176 BLOOD FLOW TO THE CORPUS LUTEUM AND PREOVULATORY FOLLICLE AFTER OVULATION INDUCTION DURING FIRST VERSUS SECOND WAVE IN WATER BUFFALO

Abstract

The objective of the study was to compare the blood flow to the corpus luteum (CL) and the preovulatory follicle in dairy buffalo (Bubalus bubalis) when ovulation was induced during the first (low to increasing progesterone levels) versus the second (luteal progesterone levels) follicular wave. We hypothesised that the wall of the first-wave dominant follicle will be less vascular compared with that of the second-wave follicle. The study was conducted during the summer months in Punjab, India. Ovulation was synchronized with prostaglandin F2α (PGF) IM followed by gonadotropin-releasing hormone (GnRH) IM 48 h later (Day 0) and buffaloes were randomised to first wave (FW; n = 6) and second wave (SW; n = 7) groups. FW group was given PGF on Days 6.5 and 7, and GnRH on Day 9.5 followed by AI (14–16 h after GnRH). The SW group was given GnRH on Day 7 (to induce ovulation of first-wave dominant follicle without luteolysis and synchronous emergence of next wave), PGF on Days 13.5 and 14, GnRH on Day 16.5 followed by artificial insemination. Transrectal colour Doppler ultrasonography (MyLab5 Vetwith 7.5 MHz transducer, Esaote S.p.A, Genoa, Italy) was performed daily and 20-s cineloops of each ovary were recorded under standardized gain controls. Images from the cineloops were processed using Fiji (ImageJ, National Institutes of Health, Bethesda, MD, USA) to calculate the area of blood flow (coloured area = vascular area, grey scale area = tissue area, and their ratio) for the preovulatory follicle (on the day before ovulation) and luteal tissue (on the day of PGF injection and 4 days post-ovulation). Data were analysed by t-test from the animals that ovulated one day before (n = 3) or the day of AI (n = 6) and had a functional CL at day 5 post-AI (FW n = 5, SW n = 4). FW follicles ovulated on 8.6 ± 0.3 days from wave emergence compared with SW follicles on 10.0 ± 0.6 days (P < 0.05) but were similar in size (i.e. follicular area on the day before ovulation did not differ between groups; P = 0.5). There was no difference in the blood flow area in the wall of preovulatory follicles (P = 0.4). Vascular area of follicles was strongly correlated with their diameter (r = 0.87). Follicles >13.5 mm in diameter had more blood flow in their wall than smaller follicles (P < 0.01). FW had a tendency (P = 0.07) for smaller luteal area on the day of PGF treatment (FW = 171 ± 24 mm²; SW = 332 ± 81 mm²) and tended (P = 0.06) to have less vascular area in the CL compared to SW group (FW = 30 ± 6 mm²; SW = 67 ± 17 mm²). There was no difference (P = 0.5) between the groups for vascular to CL area ratio. The area of luteal tissue and blood flow to the CL at Day 4 post-ovulation did not differ between the groups (P = 0.4). The diameter of the preovulatory follicle (11.6–15.7 mm) was not correlated with the cross-sectional area of developing CL at Day 4 post-ovulation (r = 0.09). In conclusion, vascularity to preovulatory follicles originating from the first wave v. second wave did not differ and preovulatory follicles ≥13.5 mm were more vascular than smaller follicles.

Research was funded by NSERC; the first author was funded by scholarships from WCVM and GADVASU.