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Anne-Laure Nivet, Audrey Bunel, Rémi Labrecque, Josée Belanger, Christian Vigneault, Patrick Blondin, and Marc-André Sirard

Combinations of genetic, environmental, and management factors are suspected to explain the loss in fertility observed for over 20 years in dairy cows. In some cases, IVF is used. When compared with in vivo embryo production, IVF resulted in low success rates until the FSH coasting process (FSH starvation after superstimulation) was introduced in 2002. Increased competence associated with FSH withdrawal of aspirated oocyte for in vitro maturation and IVF has not been optimized nor explained yet. The goal here was to determine and characterize the optimal oocyte competence acquisition window during the coasting period by determining blastocyst rates and follicular cohort development. Commercial milking cycling cows (n=6) were stimulated with 3 days of FSH (6×40 mg NIH Folltropin-V given at 12 h intervals) followed by a coasting period of 20, 44, 68, or 92 h. Each animal was exposed to the four conditions and served as its own control. At the scheduled time, transvaginal aspirations of immature oocytes were performed followed by IVF of half the oocytes. The outcomes were as follows: i) FSH coasting was optimal at a defined period: between 44 and 68 h of coasting; ii) The best estimated coasting duration was ∼54±7 h; iii) Under these conditions, the best statistical blastocyst rate estimation was ∼70%; iv) Between 44 and 68 h of coasting, follicle size group proportions were similar; v) Follicle diameter was not linearly associated with competence. In conclusion, coasting duration is critical to harvest the oocytes at the right moment of follicular differentiation.

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Ying Zhang, Camila Bruna de Lima, Rémi Labrecque, and Marc-Andre Sirard

Subfertile bulls may cause huge economic losses in dairy production since their semen could be used to inseminate thousands of cows by artificial insemination. This study adopted whole-genome enzymatic methyl sequencing and aimed to identify candidate DNA methylation markers in bovine sperm that correlate with bull fertility. Twelve bulls were selected (High Bull Fertility = 6; Low Bull Fertility = 6) based on the industry’s internally used Bull Fertility Index (BFI). After sequencing, a total of 450 CpG had a DNA methylation difference higher than 20% (q<0.01) had been screened. The sixteen most significant differentially methylated regions(DMRs) were identified using a 10% methylation difference cut-off (q<5.88x10-16). Interestingly, most of the differentially methylated cytosines (DMCs) and DMRs were distributed on the X and Y chromosomes, demonstrating that the sex chromosomes play essential roles in bull fertility. Additionally, the functional classification showed that the beta-defensin family, zinc finger protein family, and olfactory and taste receptors could be clustered. Moreover, the enriched G protein-coupled receptors such as neurotransmitter receptors, taste receptors, the olfactory receptor family, and ion channels indicated that the acrosome reaction and capacitation processes are pivotal for bull fertility. In conclusion, this study identified the sperm-derived bull fertility-associated DMRs and DMCs at the whole genome level, which could complement and integrate into the existing genetic evaluation methods, increasing our decisive capacity to select good bulls and explain bull fertility better in the future.