Effects of silver nanoparticles prenatal exposure on rat offspring development.

Many types of nanocomposites employed in food packaging are based on silver nanoparticles (AgNP) because of their antibacterial properties, which can increase food shelf-life. As the commercialization of AgNP products has been expanding, the released of such nanoparticles in the environment has caused enormous concern, once they can pose potential risks to the environment and human beings. For instance, exposure of the maternal environment to nanomaterials during pregnancy may impact the health of the dam, fetus and offspring. In this context, here we investigated the effects of prenatal exposure of AgNP on the pregnancy outcomes of dams and postnatal development of their offspring. Pregnant Wistar rats were exposed to distinct AgNP concentrations (0, 1, 3 and 5 µg/kg/day) from beginning to the end of pregnancy. At parturition, newborns were observed regarding clinical signs of toxicity and survival rate. The offspring was examined by evaluating developmental endpoints. A delay in time for vaginal opening and testes descent were detected in the offspring exposed to AgNP during embryonic development. Our results indicate that prenatal exposure to AgNP can compromise neonatal rats' postnatal development, especially the reproductive features.

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age.

In the last decades, selection for improved prolificacy has resulted in higher litter sizes and has thereby increased the proportion of low birthweight (LW) piglets. It is well documented that LW piglets have lower growth performance, muscle accretion and poor carcass quality. However, little is known about the relations of birthweight with subsequent reproductive performance in gilts. This study investigated the effects of birthweight on reproductive tract and ovarian follicle development in 150-day-old gilts. Twenty eight female pigs of different birthweight ranges (high-HW: 1.8-2.2 kg; low-LW: 0.8-1.2 kg) from higher parity commercial sows were reared until 150 days of age, and their body weights were recorded at weaning, end of nursery and end of the grower-finisher phase. The animals were killed and their reproductive tracts collected for biometrical and histomorphometrical analysis. LW gilts showed significantly lower body weights and growth rates during all phases of production compared to their HW counterparts (p < .01). Most biometrical measurements of the reproductive tract were similar between the experimental groups, except vaginal length and the gonadossomatic index (relative ovarian weight), which were affected by birthweight class (p < .05). LW females also showed fewer medium size (3-5 mm; p < .01) ovarian follicles, pre-antral follicles (p < .07) and more atretic follicles per ovarian cortex area (p < .05). Therefore, besides the effects on post-natal growth performance, birthweight affects vaginal length and the follicular dynamics process, which may impair the reproductive performance of replacement gilts.

In utero and lactational exposure to fipronil in female rats: Pregnancy outcomes and sexual development.

Fipronil, a phenylpyrazole insecticide, is used in agriculture, veterinary medicine, and public health. Because this insecticide is considered a potential endocrine disruptor, the aim of this study was to examine the influence of perinatal exposure to fipronil on neonatal female reproductive system development. Pregnant rats were exposed (via gavage) daily to fipronil (0.03, 0.3, or 3 mg/kg) from gestational day 15 to day 7 after birth, and effects on the reproductive functions assessed on postnatal day (PND) 22. No signs of maternal toxicity were observed during daily treatment with fipronil. Perinatal exposure to the highest dose of fipronil (3 mg/kg) delayed the age of vaginal opening (VO) and first estrus without markedly affecting the anogenital distance (AGD). Further, exposure to 0.3 mg/kg fipronil produced a significantly shorter estrus cycle and reduced number of cycles during the period of evaluation. However, the other reproductive parameters analyzed, including fertility, hormone levels, sexual behavior, and histology of ovaries and uterus, displayed no marked alterations. In this experimental model, fipronil interfered with development of neonatal female reproductive system as evidenced by delay in VO and estrus cycle alterations without apparent significant effects on fertility. Further studies are needed to identify the mechanisms of action associated with the observed female reproductive system changes.

Human vaginal histology in long-term users of the injectable contraceptive depot-medroxyprogesterone acetate.

OBJECTIVES: Apparently, depot-medroxyprogesterone acetate (DMPA) increases a woman's risk of acquiring HIV. The objective of this study was to test whether the vaginal mucosal thickness and Langerhans cell counts were significantly different in long-term DMPA users compared with women users of an intrauterine device (IUD) who had never used DMPA. STUDY DESIGN: Cross-sectional study. Twenty-three DMPA users were matched with 23 nonusers controlled for age, body mass index (BMI; kg/m²), and duration of contraceptive use. Four groups of women were evaluated according to the duration of DMPA use: >1, <5; ≥5, <10; ≥10, <15 or ≥15 years. Estradiol (E2) levels were compared between the two groups. Histologic sections of vaginal mucosal biopsies were evaluated to measure the mean epithelial thickness and S100 immunostained sections were used to count the number of Langerhans cells/mm. RESULTS: Mean (±S.D.) E2 levels were significantly lower in DMPA users (39.4±26.6 pg/mL) compared with nonusers (102.6±60.3 pg/mL) despite similar ages (42.3±7.4 and 42.4±7.4 years, respectively). Mean (±S.D.) vaginal thickness was 232.6±108.1 and 229.7±112.9 in DMPA users and nonusers, respectively. There were no differences in vaginal thickness or Langerhans cell count/mm between users and nonusers even after controlling for DMPA duration of use. CONCLUSIONS: Vaginal epithelial thinning or Langerhans cell count was not different between long-term DMPA users and copper-IUD users who had never used DMPA. IMPLICATIONS: No differences were found in vaginal epithelial thickness or in Langerhans cell count between long-term users of the injectable contraceptive DMPA and nonusers.

Temporal window in which exposure to estradiol permanently modifies ovarian function causing polycystic ovary morphology in rats.

OBJECTIVE: To investigate the developmental window in which E(2) exposure produces irreversible changes in ovarian function resulting in polycystic ovary. DESIGN: Basic experimental study. SETTING: University animal laboratory. ANIMAL(S): Thirty Sprague-Dawley rats were administered a single E(2) valerate dose (10 mg/kg of weight) at 1, 7, 14, 21, or 30 days of age. Control rats were injected with the vehicle at 1 day of age. All rats were sacrificed at 6 months of age. INTERVENTION(S): Observation of vaginal opening, estrous cyclicity by vaginal smears, and ovarian morphometry in the 6-month-old rat. MAIN OUTCOME MEASURE(S): Measurement of ovarian noradrenaline by high-performance liquid chromatography coupled with electrochemical detection, serum levels of LH by enzyme-linked immunoassay, P, androstenedione, and E(2) by enzyme immunoassay. RESULT(S): Rats exposed to E(2) at 1, 7, or 14 days of life did not show estrual cycling activity and maintained a polycystic ovary (PCO) condition throughout the entirety of the study. However, if the exposure to E(2) occurred after postnatal day 21, the PCO-induced condition was reversible. In rats that developed a permanent PCO condition, we observed significant effects of E(2) on ovarian morphology if exposure occurred on postnatal day 1 and a presumable effect on the hypothalamus if the exposure occurred between postnatal days 1 and 14. CONCLUSION(S): Our findings suggest that in rats, the most sensitive period for the promotion of an irreversible PCO morphology by estrogenic compounds is during neonatal early follicular development.

Effects of arsenic exposure during the pre- and postnatal development on the puberty of female offspring.

UNLABELLED: The (As) arsenic exposure is a risk factor for causing disturbances in the endocrine organs. OBJECTIVE: To evaluate if sub-chronic As exposure during the pre- and postnatal development causes disturbances in the puberty. Moreover, determine adverse effects of As on the ovarian follicle and adrenocortical cell maturation. METHODS: Females adult Wistar rats were exposed to sodium arsenite at 3 ppm calculated as As in drinking water from mating, gestation. Following the birth, the female offspring continued exposured to As via lactation. Weaned pups received the same As treatment as mothers, until they were 1-4 months (mo) old. At these ages, blood sampling and tissue harvest were done. The tissues were fixed in situ with 4% paraformaldehyde in phosphate buffer. After the perfusion the ovaries, uterus, adrenal glands were harvested, dissected out, weighted. The ovaries and the adrenal glands were processed to paraffin and sectioned at 5 µM and stained with hematoxylin and eosin for light microscopy. STATISTICAL ANALYSIS: Comparisons between groups were made by unpaired t-test or nonparametric Mann-Whitney test as appropriate. RESULTS: 100% As treated rats at 1 mo of age were at diestrous stage, with low estradiol E2. As treatment caused disturbances in the morphology of the ovarian cell consisting in DNA damage evidenced by picknotic chromatin, cariorexis, significant cytoplasmic vacuolization and also vasculature damaged. Arrest in follicle maturation was also present. CONCLUSIONS: We found that the onset of puberty in the As treated rats was 1 mo delayed since vagina was still closed, the vaginal smear showed that they were at diestrus stage with plasma low E2 levels.

Neonatal exposure to estradiol valerate programs ovarian sympathetic innervation and follicular development in the adult rat.

A single injection of estradiol valerate (EV) to 14-day-old rats (when the ovarian follicle population has been already established) disrupts cyclicity, increases the activity of key enzymes of androgen biosynthesis, and develops polycystic ovary by a causally related increase in ovarian noradrenaline (NA). The current study examined an early window of ovarian development to look for a specific stage of development at which estradiol can induce such changes in sympathetic activity and follicular development. A single dose of EV applied to rats before the first 12 h of life rapidly increases (after 24 h) the ovarian expression of nerve growth factor (Ngfb) and p75 low-affinity neurotrophic receptor (Ngfr) mRNAs. When adults, rats presented early vaginal opening, disrupted cyclicity, appearance of follicular cyst, absence of corpus luteum, and infertility. Total follicles decreased, mainly due to a reduced number of primordial follicles, suggesting that estradiol acts in the first stages of folliculogenesis, when primordial follicles are organizing. These changes paralleled a 6-fold increase in NA concentration. No changes in NA content were found in the celiac ganglia, suggesting a local, non-centrally mediated effect of estradiol. Surgical section of the superior ovarian nerve (the main source of sympathetic nerves to the ovary) to rats neonatally treated with EV decreased intraovarian NA, delayed vaginal opening, and blocked the development of follicular cyst and that of preovulatory follicles. Therefore, we can conclude that early exposure to estradiol permanently modifies ovarian sympathetic activity and causes profound changes in follicular development, leading to the polycystic ovary condition.

Pre- and postnatal toxicity of the commercial glyphosate formulation in Wistar rats.

Glyphosate is the active ingredient and polyoxyethyleneamine is the surfactant present in the herbicide Roundup formulation commercialized in Brazil. The aim of this study was to assess the reproductive effects of glyphosate-Roundup on male and female offspring of Wistar rats exposed during pregnancy and lactation. Dams were treated orally with water or 50, 150 or 450 mg/kg glyphosate during pregnancy (21-23 days) and lactation (21 days). These doses do not correspond to human exposure levels. The results showed that glyphosate-Roundup did not induce maternal toxicity but induced adverse reproductive effects on male offspring rats: a decrease in sperm number per epididymis tail and in daily sperm production during adulthood, an increase in the percentage of abnormal sperms and a dose-related decrease in the serum testosterone level at puberty, and signs of individual spermatid degeneration during both periods. There was only a vaginal canal-opening delay in the exposed female offspring. These findings suggest that in utero and lactational exposure to glyphosate-Roundup may induce significant adverse effects on the reproductive system of male Wistar rats at puberty and during adulthood.

Reproductive evaluation of two pesticides combined (deltamethrin and endosulfan) in female rats.

Data from in vitro studies suggest that the pesticides deltamethrin (D) and endosulfan (E) exert estrogen-like effects. There is concern that interaction between weakly estrogenic compounds can increase their estrogenic potency. The aim of the present study was to determine estrogenic activity in an animal model and the possible female reproductive adverse effects of these pesticides combined. Wistar rats received daily (po), from day 6 of pregnancy to day 21 of lactation, deltamethrin and endosulfan concomitantly: D: 2.0 mg/kg+E: 1.5 mg/kg, or D: 3.0 mg/kg+E: 2.0 mg/kg, or D: 4.0 mg/kg+E: 3.0mg/kg. Some offspring also were exposed directly after weaning. Maternal and reproductive outcome data were assessed. An uterotrophic assay to screen in vivo estrogenic activity of D+E was also performed. A group of female offspring was analyzed for vaginal opening (VO), first estrus, estrous cycle regularity, and weights of the uterus and ovaries. No signs of maternal toxicity were detected. Results from the uterotrophic assay indicate absence of in vivo estrogenic activity of D+E. No significant variations in reproductive endpoints of females were observed. These results suggest that administration of D+E does not pose a reproductive hazard to female rats exposed during critical periods of development, indicating that the combination does not exert estrogen-like effects in vivo or is not delivered to target organs.

Puberty onset in the female offspring of rats submitted to protein or energy restricted diet during lactation.

This study aims to determine the effects of maternal protein and energy malnutrition during lactation on the linear growth, body weight and onset of puberty of the female offspring. At parturition, dams were randomly assigned to the following groups: (C) control group, with free access to a standard laboratory diet containing 23% protein; (PR) protein-restricted group, with free access to an isoenergy and protein-restricted diet containing 8% protein; and (ER) energy-restricted group, receiving standard laboratory diet in restricted quantities. After weaning, the female pups had free access to standard laboratory diet. From day 30 onwards, the pups were inspected daily for vaginal opening. Cyclic stages of the ovaries were studied by daily vaginal smears after vaginal opening until day 40 when all animals were sacrificed with pentobarbital. From day 4 after birth until day 40, body weight and linear growth in the PR and ER rats were significantly lower than in controls (p < 0.001). In spite of the significant (p<0.05) delayed in the vaginal opening in PR and ER rats, the first estrous cycle occurred at the same time of vaginal opening in all groups. The PR and ER rats exhibited a lower uterine (PR = 42%, ER = 40%, p < 0.001) and ovarian (PR = 26%, ER=19%, p < 0.05) absolute weight and uterus relative weight (PR = 27%, ER = 22%, p < 0.05). Our data showed that maternal protein and energy malnutrition during lactation leads to growth retardation and delayed on the onset of puberty in female pups, with vaginal opening and estrous cycle occurring at the same time.

Alcohol treatment during lactation produces an advance in the onset of puberty in female rats.

It has been described that alcohol treatment after weaning produces a delay in the onset of puberty and a decrease in the body weight of female rats; however, during development, there are periods with different sensitivity to endogenous and exogenous substances. In this study, two daily doses of 2.5 g/kg of ethanol each administered to female pups during days 13-18 of postnatal age produced an advance in the age at vaginal opening but induced no effect on the body weight; however, the onset of sexual behavior was not advanced. Fertility and reproduction measures were not significantly impaired by this treatment. It is supported that, in this period, alcohol can produce different effects - even opposite to those described in other developmental stages - which seems to represent a critical period for alcohol action.

Antioestrogenic effect of trans-dehydrocrotonin, a nor-clerodane diterpene from Croton cajucara Benth. in rats.

This study examined trans-dehydrocrotonin (t-DCTN), a nor-clerodane diterpene isolated from the Brazilian medicinal plant Croton cajucara Benth., for a possible antioestrogenic activity using immature rats as a model system for bioassay of oestrogen, and for an antiimplantation effect in regularly cycling rats of proven fertility. In the antioestrogen test, t-DCTN (25 and 50 mg/kg) effectively prevented oestrogen-induced increases of uterine wet weights. In addition, the vaginal openings provoked by oestrogen were completely prevented by t-DCTN. However, blastocyst-implantation was only insignificantly affected in t-DCTN pretreated animals. These results suggest that t-DCTN may be an antioestrogen and warrants further studies with regard to its mechanism of action.