Comparison of antral and preantral ovarian follicle populations between Bos indicus and Bos indicus-taurus cows with high or low antral follicles counts.

The objective was to compare populations of antral and pre-antral ovarian follicles in Bos indicus and Bos indicus-taurus cows with high and low antral follicle counts. Nelore (Bos indicus, n = 20) and Nelore X Angus (1/2 Bos indicus-taurus, n = 20) cows were subjected to follicular aspiration without regard to the stage of their oestrous cycle (day of aspiration = D0) to remove all follicles ≥3 mm and induce growth of a new follicular wave. Ovaries were examined by ultrasonography on D4, D19, D34, D49 and D64, and antral follicles ≥3 mm were counted. Thereafter, cows were assigned to one of two groups: high or low antral follicular count (AFC, ≥30 and ≤15 antral follicles, respectively). After D64, ovaries were collected after slaughter and processed for histological evaluation. There was high repeatability in the numbers of antral follicles for all groups (range 0.77-0.96). The mean (±SD) numbers of antral follicles were 35 ± 9 (Bos indicus) and 38 ± 6 (Bos indicus-taurus) for the high AFC group and 10 ± 3 (Bos indicus) and 12 ± 2 (Bos indicus-taurus) follicles for the low AFC. The mean number of preantral follicles in the ovaries of Bos indicus-taurus cows with high AFC (116 226 ± 83 156 follicles) was greater (p < 0.05) than that of Bos indicus cows (63 032 ± 58 705 follicles) with high AFC. However, there was no significant correlation between numbers of antral and preantral follicles.

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.

MALDI-MS lipid profiles of oocytes recovered by ovum pickup from Bos indicus and 1/2 indicus × taurus with high vs low oocyte yields.

The aim of the present study was to compare the lipid profile in oocytes of indicus and 1/2 indicus × taurus cows with high and low antral follicle count (AFC)/oocyte yields. After an OPU procedure (D0), antral follicles ≥3 mm were counted by ultrasonography (D4, 19, 34, 49, 64), and cows were assigned to groups with either high AFC (≥30 follicles; indicus, NH group; 1/2 indicus × taurus, AH group) or low AFC (≤15 antral follicles; indicus, NL group; 1/2 indicus × taurus, AL group). The lipid profiles of the oocytes were determined by MALDI-MS. For GI, GII and GIII oocytes, the indicus samples tend to cluster separately from the 1/2 indicus × taurus samples. The lipid species [PC (P-38:5) + H](+) and/or [PC (P-36:2) + Na](+) , [PC (38:2) + H](+) , [PC (38:5) + Na](+) and [TAG (60:8) + NH(4) ](+) were more abundant in indicus (NH and NL groups) than 1/2 indicus × taurus. The higher lipid content in the indicus oocytes likely reflects differences in the rate of lipid metabolism and may contribute to oocyte competence and embryo development.

Correlation between phenotype, genotype and antral follicle population in beef heifers.

The present study was performed in indicus-taurus heifers 1) to determine if the antral follicle count (AFC) exhibits repeatability from puberty to yearling age and 2) to evaluate whether the phenotypic and genotypic parameters used in genetic improvement programs are correlated with AFC. In study I, Braford heifers (3/8 Nelore x 5/8 Hereford, n = 137) were serially examined by ultrasonography (with 60-day intervals) from weaning (9 ± 1 mo of age) to yearling ages (20 ± 1 mo of age) to monitor the numbers of antral follicles. In study II, the AFC of animals from experiment I and contemporary (same farm, considered at same age and kept under same conditions heifers n = 270 18-24 months) was correlated with the records of a genetic selection program using four statistical models with different covariates: i) model 1 considered effects of contemporary group and covariates age, weight gain from birth to weaning and visual scores for carcass traits at weaning, ii) model 2 covered contemporary group, age, weight gain from weaning to yearling and visual scores for carcass traits at yearling. The effects, variables and covariates of models 1 and 2 were combined to form model 3. Model 4 included the model 3 with addition of paternal effect. In study I, AFC varied from 3 to 64 follicles among females but was highly repeatable (0.89-0.92) within individuals in the same group. In study II, the four models tested showed low correlations with AFC: 0.072, 0.056, 0.082 and 0.172 for models 1, 2, 3 and 4, respectively. However, the model with paternal effect provided 17% of correlation of AFC and genotypic and phenotypic characteristics. Models 1, 3 and 4 also showed that AFC in indicus-taurus heifers can be influenced by finishing precocity at weaning (P < 0.05) with a variation of three follicles. Based on these studies, AFC in heifers from weaning to yearling age is highly variable between individuals and repeatable within the same female. Additionally, there is no correlation between phenotypic or genotypic characteristics and the antral follicle population. However, AFC can be slightly affected by finishing precocity at weaning.

Dynamics of follicular growth and progesterone concentrations in cyclic and anestrous suckling Nelore cows (Bos indicus) treated with progesterone, equine chorionic gonadotropin, or temporary calf removal.

The objective of this study was to investigate the effects of eCG and temporary calf removal (TCR) associated with progesterone (P4) treatment on the dynamics of follicular growth, CL size, and P4 concentrations in cyclic (n = 36) and anestrous (n = 30) Nelore cows. Cyclic (C) and anestrous (A) cows were divided into three groups. The control group received 2 mg of estradiol benzoate via intramuscular (IM) injection and an intravaginal device containing 1.9 g of P4 on Day 0. On Day 8, the device was removed, and the animals received 12.5 mg of dinoprost tromethamine IM. After 24 hours, the animals received 1 mg of estradiol benzoate IM. In the eCG group, cows received the same treatment described for the control group but also received 400 UI of eCG at the time of device removal. In the TCR group, calves were separated from the cows for 56 hours after device removal. Ultrasound exams were performed every 24 hours after device removal until the time of ovulation and 12 days after ovulation to measure the size of the CL. On the same day as the CL measurement, blood was collected to determine the plasma P4 level. Statistical analyses were performed with a significance level of P ≤ 0.05. In cyclic cows, the presence of the CL at the beginning of protocol resulted in a smaller follicle diameter at the time of device removal (7.4 ± 0.3 mm in cows with CL vs. 8.9 ± 0.4 mm in cows without CL; P = 0.03). All cows ovulated within 72 hours after device removal. Anestrous cows treated with eCG or TCR showed follicle diameter at fixed-timed artificial insemination (A-eCG 10.2 ± 0.3 and A-TCR 10.3 ± 0.5 mm) and follicular growth rate (A-eCG 1.5 ± 0.2 and A-TCR 1.3 ± 0.1 mm/day) similar to cyclic cows (C-eCG 11.0 ± 0.6 and C-TCR 12.0 ± 0.5 mm) and (C-eCG 1.4 ± 0.2 and C-TCR 1.6 ± 0.2 mm/day, respectively; P ≤ 0.05). Despite the similarities in CL size, the average P4 concentration was higher in the A-TCR (9.6 ± 1.4 ng/mL) than in the A-control (4.0 ± 1.0 ng/mL) and C-TCR (4.4 ± 1.0 ng/mL) groups (P < 0.05). From these results, we conclude that eCG treatment and TCR improved the fertility of anestrous cows by providing follicular growth rates and size of dominant follicles similar to cyclic cows. Additionally, TCR increases the plasma concentrations of P4 in anestrous cows.

Estimate of the population of preantral follicles in the ovaries of Bos taurus indicus and Bos taurus taurus cattle.

The number of oocytes recovered from Bos taurus indicus females subjected to ovum pick-up averaged two to four times greater compared to Bos taurus taurus females. The objective of the present study was to test the hypothesis that this difference in oocyte yield was due to more preantral follicles in the ovaries of Bos indicus females. Ovaries (n = 64) from Nelore (Bos indicus) fetuses (n = 10), heifers (n = 12), and cows (n = 10), and Aberdeen Angus (Bos taurus) fetuses (n = 10), heifers (n = 12), and cows (n = 10) were cut longitudinally into halves, fixed, and processed for histological evaluation. The number of preantral follicles was estimated by counting them in each histological section, using the oocyte nucleus as a marker and employing a correction factor. The average number of preantral follicles in the ovaries of Bos indicus vs Bos taurus was (mean ± SD) 143,929 ± 64,028 vs 285,155 ± 325,195 for fetuses, 76,851 ± 78,605 vs 109,673 ± 86,078 for heifers, and 39,438 ± 31,017 vs 89,577 ± 86,315 for cows (P > 0.05). The number of preantral follicles varied greatly among individual animals within the same category, as well as between breeds. In conclusion, we inferred that the higher oocyte yield from Bos indicus females was not due to a greater ovarian reserve of preantral follicles. Therefore, mechanisms controlling follicle development after the preantral stage likely accounted for differences between Bos indicus and Bos taurus females in number of oocytes retrieved at ovum pick-up.

Large-scale in vitro embryo production and pregnancy rates from Bos taurus, Bos indicus, and indicus-taurus dairy cows using sexed sperm.

Herein we describe a large-scale commercial program for in vitro production of embryos from dairy Bos taurus, Bos indicus, and indicus-taurus donors, using sexed sperm. From 5,407 OPU, we compared the number of recovered oocytes (n = 90,086), viable oocytes (n = 64,826), and embryos produced in vitro from Gir (Bos indicus, n = 617), Holstein (Bos taurus, n = 180), 1/4 Holstein × 3/4 Gir (n = 44), and 1/2 Holstein-Gir (n = 37) crossbred cows, and the pregnancy rate of recipient cows. Viable oocytes were in vitro matured (24 h at 38.8 °C, 5% CO(2) in air) and fertilized by incubating them for 18 to 20 h with frozen-thawed sexed sperm (X-chromosome bearing) from Gir (n = 8) or Holstein (n = 7) sires (2 × 10(6) sperm/dose). Embryos were cultured in similar conditions of temperature and atmosphere as for IVM, with variable intervals of culture (between Days 2 and 5) completed in a portable incubator. All embryos were transferred fresh, after 24 to 72 h of transportation (up to 2,000 km). On average, 16.7 ± 6.3 oocytes (mean ± SEM) were obtained per OPU procedure and 72.0% were considered viable. Total and viable oocytes per OPU procedure were 17.1 ± 4.5 and 12.1 ± 3.9 for Gir cows, 11.4 ± 3.9 and 8.0 ± 2.7 for Holstein cows, 20.4 ± 5.8 and 16.8 ± 5.0 for 1/4 Holstein × 3/4 Gir, and 31.4 ± 5.6 and 24.3 ± 4.7 for 1/2 Holstein-Gir crossbred females (P < 0.01). The mean number of embryos produced by OPU/IVF and the pregnancy rates were 3.2 (12,243/ 3,778) and 40% for Gir cows, 2.1 (2,426/1,138) and 36% for Holstein cows, 3.9 (1,033/267) and 37% for 1/4 Holstein × 3/4 Gir, and 5.5 (1,222/224), and 37% for 1/2 Holstein-Gir. In conclusion, we compared oocyte yield from two levels of indicus-taurus breeds and demonstrated the efficiency of sexed sperm for in vitro embryo production. Culturing embryos during long distance transportation was successful, with potential for international movement of embryos.

Insemination of four cows per dose of frozen semen with a fixed-time artificial insemination protocol

The aim of the study was to evaluate the practicality of a 1: 4 dilution of doses of semen in fixed - time artificial insemination ( FTAI ) programs. Nelore cows (n = 803 ) were synchronized by a conventional FTAI protocol. For AI in the dilut ed group (n = 392) , 0.5 ml semen straws were thawed and 0.5 ml of extender conta ining amino acids and methylxanthine derivatives was added. The straws were fractionated into four straws of 0.25 ml each (dilution 1:4) with approximately 2.5 x 10 6 spermatozoa each and the insemination was performed in the ipsilateral horn to the ovary c ontaining the dominant follicle . In the control group (n = 411), AI was performed in the uterine body with one straw of 0.5 ml (10 x 10 6 spermatozoa) containing seme n from the same bull and batch . The conception rate was 49.2% (193/392) for the diluted gro up and 50.1% (206/411) for the control group, with 1.97 and 0.50 pregnancies per dose of semen , respectively . T he dilution of semen provide d a pregnancy index similar to the control group ; however, the technique increased the number of pregnancies per dos e , allowing for the best use of semen of high genetic value and reducing the cost per pregnancy.

Insemination of four cows per dose of frozen semen with a fixed-time artificial insemination protocol

The aim of the study was to evaluate the practicality of a 1: 4 dilution of doses of semen in fixed - time artificial insemination ( FTAI ) programs. Nelore cows (n = 803 ) were synchronized by a conventional FTAI protocol. For AI in the dilut ed group (n = 392) , 0.5 ml semen straws were thawed and 0.5 ml of extender conta ining amino acids and methylxanthine derivatives was added. The straws were fractionated into four straws of 0.25 ml each (dilution 1:4) with approximately 2.5 x 10 6 spermatozoa each and the insemination was performed in the ipsilateral horn to the ovary c ontaining the dominant follicle . In the control group (n = 411), AI was performed in the uterine body with one straw of 0.5 ml (10 x 10 6 spermatozoa) containing seme n from the same bull and batch . The conception rate was 49.2% (193/392) for the diluted gro up and 50.1% (206/411) for the control group, with 1.97 and 0.50 pregnancies per dose of semen , respectively . T he dilution of semen provide d a pregnancy index similar to the control group ; however, the technique increased the number of pregnancies per dos e , allowing for the best use of semen of high genetic value and reducing the cost per pregnancy.(AU)

Ovarian follicle reserve: emerging concepts and applications

This paper presents new concepts in the study of folliculogenesis and describes some of the current applications to reproductive biotechnology. The importance of better understanding this issue is addressed both for basic and applied research. After a brief review of the basic conceptions of the origin, formation, and growth of follicules according to stablished concepts, some controversial points, as the postnatal production of the follicles and the role of multioocyte follicles, are discussed. The importance of the ovarian follicular reserve is considered for fertility and reproductive parameters, as well as some questiones about the presence of multioocyte follicles in adult ovaries. Finally, some future prospects are proposed.

Ovarian follicle reserve: emerging concepts and applications

This paper presents new concepts in the study of folliculogenesis and describes some of the current applications to reproductive biotechnology. The importance of better understanding this issue is addressed both for basic and applied research. After a brief review of the basic conceptions of the origin, formation, and growth of follicules according to stablished concepts, some controversial points, as the postnatal production of the follicles and the role of multioocyte follicles, are discussed. The importance of the ovarian follicular reserve is considered for fertility and reproductive parameters, as well as some questiones about the presence of multioocyte follicles in adult ovaries. Finally, some future prospects are proposed.(AU)

Reproductive performance of ewes treated with an estrus induction/synchronization protocol during the spring season

The aim of this study was to evaluate the reproductive performance of ewes treated with eCG and an exogenous progestagen protocol during the spring season. Forty - eight mixed - breed wool and hair ewes ( body condition score of 2.8 ± 0.5 and 41 ± 3 kg ) were randomly assigned into two groups ( n = 24/group), which received (G - Sync) or not (G - Control) an intravaginal device ( Day 0) containing 60 mg of medroxyprogesterone acetate. On D ay 7, ewes of the G - Sync were injected with 300 IU of eCG and 30 ug of d - C loprostenol, im . On D ay 9, the device was removed and 12 h later males were introduced into the G - Sync and G - Control groups in a proportion of 1:6. Estrus response observation and mating were performed during D ays 10, 11 and 12 from 7 to 9 AM and 4 to 6 PM. After D ay 12, males were s eparated from females for 10 days and later reintroduced into the flock for 45 days. Estrus rates for the G - Control and G - Sync group s during D ays 10, 11 and 12 were 4 and 88 % (P < 0.05) , respectively . Pregnancy rates fr om initial mating on D ays 10, 11 and 12 were 0 (G - Control) and 4 6% (G-Sync;P < 0.05). Total pregnancy rates for the whole mating season were 50 (G-Control) and 79% (G-Sync;P< 0.05). The exogenous progestagen protocol plus eCG used for estrus induction/s ynchronization improved the pregnancy rate of mixed-breed wool and hair ewes by about 29% points at the end of the breeding season. Thus, this procedure seems to be appropriate to be implemented as part of the reproductive management of some ovine farms during the non-breeding season.(AU)

Reproductive performance of ewes treated with an estrus induction/synchronization protocol during the spring season

The aim of this study was to evaluate the reproductive performance of ewes treated with eCG and an exogenous progestagen protocol during the spring season. Forty - eight mixed - breed wool and hair ewes ( body condition score of 2.8 ± 0.5 and 41 ± 3 kg ) were randomly assigned into two groups ( n = 24/group), which received (G - Sync) or not (G - Control) an intravaginal device ( Day 0) containing 60 mg of medroxyprogesterone acetate. On D ay 7, ewes of the G - Sync were injected with 300 IU of eCG and 30 ug of d - C loprostenol, im . On D ay 9, the device was removed and 12 h later males were introduced into the G - Sync and G - Control groups in a proportion of 1:6. Estrus response observation and mating were performed during D ays 10, 11 and 12 from 7 to 9 AM and 4 to 6 PM. After D ay 12, males were s eparated from females for 10 days and later reintroduced into the flock for 45 days. Estrus rates for the G - Control and G - Sync group s during D ays 10, 11 and 12 were 4 and 88 % (P < 0.05) , respectively . Pregnancy rates fr om initial mating on D ays 10, 11 and 12 were 0 (G - Control) and 4 6% (G-Sync;P < 0.05). Total pregnancy rates for the whole mating season were 50 (G-Control) and 79% (G-Sync;P< 0.05). The exogenous progestagen protocol plus eCG used for estrus induction/s ynchronization improved the pregnancy rate of mixed-breed wool and hair ewes by about 29% points at the end of the breeding season. Thus, this procedure seems to be appropriate to be implemented as part of the reproductive management of some ovine farms during the non-breeding season.

Reproductive performance of ewes mated in the spring when given nutritional supplements to enhance energy levels

The objective of this study was to evaluate the reproductive performance of ewes mated in the spring when given nutritional supplements to enhance energy levels. Ewes were assessed for weight and body condition at the beginning and end of flushing periods. Lambing rates and rates of proliferation were also evaluated. Crossbred woolly ewes (n = 46), 36 ± 2 months of age and rated 3.0 ± 0.1 on a body condition scale that spanned from 1 to 5 were divided into four treatment groups. One group received no feed supplements (NFS), while the others were supplemented for 64 days: 21 days before and 43 days during the mating season. The latter groups were fed soybean hulls at 0.6% (S06), 0.9% (S09) and 1.2% (S12) of their body weight (BW), calculated using dry mass. Ewes were mated for 50 days with four rams by means of natural breeding in October and November, which is spring in Brazil. There was no significant difference among the treatments based on mean BW of the ewes before and after the supplementation period (P > 0.05). Similarly, no differences were found in the rates of proliferation (P > 0.05). The S09 treatment presented the highest lambing rate (82%, 9/11), while the NFS treatment correlated with the lowest rate (42%, 5/12; P < 0.05). There was a positive linear effect for body condition score as the supplementation level increased (P < 0.05). We conclude that a supplementation level of 0.9% BW seems to improve lambing rates for ewes that undergo springtime mating.

Reproductive performance of ewes mated in the spring when given nutritional supplements to enhance energy levels

The objective of this study was to evaluate the reproductive performance of ewes mated in the spring when given nutritional supplements to enhance energy levels. Ewes were assessed for weight and body condition at the beginning and end of flushing periods. Lambing rates and rates of proliferation were also evaluated. Crossbred woolly ewes (n = 46), 36 ± 2 months of age and rated 3.0 ± 0.1 on a body condition scale that spanned from 1 to 5 were divided into four treatment groups. One group received no feed supplements (NFS), while the others were supplemented for 64 days: 21 days before and 43 days during the mating season. The latter groups were fed soybean hulls at 0.6% (S06), 0.9% (S09) and 1.2% (S12) of their body weight (BW), calculated using dry mass. Ewes were mated for 50 days with four rams by means of natural breeding in October and November, which is spring in Brazil. There was no significant difference among the treatments based on mean BW of the ewes before and after the supplementation period (P > 0.05). Similarly, no differences were found in the rates of proliferation (P > 0.05). The S09 treatment presented the highest lambing rate (82%, 9/11), while the NFS treatment correlated with the lowest rate (42%, 5/12; P < 0.05). There was a positive linear effect for body condition score as the supplementation level increased (P < 0.05). We conclude that a supplementation level of 0.9% BW seems to improve lambing rates for ewes that undergo springtime mating.(AU)