Effect of IGFBP-4 during In Vitro Maturation on Developmental Competence of Bovine Cumulus Oocyte Complexes.

Insulin-like growth factors (IGFs) are essential for oocyte maturation. Their bioavailability is regulated by their respective binding proteins (IGFBPs) and proteases. IGFBP-4 blocks the biological effects of IGFs. High IGFBP-4 expression has been associated with follicle atresia. We hypothesized that IGFBP-4 affects oocyte developmental competence during maturation. Therefore, the aim of this study was to examine the effect of IGFBP-4 on the developmental rate of bovine cumulus-oocyte complexes (COCs) during in vitro embryo production. Abattoir-derived COCs were matured with rbIGFBP-4 (2000, 540, and 54 ng/mL) compared to a control. Cumulus expansion, oocyte maturation, cleavage, blastocyst, and hatching rates were evaluated. Furthermore, blastocyst gene expression of SOCS2, STAT3, SLC2A1, SLCA3, BAX, and POU5F1 transcripts were quantified using RT-qPCR. No statistical differences were detected among the groups for cumulus expansion, maturation, cleavage, blastocyst rates, or all gene transcripts analyzed. However, at day 8 and 9, the number of total hatching and successfully hatched blastocysts was lower in 2000 ng/mL rbIGFBP-4 compared to the control (day 8: total hatching: 17.1 ± 0.21 vs. 31.2 ± 0.11%, p = 0.02 and hatched blastocyst 6.7 ± 0.31 vs. 21.5 ± 0.14%, p = 0.004; day 9 total hatching 36.4 ± 0.18 vs. 57.7 ± 0.10%, p = 0.009 and hatched blastocyst 18.2 ± 0.21 vs. 38.1 ± 0.11%, p = 0.004). We concluded that high concentrations of rbIGFBP-4 might negatively affect the subsequent ability of the embryo to hatch and possibly compromise further elongation.

Effect of One-Day Delaying CIDR Administration in 5-Day Cosynch Protocol in Dairy Heifers.

Effect of one-day delaying progesterone administration at the beginning of 5-day Cosynch protocol was investigated in Holstein heifers for the first artificial insemination (AI). Heifers received a synchronized ovulation and timed AI (TAI) with CIDR inserted on day (d) 0 (CIDR-5; n = 206) or d 1 (CIDR-4; n = 192). In both group, GnRH was administered on d 0 followed by a single PGF2α injection and CIDR removal five days later from GnRH. On d 8, TAI and GnRH administration were concurrently conducted. Heifers detected in estrus up to 24 h prior to TAI were inseminated without GnRH administration. Rates of ovulation, accessory CL formation and new dominant follicle development following initial GnRH injection did not differ between groups. P/AI did not differ between CIDR-4 (44.3%, [85/192]) and CIDR-5 (51.9%, [107/206]) groups, respectively. Pregnancy per AI (P/AI) was significantly (p < 0.01) declined as heifers' age (12-13, 14, 15, 16 and17-21 months) proceeded in CIDR-4 group (55.6%, 52.1%, 37.9%, 35.7%, 32.4%) compared to those in CIDR-5 group (60.0%, 50.0%, 53.9%, 51.5%, 46.2%) respectively. In conclusion, there is no benefit for delaying CIDR administration in 5-day Cosynch protocol in dairy heifers. However, higher P/AI in CIDR-5 group in older heifers can be considered for reproductive management.

Inter- and intra-individual variability of total antioxidant capacity of bovine seminal plasma and relationships with sperm quality before and after cryopreservation.

The aim of the present study was to investigate inter- and intra-individual variability of total antioxidant capacity (TAC) in seminal plasma of bulls. In addition, relationships between TAC and glutathione peroxidase (GPx), superoxide dismutase activities (SOD), and parameters of sperm quality, respectively, were examined. Eight consecutive ejaculates were collected from nine Holstein-Friesian bulls. The percentage of plasma membrane and acrosome intact (PMAI) sperm was measured by using the FITC-PNA/PI assay, the amount of membrane lipid peroxidation (LPO) without and with stimulation(s) of LPO was quantified by using the BODIPY assay before cryopreservation and immediately (0 h) as well as 3h after thawing. The percentage of sperm with a greater DNA fragmentation index was measured by the sperm chromatin structure assay. The amount of TAC differed (P < 0.0001) between bulls but not (P > 0.05) between ejaculates within bulls. The amounts of TAC were not related (P > 0.05) to amounts of SOD and GPx, but were negatively associated with LPO 0 h (r = -0.85; P ≤ 0.01). The amounts of SOD showed positive relationships with LPO 0 h (r = 0.71; P ≤ 0.05) and LPO 3h (r = 0.80; P ≤ 0.05). In conclusion, total antioxidant activity varied among bulls, but not between ejaculates within bulls. While the amounts of antioxidative enzyme GPx was not related to sperm quality and SOD was positively related with lipid peroxidation after thawing of sperm, whereas total antioxidative capacity was negatively correlated with lipid peroxidation of cryopreserved sperm.

Effects of feeding omega-3-fatty acids on fatty acid composition and quality of bovine sperm and on antioxidative capacity of bovine seminal plasma.

The aim of the present study was to examine the effects of feeding alpha-linolenic (ALA) acid on fatty acid composition and quality of bovine sperm and on antioxidative capacity of seminal plasma. Nine bulls (ALA bulls) were fed with 800 g rumen-resistant linseed oil with a content of 50% linolenic acid and eight bulls with 400 g palmitic acid (PA bulls). Sperm quality was evaluated for plasma membrane and acrosome intact sperm (PMAI), the amount of membrane lipid peroxidation (LPO), and the percentage of sperm with a high DNA fragmentation index (DFI). Fatty acid content of sperm was determined using gas chromatography. Total antioxidant capacity, glutathione peroxidase, and superoxide dismutase activity were determined in seminal plasma. Feeding ALA increased (P < 0.05) the docosahexaenoic acid (DHA) content in bulls whereas in PA bulls did not change. PMAI increased after cryopreservation in ALA bulls as well as in PA bulls during the experiment period (P < 0.005). LPO of sperm directly after thawing did not change during the study period in ALA group, but decreased in PA group (P < 0.006). After 3h of incubation LPO increased in the ALA group (P < 0.02), while LPO did not differ between phases within groups. In conclusion, feeding of neither saturated nor polyunsaturated fatty acids affect the antioxidant levels in seminal plasma. Both saturated as well as polyunsaturated fatty acids had positive effects on quality of cryopreserved bovine sperm, although the content of docosahexaenoic acid in sperm membranes increased only in ALA bulls.