TEDE Coleção: Mestrado em Ciência Animal

Impacto do canabidiol no desenvolvimento embrionario in vitro bovino

2026-11-11T04:00:34Z

Título: Impacto do canabidiol no desenvolvimento embrionario in vitro bovino Autor: Teixeira, Cintia Pimentel Mangueira Primeiro orientador: Castilho, Anthony César de Souza Abstract: Cannabis sativa contains bioactive compounds, such as THC and CBD, which interact with the endocannabinoid system and influence female reproduction. THC crosses the placenta, affects hormones and impairs fertility, while CBD, which has therapeutic potential, can modulate inflammatory processes without directly inhibiting the reproductive axis. Animal models, such as cattle, are essential to study these effects. In this study, the effects of CBD on oocyte competence and embryonic development were analyzed in vitro, contributing to the understanding of the risks and benefits of cannabinoids in reproduction. For this purpose, bovine ovaries were collected from a local abattoir and transported to the laboratory in a thermal container at 37 °C in saline (0.9% NaCl). Antral follicles (3-8 mm in diameter) were aspirated with an 18G needle. Cumulus-oocyte complexes (CCOs) classified as grade I and II were selected and subjected to IVM using TCM 199 medium containing bicarbonate, fetal bovine serum (FBS, 10%), pyruvate (2 mM), amikacin (75 µL/mL), follicle-stimulating hormone (FSH, 20 µL/mL) and luteinizing hormone (LH, 2 µL/mL). CBD (Cannabidiol Prati-Donaduzzi, 20 mg/mL) was diluted in 0.05% dimethyl sulfoxide (DMSO). The CCOs were treated with 0.1, 1 and 10 µM CBD or not. The control group was treated with TCM 199 medium and DMSO (0.05%). The CCOs were matured for 24 hours at 38.5 °C in a humidified atmosphere. After maturation, the matured CCOs were photographed to evaluate the expansion of the cumulus cells by area. This was measured using the ImageJ program and also classified according to the score: slightly expanded (score 0), partially expanded (score 1) and fully expanded (score 2). They were also subjected to in vitro fertilization with TL Stock supplemented with BSA (6 mg/mL), pyruvate (2 µL/mL of a 100 mM solution), gentamicin (75 µg/mL), heparin (3 µg/mL) and PHE solution (penicillamine, hypotaurine, epinephrine – 44 µL/mL). After 18 hours, the cumulus cells were removed and the putative zygotes were transferred to SOF (Synthetic Oviductal Fluid) medium supplemented with SFB (2.5%) and pyruvate (2 µL/mL). On the third day, the medium was partially changed and the cleavage rates were evaluated. On the seventh day, the blastocyst rate was evaluated. In general, exposure to 10 µM CBD increased the proportion of CCOs with a value of 0 (p=0.02), indicating low expansion. Consistent with this, the expansion range of CCOs was smaller when bovine CCOs were exposed to 1 µM, and with an even more pronounced detrimental effect in COCs exposed to 10 µM CBD (p<0.0001). As a result, exposure to 10 µM CBD completely impaired early embryonic development, as evidenced by a reduction in cleavage rate (p=0.027) and a reduction in blastocyst formation (p=0.002). In conclusion, the results indicate that the addition of 10 µM CBD during IVM impairs the in vitro maturation of oocytes and consequently has a negative effect on the in vitro production of bovine embryos. Instituição: Universidade do Oeste Paulista Tipo do documento: Dissertação

Efeito bacteriostático da fibrina rica em plaquetas (FRP) e membrana de látex em bactérias Staphylococcus aureus: estudo in vitro

2026-11-11T04:00:33Z

Título: Efeito bacteriostático da fibrina rica em plaquetas (FRP) e membrana de látex em bactérias Staphylococcus aureus: estudo in vitro Autor: Ramos, Camila Roncada Primeiro orientador: Santarém, Cecília Laposy Abstract: Natural rubber latex, obtained from the rubber tree Hevea brasiliensis, has been widely used in the biomedical field due to its excellent biocompatibility. Studies have shown that latex membrane films impregnated with gentamicin sulfate exhibited efficient antimicrobial activity against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa, through the gradual release of the antibiotic. Another promising biomaterial is platelet-rich fibrin (PRF), derived from autologous blood, composed of a porous fibrin structure containing platelets, leukocytes, and growth factors, exhibiting regenerative potential in addition to antimicrobial action. Knowing that S. aureus is one of the main causes of skin and soft tissue infections, in this study we investigated the antimicrobial response, “in vitro”, in 4 groups: control (C), membrane (M), platelet-rich fibrin (PRF), and membrane + platelet-rich fibrin (M-PRF), against methicillin-sensitive and methicillin-resistant S. aureus (MSSA and MRSA). Thus, the calculation for the experiment with one replicate was: E = 11 - 3 - 0 = 8 (for each bacterium), because there were 12 total observations (three plates per treatment), four treatments, and one block. The results indicated that the interaction between the latex membrane and platelet-rich fibrin produces a synergistic effect capable of limiting the bacterial growth of MSSA, with a growth reduction percentage close to 5%, while for MRSA, the values are approximately 2 to 3%. Therefore, it was observed that while MRSA maintains more stable growth, regardless of conditions, MSSA exhibits modulated responses, including a significant inhibitory effect when exposed to the combined treatment. Instituição: Universidade do Oeste Paulista Tipo do documento: Dissertação

Exposição ambiental subcrônica a herbicida a base de glifosato: avaliação do possível efeito ao epitélio traqueal de ratos

2026-11-11T04:00:33Z

Título: Exposição ambiental subcrônica a herbicida a base de glifosato: avaliação do possível efeito ao epitélio traqueal de ratos Autor: Ribeiro, Andressa Regiane Fogaça Primeiro orientador: Nai, Gisele Alborghetti Abstract: Introduction: Glyphosate is one of the main active ingredients used for crop spraying in Brazil and worldwide. Although industry studies suggest that glyphosate is minimally toxic to non-target species, scientific studies with this herbicide point to deleterious effects resulting from poisoning and cast doubt on its safety for mammalians. Inhalation is a common form of exposure to pesticides, making the respiratory tract vulnerable to damage. Objective: To evaluate the effects on the tracheal epithelium of subchronic inhalation exposure to a glyphosate-based herbicide (GBH). Materials and methods: Forty adult male and female rats were divided into four groups (males, n=5; females, n=5): a control group, nebulized with NaCl 9%, and three groups exposed to three different concentrations of GBH (low, medium, and high). The animals were nebulized five days a week for 75 days. After euthanasia, the trachea wascollected for histopathological analysis, with analysis of the following parameters: interstitial inflammatory infiltrate and type of inflammatory cell, presence of metaplasia, dysplasia, and neoplasia. Results: There was no difference in the incidence of inflammation between the groups. Only males exposed to medium and high concentrations of GBH presented cases of intense inflammation, in addition to a higher incidence of moderate inflammation compared to the control and exposed females. Animals exposed to GBH had a higher number of cases of squamous metaplasia than those in the control group, regardless of GBH concentration or sex. Animals exposed to high concentrations of GBH had the thickest tracheal epithelium, regardless of sex, but there was no statistical difference between the groups. Among male animals, those exposed to high concentrations of GBH had a lower number of goblet cells. Among females, there was a progressive decrease in the number of goblet cells as the GBH concentration increased. There was no difference in mucus concentration in the tracheal epithelium between the groups, but there was a higher incidence of acidic mucus in animals exposed to GBH. The nucleolar organizer region (NOR) count was higher in animals exposed to high concentrations of GBH. Conclusion: Data from this study show that GBH can cause damage to the tracheal epithelium, which leads to a greater risk of respiratory infections, even in cases of not very prolonged exposure. Instituição: Universidade do Oeste Paulista Tipo do documento: Dissertação

Uso de fluido ovidutal sintético (SOF) como meio único na produção in vitro de embriões (PIVE) bovinos

2026-10-08T04:05:47Z

Título: Uso de fluido ovidutal sintético (SOF) como meio único na produção in vitro de embriões (PIVE) bovinos Autor: Sampaio, Amanda Talys Primeiro orientador: Silvestre, Caliê Castilho Abstract: In vitro embryo production (IVEP) has become one of the main reproductive biotechnologies, with Brazil recognized as the world’s largest producer of embryos. IVEP consists of three main stages: in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC), which traditionally require different culture media to meet the specific metabolic demands of gametes and embryos at each stage. The aim of this study was to evaluate the efficiency of using Synthetic Oviduct Fluid (SOF) as a single medium for IVM, IVF, and IVC by analyzing cumulus cell expansion, nuclear maturation, lipid droplet content, cleavage and blastocyst production rates, and the expression of genes related to metabolism (SCD1, SREBF1 and SLC2A1), oxidative stress (GPX1, SOD1, SOD2 and CPT1B), apoptosis (CASP03, BCL2 and BAX), and embryo implantation (VEGF). Ovarian follicles obtained from a local slaughterhouse were aspirated, and cumulus–oocyte complexes (CCOs), classified as grades I to III, were selected and randomly allocated into two experimental groups: a Control group, using conventional media for IVM (TCM-199 supplemented with hormones and 10% fetal bovine serum) and IVF (TALP), and an SOF group, in which SOF-based media were used for IVM (SOF-IVM supplemented with hormones and 10% fetal bovine serum) and IVF (SOF-IVF). Cumulus cell expansion was evaluated before and after maturation. After IVM, a subset of oocytes was subjected to Sudan Black staining for lipid content assessment and Hoechst staining for nuclear maturation analysis and mitochondrial intensity analysis by MitoTracker Red CMXROs fluorescence, while the remaining oocytes were used for IVF and IVC to assess embryo development and the expression of 11 target genes in the resulting blastocysts. Oocytes matured in SOF medium showed higher nuclear maturation rates to metaphase II (p = 0.01), lower degeneration rates (p = 0.05) and lower mitochondrial intensity (p < 0.0001), although greater cumulus cell expansion was observed in the Control group (p = 0.001). The use of SOF did not affect oocyte lipid content (p = 0.39), cleavage rate (71% vs. 68.5%; p = 0.74), blastocyst production (23.6% vs. 20%; p = 0.54), or the molecular profile of embryos (p > 0.05). In conclusion, SOF used as a single culture medium throughout all stages of in vitro embryo production is a viable alternative to conventional media, maintaining oocyte competence, embryo development, and molecular markers associated with embryonic production. Instituição: Universidade do Oeste Paulista Tipo do documento: Dissertação