Validation of enzyme immunoassays for quantifying sex steroid hormones in tropical screech owls (Megascops choliba).

Androgens and estrogens are steroid hormones that regulate reproductive processes in both males and females. Monitoring plasma levels of these steroids or their metabolites present in feces, offers diagnostic support for assessing the reproductive status of animals. Immunoassays are commonly used methods for quantifying these hormones, but their protocols require species-specific validation to ensure reliability. The objective of this study was to perform analytically and biologically validation of enzyme immunoassay (EIA) kits for measuring testosterone (T), estradiol (E2), faecal androgen metabolites (fAM), and faecal estrogenic metabolites (fEM) in the tropical screech owl (Megascops choliba). Serum and fecal samples were collected from six adult females and six males both before and during breeding season, with males' gonadal activity assessed using electroejaculation (EE). The parallelism test confirmed the immunogenic similarity of the antigens in the estradiol and testosterone standards and the antigens in the serum samples and fecal extracts of M. choliba. Additionally, the EIA kits displayed nearly 100% recovery rates, and showed coefficients of variation ranging from 8% to 14% at the intra-assay level and from 10% to 16% at the inter-assay level, underscoring result reliability and consistency. In males, the highest serum T and fAM levels were recorded concurrently with the presence of spermatozoa in samples collected via EE. Although females did not exhibit oviposition events, significantly higher E2 and fEM levels were observed in August compared to May, suggesting potential seasonal variations in estrogenic hormone production. Fecal androgen and estrogen levels were significantly different between sexes in August, with males having higher fAM and females having higher fEM levels. Overall, the immunoassays validated in this study were found to be efficient in diagnosing reproductive activity in owls.

Effects of prepubertal exposure to silver nanoparticles on reproductive parameters in adult male Wistar rats.

The incidence of male reproductive pathologies, such as hypospadias, cryptorchidism, testicular cancer, and low sperm production in adulthood, is increasing and may be related to exposure to environmental contaminants. The silver nanoparticles (AgNP) are a new class of chemical compounds commonly used in both medical and nonmedical settings, and they affect development of spermatogonial stem cells in vitro. The aim of this study was to examine the adverse productive toxic effects of AgNPs in male Wistar rats exposed during the prepubertal period and sacrificed at postnatal day (PND) 53 and PND90. Growth was assessed by daily weighing. The progress of puberty in the rats was measured by preputial separation, while spermatogenesis was assayed by (1) measuring the sperm count in testes and epididymis and (2) examining the morphology and morphometry of seminiferous epithelium using stereological analysis. In addition, testosterone and estradiol levels were assayed by radioimmunoassay. The weight of the animals at PND90 did not change markedly, but growth was less in the group treated with AgNP at 50 µg/kg from PND34 to PND53. AgNP exposure produced a delay in puberty in both treated groups. Decreased sperm reserves in the epididymis and diminished sperm transit time were observed at PND53, while a reduction in sperm production occurred at PND90. The morphology of the seminiferous epithelium was markedly altered. Data demonstrated that prepubertal exposure to AgNP altered reproductive development in prepubertal male Wistar rats, as evidenced by impairment in spermatogenesis and a lower sperm count in adulthood.

Glucocorticoid metabolites in an ex situ nocturnal bird, the tropical screech owl Megascops choliba: effects of sex, activity period and inter-individual variation.

Glucocorticoids mediate physiological processes to obtain energy, presenting daily variation in basal levels that may be related to behavioural activity pattern. Identification of plasticity in the secretion of these hormones is essential to understand their effects on physiology and behaviour of wild birds and, therefore, their success in their natural or artificial environment. Serial endocrine evaluations are facilitated by implementing non-invasive methodologies that minimize possible effects of manipulation on the animal's physiological variables. However, non-invasive endocrine-behavioural studies in nocturnal birds, such as owls, are immature. The present work aimed to validate an enzyme immunoassay (EIA) to quantify glucocorticoid metabolites (MGC) in Megascops choliba as well as to evaluate differences in their production at the individual, sexual or daily level. We recorded the behaviour of nine owls during three continuous days to establish activity budget under captive conditions and aiming to correlate with daily MGC variation. The EIA proved to be effective in analytical assays and in pharmacological testing with synthetic ACTH, validating this immunoassay for the species. Additionally, individual differences in MGC production were confirmed in relation to the time of day, especially at 1700 and 2100, but not in relation to sex. During night hours, the owls showed greater behavioural activity, positively related to MGC values. Higher MGC concentrations were significantly related to greater expressions of active behaviours, such as maintenance, while lower MGC concentrations were recorded during moments of higher alertness and resting. The results presented show daily MGC variation to be inversed in this nocturnal species. Our findings can aid future theoretical studies of daily rhythm and evaluations of challenging and/or disturbing situations that result in changes in behaviour or hormonal cascades of these changes in ex situ populations of owls.

Quantitative proteomic profile analysis of thyroid dysfunction effects on seminal vesicles and repercussions on male fertility.

Hypothyroidism and thyrotoxicosis are associated with male reproductive disorders, but little is known about the influence of the thyroid hormone milieu on seminal vesicle (SV) function and metabolism. In this sense, we investigated the effects of hypothyroidism and thyrotoxicosis induced in adulthood Wistar male rats on SV function and identified new thyroid hormone targets on male reproduction regulation using novel proteomic approaches. Hypothyroidism reduces SV size and seminal fluid volume, which are directly associated with low testosterone and estradiol levels, while thyrotoxicosis increases Esr2 and Dio1 expression in the SV. We found 116 differentially expressed proteins. Hypothyroidism reduces the expression of molecular protein markers related to sperm viability, capacitation and fertilization, protection against oxidative stress and energetic metabolism in SV, while it increases the expression of proteins related to tissue damage. In conclusion, thyroid dysfunction in the adult phase impairs several morphological, molecular and functional characteristics of SV.

Additional effects using progestins in mares on levels of thyroid hormones and steroids in neonates.

The risk of pregnancy loss in mares leads to the use of exogenous hormones to help pregnancy maintenance. The objective was to evaluate the proportion of thyroid hormones and steroids in neonates, in the following postpartum period, born to mares fed with synthetic progesterone and to verify the existence of a correlation between the level of progesterone between mother and neonate. Twenty-seven mares and their foals were used. The animals were divided into 5 experimental groups: group 1 (control, without hormonal supplementation), group 2 (random samples fed to 120 days of pregnancy with long-term progesterone), group 3 (mares fed with short-term progesterone as of 280.º day of pregnancy), group 4 (mares fed with long-term progesterone as of 280.º day of pregnancy) and group 5 (mares fed with synthetic hormone [altrenogest] as of 280.º day of pregnancy). The animal's blood collection took place immediately after parturition for the hormonal measurement. The hormones measured in neonates were total T3, free T4, TSH, progesterone and cortisone. In mares, only levels of progesterone. The groups of neonates showed no difference on levels of total T3, free T4, TSH and progesterone. There was no difference on levels of progesterone in mares among the groups. Neonates from groups 4 and 5 had higher and lower cortisone levels, respectively. No neonate showed clinical change. There was also no correlation between levels of progesterone in mares and foals. Thus, hormonal supplementation with long-term progesterone as of 280 days of pregnancy leds to an increase in the neonate's cortisone levels, in the meantime, supplementation with altrenogest as of 280 days of pregnancy caused a decrease on cortisone levels in foals, despite clinical signs have not been observed on these animals.

Glyphosate impairs male offspring reproductive development by disrupting gonadotropin expression.

Sexual differentiation in the brain takes place from late gestation to the early postnatal days. This is dependent on the conversion of circulating testosterone into estradiol by the enzyme aromatase. The glyphosate was shown to alter aromatase activity and decrease serum testosterone concentrations. Thus, the aim of this study was to investigate the effect of gestational maternal glyphosate exposure (50 mg/kg, NOAEL for reproductive toxicity) on the reproductive development of male offspring. Sixty-day-old male rat offspring were evaluated for sexual behavior and partner preference; serum testosterone concentrations, estradiol, FSH and LH; the mRNA and protein content of LH and FSH; sperm production and the morphology of the seminiferous epithelium; and the weight of the testes, epididymis and seminal vesicles. The growth, the weight and age at puberty of the animals were also recorded to evaluate the effect of the treatment. The most important findings were increases in sexual partner preference scores and the latency time to the first mount; testosterone and estradiol serum concentrations; the mRNA expression and protein content in the pituitary gland and the serum concentration of LH; sperm production and reserves; and the height of the germinal epithelium of seminiferous tubules. We also observed an early onset of puberty but no effect on the body growth in these animals. These results suggest that maternal exposure to glyphosate disturbed the masculinization process and promoted behavioral changes and histological and endocrine problems in reproductive parameters. These changes associated with the hypersecretion of androgens increased gonadal activity and sperm production.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs.

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.

Puberty and oestral cycle length in captive female jaguars Panthera onca.

Habitat loss and fragmentation have been leading jaguars to constant conflicts with humans, and as a result, jaguar populations have been declining over the last decades. Captive breeding is often a tool for species conservation, and it is not different for jaguars. However, success is far from optimal due to the lack of basic knowledge about species' reproductive biology. In the present study, we assessed gonadal hormonal profiles of natural oestral cycles and puberty and compared our data to those of other wild felids. We collected faecal samples from two to seven times per week for 18 months from two adults and three pre-pubertal females. We defined baseline levels for progestins and oestrogens in order to estimate oestrous cycle length and age at puberty. We compared our data with 16 other species through generalized linear model, using weight and genus as two explanatory variables. Cycle length was 38.28 ± 2.52 days, ranging from 25 to 44 days, while sexual maturity was attained within 22 months. Due to our analysis of both hormonal and behavioural data, there is a variation between this research from other studies that employed only behavioural observations. Such difference may be caused by the absence of behavioural oestrous at the peri-pubertal period. When compared to wild felids of similar size, puberty and oestral cycle durations of the jaguar females fell within the same range. Our modelling showed that age at maturity was influenced mostly by size and only Leopardus partially explained the observed variation. Conversely, oestral cycle length did not differ among genera or size categories. Our study adds to the body literature in the reproductive endocrinology of wild felids, and because female gametes are more challenging to collect and preserve, a strong understanding on the female reproductive physiology is essential to assisted reproduction and wild population viability assessment.

First assessment of hoary fox (Lycalopex vetulus) seasonal ovarian cyclicity by non-invasive hormonal monitoring technique.

Reproduction is key to species survival, and reproductive physiology represents a high priority investigative area for conservation biology, as it provides a basic understanding of critical life-history traits, information that is helpful for the establishment of management strategies. Here, we generated knowledge about the reproductive endocrinology of the hoary fox (Lycalopex vetulus), a small canid (2.5-4 kg) endemic to open areas of the Brazilian Cerrado and listed in the Brazilian National List of Endangered species. Specifically, we utilized non-invasive hormone monitoring methods to assess oestrogen and progestagen metabolites from eight female hoary foxes housed in five zoological institutions in the state of São Paulo-Brazil. We observed the elevations of oestrogen and progestagen metabolites between July and September in six of the eight females. No significant evidence of ovarian activity was observed during other months. Two females, who shared the same enclosure, did not show a pattern of reproductive cyclicity. Based on these characteristics, we concluded that captive hoary foxes are seasonal monoestric, with the beginning of the oestrus cycle occurring mainly in July followed by 2 months of the luteal phase when conception does not occur. We suggest the dosage of faecal metabolites of estradiol and progesterone could be used to differentiate the reproductive period from a non-reproductive period in Lycalopex vetulus females, providing relevant information about their reproductive biology that may contribute to species conservation and management strategies, such as increased ex situ reproductive success.

Imbalanced testicular metabolism induced by thyroid disorders: New evidences from quantitative proteome.

Thyroid dysfunctions, such as hypothyroidism and hyperthyroidism, are the second most prevalent endocrinopathies and are associated to reproductive disorders in men. Several genes are differentially modulated by thyroid hormones in testes and imbalances in thyroid hormone levels are also associated to alterations on sperm functionality. Imbalances on antioxidant defense mechanism and stress oxidative have been pointed out as the main factors for the impairments on male reproductive function. To clarify this issue, we investigated the expression and activity of antioxidant enzymes in testis, followed by their proteomic profile in attempt to characterize the mechanisms involved in the alterations induced by hypo- or hyperthyroidism in adult male rats. Hypothyroidism reduced the Gsr transcript expression and the activity of CAT and GSR enzymes, while the hyperthyroidism reduced the Gpx4 var2 transcript expression. Among 1082 identified proteins, 123 and 37 proteins were downregulated by hypothyroidism compared to euthyroid and hyperthyroid condition, respectively, being 36 proteins commonly reduced in both comparisons and one exclusively in hypo-hyperthyroidism comparison. A network containing 29 nodes and 68 edges was obtained in protein-protein interaction analysis and the functional enrichment analysis of differentially expressed proteins revealed significant alterations for several functions in hypo-euthyroid and hypo-hyperthyroid comparisons, such as ATP metabolic process, coenzyme binding, sperm part, peroxiredoxin activity, mitochondrial protein complex, intramolecular oxidoreductase activity, binding of sperm to zona pellucida, glutathione transferase activity, response to testosterone. Thus, there is a correlation between thyroid disorders and impaired antioxidant defense mechanism, resulting in reproductive dysfunctions, as infertility, mainly observed in hypothyroidism.

Noninvasive monitoring of androgens in male Amazonian manatee (Trichechus inunguis): biologic validation.

The Amazonian manatee (Trichechus inunguis) is endemic in the Amazonian basin and is the only exclusively fresh water sirenian. Historically hunted on a large scale, this species is now considered endangered, and studies on the reproductive physiology are critical for the improvement of reproductive management of captive and wild populations of manatees. The aim of this study was to verify the viability of androgen measurement in saliva, lacrimal, urine, and fecal samples of the Amazonian manatee by conducting a hormone challenge. Two adult male manatees (A-1 and A-2) were submitted to an experimentation protocol of 12 day (D1 to D10). On D0, the animals received an intramuscular injection of gonadotropin-releasing hormone (GnRH)-analogue. Salivary, lacrimal, urinary, and fecal samples were collected daily (between 0800 hours and 0900 hours) and frozen at -20 degrees C until assayed. Fecal samples were lyophilized, extracted with 80% methanol, and diluted in buffer before the radioimmunoassay (RIA). Urine samples underwent acid hydrolysis and were diluted in depleted bovine serum. Salivary and lacrimal samples were assayed without the extraction step. Hormonal assays were conducted with a commercial testosterone RIA kit. An androgen peak (> median + 2 interquartile range [IQR]) was observed in all matrices of both animals, although it was less prominent in the lacrimal samples of A-2. However, the fecal androgen peak (A-1 peak = 293.78 ng/g dry feces, median [IQR] = 143.58 [32.38] ng/g dry feces; A-2 peak = 686.72 ng/g dry feces, median [IQR] = 243.82 [193.16] ng/g dry feces) occurred later than urinary (A-1 peak = 648.16 ng/mg creatinine [Cr], median [IQR] = 23.88 [30.44] ng/mg Cr; A-2 peak = 370.44 ng/mg Cr, median [IQR] = 113.87 [117.73] ng/mg Cr) and salivary (A-1 peak = 678.89 pg/ml, median [IQR] = 103.69 [119.86] pg/ml; A-2 peak = 733.71 pg/ml, median [IQR] = 262.92 [211.44] pg/ml) androgen peaks. These intervals appear to be correlated with the long digesta passage time in this species. The salivary and urinary peaks were closely associated. These results demonstrate that androgen concentrations in saliva, urine, or feces samples reflect reliably physiologic events and are a powerful tool for noninvasive reproductive monitoring of Amazonian manatees.

Measurement of thyroid hormones (thyroxine, T4; triiodothyronine, T3) in captive nondomestic felids.

The aim of this research was to obtain basic values for the evaluation of thyroid function in nondomestic felids. Serum thyroid hormone concentrations (thyroxine, T4; triiodothyronine, T3) were measured by radioimmunoassay in 145 cats, representing nine species of captive nondomestic felids: jaguar (Panthera onca), n = 49; puma (Puma concolor), n = 10; ocelot (Leopardus pardalis), n = 22; oncilla (Leopardus tigrinus), n = 12; geoffroy (Oncifelis geoffroyi), n = 4; jaguarundi (Herpailurus yaguarondi), n = 8; margay (Leopardus wiedii), n = 7; lion (Panthera leo), n = 26; and tiger (Panthera tigris), n = 7. For each species, mean +/- SEM of T3 and T4, respectively, were as follows: jaguar, 0.56 +/- 0.03 and 9.7 +/- 0.8 ng/ml; puma, 0.67 +/- 0.04 and 11.2 +/- 1.2 ng/ml; ocelot, 0.48 +/- 0.03 and 13.8 +/- 1.5 ng/ml; oncilla, 0.43 +/- 0.03 and 10.0 +/- 1.6 ng/ml; geoffroy, 0.44 +/- 0.04 and 8.0 +/- 0.16 ng/ml; jaguarundi, 0.7 +/- 0.03 and 5.0 +/- 1.0 ng/ml; margay, 0.48 +/- 0.04 and 12.2 +/- 2.3 ng/ml; lion, 0.43 +/- 0.02 and 5.7 +/- 2.6 ng/ml; and tiger, 0.66 +/- 0.03 and 12.6 +/- 0.9 ng/ml. Within species, T3 and T4 concentrations did not differ (P > 0.05) between males and females.

New insights for male infertility revealed by alterations in spermatic function and differential testicular expression of thyroid-related genes.

The impact of thyroid hormone (TH) disorders on male reproductive biology has been a controversial issue for many years. Recently, we reported that hypothyroid male rats have a disruption of the seminiferous epithelium, which may compromise spermatogenesis. To improve the understanding of the reproductive pathogenesis of hypothyroidism and hyperthyroidism, male Wistar rats that developed these dysfunctions in adulthood were used as an experimental model. We evaluated the sperm production, reserves, transit time, morphology, and functionality (acrosome integrity, plasma membrane integrity, and mitochondrial activity), and the testicular expression of the TH receptors (Thra1 and Thra2, Thrb1, and Thrb2), deiodinases (Dio2 and Dio3), and the Mct8 transporter (Slc16a2) were assessed by reverse transcription followed by real-time quantitative PCR (RT-qPCR). The results were evaluated statistically by ANOVA and Tukey HSD test (P < 0.05). Hypothyroidism decreased the total and daily sperm productions and increased the sperm transit time through the epididymis, while the sperm functionality was reduced in both thyroid dysfunctions. Regarding the modulation of gene expression in the testis, hypothyroidism increased the expression of Thra1 and decreased the expression of Dio3, and hyperthyroidism increased the expression of Slc16a2. The observed alterations in spermatic production and function and in the expression of the TH receptor, deiodinase, and the TH transporter are suggestive of TH participation in spermatogenesis in adulthood.

Immunohistochemical determination of estrogen receptor-alpha in vaginal and tumor tissues of healthy and TVT-affected bitches and their relation to serum concentrations of estradiol-17beta and progesterone.

One of the most frequent canine neoplasms is the transmissible venereal tumor (TVT), which affects the male and the female genital tract. The objective of this study was to determine (immunohistochemically) estrogen receptor (ER-alpha) expression in vaginal tissue of healthy bitches and in the vaginal and neoplastic tissues of TVT-affected bitches. Fifty-eight bitches were divided into two groups: tumor group (TVT) and control group (healthy). Canine estrous cycle stages were determined by means of exfoliative vaginal cytology, hormone assays, and macroscopic appearance of ovaries. Samples from vaginal and neoplastic tissues were obtained under general anesthesia, fixed in 10% buffered formaldehyde, embedded in paraffin, and sectioned. Anestrus, proestrus and estrus control females had higher ER-alpha expression than diestrus bitches. Within the tumor group, diestrus bitches had significantly higher ER-alpha expression. Although some samples had expression in the endothelium of blood vessels, no ER-alpha expression was observed in neoplastic tissues. In conclusion, vaginal tissue of tumor and control bitches, under different distinct steroid influences, had different ER-alpha expression, whereas ER-alpha expression was not present in neoplastic tissues.

Effect of a new device for sustained progesterone release on the progesterone concentration, ovarian follicular diameter, time of ovulation and pregnancy rate of ewes.

This study evaluated the effectiveness of a new progesterone intravaginal device (DPR) in ewes through four experiments: Experiment 1 compared the circulating progesterone concentration of ovariectomized ewes that received either a new or a re-used DPR. Experiment 2 compared the progesterone concentration between DPR-estrous-synchronized ewes and naturally estrous-cycling ewes. Experiment 3 evaluated the effect of new and re-used DPRs on ovarian follicular dynamics and time of ovulation of estrous cycling ewes. Experiment 4 compared the pregnancy rate after the use of a DPR and Controlled Internal Drug Releasing Device (CIDR). The mean concentration of progesterone released by the DPR device during its first use (New Group: 5.1 ± 0.5 ng/ml) was greater than that during the second use (Re-used Group: 2.4 ± 0.3 ng/ml). There was no difference between the animals that received DPR devices for first and second use in terms of ovulatory follicle diameter, follicular wave emergence day for ovulatory follicle and period of ovulatory wave of ovarian follicular development. However, there was a significant difference between groups regarding the time between DPR device removal and first ovulation (New Group: 71.7 ± 2.5h and Re-Used Group: 63.9 ± 2.7h). Pregnancy rates were similar between ewes with DPR and CIDR devices. It was concluded that DPR is effective in increasing and maintaining progesterone concentrations, controlling follicular dynamics, promoting synchronized times of ovulation from healthy follicles, promoting development of a competent corpus luteum and when used results in pregnancy rates similar to that with use of the CIDR.

Hypothyroidism in adult male rats alters posttranscriptional mechanisms of luteinizing hormone biosynthesis.

BACKGROUND: Studies in men are not consistent regarding the effects of thyroid hormone on the production of gonadotropins. In hypothyroidism consequent to diverse causes, an increase or no change in serum luteinizing hormone (LH) have been reported. The attempt to explain the mechanisms involved in this pathology using rats as an experimental model also seems to repeat this divergence, since hypothyroidism has been shown to induce hypogonadotropic hypogonadism, a hypergonadotropic state, or not to affect the basal levels of LH. Notably, the promoter region of the gene encoding the Lh beta subunit and GnRH (gonadotropin-releasing factor) does not contain a thyroid responsive element. Therefore, we investigated the hypothesis that, in male rats, posttranscriptional mechanisms of LH synthesis are altered in hypothyroidism. We also attempted to determine if hypothyroidism directly affects testicular function in male rats. METHODS: Male Wistar rats, 60 days old, were thyroidectomized or sham-operated. After 20 days, they were decapitated, and the pituitaries were collected and analyzed for Lh mRNA, LH content, poly(A) tail length, and polysome profile. The testes were collected and analyzed for Lh receptor mRNA, LH receptor content, and histology using morphometric analyses. The testis, epididymis, seminal vesicle, and ventral prostate were weighed, and serum concentrations of LH, testosterone, thyrotropin (TSH), and triiodothyronine (T3) were measured. RESULTS: Hypothyroidism was associated, in the pituitary, with an increase in Lh mRNA expression, a reduction in Lh mRNA poly(A) tail length, a reduction in the number of LH transcripts associated with polysomes. Pituitary LH was decreased but serum LH was increased from 102 to 543 pg/mL. Despite this, serum testosterone concentrations were decreased from 1.8 to 0.25 ng/mL. A decreased germinative epithelium height of the testes and a reduced weight of androgen-responsive tissues were observed (ventral prostrate: 74 vs. 23 mg/100 g body weight [BW]; seminal vesicle undrained: 280 vs. 70 mg/100 g BW; and seminal vesicle drained: 190 vs. 60 mg/100 g BW). CONCLUSIONS: Hypothyroidism in adult male rats has dual effects on the pituitary testicular axis. It alters posttranscriptional mechanisms of LH synthesis and probably has a direct effect on testicular function. However, these data suggest the possibility that reduced LH bioactivity may account in part for impaired testicular function.

Herbicide metolachlor causes changes in reproductive endocrinology of male wistar rats.

S-metolachlor is a chloroacetanilide herbicide widely used in the agriculture to control weeds and was demonstrated that it increases the activity of the aromatase enzyme in cell cultures, which may culminate as endocrine disruption action in vivo. To investigate this hypothesis, prepubertal Wistar male rats were exposed to metolachlor (5 or 50 mg/kg/day, NOEL for reproductive toxicity: 23.5-26.0 mg/kg/day) from PND23 (postnatal day) to PND53. During this period, the growth of the animals and the age and weight at puberty were recorded. In PND53, tissues were collected and the analysis of LH, FSH, testosterone, dihydrotestosterone (DHT), estradiol serum concentrations, morphometric evaluation of the seminiferous epithelium, and weight of the testes and the seminal vesicle (undrained and drained) was performed (Statistical difference: P < 0.05). Metolachlor caused an increase in serum concentrations of testosterone, estradiol, and FSH and a reduction in DHT but did not alter the LH. There were also observed a higher amount of fluid in the seminal vesicles, precocious puberty, and changes in morphology of the seminiferous epithelium of treated animals. We demonstrated in this paper that prepubertal exposure to S-metolachlor caused changes in reproductive endocrinology of male rats.

Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro / Reproductive endocrine and socio-sexual behavior study of black-tufted-marmoset (Callithrix penicillata) kept in captivity

A comunicação do estado reprodutivo nos primatas da família Callithrichidae, depende principalmente dos comportamentos sócio-sexuais como um sistema de sinalização primário, uma vez que nestas espécies a ovulação não é percebida pelos machos. Neste trabalho, os padrões de comportamentos sócio-sexuais foram analisados em conjunto com as concentrações de metabólitos fecais dos esteróides sexuais progesterona (MFP), estradiol (MFE) e testosterona (MFT) em casais cativos de Sagüi-de-tufos-pretos (Callithrix penicillata), nas diferentes fases do ciclo ovariano. O grupo estudado era composto por quarto casais adultos, mantidos no Centro de Reabilitação de Animais Selvagens da prefeitura de São Paulo. Os padrões comportamentais foram registrados pelo método de amostragem focal por intervalo de tempo a cada 30 segundos, cinco vezes por semana, totalizando 14.400 registros por animal. A mensuração das concentrações de metabólitos fecais dos esteroides sexuais foram realizados pelo método de enzima imunoensaio (EIE). Os resultados obtidos dessas concentrações possibilitaram a determinação endócrina das fases do ciclo ovariano (folicular e luteal) e de suas respectivas durações, assim como a determinação da fase periovulatória. Foram caracterizados 31 ciclos ovarianos completos, com duração de 24,3±4,1 dias (média ±DP), sendo que a fase folicular compreendeu 13,04±4,8dias e a fase lútea 11,2±4,2 dias. Os comportamentos sócio-sexuais (marcação por cheiro, cheirar genitália, catação e apresentação sexual) e a variável "proximidade" mostraram-se significativamente mais prevalentes na fase periovulatória do que nas demais fases do ciclo. Não houve alteração das concentrações de MFT dos machos ao longo de todo o período estudado. A análise conjunta das concentrações de metabólitos fecais de esteróides sexuais e dos comportamentos sócio-sexuais possibilitou um melhor entendimento das relações endócrino-comportamentais e reprodutivas de C. penicillata.

The communication of the female reproductive status in Callithrichidae relies mainly on the socio-sexual behavior, as generally the ovulation is concealed in this primate family by a primary signaling system. In this study the socio-sexual behavior patterns was analyzed in association with the concentration of fecal metabolites of sex steroid hormones progesterone (MFP), estradiol (MFE) and testosterone (MFT) in captive couples of Black-Tufted-Marmoset (Callithrix penicillata), during the different phases of the ovarian cycle. The studied group was composed of four adult couples kept in the São Paulo City Wild Animals Rehabilitation Center. The behavioral patterns were record by focal samplings, with 30 seconds intervals for each observation, five days a week, totalizing 14.400 registers per animal. The measurement of fecal metabolites of progesterone (MFP), estradiol (MFE) and testosterone (MFT) proceeded by enzyme immune assay (EIA). The results allowed to determine the duration of the ovarian cycle and to characterize three different phases (follicular, periovulatory and luteal). It was possible to determine 31 complete cycles that lasted 24.3±4.1 days (Mean ± SD). The follicular and luteal phases lasted 13.04±4.8 and the luteal phase 11.2±4.2 days. The behavioral patterns (scent marking, sniff genitals, grooming and sexual presentation) were more prevalent in the periovulatory phase as the behavioral variable "proximity" as well. There were no variations in the concentration of MFT in the males during the period studied. The associated analyses of the fecal metabolite of sex steroids and the socio-sexual behaviors led to a better understanding of the factors involved in the reproduction of C. penicillata.

Effects of perinatal period administration of clomiphene citrate in sexual behavior, organ weights and hormone concentration of Wistar male and female rats / Efeito da administração do citrato de clomifeno durante o período perinatal no comportamento sexual, peso dos órgãos e concentração hormonal de ratos Wistar machos e fêmeas

The effects of prenatal exposure to clomiphene citrate in sexual behavior, organ weight and hormone concentrations of male and female rats was evaluated. The animals received four doses of clomiphene citrate 2 mg/mL each during the prenatal period (21 days of gestation DG21) on days 1 (DN1), 2 (DN2) and 3 (DN3) after the birth of the puppies. The treatment led to the development of polycystic ovaries in 70% of the females, masculinization of female sexual behavior and changes in sexual behavior of males evidenced by the reduction in the number of ejaculations. In regards to hormone levels, a decrease in the FSH levels in male offspring was observed. It was concluded that clomiphene citrate interferes with the reproductive capacity of male and female rats and female sexual orientation when prenatally administered.

Foram investigados os efeitos da exposição perinatal ao citrato de clomifeno no comportamento sexual, peso dos órgãos e concentração hormonal de ratos machos e fêmeas. Os animais receberam quatro doses de 2 mg/mL de citrato de clomifeno, no período perinatal (21 dias de gestação DG21), nos dias 1 (DN1), 2 (DN2) e 3 (DN3) após o nascimento dos filhotes. O tratamento causou desenvolvimento de ovário policístico em 70% das fêmeas, masculinização do comportamento sexual das fêmeas e alteração do comportamento sexual dos machos evidenciado pela redução no número de ejaculações. Em relação aos níveis hormonais, observou-se diminuição de FSH na prole masculina. Concluiu-se que o citrato de clomifeno interfere na capacidade reprodutiva de ratos machos e fêmeas, e na orientação sexual de fêmeas, quando administrado perinatalmente.

Eficiência da mensuração de progesterona salivar para identificar a ovulação em cadelas / Efficiency of salivary progesterone measurement to identify the ovulation in bitches

Vários autores enfatizaram a importância do monitoramento do ciclo estral em cadelas e citaram exemplos de como realizar tal procedimento. O objetivo deste estudo foiavaliar a técnica de dosagem de progesterona salivarem cadelas para identificar a ovulação nesta espécie. Para composição do grupo experimental, foram utilizadas 13 cadelas, de diferentes raças (sem raça definida, Buldogue Inglês, Bernesse Mountain Dog) e diferentes idades (11 meses a 9 anos). Amostras de sangue e saliva foram colhidas simultaneamente em todos os animais, a partir dos primeiros sinais clínicos de proestro. Amostras salivares foram obtidas com o uso de dispositivo comercial específico (Salivette®). Este método foi eficaz, visto ter tornado possível obter volume suficiente para dosagem de progesterona na grande maioria das amostras (100 μL para dosagens em duplicata). As concentrações de progesterona no soro foram determinadas pela técnica de radioimunoensaio e na saliva por enzimaimunoensaio, ambas com kits comerciais. Observou-se uma relação linear crescente e positiva entre a progesterona sérica e salivar (r=0,704; p<0,0001) em cadelas. Uma das cadelas apresentou ciclo anovulatório, no qual, durante 13 dias, as concentrações séricas de progesterona sérica se mantiveram entre 1,67 e 3,76 ng/mL e as concentrações salivares de progesterona entre 19,55 e 236,77 pg/mL. Conclui-se que as concentrações de progesterona salivarmensuradas pela técnica de enzimaimunoensaio podem ser utilizadas para avaliar a ocorrência ou ausência de ovulação em cadelas.

Several authors have emphasized the importance of monitoring estrous cycle in bitches and mentioned technique examples of how it can be done. The aim of this study was to evaluate the salivary progesterone quantification technique in order to identify ovulation in this species. To compound the experimental group, 13 bitches of different breeds (no specific breed, English Bulldog, Bernesse Mountain Dog)and different ages (from 11 months to 9 years old) were used. Blood and saliva samples were collected simultaneously in all animals, starting about the first day of proestrus clinical signs. Salivary samples were collected with a specific commercial device (Salivette®). This method was effective, since it has become possible to obtain enough volume in almost all samples to quantify salivary progesterone (100 μL for duplicate quantification). Serum progesterone was quantified by radioimmunoassay and salivary progesterone by enzyme immunoassay, both of them with commercial kits. There was an increasing, linear and positive correlation between salivary and serum progesterone (r=0.704; p<0.0001) in bitches. One of the animals had an anovulatory cycle, in which, for 13 days, seric progesterone levels were maintained between 1.67 and 3.76 ng/mL and salivary progesterone levels were maintained between 19.55 and 236.77 pg/mL. It was concluded that salivary progesterone levels measured by enzyme immunoassay is a technique that can be used to evaluate the presence or absence of ovulation in bitches.