Ovarian follicular and luteal dynamics in wapiti during the estrous cycle

The reproductive tracts of 13 mature hinds were examined daily by transrectal ultrasonography and blood samples were taken daily from October to January to characterize follicular, luteal, and endocrine dynamics in wapiti during the estrous season. Follicle development occurred in waves characterized by regular, synchronous development of a group of follicles in temporal succession to a surge in serum FSH concentration. The mean interovulatory interval was 21.3 +/- 0.1 d, but was shorter in hinds exhibiting two follicular waves than in hinds exhibiting three and four waves (P < 0.05). The interwave interval was similar among waves in two-wave cycles and the first wave of three-wave cycles. All other interwave intervals in three- and four-wave cycles were shorter (P < 0.05). The maximum diameter of the dominant follicle of the first wave was similar among two-, three-, and four-wave cycles. For all other waves in three- and four-wave cycles, the maximum diameter was smaller (P < 0.05). Corpus luteum diameter and plasma progesterone concentrations were similar between two- and three-wave cycles, but the luteal phase was longer (P < 0.05) in four-wave cycles. The dominant follicle emerged at a diameter of 4 mm at 0.4 +/- 0.1 and 0.8 +/- 0.1 d before the largest and second largest subordinate follicles, respectively. The follicle destined to become dominant was larger (P < 0.05) than the largest subordinate follicle one day after emergence, which coincided with the first significant decrease in serum FSH concentration. We concluded that the estrous cycle in wapiti is characterized by two, three, or four waves of follicular development (each preceded by a surge in circulating FSH), that there is a positive relationship between the number of waves and the duration of the cycle, and an inverse relationship between the number of waves and the magnitude of follicular dominance (diameter and duration of the dominant follicle).     

Ovarian follicular and luteal dynamics in wapiti during seasonal transitions

Transitions from the anovulatory to the ovulatory season (n=20) and ovulatory to anovulatory season (n=11), were monitored daily by transrectal ultrasonography in wapiti. In 17 of 20 observations, the first interovulatory interval (IOI) was short (9.1+/-0.3 days; mean+/-S.E.M.) compared with later in the ovulatory season (21.3+/-0.1) and the last IOI (21.2+/-0.6 days). With one exception, the short IOI were composed of only one wave of follicular development. Subsequent IOI were composed of two or three waves. Maximum diameters of the first two ovulatory follicles were similar (11.3+/-0.4 mm versus 11.3+/-0.2 mm), but both were larger (P<0.05) than the last two ovulatory follicles of the ovulatory season (10.3+/-0.3 and 10.1+/-0.4 mm). Multiple ovulations occurred in three hinds at the first ovulation of the season and in one hind at the second ovulation, but were not at any other time. Day-to-day profiles of CL diameter and plasma progesterone concentration were smaller (P<0.05) for short versus long IOI. Maximum diameter (12.8+/-0.6 mm versus 12.5+/-0.6 mm) and the diameter profile of the last CL of the season were not different from that of the previous CL. In summary, transition to regular ovulation occurred over a 3-week interval and was preceded by one short IOI (9 days). Multiple ovulations were detected only at the onset of the ovulatory season. The characteristics of the last IOI of the ovulatory season were similar to those reported during the rut. The wave pattern of follicle development was maintained throughout both fall and winter transition periods and follicular wave emergence was preceded by a surge in serum FSH concentrations. Transition to anovulation occurred over a 3-month interval and was marked by a failure of the dominant follicle to ovulate after a typical luteal phase.     

Serial ovarian ultrasonography in wild-caught wood bison (Bison bison athabascae)

The objectives of this study were to determine the feasibility of daily examination of wild-caught wood bison and to characterize the ovarian function using serial transrectal ultrasonography and blood hormone analysis. Ten 2-year-old wood bison heifers obtained from Elk Island National Park were placed in a corral adjacent to a handling system designed for restraining bison. The handling system was left open to the corral allowing the bison to explore it freely for 2 months. Active acclimation followed for a 2-week period, during which the bison were herded daily through the handling system and rewarded with whole oats. Finally, the bison were restrained in the handling system and rewarded with whole oats upon release. Once conditioned, daily transrectal examination of the ovaries was completed in 100% of attempts for 30 days (January–February) using a B-mode scanner with a 5 to 10-MHz linear array. Follicle size and numbers were recorded, and individual follicles were identified serially. Blood samples were collected daily and the serum was analyzed for FSH concentrations. Nonrandom changes were detected in the number of follicles ≥4 mm in diameter per day (P < 0.05). Each peak in follicle numbers was associated with the development of a single dominant follicle. The interval between the emergence of successive dominant follicles was 6.8 ± 0.6 days (mean ± SEM). The maximum diameter of the dominant follicle was 9.9 ± 0.4 mm. In conclusion, wild-caught wood bison were amenable to daily examination and blood sampling, and ovarian dynamics were characterized by wave-like development of anovulatory antral follicles. The demonstrated success of this approach to the study of ovarian function will be useful for characterizing the annual reproductive pattern in wood bison, which is necessary for the development of bison-specific protocols for controlling ovarian function for species conservation.     

Folliculogenesis during the transitional period and early ovulatory season in mares

Individual follicles were monitored by ultrasonography in 15 mares during the transitional period preceding the first ovulation of the year and in 9 mares during the first interovulatory interval. During the transitional period, 7 mares developed 1-3 anovulatory follicular waves characterized by a dominant follicle (maximum diameter greater than or equal to 38 mm) that had growing, static, and regressing phases. The emergence of a subsequent wave (anovulatory or ovulatory) did not occur until the dominant follicle of the previous wave was in the static phase. After the emergence of the subsequent wave, the previous dominant follicle regressed. The mean (+/- s.d.) length of the interval between successive waves was 10.8 +/- 2.2 days. Before the emergence of waves (identified by a dominant follicle), follicular activity seemed erratic and follicles did not reach greater than 35 mm. During the interovulatory interval, 6 mares developed 2 waves (an anovulatory wave and a subsequent ovulatory wave) and 3 mares developed only 1 detected wave (the ovulatory wave). The ovulatory follicle at the end of the transitional period reached 20 mm earlier (Day - 15), grew slower (2.6 +/- 0.1 mm/day; mean +/- s.e.m.) but reached a larger diameter on Day - 1 (50.5 +/- 1.1 mm) than for the ovulatory follicle at the end of the interovulatory interval (Day - 10, 3.6 +/- 0.2 mm/day, 44.4 +/- 1.0 mm, respectively; P less than 0.05 for each end point). The interval from cessation of growth of the largest subordinate follicle to the occurrence of ovulation was longer (P less than 0.05) for end of the transitional period (9.5 +/- 0.7 days) than for the end of the interovulatory interval (6.8 +/- 0.6 days). Results demonstrated the occurrence of rhythmic follicular waves during some transitional periods and the occurrence of 2 waves during some of the first oestrous cycles of the year.     

Induction of ovarian follicular wave emergence in wapiti (Cervus elaphus)

Two experiments were done to test the effects of treatments designed to electively induce ovarian follicular wave emergence in wapiti for the purpose of group synchronization. In Experiment 1, hinds were assigned randomly to three groups and given saline im (controls; n=5), 5mg of estadiol-17ss im (n=4), or 5mg estradiol-17ss plus 100mg progesterone im (n=5). In Experiment 2, hinds were assigned randomly to two groups and given no treatment (controls; n=6), or transvaginal ultrasound-guided follicle ablation (n=7). In both experiments, ovarian follicular dynamics were monitored by daily transrectal ultrasonography from Day 0 (day of treatment) to Day 9. In Experiment 1, blood samples were collected at each examination for measurement of serum concentrations of progesterone and FSH. Both experiments were conducted during the late anestrous period (July and August). The mean (+/-S.E.M.) day of wave emergence did not differ between the control and estradiol alone groups, but tended to be later in the estradiol plus progesterone group Day 4.0+/-0.7, Day 3.5+/-0.3, and Day 5.2+/-0.2, respectively; P=0.06). The interval from treatment to wave emergence was less variable in the estradiol plus progesterone group (P<0.05) and tended to be less variable in the estradiol-alone group (P=0.07) than in the control group. The day of wave emergence was more variable (P<0.05) and tended to be later (P=0.10) in the control group compared to the ablation group (Day 2.5+/-0.8 versus Day 1.4+/-0.2). All three treatments were effective in synchronizing ovarian follicular wave emergence among a group of wapiti hinds. Follicle ablation may be an alternative method for synchronization of follicular waves in estrus synchronization and superstimulatory protocols.     

Effect of progesterone on ovarian follicles, emergence of follicular waves and circulating follicle-stimulating hormone in heifers.

The hypothesis was tested that greater growth of the dominant follicle of wave 1 (first follicular wave of an interovulatory interval), compared with that of subsequent anovulatory waves, is due to lower circulating concentrations of progesterone during the growing phase of the follicle. Control heifers (n = 6) were compared with heifers (n = 6) treated with a decreasing dose of progesterone from day 0 to day 5 (ovulation = day 0). Maximum diameter (12.7 +/- 0.9 versus 15.3 +/- 0.7 mm) and mean diameter of the dominant follicle of wave 1, averaged over days, were smaller (P < 0.05) in the progesterone-treated than in the control group. Progesterone treatment did not suppress circulating follicle-stimulating hormone (FSH); but the second FSH surge was earlier, resulting in earlier emergence of wave 2 as indicated by a tendency (P < or = 0.1) for group x day interactions attributed to earlier detection of the dominant follicle and an earlier rise in the total number of follicles detected. The stated hypothesis was supported. We also tested the hypothesis that exposure to low circulating concentrations of progesterone at the end of the growing phase of the anovulatory dominant follicle of wave 1 results in continued growth and prolonged maintenance of the dominant follicle. Heifers (n = 6 per group) were given a luteolytic dose of prostaglandin F2 alpha (PGF2 alpha) on day 6 and treated with a low (30 mg day-1), physiological (150 mg day-1), or high (300 mg day-1) dose of progesterone on days 6 to 20. Continued periodic emergence of anovulatory follicular waves occurred (2.1 +/- 0.0 waves, 2.8 +/- 0.2 waves, 3.8 +/- 0.3 waves, respectively; P < 0.05) until treatment was stopped (interovulatory intervals: 26.2 +/- 1.0, 30.8 +/- 0.6 and 40.3 +/- 1.7 days, respectively; P < 0.05). Compared with the physiological dose group, the growth of the dominant follicle was inhibited to a lesser degree in the low-dose group since it grew for longer (P < 0.05) and to a larger diameter (P < 0.05), and persisted for longer (P < 0.05). Prolonged dominance of this oversized (> 20 mm) follicle was associated with delayed emergence of wave 2. The hypothesis was supported. Results also showed that the high dose of progesterone suppressed the dominant follicle more than the physiological dose when given during the growing phase, but not when given after the growing phase.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of lactational and reproductive status on ovarian follicular waves in llamas (Lama glama)

The effects of lactational status and reproductive status on patterns of follicle growth and regression were studied in 41 llamas. Animals were examined daily by transrectal ultrasonography for at least 30 days. The presence or absence of a corpus luteum and the diameter of the largest and second largest follicle in each ovary were recorded. Llamas were categorized as lactating (N = 16) or non-lactating (N = 25) and randomly allotted to the following groups (reproductive status): (1) unmated (anovulatory group, N = 14), (2) mated by a vasectomized male (ovulatory non-pregnant group, N = 12), (3) mated by an intact male and confirmed pregnant (pregnant group, N = 15). Ovulation occurred on the 2nd day after mating with a vasectomized or intact male in 26/27 (96%) ovulating llamas. Interval from mating to ovulation (2.0 +/- 0.1 days) and growth rate of the preovulatory follicle (0.8 +/- 0.2 mm/day) were not affected by lactational status or the type of mating (vasectomized vs intact male). Waves of follicular activity were indicated by periodic increases in the number of follicles detected and an associated emergence of a dominant follicle that grew to greater than or equal to 7 mm. There was an inverse relationship (r = -0.2; P = 0.002) between the number of follicles detected and the diameter of the largest follicle. Successive dominant follicles emerged at intervals of 19.8 +/- 0.7 days in unmated and vasectomy-mated llamas and 14.8 +/- 0.6 days in pregnant llamas (P = 0.001). Lactation was associated with an interwave interval that was shortened by 2.5 +/- 0.05 days averaged over all groups (P = 0.03). Maximum diameter of anovulatory dominant follicles ranged from 9 to 16 mm and was greater (P less than 0.05) for non-pregnant llamas (anovulatory group, 12.1 +/- 0.4 mm; ovulatory group, 11.5 +/- 0.2 mm) than for pregnant llamas (9.7 +/- 0.2 mm). In addition, lactation was associated with smaller (P less than 0.05) maximum diameter of dominant follicles averaged over all reproductive statuses (10.4 +/- 0.2 vs 11.7 +/- 0.3 mm). The corpus luteum was maintained for a mean of 10 days after ovulation in non-pregnant llamas and to the end of the observational period in pregnant llamas. The presence (ovulatory non-pregnant group) and persistence (pregnant group) of a corpus luteum was associated with a depression in the number of follicles detected and reduced prominence of dominant follicles (anovulatory group greater than ovulatory non-pregnant group greater than pregnant group).(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of dominant follicle aspiration and treatment with recombinant bovine somatotropin (BST) on ovarian follicular development in nelore (Bos indicus) heifers

Follicle ablation has been recognized as an efficient method of follicular wave synchronization. Treatment with recombinant bovine somatotropin (BST) has been shown to enhance follicular development in Bos taurus. This experiment assessed the effects of these treatments in Nelore (B. indicus) heifers. Eight cycling Nelore heifers were randomly assigned to 3 different treatments. On Day 2 of a synchronized cycle (Day 0 = day of ovulation), heifers assigned to Treatments 1 and 2 received 2 mL of saline, whereas heifers assigned to Treatment 3 received 320 mg of BST. On Day 5, the first-wave dominant follicle was ablated by ultrasound-guided transvaginal aspiration in heifers in Treatments 2 and 3, and all heifers received an injection of prostaglandin on Day 11. Aspiration of the dominant follicle advanced and synchronized (P < 0.05) the day of second-wave emergence (6.9 +/- 0.1 vs. 8.4 +/- 0.4) and the day of the pre-wave FSH peak (6.0 +/- 0.0 vs. 6.9 +/- 0.4), and increased FSH peak concentrations (381 +/- 21 vs. 292 +/- 30; pg/mL; P < 0.01). Recombinant bovine somatotropin treatment caused a two-fold increase in plasma insulin-like growth factor-I (IGF-I) concentrations (P < 0.001) and resulted in a 36% increase in the number of small follicles (<5 mm; P < 0.001) compared with saline-treated heifers. In summary, in agreement with previous reports on B. taurus, dominant follicle aspiration synchronized ovarian follicular development, and BST treatment increased peripheral concentrations of IGF-I in Nelore heifers. Recombinant bovine somatotropin also increased the number of small follicles, but this response appeared to be inferior to that reported for B. taurus.     

Follicular activity and hormonal secretory profile in vicuna (Vicugna vicugna)

The objective of the present study was to characterize ovarian activity in non-mated vicunas, relating ovarian structures (evaluated by transrectal ultrasonography, daily for 30 days) to changes in plasma concentrations of estradiol-17beta and progesterone. Ovarian follicular activity occurred in waves, characterized by the follicle emergence, growth and regression. The mean duration of follicular waves was 7.2+/-0.5 days (mean+/-S.E.M.), with a range of 4-11 days. The follicular growth phase averaged 3.0+/-0.2 days, the static phase 1.4+/-0.1, the regression phase 2.9+/-0.3 days, and the inter-wave interval was 4.2+/-0.3 days. The mean growth rate during the growing phase was 1.8+/-0.1mm/day, while the duration of the interval from 6mm to maximum diameter was 1.4+/-0.1 days. The mean maximum diameter of the dominant follicle was 8.4+/-0.3mm (range: 6.2-11.2) and mean diameter of the largest subordinate follicle was 5.4+/-0.1mm. There was an inverse relationship between the size of the largest follicle and the total number of follicles (r=-0.21, P=0.002). Follicle activity alternated between ovaries in 77% of the waves, with 40% of dominant follicles present in the left ovary and 60% in the right ovary. Plasma estradiol-17beta concentrations also had a wave-like pattern, varying between 12.0 and 62.8 pmol/l. Plasma progesterone concentrations remained below 5.0 nmol/l and there was no ultrasonographic evidence of ovulation during the study.     

Effects of a dominant follicle on ovarian follicular dynamics during the oestrous cycle in heifers

Two hypotheses were tested: (1) a dominant follicle causes regression of its subordinate follicles, and (2) a dominant follicle during its growing phase suppresses the emergence of the next wave. Cyclic heifers were randomly assigned to one of four groups (6 heifers/group): cauterization of the dominant follicle of Wave 1 or sham surgery (control) on Day 3 or Day 5 (day of ovulation = Day 0). Ultrasonic monitoring of individually identified follicles was done once daily throughout the interovulatory interval. The onset of regression (decreasing diameter) of the largest subordinate follicle of Wave 1 was delayed (P less than 0.01) by cauterization of the dominant follicle of Wave 1 on Day 3 compared to controls (mean onset of regression, Days 10.8 +/- 2.1 vs 4.3 +/- 0.4). Cauterization of the dominant follicle of Wave 1 on Days 3 or 5 caused early emergence (P less than 0.01) of Wave 2 when compared to controls (Day-3 groups: Days 5.5 +/- 0.4 vs 9.6 +/- 0.7; Day-5 groups: Days 7.0 +/- 0.3 vs 9.1 +/- 0.4). The results supported the two hypotheses. In addition, cauterization of the dominant follicle of Wave 1 on Days 3 or 5 increased the incidence of 3-wave interovulatory intervals.     

Emergence and deviation of follicles during the development of follicular waves in cattle

The nature of emergence and deviation of follicles during follicular waves in cattle was studied in 3 experiments by re-examining data from previous projects. Wave emergence was defined as the day or examination (when more than 1 examination per day) the future dominant follicle was 4 mm (Day 0 or Examination 0). Deviation was defined as the beginning of the greatest difference in growth rates between the 2 largest follicles and between 2 consecutive examinations. The search for deviation in an individual wave was done retrospectively from the examination with the maximum diameter of the second largest follicle. In Experiment 1, follicles were assessed ultrasonically for 28 waves every 8 h. The number of examinations that encompassed the emergence of all growing 3-mm follicles was 10.0 +/-0.5 (mean +/-SEM; equivalent to 3.3 d) and extended from mean Examination -3.1 +/-0.3 to mean Examination 6.0 +/-0.6. A mean of 24 growing 3-mm follicles was found, and the maximal attained diameters were 4 mm (46%), 5 mm (25%), and >/=6 mm (29%). More (P<0.05) 3-mm follicles at Examinations -2 and -1 grew to >/=6 mm than to 4 or 5 mm, whereas more 3-mm follicles at Examinations 2 to 6 grew to only 4 mm. On average, the future dominant follicle appeared as a 3-mm follicle (Examination -2.1 +/-0.2) 6 and 10 h earlier (P<0.03) than for the largest (Examination -1.4 +/-0.3) and second-largest (Examination -0.8 +/-0.4) future subordinates, respectively. This result supported the hypothesis that the future dominant follicle has, on the average, an early developmental advantage. In Experiment 2 (n=33 waves), data were normalized to the day at the beginning of deviation (Day 2.8 +/-0.2) when the mean diameters of the dominant and largest subordinate follicle were 8.5 +/-0.2 mm and 7.2 +/-0.2 mm, respectively. This result suggests that the follicle selected to become dominant, as manifested by deviation, is the first follicle to develop to a decisive stage. In Experiment 3 (n=19 waves), FSH concentrations were lower (P<0.05) on the day at the beginning of deviation (8.5 +/-0.5 ng/ml) than on the day before (10.1 +/-0.8 ng/ml), with no continuing decrease after deviation. This temporal result suggests that the attainment of approximate basal levels of FSH is a component of the deviation mechanism.     

Temporal associations among ovarian events in cattle during oestrous cycles with two and three follicular waves

For 18 two-wave interovulatory intervals in heifers, the follicular waves were first detected on Days -0.2 +/- 0.1 and 9.6 +/- 0.2, and for 4 three-wave intervals on Days -0.5 +/- 0.3, 9.0 +/- 0.0 and 16.0 +/- 1.1 (ovulation is Day 0). The day-to-day mean diameter profile of the dominant follicle of the 1st wave and the day of emergence of the 2nd wave were not significantly different between 2-wave and 3-wave intervals. There were no indications, therefore, that events occurring during the first half of the interovulatory interval were associated with the later emergence of a 3rd wave. The dominant ovulatory follicle differed significantly (P less than 0.05 at least) between 2-wave and 3-wave intervals in day of emergence (Day 9.6 +/- 0.2 and 16.0 +/- 1.1), length of interval from emergence of follicle to ovulation (10.9 +/- 0.4 and 6.8 +/- 0.6 days), and diameter on day before ovulation (16.5 +/- 0.4 and 13.9 +/- 0.4 mm). The mean length of 2-wave interovulatory intervals (20.4 +/- 0.3 days) was shorter (P less than 0.01) than for 3-wave intervals (22.8 +/- 0.6 days). The mean day of luteal regression for 2-wave and 3-wave intervals was 16.5 +/- 0.4 and 19.2 +/- 0.5 (P less than 0.01). For all intervals, luteal regression occurred after emergence of the ovulatory wave, and the next wave did not emerge until near the day of ovulation at the onset of the subsequent interovulatory interval. In conclusion, the emergence of a 3rd wave was associated with a longer luteal phase, and the viable dominant follicle present at the time of luteolysis became the ovulatory follicle.     

Evaluation of an ovarian synchronization scheme for fixed-time artificial insemination in wapiti

The ovarian response to an empirically derived treatment protocol used commercially for fixed-time insemination in wapiti (Cervus elaphus) was evaluated by transrectal ultrasonography in hinds during transition into the ovulatory season. On September 29, hinds (n=7) were given an intravaginal progesterone-releasing device (CIDR-B, 1.9 g of progesterone) or left untreated (controls, n=9). Fourteen days later, hinds in the treated group were given 200 IU eCG and the CIDR was removed. Hinds in the control group ovulated randomly over a 15 day period. In the treated group, five hinds ovulated 3 days after eCG treatment, one ovulated 7 days after treatment, and one failed to ovulate by November 1. All extant dominant follicles ceased growth and/or began to regress within 2 days of CIDR placement. Two waves of follicular development were detected between CIDR insertion and removal; the first emerged 5.1+/-0.5 days after CIDR insertion and the second at 11.0+/-0.7 days. Serum progesterone concentration was 0.6+/-0.5 ng/mL (range 1.0-0.3 ng/mL) before CIDR placement, remained above 6 ng/mL during CIDR placement, and fell to 0.8+/-0.9 ng/mL after CIDR removal. In the control group, maximal luteal-phase progesterone concentration was lower (1.1+/-0.1 ng/mL; P<0.05) and emergence of the first follicular wave was more variable (P=0.05) than in the treated group. The protocol to synchronize ovulation was effective in 5/7 (71%) hinds, and 4/7 (57%) became pregnant and calved. The pregnancy rate (6/9) and calving rate (5/9) was similar in the control group. In conclusion, synchronization with CIDR-B was effective; however, the protocol may be improved by shortening the interval of CIDR placement to < or = 7 days and by reducing the circulating concentrations of progesterone to physiologic concentrations (< 4 ng/mL).     

Aberrant blood flow area and plasma gonadotropin concentrations during the development of dominant-sized transitional anovulatory follicles in mares

Color Doppler transrectal ultrasound was used to evaluate blood flow area in the wall of dominant anovulatory follicles versus ovulatory follicles in mares during the transition between anovulatory and ovulatory seasons. Daily examinations were done in 11 control mares toward the end of the anovulatory season. In 13 separate mares, follicular fluid was collected from 30-mm follicles, and blood flow areas from control mares were used as a basis for designating the sampled follicle as either anovulatory or ovulatory. Blood flow area in the controls ranged from 0.18 to 0.35 cm(2) in six mares on the day of a 30-mm anovulatory follicle and from 0.25 to 0.86 cm(2) in 11 mares on the day of a 30-mm ovulatory follicle; the ranges did not overlap except for one follicle. In the controls, mean blood flow area was lower (P < 0.05) in the anovulatory group than in the ovulatory group for each day beginning with the first Doppler examination at 25 mm. For plasma LH in controls, an effect of follicle group (P < 0.0001) and an interaction (P < 0.0001) of group by day reflected lower (P < 0.05) concentrations in the anovulatory group on Days -6, -2, and 5-8 (Day 0 = 30-mm follicle). For plasma FSH, an interaction (P < 0.0001) reflected higher (P < 0.05) concentrations in the anovulatory group on Days -3 and 1-4. More (P < 0.05) statistically identified FSH surges occurred in the anovulatory group during Days -7 to 8. In the sampled mares, follicular-fluid concentrations of estradiol, free insulin-like growth factor-1, inhibin-A, and vascular endothelial growth factor were lower (P < 0.05) in 30-mm designated anovulatory follicles than in 30-mm designated ovulatory follicles. Results were interpreted as follows: 1) The future anovulatory dominant-sized follicle developed under an LH deficiency, 2) the LH deficiency led to reductions in blood flow area and in concentrations of follicular-fluid factors, and 3) the reduction in follicle production of FSH suppressors resulted in higher plasma FSH concentrations.     

Genetic influences on reproduction of female red deer (Cervus elaphus) (2) seasonal and genetic effects on the superovulatory response to exogenous FSH

This study evaluated the influences of seasons and genotype on the superovulatory response to a standardised oFSH regimen in red deer (Cervus elaphus scoticus) and its hybrids with either wapiti (C.e. nelsoni) or Père David's (PD) deer (Elaphurus davidianus). Adult red deer (n=9), F(1) hybrid wapiti x red deer (n=6), and maternal backcross hybrid PD x red deer (i.e., 14 PD hybrid; n=9) were kept together in the presence of a vasectomised stag for 13 months. At 6 weekly intervals, all hinds received a standardised treatment regimen used routinely to induce a superovulatory response in red deer hinds, with 10 consecutive treatments spanning an entire year. This involved synchronisation with intravaginal progesterone devices and delivery of multiple injections of oFSH (equivalent to 72 units NIH-FSH-S(1)). Laparoscopy to assess ovarian response was performed 6-7 days after the removal of the devices. Both season and genotype had significant effects on ovulation rate (OR) and total follicular stimulation (TFS) (P<0.05). For all the three genotypes, ovarian responses were highest from March to November (breeding season) and lowest in the period from December to January, inclusive. Mean OR for red deer hinds ranged from 3.7 to 1.8 during the breeding season, with no observable trend. All red deer hinds were anovulatory during December and January. A similar pattern occurred for 14 PD hybrids, although mean OR during the breeding seasons were twofold lower than for the red deer. For F(1) wapiti hybrids, the first two treatments in March and April resulted in the highest mean OR observed (15.6 and 11.7, respectively). Thereafter, mean values ranged between 6.3 and 4.7 for the remainder of the breeding season. Furthermore, mean OR of 3.0 and 0.5 were recorded in December and January, respectively. For the red deer and F(1) wapiti hybrids, between-hind variation in OR was not randomly distributed across the treatment dates, indicating that the individuals varied significantly in their ability to respond to oFSH, at least within a given season.In conclusion, the study has shown that relative to red deer, F(1) wapiti hybrid hinds exhibit a higher sensitivity to oFSH, whereas 14 PD hybrid hinds have a lower sensitivity. However, individual variation within genotype was very marked. A seasonal effect was apparent for all genotypes, although some F(1) wapiti hybrid hinds exhibited ovulatory responses throughout the year.     

Ovarian follicular wave synchronization and superstimulation in prepubertal calves

Two experiments were designed to artificially alter the follicular wave pattern in calves to determine if the mechanisms controlling the well-ordered pattern of follicular growth in adults are extant in prepubertal animals as well. Experiment 1 was designed to test the hypothesis that follicle ablation in a random group of calves will induce synchronous emergence of a new follicular wave which is not different from a spontaneous wave. Experiment 2 was designed to test the hypothesis that ovarian superstimulatory response in calves is enhanced when treatment is initiated before rather than after the time of selection of the dominant follicle. In Experiment 1, 6-month-old calves were assigned randomly to an ablation group (n = 10) and a control group (no ablation, n = 10). Follicle ablation was accomplished by transvaginal ultrasound-guided needle aspiration of all follicles > or = 4 mm in diameter. Blood samples were taken and ovarian changes were monitored daily. A rise (P < 0.01) in mean plasma FSH concentration was detected 24 h after follicle ablation (1.51 ng/ml in the ablation group and 0.93 ng/ml in the control group). Wave emergence was detected earlier (P < 0.01) and with less variation (P < 0.0001) in the ablation group than the control group (1.2 +/- 0.1 vs 4.0 +/- 0.7 d). Characteristics of the induced wave were not different from those of the spontaneous wave. In Experiment 2, 7-month-old calves were assigned randomly to a pre-selection group in which superstimulation treatment was initiated at the time of wave emergence (1 d after follicle ablation, n = 11), or to a post-selection group in which superstimulation treatment was initiated after selection of a dominant follicle (4 d after follicle ablation, n = 11). Superstimulation treatment consisted of 30 mg of FSH im twice daily for 3 d. Ultrasound-guided transvaginal follicle ablation was used to synchronize follicle wave emergence at the outset of the experiment. The mean diameter of the largest follicle at the start of superstimulation treatment was 3.2 versus 8.5 mm in the pre- and post-selection groups, respectively (P < 0.001). The day after the last treatment, the number of follicles > or = 3 mm in diameter was greater (P < 0.002) in the pre-selection group than in the post-selection group (19.3 +/- 1.7 versus 11.3 +/- 1.3). In summary, ultrasound-guided follicle ablation resulted in synchronous wave emergence in a random group of calves, and superstimulation treatment initiated at the time of wave emergence (pre-selection group) resulted in the growth of more follicles than treatment initiated later (post-selection group). Mechanisms involved in the control of follicle recruitment, selection, and suppression are extant in calves, similar to those found in adults.     

Concentrations of inhibin,progesterone and oestradiol in fluid from dominant and subordinate follicles from mares during spring transition and the breeding season

Dominant and subordinate follicles were collected from mares on the day after the dominant follicle reached 30 mm in diameter, to investigate regulation of folliculogenesis during spring transition and the breeding season. Concentrations of oestradiol-17beta, progesterone and inhibin A, but not inhibin isoforms with pro- and alpha C-immunoreactivity, were significantly higher in preovulatory follicles than in dominant anovulatory transitional follicles. Steroidogenic activity was regained gradually in the dominant follicles of successive anovulatory waves through spring transition. The dominant follicles, during both spring transition and cyclicity, contained higher concentrations of oestradiol, progesterone and inhibin A, but not inhibin pro- and alpha C-isoforms, than subordinate follicles. The results indicate that high follicular levels of oestradiol, progesterone and inhibin A are associated with continued follicle growth and ovulation. The low concentrations of oestradiol and progesterone in transitional follicles indicate that the deficiency in steroidogenesis exists early in the steroidogenic pathway. The similarity in patterns of follicular hormones in spring transition and during cyclicity strongly suggests that the mechanism of dominance is the same in both types of follicle.     

Effect of exogenous LH pulses on the fate of the first dominant follicle in postpartum beef cows nursing calves

Prolonged postpartum anoestrus in beef cows is due to failure of early dominant follicles to ovulate. It is hypothesized that this failure to ovulate is due to inadequate LH pulse frequency. The objective of this study was to determine whether administration of hourly LH pulses would cause the first dominant follicle to ovulate. In Expt 1, 16 cows received either saline (n = 8) or porcine LH (pLH; 50 micrograms h-1; n = 8) as hourly pulses for 3-5 days from the second day of dominance of the first dominant follicle (day 0). In Expt 2, 21 cows received either saline (n = 7), or 50 micrograms pLH (n = 7) or 100 micrograms pLH (n = 7) as hourly pulses for 3 days. Appropriate ovarian scanning and assays of blood samples were carried out. In Expt 1, the number of dominant follicles that underwent atresia was not affected by increasing the number of LH pulses, but the duration of dominance (days) of the first and second dominant follicles and maximum size (mm) of the second dominant follicle were increased (P < 0.05). Oestradiol concentrations were higher (P < 0.05) in cows given hourly pLH pulses (3.1 +/- 1.2 pg ml-1) compared with controls (1.2 +/- 0.2 pg ml-1). Four of eight treated cows had an anovulatory LH surge. The number of follicle waves to first ovulation was not different (P < 0.05) between control (4.6 +/- 0.9) and pLH treated cows (3.9 +/- 0.5). In Expt 2, four of seven cows given pulses of 100 micrograms pLH h-1 ovulated the first dominant follicle, and the interval from calving to first ovulation was decreased (P < 0.05). In the remaining three cows, the duration of dominance of the first dominant follicle was increased (P < 0.005), the maximum size of the first dominant follicle was greater (P < 0.05), and the interval (days) from the start of infusion to new wave emergence was greater (P < 0.05) compared with cows that failed to ovulate in either the 50 micrograms pLH h-1 or control treatments. In conclusion, hourly pulses of pLH from day 1 after dominance of the first dominant follicle in postpartum beef cows can either prolong dominance or induce it to ovulate. This finding supports the hypothesis that LH pulse frequency is a key determinant of the fate of the dominant follicle in the early postpartum period.     

Ovarian follicular activity and hormonal profile during estrous cycle in cows: the development of 2 versus 3 waves

Most estrous cycles in cows consist of 2 or 3 waves of follicular activity. Waves of ovarian follicular development comprise the growth of dominant follicles some of which become ovulatory and the others are anovulatory. Ovarian follicular activity in cows during estrous cycle was studied with a special reference to follicular waves and the circulating concentrations of estradiol and progesterone. Transrectal ultrasound examination was carried out during 14 interovulatory intervals in 7 cows. Ovarian follicular activity was recorded together with assessment of serum estradiol and progesterone concentrations. Three-wave versus two-wave interovulatory intervals was observed in 71.4% of cows. The 3-wave interovulatory intervals differed from 2-wave intervals in: 1) earlier emergence of the dominant follicles, 2) longer in length, and 3) shorter interval from emergence to ovulation. There was a progressive increase in follicular size and estradiol production during growth phase of each wave. A drop in estradiol concentration was observed during the static phase of dominant anovulatory follicles. The size of the ovulatory follicle was always greater and produced higher estradiol compared with the anovulatory follicle. In conclusion, there was a predominance of 3-wave follicular activity that was associated with an increase in length of interovulatory intervals. A dominant anovulatory follicle during its static phase may initiate the emergence of a subsequent wave. Follicular size and estradiol concentration may have an important role in controlling follicular development and in determining whether an estrous cycle will have 2 or 3-waves.     

Aspects of ovarian follicle development throughout life

The pool of primordial follicles present in the female ovary reaches its maximum number around 20 weeks of gestational age and then decreases in a logarithmic fashion throughout life until complete depletion occurs around the age of the menopause. Reproductive life is initiated when less than 10% (0.5 million) of primordial follicles are left. The entire growth trajectory of the follicle takes at least 3 months. Follicle growth up to the antral stage occurs during fetal life and infancy. While the role of gonadotropins in early follicular development remains controversial, the last 2 weeks of development are FSH dependent. The intercycle rise in FSH and decreasing levels thereafter are crucial for recruitment of a cohort of healthy, early antral follicles and subsequent single dominant selection. Following puberty, anovulation may persist for years and this may presage the development of adult anovulatory infertility. The menopause is preceded by a period of reduced fertility. The development of reliable and sensitive markers for ovarian ageing will be the challenge of the near future.