Project description:Description of the vaginal microbiota in nulliparous ewes during natural mating and pregnancy: preliminary evidence of the male preputial microbiota influence
Project description:Hormonal protocols for cervix dilation are essential for non-surgical embryo recovery (NSER) in ewes, due to the cervical complexity of the species. However, little is known about the possible effects of these hormones on the uterine environment. Thus, this study aimed to evaluate in sheep the effect of a hormonal protocol for cervical dilation on the protein profile of the uterine fluid. After superovulation, Santa Inês ewes (n = 40) were allocated into either a treated group with estradiol benzoate (100 µg, iv) and sodium cloprostenol (0.12 mg, im) 12 h before the embryo collection plus oxytocin (100 IU, iv) 15 min before the collection procedure; or a control group, which consisted of the administration of saline solution. The recovery of the uterine lavage and embryos was performed by laparotomy. After precipitation (trichloroacetic acid) and digestion (trypsin), proteins were identified by mass spectrometry (LC-MS/MS). Overall, 1447 proteins were identified, of which 436 and 126 were found exclusively in the treated and control groups, respectively. A total of 16 proteins were highly expressed in the treated group (P < 0.05), which included glutathione transferase (related to prostaglandin biosynthesis) and osteopontin (involved in the regulation of estradiol). In the control group, five proteins were up-regulated (P < 0.05), including cofilin (linked to up-regulation of embryonic development). Based on the gene ontology, in both groups, the proteins most abundantly in the uterine fluid are related to cellular and metabolic processes, in addition to locomotion and localization in the control group. In conclusion, the hormonal protocol for cervical dilation modifies the protein profile of uterine lavage recovered from superovulated ewes after embryo collection.
Project description:In the current study, 5 multiparous ewes during the late gestation were restricted to 30% feeding level to establish an undernutrition model while another 5 ewes were fed normally as controls. All the ewes were sacrificed and pituitary samples were collected to do transcriptome sequencing to investigate the metabolic changes.
Project description:Clinical treatment protocols for infertility with in vitro fertilization-embryo transfer (IVF-ET) provide a unique opportunity to assess the human vaginal microbiome in defined hormonal milieu. Herein, we have investigated the association of circulating ovarian-derived estradiol (E2) and progesterone (P4) concentrations to the vaginal microbiome. Thirty IVF-ET patients were enrolled in this study, after informed consent. Blood was drawn at four time points during the IVF-ET procedure. In addition, if a pregnancy resulted, blood was drawn at 4-to-6 weeks of gestation. The serum concentrations of E2 and P4 were measured. Vaginal swabs were obtained in different hormonal milieu. Two independent genome-based technologies (and the second assayed in two different ways) were employed to identify the vaginal microbes. The vaginal microbiome underwent a transition with a decrease in E2 (and/or a decrease in P4). Novel bacteria were found in the vagina of 33% of the women undergoing IVF-ET. Our approach has enabled the discovery of novel, previously unidentified bacterial species in the human vagina in different hormonal milieu. While the relationship of hormone concentration and vaginal microbes was found to be complex, the data support a shift in the microbiome of the human vagina during IVF-ET therapy using standard protocols. The data also set the foundation for further studies examining correlations between IVF-ET outcome and the vaginal microbiome within a larger study population.
