This study examined the correlation between measurement of follicle growth by ultrasound, and measurement of intrafollicular ratios of oestradiol and progesterone concentrations and the serum concentrations of FSH during selection, dominance and atresia or ovulation of dominant follicles in heifers. Heifers were ovariectomized on days 0 (before LH surge), 1 (after LH surge, preovulation), 1 (postovulation), 3, 6 and 12 of the oestrous cycle. Blood samples were collected at 4–6 h intervals. After ovariectomy all follicles ≥ 5 mm were measured and follicular fluid was aspirated. Follicles were classified by size according to ultrasound (F1, largest; F2, second largest; F3, all remaining follicles ≥ 5 mm) and by the ratio of oestradiol:progesterone concentrations. During the follicular phase, a single dominant oestrogen-active follicle increased in diameter while serum concentrations of LH increased and FSH decreased (P < 0.05). On day 1 (after LH surge, preovulation), serum LH and FSH decreased to pre-surge concentrations (P < 0.0001), while follicle size and intrafollicular progesterone concentration increased and oestradiol concentration decreased (P < 0.05). A dominant nonovulatory follicle, classified as oestrogen-active on days 1, 3 and 6 and oestrogen-inactive on day 12, increased in size from day 1 to day 7 and lost dominance during days 10–12, coincident with the growth of multiple oestrogen-active follicles. The serum FSH concentration increased transiently (P < 0.05) before each new wave of dominant follicular growth. The overall correlation of ultrasound measurements of follicle diameter with measures of follicle size after ovariectomy was high. The ratio of oestradiol:progesterone concentrations, but not of size, reliably distinguished potential dominant from atretic follicles. The size of the follicle and the oestradiol concentration were not determinants of subsequent dominance during a selection phase. We conclude that: (1) ovarian follicles go through selection, dominance and atresia phases coincident with transient increases and decreases in FSH; and (2) ultrasound is an accurate measure of follicle growth, but that size alone is not a sufficient measure to ascribe dominance and both ultrasound and the intrafollicular ratio of oestradiol:progesterone concentrations are needed to monitor selection, dominance and atresia of follicles accurately.
S. J. Sunderland, M. A. Crowe, M. P. Boland, J. F. Roche and J. J. Ireland
D. O'Callaghan, A. Donovan, S. J. Sunderland, M. P. Boland and J. F. Roche
Four experiments were carried out to determine the effect of the presence of ewes and rams on the reproductive state of ewes. In Expt 1, the breeding season of ewes kept with a vasectomized ram ended later (April 18 ± 8 days; mean ± sem) than that of ewes isolated from rams (6 March ± 7 days; P < 0.01). In Expt 2, the end of the breeding season was later (5 May ± 6 days; P < 0.05) and the onset of the next breeding season earlier (29 September ± 2 days; P < 0.001) in ewes maintained with rams, compared with ewes isolated from rams (14 April ± 7 days and 1 November ± 2 days, respectively). There was no difference in the timing of, or variation in, reproductive transitions between ewes maintained either as individuals or in groups. In Expt 3, all ewes exposed to artificial short days from the date of the winter solstice and interrupted with 35 long days in spring resumed cyclicity (median date, 7 September; range, 59 days). Most ewes (seven of nine) exposed to short days from the date of the winter solstice and isolated from other ewes did not resume cyclicity in the following 11 months. In contrast, all ewes resumed cyclicity (median date, 19 October; range, 144 days) when exposed to short days but housed in social contact with other ewes that became reproductively active in early September; however, the onset of cyclicity was later than in ewes exposed to long days (P < 0.01). In Expt 4, the number of LH pulses per 6 h in ewes exposed to rams was higher (P < 0.001) and the time of first ovulation earlier (16 August ± 5 days; P < 0.05) than it was in ewes that were isolated from rams and exposed to either oestrous or anoestrous ewes. We conclude that there was a chronic stimulus from rams to ewes that increased the duration of the breeding season and decreased anoestrus. There was no acute effect of introduction of oestrous ewes to anoestrous ewes on LH pulse frequency and time of first ovulation of the breeding season under the natural photoperiod, and the onset of the breeding season of housed anoestrous ewes exposed to a constant photoperiod was advanced by housing them with cyclic ewes. These results highlight a role for social or other animal-related stimuli in seasonal reproduction in ewes.
A. R. Scanlon, S. J. Sunderland, T. L. Martin, D. Goulding, D. O'Callaghan, D. H. Williams, D. R. Headon, M. P. Boland, J. J. Ireland and J. F. Roche
Two experiments were conducted in cyclic beef heifers to determine whether active immunization against bovine inhibin α 1–26 Gly-Tyr (bINH) affected follicular dynamics, hormone concentration or ovulation rate. In Expt 1, heifers (n = 9) were actively immunized against bINH conjugated to human α globulins (HAG) using bisdiazotized benzidine in non-ulcerative Freund's adjuvant (NUFA; primary on day 0; booster injections on days 53, 84 and 116 using conjugated bINH and on days 176 and 366 using unconjugated bINH; ten heifers were used as controls). Ovaries were examined daily using ultrasound scanning (days 70–155 and 384–391) and corresponding blood samples were collected for bINH antibody titre, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and oestradiol determinations. Four treated and four control heifers were injected with 10 μg gonadotrophin-releasing hormone (GnRH) on day 386 (day 2 of the oestrous cycle). Although bINH-immunized heifers had variable antibody titres ranging from 4 to 50% I125-labelled bINH bound to serum diluted 1:2000, ovulation rate was unaffected. In oestrous cycles with three dominant follicles, the ovulatory follicles grew faster (2.5 ± 0.2 versus 1.6 ± 0.3 mm day−1; mean ± sem), had shorter durations of growth (5.7 ± 0.8 versus 9.6 ± 1.6 days) and duration of detection (7.5 ± 0.8 versus 12.0 ± 2.4 days) in immunized heifers. Mean concentrations of FSH, LH and oestradiol were unaltered in most cases during oestrous cycles in bINH-immunized compared with control heifers. There was no significant difference in the percentage increase in FSH or LH, after GnRH injection, between control and immunized heifers. As ovulation rate was unaltered in the first experiment, a second similar study was designed using a different immunization protocol. In Expt 2, heifers were immunized with bINH conjugated to human serum albumin using glutaraldehyde with the following doses: 0.0 (control; n = 7), 0.33 (n = 7), 1.0 (n = 8) and 3.0 (n = 7) mg. Three booster immunizations were given 33, 66 and 209 days after primary immunization. Immunization increased the number of oestrous cycles with multiple ovulations (42 of 132 (32%) oestrous cycles examined) compared with controls (1 of 30 (3.3%) oestrous cycles examined). Neither titre nor ovulation rate was affected by dose of bINH used. In summary, following bINH immunization, ovulation rate was not increased despite changes in follicular dynamics in Expt 1, but was increased in 32% of oestrous cycles in Expt 2. We conclude that immunization protocols can affect responsiveness of heifers to bINH immunization, and that immunization against inhibin can increase ovulation rate.