Guilherme Batista dos Santos

Presynchronization by induction of a largest follicle using a progesterone device in GnRH-based-ovulation synchronization protocol in crossbred dairy cows

The objective was to compare the fertility of dairy cows using a presynchronization protocol by induction of a largest follicle using a progesterone intravaginal device prior to an Ovsynch protocol (P4synch) with the Double-Ovsynch in lactating dairy cows. Lactating Bos indicus x Bos taurus crossbred cows (n = 440) were randomly allocated to one of two treatments: (I) Double-Ov (n = 228), GnRH (D-17), PGF2α 7 days later (D-10) and GnRH 3 days later (D-7) followed by an Ovsynch protocol 7 days later (GnRH on D0, PGF on D7, GnRH on D9); (II) P4synch (n = 212), insertion of a sustained release progesterone intravaginal device (D-10), 10 days later (D0) an Ovsynch protocol was initiated (GnRH on D0, PGF on D7, GnRH on D9) with progesterone device withdrawal on Day 7. All cows were artificially inseminated (TAI) 16 h after the second GnRH of the Ovsynch protocol and pregnancy diagnosis was performed by transrectal ultrasonography 30 and 60 days after TAI. A subset of cows (n = 52 for Double-Ov and n = 50 for P4synch) ultrasonography was performed on days 0, 7, 9 and 24 of the experimental period. There were no differences among treatments on presynchronization rate [presence of a follicle>12 mm on D0, Double-Ov 94.2% (49/52) and P4synch 92.0% (46/50); P = 0.66], follicular diameter on the 1st GnRH (Double-Ov 17.2 ± 0.7 mm e P4synch 18.6 ± 0.9 mm; P = 0.28), ovulation rate to the 1st GnRH [Double-Ov 86.3% (44/51) and P4synch 81.2% (39/48); P = 0.50], synchronization rate [presence of a follicle>12 mm on D9; Double-Ov 84.6% (44/52) and P4synch 86.0% (43/50); P = 0.84], follicular diameter on the 2nd GnRH (Double-Ov 17.5 ± 0.6 mm and P4synch 18.0 ± 0.5 mm; P = 0.48), ovulation rate to the 2nd GnRH [Double-Ov 90.9% (40/44) and P4synch 86.0% (37/43); P = 0.48] and CL diameter on D24 (Double-Ov 27.9 ± 0.7 mm and P4synch 29.4 ± 0.9 mm; P = 0.19). Corpus luteum presence on D0 was different (P = 0.03) among treatments [Double-Ov 57.7% (30/52) and P4synch 36.0% (18/50)]. There was no difference (P = 0.85) among the pregnancy per AI on day 30 [Double-Ov 39.0% (89/228) and P4synch 40.1% (85/212)], on day 60 [Double-Ov 34.8% (79/227) and P4synch 38.7% (82/212); P = 0.41] and pregnancy loss [Double-Ov 10.2% (9/88) and P4synch 3.5% (3/85); P = 0.08]. The presynchronization by induction of a largest follicle using a sustained release progesterone device prior to Ovsynch yielded similar results compared with the Double Ovsynch protocol on follicular development patterns and on the fertility of lactating dairy cows.

Subclinical mastitis interferes with ovulation, oocyte and granulosa cell quality in dairy cows

The objective was to evaluate the effect of mastitis by somatic cell count (SCC) on follicular growth, ovulation, oocytes and cumulus cells quality and on the concentration and size of exosomes in follicular fluid of dairy cows. In the study, crossbred cows (Bos taurus - Holstein x Bos indicus - Gir) were classified for analysis as Control (SCC<200.000 cells/mL) and Mastitis (SCC>400.000 cells/mL) groups. In experiment 1 (follicular dynamics), cows (n = 57) were submitted to ultrasound evaluations every 24 h, from progesterone-releasing-intravaginal-device (PRID) removal (D8) until 48 h later (D10). Thereafter, evaluations were performed every 12 h, until ovulation or up to 96 h after PRID removal. In experiment 2 (oocyte, cumulus complexes, and follicular fluid evaluation), cows (n = 26) were submitted to follicular aspiration (OPU) for oocyte quality and cumulus cells transcript evaluation. The amount of cumulus complexes transcripts (BCL2, BAX, PI3K, PTEN, FOXO3) was determined by Real-Time Polymerase Chain Reaction. Moreover, seven days after the OPU session, the dominant follicle was aspirated. Exosomes were isolated from the follicular fluid for evaluation of particle size and concentration. Ovulation rate [Control 77.4% (24/31) and Mastitis 57.7% (15/26); P = 0.09] and viable oocytes rate [Control 59.1% (130/220) and Mastitis 41.9% (125/298); P = 0.01] were higher in Control animals. Additionally, there was a greater number of degenerate oocytes [Control 6.7 ± 1.2 and Mastitis 13.3 ± 5.5; (P = 0.001)] in subclinical mastitis cows. There was greater abundance (P = 0.003) of BAX cumulus cell transcripts and exosome mean (P = 0.03) and mode (P = 0.02) was smaller in subclinical mastitis cows. In conclusion, ovulation rate, oocyte quality, and exosome diameter were smaller in cows with SCC>400.000 cells/mL.

Molecular and endocrine factors involved in future dominant follicle dynamics during the induction of luteolysis in Bos indicus cows

The growth profiles of the future dominant follicle (DF) and subordinate follicle (SF) and the gene expression of the granulosa cells during luteolysis induction in Bos indicus cows were evaluated. Forty cows were synchronized with a progesterone and estradiol based protocol. After synchronization, cows with a corpus luteum (CL) were evaluated by ultrasonography every 12 h, beginning at eight days post ovulation. Cows identified with a follicle of at least 6.0 mm in diameter in the second wave were split into two groups (BD-before follicular deviation and AD-after follicular deviation. In the BD group cows received 500 μg of cloprostenol (a synthetic analogue of prostaglandin F2α) when the DF reached a mean diameter of 7.0 mm (6.5-7.5 mm). In the AD group, cows received 500 μg of cloprostenol when the DF reached a mean diameter of 8.0 mm (7.5-8.5 mm). Cows in both groups were submitted to aspiration of the DF at 96 and 72 h after prostaglandin was given. Follicular aspirations were performed to quantify IGF1R, LHR and PAPPA transcripts in the granulosa cells. The diameter of the DF at the moment of prostaglandin administration (P = 0.001) and the growth rate of the SF (P = 0.05) were greater in the AD group. There was greater abundance of LHR transcripts in BD cows (P = 0.04). The remaining variables tested were similar between the experimental groups (P > 0.05). In conclusion, the induction of luteolysis before follicular deviation does not interfere with dominant follicle dynamics. However, it causes granulosa cell LHR down regulation.