Katia Cristina Silva-Santos

Antral Follicle Populations and Embryo Production – In Vitro and In Vivo – of Bos indicus–taurus Donors from Weaning to Yearling Ages

Interest in indicus-taurus cattle has been increasing, as these animals are likely to present the best characteristics of Zebu and European bovine breeds. The aim of this study was to compare the embryo production of indicus-taurus donors with high vs low antral follicle counts obtained by ovum pickup/in vitro production (OPU/IVP) and superovulation (SOV)/embryo collection. Braford females at weaning age (3/8 Nelore × 5/8 Hereford, n = 137, 9 ± 1 month old) were subjected to six serial ovarian ultrasonographs and were assigned to two groups according to the number of antral follicles ≥ 3 mm as follows: G-High antral follicular count (AFC, n = 20, mean ≥ 40 follicles) and G-Low AFC (n = 20, mean ≤ 10 follicles). When the females (n = 40) reached 24 months of age, they were subjected to both OPU/IVP and SOV/embryo collection. The average number of follicles remained highly stable throughout all of the ultrasound evaluations (range 0.90-0.92). The mean number of COCs recovered (36.90 ± 13.68 vs 5.80 ± 3.40) was higher (p < 0.05) for females with high AFC, resulting in higher (p < 0.05) numbers of total embryos among females with high vs low AFC (6.10 ± 4.51 vs 0.55 ± 0.83). The mean number of embryos per collection was also higher (p < 0.05) for G-High vs G-Low (6.95 ± 5.34 vs 1.9 ± 2.13). We conclude that a single ultrasound performed at pre-pubertal ages to count antral follicles can be used as a predictor of embryo production following IVP and SOV/embryo collection in indicus-taurus females.

High numbers of antral follicles are positively associated with in vitro embryo production but not the conception rate for FTAI in Nelore cattle.

The objective was to compare the conception rates for FTAI and in vitro embryo production between Nelore cows with different antral follicle counts (AFC=number of follicles ≤3mm in diameter in the ovaries). Nelore cows (n=547) were subjected to ovulation synchronization. Randomly during the estrous cycle (D0), cows received an intravaginal device containing 1.9g P4 (CIDR(®)) and 2mg BE (Estrogin(®)), IM. When the device was removed (D8), the cows received 500μg PGF2α (Ciosin(®)), 300IU eCG (Novormon(®)) and 1mg EC (ECP(®)), IM. All cows were inseminated 48h after P4 device removal. Antral follicles ≥3mm were counted using an intravaginal microconvex transducer (D0), and the cows were assigned to high (G-High, ≥25 follicles, n=183), intermediate (G-Intermediate, 16-20 follicles, n=183) or low AFC groups (G-Low, ≤10 follicles, n=181). In another experiment, COCs were retrieved by OPU from Nelore cows (n=66), which were assigned to groups according to oocyte production: G-High (n=22, ≥40 oocytes), G-Intermediate (n=25, 18-25 oocytes) or G-Low (n=19, ≤7 oocytes). All COCs from the same cow were cultured individually (maximum of 25 COCs per drop) and then in vitro fertilized using thawed frozen sperm (2×10(8)/dose) from a Nelore sire of known fertility. The data were analyzed using a Kruskal-Wallis and a Chi-square test (P≤0.05). There was no difference in the conception rates after FTAI between Nelore cows with high, intermediate or low AFC (51.9 vs. 48.6 vs. 58.6%). The number of viable embryos was 18.4±6.7 (G-High), 6.1±3.6 (G-Intermediate) and 0.6±0.7 (G-Low; P<0.05). Therefore, AFC had no influence on the conception rates for FTAI; however, Nelore cows with high oocyte production exhibited better in vitro embryo production.

Ovarian follicular dynamics and conception rate in Bos indicus cows with different antral follicle counts subjected to timed artificial insemination

Two experiments in Nelore cows subjected to timed artificial insemination (TAI) were designed to compare the influence of the antral follicle count (AFC/AFCs) on (1) ovarian follicular dynamics and (2) conception rates. First, multiparous cows with high (≥45 follicles; n=43) or low (≤15 follicles; n=32) AFCs were selected to undergo a TAI protocol to monitor ovarian follicular dynamics. Second, the AFCs of 962 cows also subjected to TAI were determined and classified as groups of high (G-high; ≥45 follicles; n=194), intermediate (G-intermediate; ≥20≤40 follicles, n=397) or low (G-low; ≤15 follicles; n=243) AFCs. In study I, the ovarian measurements (diameter, perimeter and area) were greater (P≤0.05) and there was a greater consistency in number of antral follicles during the period of synchronization for TAI in the high than low group. Effects of the AFC and ultrasonic evaluation interval on the follicular diameter were observed (P≤0.05); however, there was no interaction (P>0.05). Dominant follicles had greater diameters (P≤0.05) in the G-low than in the G-high at D4 (7.3±2.2 vs. 6.2±1.4mm, respectively, P=0.06), D8 (11.2±1.8 vs. 9.5±1.8mm, respectively), D9 (12.3±1.7 vs. 10.6±1.7mm, respectively), and D10 (13.4±1.3 vs. 12.2±1.8mm, respectively), as well as greater estimated diameters of ovulatory follicles (14.4±1.5 vs. 13.4±2.1mm, respectively, P=0.08). In study II, the cows with fewer AFCs had greater (P≤0.05) conception rates (61.7%) than the cows with the intermediate (52.9%) and greater (49.5%) AFCs. Nelore cows with fewer AFCs subjected to synchronization of time of ovulation had a larger follicular diameter and a greater conception rate than the groups with intermediate and a greater AFCs.

Effects of progestagen exposure duration on estrus synchronization and conception rates of crossbreed ewes undergoing fixed time artificial insemination.

Synchronization of estrus and ovulation are of paramount importance in modern livestock improvement programs. These methods are critical for assisted reproduction technologies, including artificial insemination and embryo transfer, that can increase productivity. In the current study, subcutaneous implants containing norgestomet were placed for long (14 days), medium (9 days), and short (5 days) periods of time in 70 crossbred ewes undergoing fixed-time artificial insemination. The resulting effects on estrus synchronization and conception rates were subsequently evaluated. Among the synchronized ewes, 85.7% (60/70) underwent estrus over a period of 72 h after progestagen treatment ceased. The shortest mean interval between withdrawal of the device and onset of estrus (34.2 ± 8.9 h) was observed in the G14 days of P4 group (p < 0.05). The conception rate of the G14 days of P4 group was statistically higher than that of the other groups (83.3% vs. 60.9% vs. 47.8%; p < 0.05). In conclusion, 14 days of norgestomet treatment produced higher conception rates and a greater number of pregnancies at the beginning of the breeding season.

The correlation between the number of antral follicles and ovarian reserves (preantral follicles) in purebred Bos indicus and Bos taurus cows

The objectives of this study were to compare populations of preantral follicles between purebred Bos indicus and Bos taurus cows with high or low antral follicle counts (AFC) and to correlate the number of preantral follicles with the population of antral follicles. Nelore (Bos indicus, n=100) and Angus (Bos taurus, n=100) cow ovaries were collected at abattoirs and examined using ultrasonography. Antral follicles ≥3mm were counted, and the cows ovaries were assigned to high (G-High) or low (G-Low) AFC groups based on the mean number (±1 SD) of ovarian antral follicles: Bos indicus with high AFC (≥57 follicles, n=8) or low AFC (≤21 follicles, n=8) and Bos taurus with high (≥45 follicles, n=10) or low AFC (≤13 follicles, n=10). The ovaries were processed, and the number of preantral follicles was estimated. Between-groups comparisons were performed using a Kruskal-Wallis test, and the correlation between preantral and antral follicles was evaluated using a Pearsons correlation test (P≤0.05). A large variation in the number of preantral follicles was observed among the animals. Although there was a correlation between the population of preantral follicles and the number of antral follicles, there was no difference between the mean number of preantral follicles in the Bos indicus G-High (48,349±30,149) and G-Low groups (33,037±31,710) or between the Bos taurus G-High (35,050±36,060) and G-Low groups (30,481±43,360). Therefore, the preantral follicle population did not differ between purebred Bos indicus and Bos taurus cattle with high or low AFC but was correlated with the number of antral follicles. In addition to the large within-groups variation in the number of preantral follicles, some cows with high AFC had lower populations of preantral follicles compared to the low AFC group, and the highest population of preantral follicles was observed in both Bos indicus and Bos taurus with low AFC.