 23), and gestationally age-matched with samples from control mares (n = 34). Mares with moderate-severe ascending placentitis had increased (P = 0.03) plasma 20α-DHP concentration and reduced (P = 0.03) DHP/20α-DHP ratio when compared to control mares. Our results suggest that the early events of ascending placentitis detected by ultrasonographic findings include hormonal alterations of feto-placental metabolism measurable in the mare's circulation, yet without obvious systemic immunological and inflammatory changes. Additional studies are warranted to further assess how hormonal markers and cutoff values could guide decisions for timely therapeutic intervention.

Impairment of oocyte maturation as a mechanism of decreased fecundity in Japanese medaka (Oryzias latipes) exposed to the brominated flame retardant, 1,2,5,6-tetrabromocyclooctane (TBCO)

Inhibition of oocyte maturation is an understudied mechanism by which chemical stressors can impair fecundity of female fishes. The primary objective of the present study was to develop an assay to assess oocyte maturation disruption by chemical stressors in Japanese medaka (Oryzias latipes). First, an in vitro assay to assess maturation inducing hormone (MIH)-stimulated oocyte maturation in zebrafish was validated for use with Japanese medaka. Next, using the brominated flame retardant, 1,2,5,6-tetrabromocyclooctane (TBCO), which previously was shown to decrease fecundity of Japanese medaka and inhibit oocyte maturation in zebrafish, effects on oocyte maturation were quantified using in vitro and in vivo exposure. Adaptation of the protocol for in vitro MIH-stimulated maturation of stage IV oocytes from zebrafish was successful in inducing greater than 80% of stage IX oocytes from female Japanese medaka to mature. To assess effects of in vitro exposure, stage IX oocytes were exposed to 0, 2, 20, and 200 μg/L of TBCO, followed by exposure to MIH. The in vitro exposure caused a significant decrease in maturation of oocytes exposed to 20 and 200 μg/L of TBCO. To assess effects of TBCO on fecundity and oocyte maturation following in vivo exposure, sexually mature fish were fed a control, 100 μg/g, or 1000 μg/g concentration of TBCO-spiked fish food for 21 days, where fecundity was measured daily, and following the exposure, stage IX oocytes were excised to assess MIH-stimulated maturation. Fecundity and oocyte maturation were significantly decreased at either concentration of TBCO. Plasma concentrations of 17β-estradiol (E2) and hepatic abundances of transcripts of vitellogenin (vtgI and vtgII) were quantified, but there were no significant differences between treatments. Results suggest that inhibition of oocyte maturation is a mechanism by which TBCO decreases fecundity, and that in vitro assays of oocyte maturation might be predictive of fecundity in this species.

Putative adverse outcome pathway development based on physiological responses of female fathead minnows to model estrogen versus androgen receptor agonists

Several adverse outcome pathways (AOPs) have linked molecular initiating events like aromatase inhibition, androgen receptor (AR) agonism, and estrogen receptor (ER) antagonism to reproductive impairment in adult fish. Estrogen receptor agonists can also cause adverse reproductive effects, however, the early key events (KEs) in an AOP leading to this are mostly unknown. The primary aim of this study was to develop hypotheses regarding the potential mechanisms through which exposure to ER agonists might lead to reproductive impairment in female fish. Mature fathead minnows were exposed to 1 or 10 ng 17α-ethynylestradiol (EE2)/L or 10 or 100 µg bisphenol A (BPA)/L for 14 d. The response to EE2 and BPA was contrasted with the effects of 500 ng/L of 17β-trenbolone (TRB), an AR agonist, as well as TRB combined with the low and high concentrations of EE2 or BPA tested individually. Exposure to 10 ng EE2/L, 100 µg BPA/L, TRB, or the various mixtures with TRB caused significant decreases in plasma concentrations of 17β-estradiol. Exposure to TRB alone caused a significant reduction in plasma vitellogenin (VTG), but VTG was unaffected or even increased in females exposed to EE2 or BPA alone or, in most cases, in mixtures with TRB. Over the course of the 14-d exposure, the only treatments that clearly did not affect egg production were 1 ng EE2/L and 10 µg BPA/L. Based on these results and knowledge of hypothalamic–pituitary–gonadal axis function, we hypothesize an AOP whereby decreased production of maturation-inducing steroid leading to impaired oocyte maturation and ovulation, possibly due to negative feedback or direct inhibitory effects of membrane ER activation, could be responsible for causing adverse reproductive impacts in female fish exposed to ER agonists.

Diagnosis of imminent placentitis in mares

Pregnancy loss and neonatal mortality have a significant economic impact on the equine industry. Placentitis is a common cause of abortion, stillbirths, and neonatal infections. Early diagnosis of placentitis allows prompt treatment and increases the chances of a pregnancy reaching term. Our study was designed to identify blood parameters that are diagnostic of imminent placentitis under natural conditions. We hypothesized that differences from physiological parameters of a healthy pregnancy indicate imminent placentitis. Blood sample collections and reproductive exams were prospectively performed, monthly, between 180 to 320 days of gestation, in a cohort of mares from 2 large broodmare herds in Lexington, KY. Twelve mares were diagnosed with placentitis based on rectal ultrasonographic examination (opening at the cervix, thickened placenta, fluid collection, placental edema, and/or placental separation). Blood samples collected on the day of placentitis diagnosis were designated as early onset, and the blood samples collected in the previous reproductive exam with no ultrasonographic abnormalities of the fetoplacental unit were named as pre-onset (before placentitis diagnosis). In addition, 12 gestion and age-matched control mares carrying healthy pregnancies based on the same parameters were paired to the mares with placentitis. We measured several immunological, inflammatory, and hormonal parameters (Table 1). Values were compared between groups within the same timepoint (placentitis vs control) and between timepoints (pre-onset vs early onset) using Paired Student's T test and Mann-Whitney U test, and significance set at p < 0.05. According to the gestational age, the interval between pre-onset timepoint and the diagnosis of placentitis (early onset) was around 40 days (227±6 days and 267±5 days, respectively).No differences were found between groups and timepoints for immunological and inflammatory parameters in the maternal circulation, including serum amyloid A (SAA) and alpha-fetoprotein (AFP). Both proteins have been shown to markedly increase in maternal circulation after experimental induction of placentitis. In the hormonal analyses, no differences were found in plasma progesterone levels and estradiol-17β/progesterone ratio. Plasma estradiol-17β concentration approached statistical difference (p=0.08) in mares at early onset of placentitis (408±35 pg/mL) compared to the control group (498±35 pg/mL). In conclusion, our data showed that early events of spontaneous placentitis do not increase inflammatory and immunological parameters in the maternal circulation. In addition, plasma estradiol-17β may be further investigated as an early indicator of placental dysfunction in mares with naturally occurring placentitis. Acknowledgments: We thank the Harry M. Zweig Memorial Fund for Equine Research for supporting our study.

Plasma profile of follicle-stimulating hormone and sex steroid hormones after a single epidural administration of follicle-stimulating hormone via caudal vertebrae in Holstein dry cows.

The conventional follicle-stimulating hormone (FSH) treatment for bovine superstimulation involves multiple intramuscular injections, which is stressful for animals and onerous. We herein investigated whether a single epidural injection of porcine FSH (pFSH) can induce superovulation and peripheral concentrations of pFSH and steroid hormones after the treatment in Holstein dry cows. We intramuscularly administered pFSH twice daily to three cows for 3 days (control) or a single epidural pFSH administration (epidural). Numbers of follicles (≥10mm in diameter) at estrus and corpora lutea at luteal phase were counted by ultrasonography. Blood was sampled from 0 to 104 h after the first pFSH administration and plasma pFSH, progesterone, androstenedione, testosterone, and estradiol-17β concentrations were measured. Numbers of follicles (control: 18.3 ± 7.5, epidural: 15.7 ± 4.0; mean ± SD) and corpora lutea (control: 7.3 ± 4.2, epidural: 8.0 ± 2.6) were similar between both treatments. Plasma pFSH concentrations were higher in epidural than in control (p < 0.01). Although no significant differences were observed in progesterone, androstenedione, or estradiol-17β concentrations between the groups, testosterone concentrations were slightly lower with the epidural treatment than with the control treatment (p = 0.08). In conclusion, superovulation was induced by a single epidural injection of pFSH, which achieved higher pFSH level than the multiple injections in Holstein dry cows.

Ageing affects plasma steroid concentrations and testicular volume, echotexture and haemodynamics in rams.

The objective was to determine the effect of ageing on plasma steroid concentrations and testicular volume, echotexture and haemodynamics in Ossimi rams. Twenty-four rams were allocated, by age, into three groups: young (1year; 32.00±0.78kg, n=6); mature (2.5years; 48.00±1.02kg, n=9) and aged (7.5years; 65.00±1.10kg, n=9). Aged rams had the highest (p˂0.05) plasma testosterone concentrations 3.90±0.75ng/ml (three-fold the other groups), whereas in young rams, plasma estradiol-17β concentrations (5.85±0.34pg/ml) were approximately one-quarter (p˂0.05) of other groups. Regarding pulsed-wave Doppler indices values of the (resistive index RI; and pulsatility index PI) were lower (p˂0.05) in the mature (0.32±0.04 and 0.36±0.03, respectively) and aged (0.51±0.03 and 0.77±0.06) rams compared to young rams (0.62±0.03 and 0.98±0.07). In the young group, testicular pixel intensity was highest (p˂0.05) but values of coloured area/pixels were lowest (p˂0.05) compared to the other groups. In conclusion, these parameters indicate that aged rams (7.5years) are likely to be suitable for breeding ahead of the younger aged groups (1 and 2.5years) examined, but a final decision would need to be tempered by consideration of other parameters in the breeding soundness examination (BSE), such as physical soundness and semen characteristics. Of the parameters studied, testicular pixel intensity could be most readily incorporated in a BSE for deciding a ram's breeding efficiency.

SY4-5. Temporary hypertension as well as white coat hypertension in the first trimester as risk factors for preeclampsia

This study investigated the relationship among estrogen, placental blood flow and placental weight gain in rats treated with ketoconazole. Oral administration of ketoconazole (25 mg/kg/day) on Days 12–14 of pregnancy induced reduction of plasma estradiol-17β (E2) concentration, transient decrease in placental blood flow and increased intensity of a hypoxia-related marker in the placenta on Day 14 of pregnancy. On Day 20 of pregnancy, placental weights of ketoconazole-treated rats increased when compared to controls. Histologically, maternal sinusoidal area of the placenta decreased on Day 14 of pregnancy and the total area of maternal and fetal sinusoids increased on Day 20. All the changes disappeared by concomitant subcutaneous infusion of E2. These results indicate that ketoconazole-induced E2 deficiency causes transient decrease in placental blood flow associated with hypoxia and later placental weight gain in rats.

Follicular dynamics during the pre-ovulatory and subsequent first follicular wave stages affect the pregnancy outcome in Japanese Black cows.

The diameters of the pre-ovulatory follicles (PF) and the largest follicle during the subsequent first follicular wave (W1LF), and plasma estradiol-17β (E2) concentrations were monitored on Days 0, 1, 3, 5, and 7 (ovulation = Day 1). Pregnancy was diagnosed on Day 30. Cows were classified into two groups according to the location of the dominant follicle ipsilateral (IG) or contralateral (CG) to the corpus luteum on Day 7. From Days 3 to 7, some follicles that had been determined as the subordinate in the previous examination exceeded the W1LF located in the opposite ovary in terms of the diameter. These were defined as switching (SW), whereas others were defined as non-switching (NSW). The diameter of PF was significantly smaller in pregnant (P) animals than in non-pregnant (NP) animals. The plasma E2 concentration on Day 0 was significantly higher in P animals than in NP animals and tended to be higher in NSW than in SW. In addition, plasma E2 concentrations around Days 3 to 7 tended to be higher in P animals of NSW than in NP animals of SW. The conception rates did not differ between IG and CG but were significantly higher in NSW than in SW. In the IG group, the conception rate tended to be higher in NSW than in SW.

Re-evaluation of reproductive cycle and fecundity of finetooth sharks Carcharhinus isodon (Valenciennes 1839) from the Northwest Atlantic Ocean, with new observations on ovarian cycle and reproductive endocrinology of biennially reproducing sharks.

To provide updated information on life history for improved fishery management, the reproductive cycle of the finetooth shark Carcharhinus isodon from Northwest Atlantic (NWA) populations was investigated by examining temporal changes in morphology and histology of reproductive organs. Changes in plasma concentrations of gonadal sex hormones in relation to reproductive stage were also examined. Increases in testis width, epididymis head width, plasma testosterone concentrations and occurrence of mature spermatozoa were observed in male sharks between December and April, suggesting a seasonal pattern in reproduction that culminates with copulatory activity in May. Increases in maximum follicular diameter, oviducal gland width, plasma 17β-estradiol concentrations and occurrence of vitellogenic follicles were observed in non-pregnant female sharks during the same time period along with the occurrence of newly pregnant females in May, demonstrating strong synchronicity between male and female reproductive cycles. Pregnant females bearing full-term embryos were also observed in May, indicating that parturition occurs between mid-May and early June and gestation requires 12 months. Only transient temporal changes in follicle size and oviducal gland width were observed in pregnant females, indicating that reproductive periodicity is biennial; nonetheless, a single female exhibiting signs of concurrent vitellogenesis and pregnancy was observed. Mean brood size ± S.D. was 3.9 ± 0.9 offspring/female. Fecundity was not significantly correlated with female size, in part due to an unexpectedly high rate of early embryo mortality, which occurred in 11% of pregnant females, and was more common in larger individuals. Changes in ovarian activity during mid-pregnancy were observed, suggesting possible roles for the ovary in regulating some aspects of early to mid-gestation. This study confirms that earlier characterizations of the reproductive cycle and fecundity in NWA finetooth sharks remain valid for use in fishery management. This study also highlights unusual features of finetooth shark pregnancy (e.g., early embryo death, mid-pregnancy changes in ovarian function) that may have broader relevance to understanding elasmobranch reproduction.

Prenatal maternal vitamin D deficiency sex-dependently programs adipose tissue metabolism and energy homeostasis in offspring.

In utero environment is crucial to ensure normal development of the fetus and to program metabolic health throughout the life. Beside macronutrients, the role of micronutrients, including vitamin D, begins to be explore. The aim of this study was to decipher the impact of maternal vitamin D deficiency (VDD), in normal and high-fat (HF) diet context, on adipose tissue metabolism and energy homeostasis in offspring, considering sex-specific responses. Body weight, energy expenditure, and spontaneous activity was differential impacted in juvenile male and female offspring born from VDD mice. In adulthood, a HF diet combined with maternal VDD disrupted glucose homeostasis and adiposity in male offspring but not in females. Such phenotypes were associated to different transcriptomic profiles in adipose tissue, which could be related to differential modulation of plasma 17β-estradiol concentrations. Thus, maternal VDD sex-dependently modulated metabolic fate of the offspring, especially when associated with HF diet in adulthood.

Impact of herbicide pretilachlor on reproductive physiology of walking catfish, Clarias batrachus (Linnaeus).

Herbicide pretilachlor is widely used in paddy fields to control annual weeds. The present study has been carried out in walking catfish, Clarias batrachus, to evaluate the impact of herbicide pretilachlor on reproductive physiology after chronic exposure. Based on the median lethal concentration value (96 h), fish were exposed to three nominal test concentrations of pretilachlor ((SL-I (1/20th LC50), SLII (1/15th LC50), and SL-III (1/10th LC50)) for 30, 45, and 60 days after which plasma sex steroid profile, plasma vitellogenin concentration, and gonadal aromatase activity were analyzed in both sexes. Plasma concentration of testosterone decreases in herbicide-exposed male fish. Significant increase in plasma 17β-estradiol, plasma vitellogenin concentration, and gonadal aromatase activity were observed in herbicide-exposed male fish. All these alterations in reproductive parameters in male fish are dependent on concentration and exposure duration of herbicide. On the other hand, significant decrease in plasma concentration of testosterone was observed in female fish which was also dependent on concentration and exposure duration of herbicide. No significant changes in plasma 17β-estradiol concentrations, plasma vitellogenin concentration, and gonadal aromatase activity were observed in female fish. Above findings clearly suggested that herbicide pretilachlor acts as endocrine disruptor in fish and affects overall reproductive physiology of fish, but its ability to induce reproductive toxicity in male and female differs considerably.

Plasma estradiol-17β, cortisol, and insulin concentrations and serum biochemical parameters surrounding puberty in Japanese Black beef bulls with normal and abnormal semen

The associations of semen abnormalities with circulating hormones (estrogens, glucocorticoid, insulin) and common biochemical parameters are unclear in beef bulls. We compared plasma concentrations of estradiol-17β, cortisol, and insulin and serum biochemical parameters surrounding puberty in Japanese Black beef bulls (n = 96) with normal post-thaw or abnormal semen (fresh and frozen). Blood samples were collected monthly from 4 to 24 months of age (n = 50) for the assays of plasma estradiol, cortisol, and insulin and every 3 months from 6 to 21 months of age (n = 92) for the serum biochemical analyses. Semen was collected weekly from 12 months until at least 18 months of age. Fresh semen was evaluated for semen volume, sperm progressive motility, concentrations, and morphological defects. The normal fresh semen was frozen by a standard method and examined for post-thaw sperm motility and fertility, which were evaluated for rates of transferable embryos. Bulls were classified as having either normal fresh semen or abnormal fresh semen (when at least one of the above test items was abnormal for 6 months). The normal fresh semen was categorized as having either normal post-thaw semen or low fertility post-thaw semen. The abnormal fresh semen was categorized as having sperm morphological defects, low motility, or morphological defects plus low motility. Plasma cortisol concentrations in the abnormal fresh semen group were higher than those of the normal fresh semen group (p < 0.0001). Plasma estradiol-17β and insulin concentrations in the low-fertility post-thaw semen group were lower than those of the normal post-thaw semen group (p < 0.0001). Serum aspartate aminotransferase and magnesium concentrations were greater for the abnormal fresh semen group vs. the normal fresh semen group (p < 0.005). These results suggest that fresh semen abnormality in pubertal beef bulls might be associated with increased circulating aspartate aminotransferase, magnesium and cortisol. Low-fertility post-thaw semen could have been involved with the lower peripheral estradiol and insulin levels in beef bulls.

Sleep deprivation decreases the reproductive capacity by affecting the arrival of morulas in the uterus.

A previous animal study by our group found that sleep deprivation during preimplantation was associated with decreased pregnancy maintenance. Given its impact on human society, we aimed in the current study to assess whether sleep deprivation affects blastocyst gene expression and/or the implantation process. For this, pregnant mice (gestational day 0 [GD 0]) were assigned into paradoxical sleep deprivation (SD, 72 hr; multiple platform method) and, a control (CT) group. Animals were euthanized on GD 3.5 and blood, uterus (embryos) and fallopian tube were collected. Then, 89% of CT presented blastocysts in the uterus versus 25% from SD group. Compared to CT, SD presented lighter relative uterus weight, increased plasma concentrations of corticosterone and testosterone, decreased concentrations of progesterone and luteinizing hormone, but no statistical differences in plasma concentrations of 17β-estradiol and follicle stimulating hormone. There were no differences in uterus and blastocyst gene expression related to embryo implantation and development, and no alteration in blastocysts global DNA methylation. Considering this, the decreased pregnancy maintenance after sleep deprivation seems not to be associated with implantation losses or developmental problems related to the blastocysts. It is likely that complications in morula development and/or its movement through the fallopian tubes affect the pregnancy rate, since only 25% of SD females presented a blastocyst on the GD 3.5. In fact, three out of four females without blastocysts in the uterus presented morula in the fallopian tubes due to a phase delay. Additionally, we suggest that the observed hormonal changes may play a role in this outcome.

Annual changes in plasma progesterone and estradiol-17β concentrations compared to pituitary-adrenal axis activity in the female goat reared under arid environment

ABSTRACT This study investigates the relationships between the pituitary-adrenal axis and the ovarian activity in the cyclic goat reared under arid environment. Plasma hormone levels are assessed on blood samples collected weekly at 9:00 a.m., during one year. The ovarian activity is characterized by an anestrous period from mid-March to mid-July and a breeding period during the rest of the year. Adrenal histomorphometry and immunolocalization of Estrogen Receptors α (ERα) and Progesterone Receptors (PR) were performed on adrenal glands collected during each season. During the anestrous period, all adrenal compartments appeared enlarged, as did cellular and nuclear areas. ERα and PR immunostaining were identified in all the adrenal gland zonae with more intensity for the ERα. ERα immunostaining is greater in zona fasciculata whereas PR immunostaining is more intense in zona fasciculata and zona reticularis during the breeding season. Plasma ACTH shows no difference between the two seasons, while cortisol rises slightly during the anestrous period. There is a negative but not significant correlation between progesterone levels with plasma ACTH and cortisol. The pituitary-adrenal axis is activated essentially at the anestrous onset and at the ovarian resumption which represents stressful events, but it varies slightly between these two seasons in this goat.

A model of pre-pubertal broiler breeder estradiol-17β levels predicts advanced sexual maturation for birds with high body weight or short juvenile day-length exposure

ABSTRACTAs broiler breeders face increased reproductive challenges specifically related to overfeeding, a clear understanding of the physiological effects of BW and rearing photoperiod on reproductive development is needed. The objective was to use mathematical models to compare plasma estradiol-17β (E2) concentration to characterize the effect of BW and rearing photoperiod on E2 levels. A 2 × 3 factorial arrangement of treatments was used. Hens (n = 180) were fed with a precision feeding system to allocate feed individually to achieve the breeder-recommended BW curve (Standard) or to a BW curve reaching the 21 wk target at 18 wk (High). Hens were on 8L:16D, 10L:14D, or 12L:12D photoschedules during rearing and were photostimulated at 21 wk. Age at first egg (AFE) was recorded. Plasma E2 levels were determined weekly between week 20 and 28. Two modified Gompertz models described E2 level as a function of (a) chronological or (b) physiological (relative to AFE) age. Timing of E2-inflection point was compared between models and treatments. Differences were reported as significant at P ≤ 0.05. The chronological age model inferred that High BW reduced the duration between the E2-inflection point and AFE, whereas the physiological age model inferred that High BW only reduced the duration between photostimulation and the E2-inflection point. Hens on the Standard BW treatment had a longer period between photostimulation and the E2-inflection point compared to hens on the High-BW treatment (11.03 vs. 1.50 wk, respectively, based on physiological age). Hens on the 12L:12D photoschedule had a longer period between photostimulation and the E2-inflection point compared to hens on the 8L:16D or 10L:14D photoschedule, both in the Standard and High BW (28.91 vs. 1.78 and 2.40 wk, 2.65 vs. 0.93 and 0.94 wk, respectively, based on physiological age). The described methodology and results provide quantitative insight into E2 dynamics and serves as a model for future endocrinological studies in poultry reproduction.

Monitoring follicular dynamics using ultrasonography in captive brown bears (Ursus arctos) during the breeding season

Artificial insemination (AI) may be a useful tool in the reproductive management of endangered animals, including bears. To establish an AI program for bears, we investigated follicular dynamics using weekly transrectal ultrasonography in six captive brown bears. Along with ultrasonography, we monitored plasma progesterone (P4) and estradiol-17β (E2) concentrations. Furthermore, two bears were administered a gonadotropin releasing hormone (GnRH) agonist to induce ovulation on the first day on which the largest follicle reached more than 10.0 mm in diameter. Brown bears showed two patterns of follicular development in the early and late periods of the breeding season. In the early period (May to mid-June), multiple follicular waves were observed; namely, many follicles developed, and the largest follicles grew to less than 6.0 mm in diameter then regressed. In the late period (mid-June to July), one or two follicles grew to greater than 6.0 mm in diameter and developed as dominant follicles. Moreover, the growth rate of the largest follicle in the late period was faster than that in the early period of the breeding season. One bear with a follicle of 13.1 mm ovulated spontaneously, and one bear ovulated when the follicle was 10.2 mm in diameter after GnRH agonist treatment. Plasma E2 concentrations increased and showed peaks five to seven days before the largest follicles reached their maximum size. Plasma P4 concentrations increased on the day the corpus luteum could be detected using ultrasonography. This is the first study that showed there are two patterns of follicular development in brown bears. Furthermore, the largest follicle reaching greater than 10.0 mm in diameter could be an indicator of the appearance of ovulatory follicles.

Effects of 17α-ethynylestradiol (EE2) on the immune system of Juvenile European sea bass with a special focus on B and T cells

Synthetic progesterones and estrogens are broadly used bioactive pharmaceutical agents and have been detected in aquatic environments. In the present study, we investigated the combined reproductive effects of megestrol acetate (MTA) and 17α-ethinylestradiol (EE2) on zebrafish. Adult zebrafish were exposed to MTA (33, 100 or 333 ng/L), EE2 (10 ng/L) or a mixture of both (MTA + EE2: 33 + 10, 100 + 10 or 333 + 10 ng/L) for 21 days. Results demonstrated that egg production was significantly reduced by exposure to 10 ng/L EE2, but not MTA. However, a combined exposure to MTA and EE2 caused further reduction of fish fecundity compared to EE2 exposure alone, suggesting an additive effect on egg production when EE2 is supplemented with MTA. Plasma concentrations of 17β-estradiol and testosterone in the females and 11-ketotestosterone in the males were significantly decreased in the groups exposed to EE2 or MTA alone compared with the solvent control, and the plasma concentrations of the three hormones were further reduced in the co-exposure groups relative to the MTA exposure group, but not the EE2 exposure group. These data indicate that the inhibitory effects on plasma concentrations in the co-exposures were predominantly caused by EE2. Furthermore, exposure to MTA and EE2 (alone or in combination) led to histological alterations in the ovaries (decreased vitellogenic/mature oocytes), but not in the testes. This study has important implications for environmental risk assessment of synthetic hormones that are concurrently present in aquatic systems.

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Is the effect of fluoxetine on plasticity related to serotonin transporter or tropomyosin-related receptor kinase B?

The associations of semen abnormalities with circulating hormones (estrogens, glucocorticoid, insulin) and common biochemical parameters are unclear in beef bulls. We compared plasma concentrations of estradiol-17β, cortisol, and insulin and serum biochemical parameters surrounding puberty in Japanese Black beef bulls (n = 96) with normal post-thaw or abnormal semen (fresh and frozen). Blood samples were collected monthly from 4 to 24 months of age (n = 50) for the assays of plasma estradiol, cortisol, and insulin and every 3 months from 6 to 21 months of age (n = 92) for the serum biochemical analyses. Semen was collected weekly from 12 months until at least 18 months of age. Fresh semen was evaluated for semen volume, sperm progressive motility, concentrations, and morphological defects. The normal fresh semen was frozen by a standard method and examined for post-thaw sperm motility and fertility, which were evaluated for rates of transferable embryos. Bulls were classified as having either normal fresh semen or abnormal fresh semen (when at least one of the above test items was abnormal for 6 months). The normal fresh semen was categorized as having either normal post-thaw semen or low fertility post-thaw semen. The abnormal fresh semen was categorized as having sperm morphological defects, low motility, or morphological defects plus low motility. Plasma cortisol concentrations in the abnormal fresh semen group were higher than those of the normal fresh semen group (p < 0.0001). Plasma estradiol-17β and insulin concentrations in the low-fertility post-thaw semen group were lower than those of the normal post-thaw semen group (p < 0.0001). Serum aspartate aminotransferase and magnesium concentrations were greater for the abnormal fresh semen group vs. the normal fresh semen group (p < 0.005). These results suggest that fresh semen abnormality in pubertal beef bulls might be associated with increased circulating aspartate aminotransferase, magnesium and cortisol. Low-fertility post-thaw semen could have been involved with the lower peripheral estradiol and insulin levels in beef bulls.

Body condition and reproductive status of chum salmon during homing migration in Otsuchi Bay, Japan, after the 2011 Great East Japan Earthquake and Tsunami

To understand how the earthquake and tsunami on March 11 2011 affected homing chum salmon Oncorhynchus keta in Otsuchi Bay, Japan, we monitored the body condition and reproductive status at the mouth of the bay in October, November, December, and January from 2012 to 2016. The average fork length in 2014 was larger than that in 2012 and 2015, due to the high proportion of 5- and 6-year-old salmon and the low proportion of 3- and 4-year-old salmon, possibly because of the decrease in the salmon fry released by the hatchery program after the disaster. Plasma estradiol-17β (E2) concentrations in females were high in October and November and declined in December and January, whereas plasma 11-ketotestosterone concentrations in males were consistently high throughout the sampling period. Plasma 17α,20β-dihydroxy-pregnen-3-one (17α,20β-P) increased in both sexes in December and/or January. These changes were consistently observed from 2012 to 2016; however, the rise in 17α,20β-P and decline in E2 levels in female salmon in December were conspicuous in 2013 and 2014, reflecting changes in age composition. These results suggest that the earthquake and tsunami temporarily affected the age composition and the physiological condition of chum salmon returning to Otsuchi Bay.