Postimplantation Loss Research Articles

Assessment of reproductive impact of the aerial parts of Caralluma dalzielii N. E. Br in female Wistar rats

Caralluma dalzielii N. E. Brown belonging to family Asclepiadaceae, is a popular cactus-shaped plant native to East Africa. The aerial parts are used traditionally for treating various diseases including infertility. The present study evaluated the effects of the aqueous extract of the aerial parts of Caralluma dalzielii (AECD) on reproductive performance of female Wistar rats. Adult female virgin rats were allotted into four major groups namely pre-conception, post-conception, implantation site and ovariectomized rats' groups. Each group was subdivided into 4 groups and treated orally with 125, 250, 500 mg/kg of AECD or distilled water (vehicle). In the pre-conception and post-conception groups, litter sizes, pups' weights, deformities, gestation length and reproductive indices were determined. Number of implantation sites and weights of embryos were assessed in the implantation site group while in the ovariectomised rats' group, uterine weights were determined. AECD produced no difference in litter size and reproductive indices in pre-conception group while in post-conception group the litter size at 500 mg/kg was significantly (p < 0.05) reduced compared to the control. Post-implantation loss index was high, and the other reproductive indices were reduced at 500 mg/kg. Whereas at the dose of 125 mg/kg, post-implantation loss index was reduced, and litter size was increased when compared to the control group. At 500 mg/kg, AECD caused a significant (p < 0.05) decrease in the number of implantation sites and weight of embryos while at 125 mg/kg the implantation sites increased. A significant (p < 0.05) increase in the uterine weight in the ovariectomised rats' group was observed at all dose levels. Our study provides scientific evidence that supports the traditional use of AECD in the treatment of infertility. At a lower dose, AECD acts by increasing the number of implantation sites and litter size of animals but at a higher dose, it may be embryotoxic. AECD increases uterine wet weight in ovariectomised rats suggesting that the plant may be oestrogen-like.

Prenatal developmental toxicity study of an alkaloid-free Ageratum conyzoides extract powder in rats by oral administration

A prenatal developmental toxicological study was conducted to evaluate the safety of an alkaloid-free Ageratum conyzoides extract powder administration on pregnant female Wistar rats and on the development of the conceptus in accordance with OECD test guideline (no. 414). Pyrrolizidine alkaloids (PAs) naturally present in A. conyzoides have been shown to induce toxicity in past studies, particularly towards hepatic cells. Therefore our test item preparation of A.conyzoides extract (aerial part of the plant) consisted of the removal of PAs. There were no treatment related adverse effects found during maternal examinations (body weights, food consumption, numbers of pregnant and non-pregnant female rats, endocrine evaluation, gravid uterine weights, and number of corpora lutea), maternal/fetal examinations (numbers of implantation sites, pre-and post-implantation loss (%), dead and live fetuses (%), resorption sites), or fetal examinations (litter size and weights, number of fetuses, sex ratio, or external, visceral, and skeletal variations and malformations) in the Ageratum conyzoides extract powder groups at doses of 500, 1000 and 2000 mg/kw bw/day compared to vehicle control group. The no observed adverse effect level (NOAEL) determined for both maternal and developmental toxicity was 2000 mg/kg bw/day, which was the highest dose tested.

Pregnancy Outcome in Female Rats Exposed to Bispyribac Sodium and Ameliorative Effect of Nigella Sativa Oil

Bispyribac sodium is commonly used as a herbicide worldwide. Few studies had investigated the health impact of exposure to it. Redox homeostasis has a great role during organogenesis through in cell growth, differentiation, and death. The aim of this study is to investigate the redox homeostasis imbalance and teratogenic effect of BPS in female rats exposed to bispyribac sodium (BPS) during organogenesis and evaluates the protective role of Nigella Sativa Oil (NSO). Pregnant female rats were grouped into four groups, negative and positive (NSO) control groups. While the others, BPS with and without NSO treatment groups. Results showed that BPS exposure during the pregnancy led to maternal and fetotoxicity as evidenced by preimplantation, post-implantation loss, reduction in placenta weight, reduction in fetal weight, fetal length, skeletal anomalies (incomplete ossification, misaligned sternbrae), visceral malformation (anophthalmia, microphthalmia, internal hydrocephaly, and microcephaly) and oxidative stress in dams serum. NSO treatment exhibited amelioration in the aforementioned maternal and fetotoxicity against BPS exposure.

Testosterone recuperates deteriorated male fertility in cypermethrin intoxicated rats.

The present study investigates the protective effects of testosterone against reproductive toxicity induced by cypermethrin (50mg/kg body weight) in rats. Significant reduction in the testicular and accessory sex organ weights were observed in cypermethrin-treated rats over controls. Cypermethrin intoxication significantly reduced testicular daily sperm count, epididymal sperm count, sperm motility, sperm viability and HOS-tail coiled sperm accompanied by significant reduction in the activity levels of testicular steroidogenic enzymes such as 3β- and 17β- hydroxysteroid dehydrogenases in rats as compared to controls. Further, qPCR studies indicated that the mRNA expression levels of steroidogenic acute regulatory protein (StAR) significantly decreased in cypermethrin-treated rats over controls. Molecular docking analysis indicated that the binding affinity of cypermethrin (-11.2kcal/mol) towards StAR protein was greater as compared to its natural ligand, cholesterol (-8.2kcal/mol) suggesting improper cholesterol channeling across the testis. Significant reduction in the circulatory levels of testosterone was also recorded in cypermethrin-exposed rats. An increase in pre- and post-implantation loss was observed in rats cohabited with cypermethrin-treated rats. On the other hand, testosterone (4.16mg/kg body weight) treatment ameliorated cypermethrin-induced reprotoxic effects in rats. To conclude, cypermethrin-induced deterioration of suppressed reproductive performance in male rats could be linked to its antiandrogenic effects and on the other hand, testosterone-mediated protection of male reproductive health in cypermethrin-treated rats at least in part occurs via restoration of testosterone biosynthesis, spermatogenesis and sperm maturation events.

Un-riped fruit pods of Prosopis cineraria (L.) Druce ameliorates Cisplatin therapy-induced partial testicular atrophy in male Wistar rats

Ethnopharmacological relevanceProsopis cineraria (L.) Druce is a plant that is widely found in dry parts of India. The unripe fruit pod has a very specific traditional claim of treating male infertility and increasing sperm volume and count. AimThe present work was endeavored to investigate the long-standing traditional claim of P. cineraria on meliorating male fertility. The study focussed on cancer therapy-induced male infertility and curative effect of the extract with an appraisal on any possible revitalizing effects on sperm count, morphology, motility, and viability combined with hormonal and histopathological investigations. Materials and methodsMale Wistar rats were used for the study. Two different doses of 400 mg/kg/d and 800 mg/kg/d (both p.o.) of the Hydroalcoholic extract were chosen as test dose while Clomiphene (25 mg/kg/d; p.o.) treatment served as standard treatment. Animals were initially injected with cisplatin (1 mg/kg/d; i.p.) for 15 days and the drug treatment was begun at the 16th day and continued till 43rd day (28 days treatment). Later all male animals got cohabited with female animals in the ratio 1:3. On confirmation of mating, female animals were isolated. Male animals were euthanized on batches. Testis and epididymis were weighed and homogenized. Sperm count, motility, morphology, viability, and headcount. The serum collected was evaluated for serum FSH, LH, and testosterone levels. On day Gestational day 15, gravid uterus observations were calculated to evaluate male and female fertility parameters. ResultsThere were statistically significant improvements (p < 0.001) in sperm motility, sperm count, sperm viability, and improved morphological features. The same pace was also noticed in testosterone, FSH and LH levels in serum and LPO, CAT, GSH, GPx and SOD in testicular tissues. The extract treated male animals produced better and healthy litter compared to cisplatin-treated animals with less pre- and post-implantation loss. ConclusionConsolidating the results seen, the extract ameliorated the testicular toxicity caused by cisplatin in a dose-dependent manner. Further insight and evaluation of the phytochemicals of the pods should be performed to bring up commercial viability.

Maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin suppresses male reproductive functions in their adulthood.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental contaminant in the environment. The developmental period is more sensitive to TCDD and there is a possibility that maternal exposure to TCDD may affect in adulthood. Adult female Wistar rats were exposed to 0.5, 1.0, and 2.0 µg/kg TCDD during the critical stage of organogenesis, that is, on GD15. The results revealed a significant decrease in indices of reproductive organ weight in adult male rats exposed to prenatal TCDD, and dose-dependent reduction in epididymal sperm reserves, percent motile, and viable sperm with an increase in percent morphological abnormal sperm. Polymerase chain reaction analysis revealed downregulated expression levels of steroidogenic markers such as steroidogenic acute regulatory, cholesterol side-chain cleavage, and 3β- and 17β-hydroxysteroid dehydrogenase (HSDs) in experimental rats. Immunofluorescence sections portrayed reduced distribution of 3β- and 17β-HSD proteins in testes of experimental rats. Furthermore, spermatogenic markers (acid phosphatase, alkaline phosphatase, lactate dehydrogenase, and sorbitol dehydrogenase) were significantly altered in the testes. Serum levels of testosterone, follicle stimulating hormones, and luteinizing hormone were significantly decreased. Testicular levels of hydrogen peroxide and lipid peroxidation were significantly elevated with a decline in superoxide dismutase, catalase, glutathione peroxidase activities, and total thiol levels. Moreover, histological and morphometric examination of testicular cross-sections depicted degenerative changes. Male fertility assessment in adult rats revealed a significant decrease in mating index, fertility index, and mean number of pre- and postimplantations with an increase in pre- and postimplantation losses in rats cohabited with in utero TCDD-exposed adult males. In conclusion, the findings of this study provided clear evidence that maternal exposure to TCDD during the critical stage of development results in suppressed reproductive health in adulthood.

Malaysian propolis and metformin mitigate subfertility in streptozotocin-induced diabetic male rats by targeting steroidogenesis, testicular lactate transport, spermatogenesis and mating behaviour.

Diabetes mellitus is one of the risk factors for male subfertility/infertility. Malaysian propolis is reported to decrease hyperglycaemia in diabetic state. The present study investigated the protective effect of Malaysian propolis on diabetes-induced subfertility/infertility. Additionally, its combined beneficial effects with metformin were investigated. Forty adult male Sprague Dawley rats were randomly assigned into five groups, namely normal control, diabetic control, diabetic+Malaysian propolis (300mg/k.g. b.w.), diabetic+metformin (300mg/kg b.w.) and diabetic+Malaysian propolis+metformin. Diabetes was induced using a single intraperitoneal injection of streptozotocin (60mg/kg b.w.) and treatment lasted for 4weeks. During the 4th week, mating behavioural experiments were performed using sexually receptive female rats. Thereafter, fertility parameters were assessed in the female rats. Malaysian propolis increased serum and intratesticular free testosterone levels, up-regulated the mRNA levels of AR and luteinizing hormone receptor, up-regulated the mRNA and protein levels of StAR, CYP11A1, CYP17A1, 3β-HSD and 17β-HSD in the testes of diabetic rats. Furthermore, Malaysian propolis up-regulated testicular MCT2, MCT4 and lactate dehydrogenase type C mRNA levels, in addition to improving sperm parameters (count, motility, viability and normal morphology) and decreasing sperm nDNA fragmentation in diabetic rats. Malaysian propolis improved mating behaviour by increasing penile guanosine monophosphate levels. Malaysian propolis also improved fertility outcome as seen with decreases in pre- and post-implantation losses, increases in gravid uterine weight, litter size per dam and foetal weight. Malaysian propolis's effects were comparable to metformin. However, their combination yielded better results relative to the monotherapeutic interventions. Malaysian propolis improves fertility potential in diabetic state by targeting steroidogenesis, testicular lactate metabolism, spermatogenesis and mating behaviour, with better effects when co-administered with metformin. Therefore, Malaysian propolis shows a promising complementary effect with metformin in mitigating Diabetes mellitus-induced subfertility/infertility.

Absence of developmental and reproductive toxicity in animals exposed to dolutegravir

The success of new antiretroviral medicines for HIV resulted in a change to guidelines of standard therapy where continuation of antiretroviral therapy is recommended to maintain the low viral load during pregnancy, thereby preventing transmission of the virus to the fetus. As a result, pregnancy related exposure to HIV medicines has increased. Understanding the safety of these medicines during pregnancy is of paramount importance to ensure health of mothers and their offspring; well-designed animal studies that evaluate the reproductive life cycle play a key role in this effort. As part of the medicine development program for dolutegravir (DTG), a series of reproductive and developmental toxicity studies were conducted using rats and rabbits. In a fertility study, where exposure to DTG occurred in female rats before mating through conception and up to implantation of the embryo, no effects on reproductive cycles, ovulation, fertility) or preimplantation embryonic growth were observed. In rat and rabbit embryo-fetal development studies, where exposure to DTG occurred during organogenesis, no malformations or other developmental abnormalities were observed. In a rat pre- and post-natal development study, where DTG exposure to the pups occurred during pregnancy and postnatally via milk, no malformations or other developmental abnormalities were observed. In these studies, no DTG-related effects occurred on fertility, embryonic (pre- and post-implantation loss, resorptions, abortions, and malformations) or fetal development where the multiples of exposure at the maximum recommended human dose were up to 27 times higher in rats or below the human exposure in rabbits.

Reproductive Toxicity of Eugenol in Wistar Rats

Objective: To analyze the effects of eugenol oral administration during pre-implantation and organogenesis periods in pregnant Wistar rats. Methodology: The animals were divided into eight experimental groups (4 groups for each period) subdivided into control groups and daily treated with eugenol at doses of 37.5, 187.5 and 375 mg/kg/day (n= 9) during pre-implantation or organogenesis phases. Results: At all tested doses of eugenol, the groups treated during the pre-implantation presented alterations in the maternal organs, increased pre-and post-implantation loss and stillbirth records. The groups treated during organogenesis also presented alterations in the maternal organs and reduction of the placental indexes, added to the skeletal alterations, in all treatment doses. There were no significant fetal visceral changes. Conclusions: Oral administration of eugenol reduced maternal reproductive capacity in the pre-implantation phase and promoted toxic effects and fetal malformations in organogenesis.

Cypermethrin exposure during perinatal period affects fetal development and impairs reproductive functions of F1 female rats

Cypermethrin (CYP) is a ubiquitously present synthetic pyrethroid insecticide. It has endocrine disrupting activities which may adversely affect reproductive development and functions of offspring if exposed during critical developmental period. The present study was undertaken to delineate the effects of CYP exposure in pregnant female rats during perinatal period on the sexual maturation, hormonal regulation, reproductive development and fertility of F1 female offspring and its molecular mechanism of action. Pregnant rats (F0) were gavaged daily with 0, 1, 10, 25 mg/kg bw/day CYP and 10 μg/kg bw/day Diethylstilbestrol (DES; positive control) from gestation day (GD) 6 to postnatal day (PND) 21. The reproductive development and function parameters were evaluated at PND 45 and 75. Reduced body weight, delayed vaginal opening, and disrupted estrous cyclicity were observed at 25 mg/kg CYP dose. CYP exposure significantly affected the reproductive organ development and their functions at all doses. Significant alterations in ovarian and uterine histology such as luteinization, reduction of primordial follicular reserves, presence of multi-oocyte follicles and thin degenerative luminal and glandular uterine epithelium were observed at adulthood. Altered circulatory steroid hormone levels and expression of ovarian and uterine steroid hormone receptors were observed at PND 75 in the F1 female offspring. Expression of HOXA10 and α-SMA which are important for uterine integrity and functions, were found to be altered at PND 75. Increased pre-implantation loss (PIL%), post-implantation loss (POL%), and reduced litter size in F1 females when cohabitated with unexposed fertile male rats were observed. Overall, perinatal exposure of pregnant rats to CYP led to significant long lasting effects on the reproductive functions of F1 female offspring. The adverse effects were passed on to F2 generation via female germ line and posed developmental anomalies. The present finding necessitates additional molecular studies to understand its trans-generational mechanism of action via female germline.

A COMPARATIVE STUDY OF THE TERATOGENIC EFFECTS OF ANTIEPILEPTIC DRUGS: LAMOTRIGINE AND LEVETIRACETAM ON ADULT ALBINO RATS

Background: Lamotrigine (LTG) and levetiracetam (LEV) are widely and increasingly prescribed second generation antiepileptic drugs for women with epilepsy during childbearing age. However, data pertaining to their congenital malformations are lacking. Aim: This study aimed to investigate and compare the teratogenic effects of LTG and LEV on normal adult female albino rats and their fetuses in order to find out the least teratogenic one of them. Materials and methods: Ninety-six adult albino rats (64 virgin females and 32 males) of almost the same range of weight and age were used. Pregnant rats were divided into 4 groups. The 1st group (control; received 1 ml of distilled water) and the 2nd group (vehicle; received 1 ml of 2% Arabic gum), while the 3rd (LTG-treated) and 4th (LEV-treated) groups received 50 mg/kg/b.wt. of LTG and 310 mg/kg/b.wt. of LEV, respectively. All substances were administered as single oral dose starting from gestational day (GD) 7 to 15. On GD 20, all dams were weighted then sacrificed and subjected to Cesarean section. The total numbers of ovarian corpora lutea and the status of all implantation sites (the total numbers of resorption sites, live and dead fetuses, and the total implantations) were counted. The gravid uteri were subsequently removed, weighted, and fetuses were delivered. The fetuses' numbers, weights, different body lengths, gross abnormalities, and skeletal malformations were recorded. Results: On GD 20, maternal weight gains and gravid uteri weights were statistically significantly lower in both treated groups than control and these differences were more significant in LTG than LEV. The percentages of pre-implantation and post-implantation losses in LTG and LEV exhibited statistically significant increase and non-significant differences compared with control, respectively, and the latter was more pronounced in LTG than LEV. As regard the numbers, weights, crown-rump lengths, biparietal diameters and head lengths of the fetuses, both treated groups showed significant differences in comparison to the control with higher abnormalities in LTG-treated than LEV-treated groups. Fetuses from treated groups revealed various gross abnormalities in the internal organs. Fetuses' skeletons examination revealed bone defects in both treated groups. Skull, vertebrae, and hind limb skeletal malformations were more prevalent in LTG-treated group, while sternum was more affected in LEV-treated group. Conclusion: Both antiepileptic drugs were teratogenic in variable degrees; however, LEV appears to be less teratogenic than LTG.

Gestational and lactational exposure to triclosan causes impaired fertility of F1 male offspring and developmental defects in F2 generation

Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol, TCS), is a broad-spectrum antimicrobial agent and extensively used in household and daily daycare products. Recently, several reports have demonstrated the endocrine disruptive action of TCS to alter the testicular steroidogenesis. However, the gestational and lactational effects of TCS exposure on F1 offspring has not been studied. Present study aimed to investigate the effect of gestational and lactational exposure to TCS on F1 male progeny and its effect on fertility. Pregnant dams (F0) were administered with different doses of TCS (0.1, 4, 40 and 150 mg/kg b. wt./day) and Diethylstilbestrol (1 μg/kg b. wt./day), as a positive control daily by subcutaneous injection during Gestation Day 6 to Postnatal Day 21. Delayed testicular descent was observed at 150 mg/kg b. wt./day dose group. Dose-dependent decrease in testosterone level, sperm count and motility was observed. Significantly decreased expression of steroid hormone receptors (AR, ERα and ERβ), StAR and aromatase were observed in F1 male rats; indicating its prolonged effect on spermatogenesis and steroidogenesis in adulthood and poor development in F2 fetuses. Further, gestational and lactational exposure to TCS has negative impact on the fertility of F1 male rats. The F1 male rats were found sub-fertile with increased (%) pre- and post-implantation loss (at 40 and 150 mg/kg b.wt./day dose) with a simultaneous decrease in litter size. The significant decrease in mean fetal weight and crown-rump length (CRL) of F2 fetuses were observed at 0.1, 4, 40 and 150 dose groups indicating impaired development of F2 fetuses caused by TCS exposure.Present study emphasizes for the first time that TCS exposure during the vulnerable developmental time point (gestation and lactation) adversely affects reproductive functions and fertility of F1 male rats, which were transmitted to F2 generations leading to reduced CRLs and weights of F2 fetuses.

Developmental and reproductive toxicity studies of BIA 10-2474

A series of regulatory studies were carried out to investigate the effects of the FAAH inhibitor BIA 10–2474 on fertility, embryo-fetal toxicity and pre- and post-natal development in rats and rabbits. Despite some reductions in sperm count in rats from 50 mg/kg, there were no major changes in male fertility up to 100 mg/kg. In female rats administered up to GD6, there were increases in pre-implantation loss at 50 and 100 mg/kg but neither post-implantation loss nor early embryonic development was affected. In contrast, when administered to female rats during pregnancy (GD6-GD17), BIA 10–2474 at 75 mg/kg/day reduced food consumption resulting in weight loss, increased post-implantation loss and reduced mean fetal body weight. In rabbits, the same maternal toxicity was seen but there were no effects in this species on post-implantation loss or fetal body weights. There were no teratological effects clearly due to BIA 10–2474 and developmental milestones and behavior of offspring were not affected. When administered during pregnancy and lactation (GD6-PND20), some post-implantation loss was seen from 20 mg/kg/day, but developmental milestones and behavior of the offspring were not affected, although males tended to have lower body weight. Based on these data the NOAEL for parental fertility was established as 50 mg/kg/day, the maternal NOAEL during pregnancy was 25 mg/kg/day in rats and developmental NOAEL was 25 and 75 mg/kg/day in rats and rabbits, respectively. When administered during post-natal development to rats the maternal NOAEL was 6 mg/kg/day. The parental reproductive NOAEL, the NOAEL for viability and growth of the F1 offspring, the F1 parental NOAEL and the F1 reproductive NOAEL were all considered to be 20 mg/kg/day.

Lentinus edodes Exposure before and after Fetus Implantation: Materno-Fetal Development in Rats with Gestational Diabetes Mellitus.

Background: The presence of β-glucans and phenolic compounds in Lentinus edodes suggests this mushroom can be used as a nutritional supplement. Two gestational conditions (before and after fetus implantation) were evaluated, and Lentinus edodes exposure was performed in diabetes mellitus rat model induced by streptozotocin in pre-clinical tests. Methods: On the 20th day of pregnancy, cesarean sections were performed. Blood was collected for biochemical, hematologic parameters and oxidative stress biomarkers. Placenta and amniotic fluid were collected, and fetuses were analyzed through morphological evaluation. Results: The mushroom did not reduce the severe hyperglycemia of the mother-concept but promoted an increase in maternal insulin levels; reduced the levels of alanine aminotransferase, and aspartate aminotransferase, triglyceride and total cholesterol; protected the animals from post-implantation losses. Liver damage induced by streptozotocin was reversed in experimental groups. Conclusions: Lentinus edodes mushroom has antioxidant properties that can minimize the damage caused by gestational diabetes mellitus.

Acute uterine effects and long-term reproductive alterations in postnatally exposed female rats to a mixture of commercial formulations of endosulfan and glyphosate

Endosulfan and glyphosate are widely used pesticides and have been associated to reproductive disorders. We examine the acute and long-term effects of postnatal exposure to commercial formulations of endosulfan (EF), glyphosate (glyphosate-based herbicide, GBH) and a mixture of both pesticides (MIX). After birth, female pups of Wistar rats received saline solution (CONTROL), EF (600 μg/kg of b.w/day), GBH (2 mg/kg of b.w/day) or a mixture (at the same doses) from postnatal day (PND) 1 to PND7. The uterine histology and expression of Hoxa10, estrogen (ERα) and progesterone (PR) receptors were evaluated on PND8. Reproductive performance was evaluated on gestational day 19. GBH and MIX rats showed an increment of 1) the incidence of luminal epithelial hyperplasia, 2) PR and Hoxa10 expression. EF modified ERα and Hoxa10 expression. During adulthood, MIX and GBH rats showed higher post-implantation losses while EF alone produced an increase of pre-implantation losses. We showed that the co-administration of both pesticides produced acute uterine effects and long–term deleterious reproductive effects that were similar to those induced by GBH alone. We consider important to highlight the necessity to evaluate the commercial pesticide mixture as a more representative model of human exposure to a high number of pesticides.

Reproductive disorders in female rats after prenatal exposure to sodium arsenite

Arsenic is a metalloid widely found in the environment in organic and inorganic forms. Exposure to inorganic arsenic forms via drinking water has been associated with an increased incidence of negative health effects, including reproductive disorders and dysfunction of the endocrine system. However, the impact of arsenic exposure on female reproductive development is still unclear. Therefore, in the present study, we evaluated the effects of prenatal exposure to arsenic on the initial sexual development and puberty onset, and in the morphology of the female reproductive organs, estrous cycle regularity and fertility parameters during adulthood. To do that, pregnant female Wistar rats were exposed to 10mg/L sodium arsenite via drinking water from gestational day (GD) 1 until GD 21 and the female offspring was evaluated in different postnatal days. Our results showed that prenatal arsenic exposure induced a decrease of litter weight and morphological masculinization in females at postnatal day 1. Moreover, these females had a delay in the age of puberty onset and alteration in estrous cycle number and length. During adulthood, females from the sodium arsenite group showed an increase in endometrium, myometrium and perimetrium areas, and an imbalance in uterine antioxidant enzyme activity. These animals also presented an increase in post-implantation loss and reabsorption number, leading to reduced viable fetus number. In conclusion, prenatal arsenic exposure in rats was able to promote female masculinization, alter sexual development and impair reproductive performance.

N-Butylparaben exposure through gestation and lactation impairs spermatogenesis and steroidogenesis causing reduced fertility in the F1 generation male rats

Parabens are class of preservatives used in vast majority of commercial products, and a potential Endocrine Disrupting Chemical (EDC). The present study was undertaken to delineate the effects of n-butylparaben on F1 male progeny exposed maternally through gestation and lactation via subcutaneous route. The F0 dams were given subcutaneous injections of n-butylparaben from gestation day (GD) 6 to postnatal day (PND) 21 with doses of 10, 100, 1000 mg/kg Bw/day in corn oil. The F1 male rats were monitored for pubertal development and sexual maturation; these were sacrificed on PND 30, 45 and 75. On PND 75, these F1 male rats were subjected for fertility assessment with unexposed female rats.A delayed testicular descent at 100 and 1000 mg/kg Bw dose and delayed preputial separation at 10 mg/kg Bw dose was observed in exposed F1 male rats. Decreased sperm count, motility and Daily Sperm Production was observed at 100 mg/kg Bw dose at PND 75. Interestingly, the sperm transit time in the epididymis was accelerated at this dose. Significant perturbed testicular expression of steroid receptors (ERα and β, AR), INSL3 and StAR genes with increased T and LH levels indicates direct effect on spermatogenesis and steroidogenesis. These F1 generation adult rats were sub-fertile with increased (%) pre- and post-implantation loss at 100 and 1000 mg/kg Bw/day dose. This is the first report on n-butylparaben highlighting the involvement of testicular leydig cells with accelerated sperm transit time leading to reduced fertility in the maternally exposed F1 male rats through estrogenic/anti-androgenic action.

Low dose of flurochloridone affected reproductive system of male rats but not fertility and early embryonic development

BackgroundFluorochloridone (FLC) is a widely used herbicide, and its target organs are testes and epididymides. The Globally Harmonized System of Classification and Labelling of Chemicals classified FLC as Level 2-possibly cause fertility or fetal damage (no relevant data support). The maximum residue levels of FLC in processed crops have been reviewed in the latest European Food Safety Authority (EFSA) report in 2018. However, the toxic effect of FLC on fertility and early embryonic development is limited, and the health risk assessment of FLC needs further consideration. This study investigated the potential effects of FLC on fertility and early embryonic development in rats.MethodsOne hundred rats of each sex were divided into four groups including three FLC-treated groups at doses of 2 mg/kg, 5 mg/kg and 15 mg/kg, and a vehicle control group (0.5% (w/v) sodium carboxymethyl cellulose). Male and female rats were dosed for 9 and 2 consecutive weeks, intragastrically, prior to cohabitation and lasted throughout the mating period for males and continued until Gestation Day 7 (GD7) for females. Parameters such as weights and coefficients of reproductive organs, epididymal sperm number and motility, indexes of copulation, fecundity and fertility indexes, mating period, estrous cycle, corporalutea number, implantations, live, dead and resorbed fetuses, preimplantation loss rate, and postimplantation loss rate were observed in this study.ResultsObvious toxicity of male reproductive system was found at the dose of 15 mg/kg including decreases in testicular and epididymal weight, also in sperm motility rate. Whereas the increase in sperm abnormality rate was observed. However, no significant effects of FLC were found on lutea count, implantations count, fetuses count and weight, live fetuses count (rate), dead fetuses count (rate), resorbed fetuses count (rate), placentas weight, fetuses gender, preimplantation loss (rate) and postimplantation loss (rate). Furthermore, FLC had no adverse effects on fertility and early embryonic development in rats.ConclusionThe no-observable-adverse-effect level (NOAEL) of FLC on fertility and early embryonic development in rats was considered to be 5 mg/kg/day.

Allogeneic murine pregnancy models for assessing the developmental effects of immune-stimulating antibodies: Challenges in reproducibility.

Literature suggests that murine allogeneic pregnancy models are an alternative approach for evaluating the developmental toxicity of immune-stimulating agents. In this study, multiple syngeneic and allogeneic murine pregnancy models were used to assess the potential embryo-fetal effects of four different murine antibodies (IgG1 or IgG2 ) that activate the immune system by binding to T-cell receptors (PD-L1, LAG-3, and GITR). The pregnancy models were generated by within and between matings of five different inbred strains of mice (CBA/CaJ, DBA/2J, BALB/c, C57BL/6, and CBA/J). The antibodies were administered every 2-3 days by intraperitoneal injection (n = 12-29/group) during gestation days 6 to 14. There were no differences in embryo-fetal endpoints between the allogeneic and syngeneic pregnancies. Additionally, treatment with the antibodies had no effect on mean postimplantation loss in either the syngeneic or allogeneic pregnancies despite confirmation of pharmacologically-relevant systemic exposures. These results suggest that allogeneic murine pregnancy models need further validation and testing before they can be reliably used as an alternative approach for assessing the developmental effects of agents that stimulate the immune system.

Effects of subchronic exposure to Caryocar brasiliense peel ethanolic extract on male reproductive functions in Swiss mice

This study evaluated the effects of subchronic exposure to Caryocar brasiliense peel ethanolic extract (CBPE) on reproductive function in male Swiss mice. CBPE was administered orally for 28 days at doses of 75, 150 and 300 mg.kg−1 bw; control group received saline. Fertility test was performed after 14 days of treatment and animals were euthanized after the end of the 28-day period for evaluation of the reproductive parameters. The tested doses produced no significant changes in plasma testosterone levels, daily sperm production, sperm concentration, sperm morphology or fertility rate. However, sperm transit time was reduced in the caput/corpus epididymis, the post-implantation loss rate increased and there were significant changes in spermatogenic dynamics at the highest dose. These results indicated that subchronic exposure to CBPE has low reproductive toxicity, but additional studies are recommended to provide evidence of long-term safety at high concentrations.