Effect of butafosfan supplementation during oocyte maturation on bovine embryo development.

Around 60-80% of oocytes maturated in vivo reached competence, while the proportion of maturation in vitro is rarely higher than 40%. In this sense, butafosfan has been used in vivo to improve metabolic condition of postpartum cows, and can represent an alternative to increase reproductive efficiency in cows. The aim of this study was to evaluate the addition of increasing doses of butafosfan during oocyte maturation in vitro on the initial embryo development in cattle. In total, 1400 cumulus-oocyte complexes (COCs) were distributed in four groups and maturated according to supplementation with increasing concentrations of butafosfan (0 mg/ml, 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml). Then, 20 oocytes per group were collected to evaluate nuclear maturation and gene expression on cumulus cells and oocytes and the remaining oocytes were inseminated and cultured until day 7, when blastocysts were collected for gene expression analysis. A dose-dependent effect of butafosfan was observed, with decrease of cleavage rate and embryo development with higher doses. No difference between groups was observed in maturation rate and expression of genes related to oocyte quality. Our results suggest that butafosfan is prejudicial for oocytes, compromising cleavage and embryo development.

Effect of yeast products supplementation during transition period on metabolic profile and milk production in dairy cows.

The aim of the current study was to assess the effect that supplementation with yeast culture plus enzymatically hydrolyzed yeast (YC-EHY) during the transition period and lactation had on the performance, somatic cell count (SCC), and metabolic profile of dairy cows. Thirty multiparous Holstein dairy cows were divided into two groups. The treatments were 0 supplementation (control) and supplementation with 28 g/cow/day of YC-EHY. The supplementation began 35 ± 5 days before the expected calving date. The cows were kept in their respective treatments for 50 days after the calving date. Body condition score (BCS), body weight, milk composition, SCC, and milk yield were assessed on a 2-weekly basis. Plasma samples were collected on days - 21st, - 7th, 0, 3rd, 7th, and weekly thereafter until 42 days postpartum and analyzed for albumin, ß-hydroxybutyrate (BHBA), urea, and non-esterified fatty acids (NEFA). There was an effect of treatment on milk yield in the supplemented animals in comparison to the control group (27.88 ± 0.98 vs 24.58 ± 0.99 kg/days, P = 0.03). There was no effect of treatment (P > 0.05) on variables like 3.5% fat-corrected milk (FCM) and energy-corrected milk (ECM), milk component (%), milk composition yield (kg/day), and SCC. There was an interaction between group × days on ECM (P = 0.04) and protein (P = 0.008). The supplementation had no effect on the analyzed metabolites and on body weight and BCS. The supplementation with YC-EHY during the transition period and lactation improved milk yield without altering the metabolic profile.

Prostaglandin F2α regulation and function during ovulation and luteinization in cows.

Although prostaglandins are important in the ovulation process, a precise role for prostaglandin F2α (PGF) has not been elucidated. This study aimed to evaluate the regulation of PGF receptor mRNA (PTGFR) in granulosa cells and the local effect of PGF on ovulation and luteinization. In Experiment 1, using samples collected in vivo before (Day 2), during (Day 3) and after (Day 4) follicular deviation, expression of PTGFR in bovine granulosa cells was more abundant in the dominant follicle after deviation than in subordinates (P < 0.05). However, the expression of PTGFR was not regulated (P = 0.1) in preovulatory follicles at different time-points (0, 3, 6, 12 and 24 h) after ovulation induction with GnRH. In Experiment 2, to assess the role of systemic PGF treatment on luteinization and vascularization of preovulatory follicles, flunixin meglumine (FM), a nonsteroidal anti-inflammatory drug, was used to inhibit endogenous prostaglandin synthesis. Cows with preovulatory follicles were induced to ovulate with GnRH (0 h) and allocated to three groups: Control, with no further treatment; FM, treated with 2.2 mg/kg FM im 17 h after GnRH treatment; and FM + PGF, treated with FM 17 h after GnRH, followed by 25 mg dinoprost tromethamine (PGF) 23 h after GnRH treatment. FM injection was able to reduce the concentration of PGF in the follicular fluid (FF) (P < 0.001). However, contrary to our hypothesis, color Doppler ultrasound evaluations revealed decreased vascular flow in FM + PGF group (P < 0.05), and no effect of the treatments on intrafollicular P4 and E2 concentrations 24 h after GnRH. The prostaglandin metabolite (PGFM) concentrations in the FF were greater in cows receiving systemic PGF (P < 0.001), which prompted us to further check its role on ovulation. Therefore, in Experiment 3, in a final attempt to demonstrate the local effect of PGF on ovulation, cows with preovulatory follicles received an intrafollicular injection (IFI) of PBS (Control) or 100 ng/mL purified PGF (PGF group). PGF treatment did not affect the time of ovulation after IFI (66 ± 6.4 and 63 ± 8.5 h for control and PGF, respectively; P > 0.05), further suggesting that it has no direct effect in the ovulatory process. Based on our findings, we concluded that FM decreased PGF synthesis within the follicle, whereas PGF treatment decreased follicular vascularization. In addition, the in vivo model of intrafollicular injection evidenced that PGF alone is not able to locally induce ovulation.

Intrafollicular lipopolysaccharide injection delays ovulation in cows.

The aim of this study was to evaluate effects of intra-follicular (i.f.) treatment with lipopolysaccharide (LPS) on follicular and luteal development in cows. There were 18 non-lactating cows assigned to two groups to address this aim: control group (n = 9), which received an i.f. injection of saline; and LPS group (n = 9), which received an i.f. injection of 1 µg of LPS per mL of follicular fluid. Cows were treated with an intravaginal P4 releasing device (IVD) and estradiol benzoate on D0. On D4 and D5 cows were treated with cloprostenol sodium and on D7 the IVD was removed. At 12 h after IVD removal, cows were administered the i.f. injection of LPS or saline. After administration of these treatments, follicular development was evaluated every 12 h until ovulation. The LPS treatment increased blood flow in pre-ovulatory follicles (P = 0.05). Follicle growth was reduced by LPS injection (P < 0.02) resulting a longer period to the time of ovulation for cows in the LPS than control group (P = 0.03). The percentage of cows having ovulations was less for the LPS than control group (P = 0.03). The diameter of the CL, CL blood flow and P4 concentrations 5 and 12 days after ovulation did not differ between groups (P> 0.05). In conclusion, intra-follicular treatment with LPS resulted in a decreased rate of follicle growth, delayed timing of ovulations and a lesser number of cows having ovulations.

Hipomagnesemia subclínica em vacas leiteiras durante o período de transição: ocorrências hormonais e metabólicas

O objetivo, com este estudo, foi investigar as conseqüências da hipomagnesemia subclínica sobre parâmetros metabólicos de vacas leiteiras. Amostras de sangue foram coletadas de 12 animais, a cada dois dias, entre os dias 22 pré e 22 pós-parto, para a realização de análises sanguíneas e determinação do perfil metabólico. Os animais foram categorizados de acordo com os níveis séricos de magnésio: Grupo Hipomagnesêmicas (n=5), com níveis abaixo de 1,8 mg/dL em ao menos dois dias consecutivos, e o Grupo Controle (n=7), com níveis acima de 1,8 mg/dL em todo o período. O grupo hipomagnesêmicas apresentou níveis de glucagon maiores nos dias 10, 18 e 20, e a taxa Glucagon/Insulina foi maior nos dias 6, 8, 10, 12 e 16 após o parto. As concentrações de aspartato amino transferase foram maiores no grupo hipomagnesêmicas nos dias 0, 6, 8, 10, 12 e 14 pós-parto. Os resultados indicaram que níveis reduzidos de magnésio no periparto podem estar relacionados com níveis elevados de aspartato amino transferase e de glucagon. Em geral, a hipomagnesemia subclínica não altera os níveis dos indicadores do metabolismo energético, mas os resultados demonstraram que as vacas com hipomagnesemia apresentaram maior taxa de glucagon/insulina, demonstrando um maior desafio para manter os níveis glicêmicos.