Effects of FGF10 on bovine oocyte meiosis progression, apoptosis, embryo development and relative abundance of developmentally important genes in vitro

Contents Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion-related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22h in TCM-199 supplemented with 0, 2.5, 10 or 50ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5ng/ml FGF10 increased (p<0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real-time RT-PCR showed a tendency of 50ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.

In vitro effect of calcium-containing prescription-strength fluoride toothpastes on bovine enamel erosion under hyposalivation-simulating conditions

Purpose: To evaluate the ability of calcium-containing prescription-strength fluoride (F) toothpastes in preventing enamel erosion under low salivary flow simulating conditions. Methods: Enamel and dentin bovine specimens were assigned to the following groups: A - placebo; B - 1,100 ppm F/NaF (Aquafresh Advanced); C - 5,000 ppm F/NaF (Prevident 5000 Booster); D - 5000 ppm F/NaF+calcium sodium phosphosilicate (Topex Renew); and E - 5,000 ppm F/NaF+tri-calcium phosphate (Clinpro 5000). Specimens were positioned in custom-made devices, creating a sealed chamber on the surface, connected to peristaltic pumps. Citric acid was injected into the chamber for 2 minutes, followed by artificial saliva (0.05 ml/minute), for 60 minutes, 4x/day, for 3 days. Aquafresh was also tested under normal salivary flow (0.5 ml/minute), as reference (Group F). Specimens were exposed to the toothpastes for 2 minutes, 2x/day. After cycling, surface loss (SL) and concentration of loosely- and firmly-bound F were determined. Data were analyzed by ANOVA. Results: Group A (placebo) presented highest surface loss (SL), while Group F had the lowest, for both substrates. For enamel, none of the dentifrices differed from Group B or among each other. For dentin, none of the dentifrices differed from Group B, but Group E showed greater protection than Group C. Group E presented the highest F concentrations for both substrates, only matched by Group D for firmly-bound fluoride on enamel. All fluoridated dentifrices tested reduced SL, with no additional benefit from higher F concentrations. Some formulations, especially Clinpro 5000, increased F availability on the dental substrates, but no further erosion protection was observed.

Bovine cysticercosis in slaughtered cattle as an indicator of Good Agricultural Practices (GAP) and epidemiological risk factors

This study focused on estimating the economic losses resulting from cysticercosis at beef cattle farms that supply an export slaughterhouse located in the state of Sao Paulo, Brazil, and to identify the epidemiological risks factors involved in the disease to ascertain if these farms adopt Good Agricultural Practices (GAP). To this, we used data recorded in 2012 by Brazil's Federal Inspection Service (SIP) on the daily occurrence of the disease, according to the farm from which the animals originated. In addition, the associated risk factors were determined based on a case-control study at 48 farms. Cysticercosis was detected in 2.26% (95% CI 2.2-2.33) of the 190,903 bovines supplied by 556 farms in the following four states: 2.92% (95% CI 2.83-3.03) in Sao Paulo, 1.81% (95% CI 1.71-1.93) in Minas Gerais, 0.71% (95% CI 0.6-0.82) in Goias and 1.11% (95% CI 0.79-1.57) in Mato Grosso do Sul, with significant differences in the epidemiological indices of these states. Cysticercosis was detected at 58.45% (95% CI 54.36-62.55) of the farms of this study, representing estimated economic losses of US$312,194.52 for the farmers. Lower prevalence of this disease were found at the farms qualified for exports to the European Union, indicating a statistically significant difference from those not qualified to export to Europe. The access of cattle to non-controlled water sources, as well as sport fishing activities near the farms, was identified as risk factors. Cysticercosis causes considerable losses in Brazil's beef supply chain, with lower prevalence appearing only at farms qualified to export to the European Union. As for the access of cattle to non-controlled water sources, this is an indication that GAP are not implemented by some farms, demonstrating the violation of international agreements by the industry and the farms. (C) 2015 Elsevier B.V. All rights reserved.

Paradoxical effects of bovine somatotropin treatment on the ovarian follicular population and in vitro embryo production of lactating buffalo donors submitted to ovum pick-up

The aim of the present study was to evaluate the effect of bovine somatotropin (bST; 500 mg) administration on lactating buffalo donors submitted to two different ovum pick-up (OPU) and in vitro embryo production schemes with a 7 or 14 d intersession OPU interval. A total of 16 lactating buffalo cows were randomly assigned into one of four experimental groups according to the bST treatment (bST or No-bST) and the OPU intersession interval (7 or 14 d) in a 2 x 2 factorial design (16 weeks of OPU sessions). The females submitted to OPU every 14d had a larger (P < 0.001) number of ovarian follicles suitable for puncture (15.6 +/- 0.7 vs. 12.8 +/- 0.4) and an increased (P = 0.004) number of cumulus-oocyte complexes (COCs) recovered (10.0 +/- 0.5 vs. 8.5 +/- 0.3) compared to the 7 d interval group. However, a 7 or 14 d interval between OPU sessions had no effect (P = 0.34) on the number of blastocysts produced per OPU (1.0 +/- 0.1 vs. 13 +/- 0.2, respectively). In addition, bST treatment increased (P < 0.001) the number of ovarian follicles suitable for puncture (15.3 +/- 0.5 vs. 12.1 +/- 0.4) but reduced the percentage (18.9% vs. 10.9%; P = 0.009) and the number (1.4 +/- 0.2 vs. 0.8 +/- 0.1; P = 0.003) of blastocysts produced per OPU session compared with the non-bST-treated buffaloes. In conclusion, the 14d interval between OPU sessions and bST treatment efficiently increased the number of ovarian follicles suitable for puncture. However, the OPU session interval had no effect on embryo production, and bST treatment reduced the in vitro blastocyst outcomes in lactating buffalo donors.

Molecular detection and phylogenetic analysis of bovine astrovirus in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Plasma Steroid Dynamics in Late- and Near-term Naturally and Artificially Conceived Bovine Pregnancies as Elucidated by Multihormone High-resolution LC-MS/MS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of an untyped strain of bovine respiratory syncytial virus in a dairy herd

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of cell cycle synchronization on the accuracy of murine and bovine embryo sex determination

Different cell cycle synchronization methods were used to increase the mitotic index and accuracy of sex determination in murine and bovine embryos. For sexing purposes, colchicine treatment for 2, 4, 6 and 8 h and the FdU-thymidine-colchicine combination were tested in murine embryos. The best results were obtained with colchicine treatment for 8 h (96.88% accuracy) and with FdU-thymidine-colchicine (97.22% accuracy). Mitotic indexes differed significantly between the 2 treatments (21.71% for colchicine and 32.95% for FdU-thymidine-colchicine). For sex identification of murine and bovine demi-embryos, both treatments were demonstrated to be equally effective (nearly 90%). The mitotic index for the FdU-treated murine demi-embryos (19.04%) was higher than the one obtained for the 8-h colchicine treatment (15.62%).

Penetration of 35% hydrogen peroxide into the pulp chamber in bovine teeth after LED or Nd:YAG laser activation

This aim of the present study was to evaluate the pulp chamber penetration of 35% hydrogen peroxide activated by LED (light-emitting diode) or Nd:YAG laser in bovine teeth, after an in-office bleaching technique. Forty-eight bovine lateral incisors were divided into four groups, acetate buffer was placed into the pulp chamber and bleaching agent was applied as follows: for group A (n = 12), activation was performed by LED; for group B (n = 12), activation was performed by Nd:YAG laser (60 mJ, 20 Hz); group C (n = 12) received no light or laser activation; and the control group (n = 12) received no bleaching gel application or light or laser activation. The acetate buffer solution was transferred to a glass tube and Leuco Crystal Violet and horseradish peroxidase were added, producing a blue solution. The optical density of this solution was determined spectrophotometrically and converted into microgram equivalents of hydrogen peroxide. The results were analysed using ANOVA and Tukey's test (5%). It was verified that the effect of activation was significant, as groups activated by LED or laser presented greater hydrogen peroxide penetration into the pulp chamber (0.499 +/- 0.622 microg) compared with groups that were not (0.198 +/- 0.218 microg). There was no statistically significant difference in the penetration of hydrogen peroxide into the pulp chamber between the two types of activation (LED or laser). The results suggest that activation by laser or LED caused an increase in hydrogen peroxide penetration into the pulp chamber.

Immunohistochemical approach to the pathogenesis of clinical cases of bovine Herpesvirus type 5 infections

Meningoencephalitis by Herpesvirus type 5 (BoHV-5) in cattle has some features that are similar to those of herpetic encephalitis in humans and other animal species. Human Herpesvirus 3 (commonly known as Varicella-zoster virus 1), herpes simplex viruses (HSV), and equid Herpesvirus 1 (EHV-1) induce an intense inflammatory, vascular and cellular response. In spite of the many reports describing the histological lesions associated with natural and experimental infections, the immunopathological mechanisms for the development of neurological disorder have not been established. A total of twenty calf brains were selected from the Veterinary School, University of São Paulo State, Araçatuba, Brazil, after confirmation of BoHV-5 infection by virus isolation as well as by a molecular approach. The first part of the study characterized the microscopic lesions associated with the brain areas in the central nervous system (CNS) that tested positive in a viral US9 gene hybridization assay. The frontal cortex (Fc), parietal cortex (Pc), thalamus (T) and mesencephalon (M) were studied. Secondly, distinct pathogenesis mechanisms that take place in acute cases were investigated by an immunohistochemistry assay. This study found the frontal cortex to be the main region where intense oxidative stress phenomena (AOP-1) and synaptic protein expression (SNAP-25) were closely related to inflammatory cuffs, satellitosis and gliosis, which represent the most frequently observed neurological lesions. Moreover, MMP-9 expression was shown to be localized in the leptomeninges, in the parenchyma and around mononuclear infiltrates (p < 0.0001). These data open a new perspective in understanding the role of the AOP-1, MMP-9 and SNAP-25 proteins in mediating BoHV-5 pathogenesis and the strategies of host-virus interaction in order to invade the CNS.

Estimation of the diagnostic accuracy of the glyco-C and US9 gene-based polymerase chain reaction technique for the detection of bovine Herpesvirus type 5 DNA in decomposed brain suspension from a slaughter house using Bayesian analysis, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of incorporation of remineralizing agents into bleaching gels on the microhardness of bovine enamel in situ

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prevalence survey of selected bovine pathogens in water buffaloes in the north region of Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Different extenders in the cryopreservation of bovine epididymal spermatozoa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of sex on basal and dickkopf-1 regulated gene expression in the bovine morula

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)