Chemical gasification: An alternative approach to in vitro maturation of bovine oocytes.

This study aimed to evaluate the effect of chemical gasification and HEPES as alternative systems to pH control during in vitro maturation on bovine oocytes competence. Groups of 20 bovine cumulus oocytes complexes (COCs) were randomly distributed and cultured for 24 h in one of the following experimental groups: (i) chemical reaction (ChRG) system: CO2 generated from sodium bicarbonate and citric acid reaction (ii) culture media TCM-HEPES (HEPES-G); and (iii) control group (CNTG) in conventional incubator. After in vitro maturation (IVM), the COCs were in vitro fertilized (IVF), and in vitro cultivated (IVC) in a conventional incubator. We evaluated oocyte nuclear maturation, cleavage and blastocyst rates, in addition to the relative mRNA expression of BAX, BMP-15, AREG and EREG genes in oocytes and cumulus cells. The proportion of oocytes in metaphase II was higher in CNTG and ChRG (77.57% and 77.06%) than in the HEPES-G (65.32%; p = .0408 and .0492, respectively). The blastocyst production was similar between CNTG and ChRG (26.20% and 28.47%; p = .4232) and lower (p = .001) in the HEPES-G (18.71%). The relative mRNA expression of BAX gene in cumulus cells was significantly higher (p = .0190) in the HEPES-G compared to the CNTG. Additionally, the relative mRNA expression of BMP-15 gene was lower (p = .03) in oocytes from HEPES-G compared to the CNTG. In conclusion, inadequate atmosphere control has a detrimental effect on oocyte maturation. Yet, the use of chemical gasification can be an efficient alternative to bovine COCs cultivation.

Rumen-protected methionine supplementation alters lipid profile of preimplantation embryo and endometrial tissue of Holstein cows.

Our objective is to evaluate the effects of feeding rumen-protected Met (RPM) throughout the transition period and early lactation on the lipid profile of the preimplantation embryos and the endometrial tissue of Holstein cows. Treatments consisted of feeding a total mixed ration with top-dressed RPM (Smartamine® M, Adisseo, Alpharetta, GA, United States; MET; n = 11; RPM at a rate of 0.08% of DM: Lys:Met = 2.8:1) or not (CON; n = 9, Lys:Met = 3.5:1). Endometrial biopsies were performed at 15, 30, and 73 days in milk (DIM). Prior to the endometrial biopsy at 73 DIM, preimplantation embryos were harvested via flushing. Endometrial lipid profiles were analyzed using multiple reaction monitoring-profiling and lipid profiles of embryos were acquired using matrix assisted laser desorption/ionization mass spectrometry. Relative intensities levels were used for principal component analysis. Embryos from cows in MET had greater concentration of polyunsaturated lipids than embryos from cows in CON. The endometrial tissue samples from cows in MET had lesser concentrations of unsaturated and monounsaturated lipids at 15 DIM, and greater concentration of saturated, unsaturated (specifically diacylglycerol), and monounsaturated (primarily ceramides) lipids at 30 DIM than the endometrial tissue samples from cows in CON. In conclusion, feeding RPM during the transition period and early lactation altered specific lipid classes and lipid unsaturation level of preimplantation embryos and endometrial tissue.

Effects of rumen-protected methionine and choline supplementation on vaginal discharge and uterine cytology of Holstein cows.

Methionine is one of the most limiting amino acids in dairy diets and low feed intake around the time of calving could lead to decreased synthesis of phosphatidylcholine. An alternative pathway for phosphatidylcholine is to have choline as a precursor. The objective of this study was to determine the effects of feeding rumen-protected methionine and choline pre - and postpartum on reproduction of Holstein cows. Seventy-two Holstein cows were randomly assigned to four treatments from 21 days before calving to 30 days in milk (DIM): supplementation with rumen-protected methionine (MET; n = 20, received 0.08% of the dry matter (DM) of the diet/d as methionine, Smartamine M® to a Lys:Met = 2.9:1), rumen-protected choline (CHO; n = 17, received 60 g/d choline, Reassure), both rumen protected methionine and choline (MIX; n = 19, received 0.08% of the DM of the diet/d as methionine to a Lys:Met = 2.9:1 and 60 g/d choline), or no supplementation to serve as control (CON; n = 16, fed total mixed ration with a Lys:Met = 3.5:1). Cows were evaluated at 4, 7, 10, 13, 15, 17, and 30 d after calving for the presence of secretion using the Metricheck® device. On 15, 30, and 72 d after calving, the uterine endometrium of all cows was sampled using a cytological brush and streaked onto slides for analysis of the presence of polymorphonuclear neutrophils (PMN). We hypothesized that cows supplemented with methionine would have lower metricheck smell scores and lower rates of PMN than non-supplemented cows. On d 30, a treatment difference was detected using the metricheck score and smell (P < 0.04), with treatment MIX (score = 0.38) having a lower score than CHO (score = 2.11). Supplementing cows with rumen-protected methionine may have a beneficial effect on cows' uterine health.