Size Of Ovarian Follicles Research Articles

Plasma steroid levels and follicular development in the female staghorn damselfish Amblyglyphidodon curacao

Female staghorn damselfish were captured from coral reefs by divers and bloodsampled. Ovarian follicle type and size distribution were correlated with plasma levels of the ovarian steroids testosterone (T) and 17β-estradiol (E2). Ovaries contained up to three clutches of vitellogenic follicles and steroid levels were maximal when all three clutches were present.

Gonadotropin Surge Induces Two Separate Increases in Messenger RNA for Progesterone Receptor in Bovine Preovulatory Follicles1

In mice deficient in progesterone receptor (PR), follicles of ovulatory size develop but fail to ovulate, providing evidence for an essential role for progesterone and PR in ovulation in mice. However, little is known about the expression and regulation of PR mRNA in preovulatory follicles of ruminant species. One objective of this study was to determine whether and when PR mRNA is expressed in bovine follicular cells during the periovulatory period. Luteolysis and the LH/FSH surge were induced with prostaglandin F(2alpha) and a GnRH analogue, respectively, and the preovulatory follicle was obtained at 0, 3.5, 6, 12, 18, or 24 h after GnRH treatment. RNase protection assays revealed a transient increase in levels of PR mRNA, which peaked at 6 h after GnRH and declined to the time 0 value by 12 h and a second increase at 24 h. The second objective was to investigate the mechanisms that regulate PR mRNA expression through in vitro studies on follicular cells of preovulatory follicles obtained before the LH/FSH surge. Theca and granulosa cells were isolated and cultured with or without a luteinizing dose of LH or FSH, progesterone, LH + progesterone, or LH + antiprogestin (RU486). Levels of PR mRNA increased in a time-dependent manner in granulosa cells cultured with LH or FSH and in theca cells cultured with LH, peaking at 10 h of culture. In contrast, progesterone (200 ng/ml) did not upregulate mRNA for its own receptor, and neither progesterone nor RU486 affected LH-stimulated PR mRNA accumulation. Furthermore, RU486 completely blocked LH-stimulated expression of oxytocin mRNA, indicating that PR induced by LH in vitro is functional. These results show that the gonadotropin surge induces a rapid and transient increase in expression of PR mRNA in both theca and granulosa cells of bovine periovulatory follicles followed by a second rise close to the time of ovulation and that the first increase in PR mRNA can be mimicked in vitro by gonadotropins but not by progesterone. These results suggest multiple and time-dependent roles for progesterone and PR in the regulation of periovulatory events in cattle.

Ovarian Structures and Circulating Steroids in Heifers and Lactating Cows in Summer and Lactating and Dry Cows in Winter

Two experiments compared follicular and luteal development and circulating steroid concentrations from induced luteolysis to ovulation in lactating Holstein cows (n=27; 40.0±1.5kg milk/day) vs. nulliparous heifers (n=28; 11 to 17 mo-old) during summer (Experiment 1), and in lactating (n=27; 45.9±1.4kg milk/d) vs. dry cows (n=26) during winter (experiment 2). All females received PGF2α 6 d after ovulation and were monitored until next ovulation by daily ultrasound and assay of serum progesterone (P4) and estradiol (E2). Every female was used two or three times. In Experiment 1, lactating cows had high incidence of multiple ovulation (63.5%) compared with heifers (1.3%). Among single ovulators, there was no difference in maximal size of ovulatory follicles between lactating cows and heifers (15.8 vs. 16.5mm, respectively). However, lactating cows had lower peak serum E2 (8.6 vs. 12.1pg/ml), took longer to ovulate after luteolysis (4.6 vs. 3.8 d), developed more luteal tissue volume (7293.6 vs. 5515.2mm3), and had lower serum P4 on d 6 after ovulation (2.0 vs. 3.0ng/ml) than heifers (data included multiple ovulators). In experiment 2, multiple ovulations were similar between lactating and dry cows (17.9 vs. 17.2%, respectively). Peak serum E2 was also similar between lactating and dry cows (7.6 vs. 8.5pg/ml) although lactating cows had larger ovulatory follicles (18.6 vs. 16.2±0.4mm). Lactating cows took longer to ovulate (4.8 vs. 4.2 d), developed more luteal tissue (7599 vs. 5139±468mm3), but had similar serum P4 (2.2 vs. 1.9ng/ml) compared with dry cows. Therefore, lactating cows had similar or lower circulating steroid concentrations than dry cows or heifers, respectively, despite having larger ovarian structures.

Ovarian follicular development and endocrine function in non-prolific Western White Face ewes with early or late onset of seasonal anestrous

The aim of this study was to see if there were differences in antral follicle development and endocrine profiles among non-prolific Western White-Faced ewes entering anestrous early (January–February; n=6) or late (March; n=6), using daily transrectal ultrasonography of ovaries and hormone measurements, over a 17-day period during mid-anestrous (May). The ovarian follicles were measured and categorized by size (1-2, 2, 3, 4 and ≥5 mm in diameter). For determination of the patterns of FSH and LH secretion, a 12 h intensive bleed, with blood samples taken every 12 min, was carried out 2 days after the end of the scanning period. A bolus injection of GnRH (250 μg per ewe) was given 6 h into the period of intensive blood collection. The ewes entering anestrous early had significantly lower numbers of 2 and ≥5 mm follicles when compared with ewes entering anestrous late. Similar results were observed for the numbers of all follicles >1 mm in size, total and maximum follicle diameters of all follicles ≥2 mm in size, and the number of ≥5 mm follicles growing by ≥1 mm each day. In contrast, significantly more 4 mm follicles were recorded in ewes entering anestrous early. Daily serum concentrations of FSH and estradiol as well as the peak concentration increases in daily serum FSH concentrations and duration of successive estradiol fluctuations, were greater ( P<0.05) in the early compared to late anestrous ewes. Various parameters of the GnRH-induced surges of FSH and LH did not differ ( P>0.05) between ewes entering anestrous early or late. We concluded that the timing of the onset of seasonal anestrous in Western White-Faced ewes affected the number and size of ovarian antral follicles as well as circulating levels of FSH and estradiol, but not pituitary responsiveness to GnRH at mid-anestrous.

Reproductive ecology and abundance of the sand tiger shark, Carcharias taurus, from the southwestern Atlantic

This study analyses the sexual activity and segregation of sand tiger sharks, Carcharias taurus , from Anegada Bay (Argentina). Reproduction-linked movements along the South American Atlantic coast were inferred from data from several SW Atlantic localities. Male sand tigers (n=162) matured at 193 cm total length (L T ). Females (n=77) matured between 218 and 235 cm L T . These figures are similar to those from other populations, although size-at-maturity of males was slightly different from South African and Australian populations. In females, the size of ovarian follicles was positively correlated with gonadosomatic index and negatively correlated with hepatosomatic index, while the liver was significantly larger than in males. Sand tiger sharks were present in Anegada Bay from December to April. Males were significantly more abundant than females (2:1). Significant differences in reproductive condition through time were observed in males. During January and February males had seminal vesicles full of spermatozeugmata but by March and April the vesicles were empty. As the proportion of males with a lighter colouration peaked from January to March, it is very likely that mating takes place during January and February. The skewed sex-ratio during the mating season indicates a possible strong competition for mates among males, as observed in captivity. Males, females and some juveniles occur in Argentinean and Uruguayan waters, where mating takes place. Pregnant females occur in subtropical waters of southern Brazil, where they give birth. Given that pregnant and non-pregnant females occur at the same time in different zones, we suggest that the female reproductive cycle is biennial. Striking differences among migratory patterns of sand tiger sharks from the SW and NW Atlantic and South Africa were observed. Copyright 2002 International Council for the Exploration of the Sea. Published by Elsevier Science Ltd. All rights reserved .

Dose‐dependent effects of melengestrol acetate (MGA) on plasma levels of estradiol, progesterone and luteinizing hormone in cycling heifers and influences on oestrogen residues in edible tissues

Melengestrol acetat (MGA) is widely used as a growth promoting feed additive in cattle breeding in the USA and several other non‐European countries. To explore the physiological effects of MGA four heifers were fed during 8 weeks with 0.5 mg MGA daily as registered in the USA and two heifers each received 0, 1.5 or 5 mg/day, respectively. Plasma samples were collected twice a week and concentrations of MGA, progesterone (P4) and estradiol‐17β (E2‐17β) were quantified. The pulsatile secretion of luteinizing hormone (LH) was investigated in 6‐hour profiles before and during treatment. After slaughter the reproductive organs were examined and oestrogen residues in edible tissues were measured. Four days after the beginning of MGA feeding MGA concentrations in plasma reached levels of 30 and 100–400 pg/mL depending on the dose received. Three weeks after the beginning of MGA feeding P4 plasma concentrations had dropped to base levels below 0.3 ng/mL in all three treatment groups. Mean plasma E2‐17β levels increased in physiological range from 1 to 5 pg/mL during 0.5 mg MGA/day feeding with many acyclic peaks. Overdosed MGA decreased E2 levels and suppressed cyclic peaks. Number and size of ovarian follicles were not altered by any treatment. Mean LH levels and pulse frequencies increased significantly during labelled treatment (0.5 mg/day), while higher doses had reducing effects. The development of corpus luteum was suppressed. E2‐17β residues in fat increased about 300% following labelled MGA treatment.

Effects of the Persistent Dominant Follicle on the Ability of Follicle Stimulating Hormone to Induce Follicle Development and Ovulatory Responses

Three experiments were conducted to evaluate the effect of an induced first wave persistent dominant follicle on folliculogenesis and ovulatory responses induced by FSH. On d 6 of asynchronized estrous cycle (d 0=estrus), cows were treated with a Syncromate-B implant and two injections of PGF2α (25mg, 0700h; 15mg, 1900h, i.m.). Cows in the control group retained a first-wave persistent dominant follicle, but in the aspirated group, the first-wave dominant follicle was removed via transvaginal aspiration on d 10 (d 0=estrus). Beginning on d 12, cows received 32mg of FSH-P i.m. in decreasing doses at 12-h intervals over a 4-d period. On d 15, the Syncromate-B implant was removed, and cows were ovariectomized (experiment 1, n=8) or inseminated (experiment 2, n=11) at 10 and 22h after the onset of estrus. Cows in experiment 3 received a used controlled intravaginal drug releasing (CIDR) device and two injections of PGF2α (25mg, 0700h; 15mg, 1900h; i.m.) on d 6. On d 8, the first-wave dominant follicle was aspirated (n=6) or left intact (n=5), and FSH treatment was initiated (20mg of Folltropin in decreasing doses at 12-h intervals over a 4-d period), and on d 10 the used CIDR device was removed from all cows. Ovarian follicle size and number were examined daily by ultrasonography from d 5 of the estrous cycle. The persistent dominant follicle increased in size from 10.7mm on d 5 to 15.4mm on d 10 (experiments 1 and 2), and from 9mmon d 5 to 20.4mmon d 11 (experiment 3). From d 11 to 14, the number of class 1 (2 to 5mm) follicles was lower in the aspirated group than in the control group; the number of class 2 (6 to 9mm) follicles was higher on d 12 and 13 for the aspirated group (experiments 1 and 2). The number of class 3 (≥10mm) follicles was higher in the aspirated group on d 14 to 16, but the same on d 17. Ovarian and embryo responses to superovulation did not differ between groups. In experiment 3, the numbers of class 1, 2, and 3 follicles, as well as ovarian and embryo responses following ovulation did not differ between groups. Initiation of exogenous FSH treatment appears to override any systemic inhibitory effect that a persistent dominant follicle may be exerting at the pituitary and possibly the ovary.

Effect of short-term calf removal at three stages of a follicular wave on fate of a dominant follicle in postpartum beef cows.

The hypothesis that ovulation in response to short-term (48 h) calf removal (CR) is dependent on the developmental stage of the dominant follicle was tested in two studies. The objective of Exp. 1 was to characterize the fate of a dominant follicle following 48-h CR on d 2, 4, or 8 of a postpartum follicular wave. Ovaries of 61 beef cows were examined daily by transrectal ultrasonography starting at d 20 to 21 postpartum. Treatments were no CR (n = 14) and CR on d 2 (n = 12), 4 (n = 16), or 8 (n = 10) of first detected follicular wave. Percentage of cows that ovulated a dominant follicle following treatment was not different among groups (P = 0.62). Maximum size of dominant follicles was larger in cows that ovulated (P = 0.002) than in cows that did not ovulate. The objectives of Exp. 2 were 1) to determine whether a follicular wave could be synchronized in anestrous cows following injection of 1 mg of estradiol benzoate (EB) and 200 mg of progesterone (P4; EB + P4); 2) to characterize the fate of dominant follicles following 48-h CR at three stages of a synchronized follicular wave; and 3) to determine whether estrous cycles of normal length followed ovulation in cows pretreated with EB + P4. Ovaries of 50 anestrous beef cows were examined daily as in Exp. 1. Treatments were sesame oil (SO) injected (i.m.) on d 25 postpartum and no CR (n = 9); EB + P4 and no CR (n = 9); EB + P4 and CR on 6 (n = 12), 8 (n = 9), or 12 (n = 11) d after injection. The EB and P4 injections were given on d 25 postpartum. Variability in day of emergence of subsequent follicular waves was lower in cows receiving EB + P4 than in SO-injected cows (P < 0.05). The percentage of cows that ovulated was not different (P = 0.16), but CR increased the percentage of cows that ovulated when groups that received EB + P4 were compared to the EB + P4 group that did not have CR (53.1 vs 11.1%, respectively; P < 0.05). Maximum diameter of dominant follicles was larger (P = 0.05) in ovulatory follicles. The luteal phase was longer (P < 0.03) in cows receiving EB + P4 injection (10.6 +/- 1.2 d) than in cows receiving SO (4.4 +/- 2.2 d). In summary, the maximum size of ovulatory follicles was greater than that of nonovulatory follicles, the ovulatory response of postpartum anestrous cows was maintained through d 8 of a follicular wave, synchronization of follicular waves was accomplished in postpartum cows using EB + P4, and the subsequent luteal phase length was increased in animals that were administered EB + P4.

Control of ovarian follicular growth and maturation by the corpus luteum and the placenta during pregnancy in sheep

Ovarian follicular growth and maturation and its control throughout pregnancy have not been described fully in sheep. Experiment 1 characterized the size and maturation (steroid production in vitro and aromatase activity) of ovarian follicles obtained at days 20, 50, 80 and 110 of pregnancy compared with those obtained at day 12 of the oestrous cycle. There was no difference in the number of small follicles (&lt; 3 mm in diameter) between cyclic and pregnant ewes, regardless of the stage of pregnancy. There was a marked reduction (P &lt; 0.01) in the number of medium follicles (3-5 mm) starting at day 80 of pregnancy. Large follicles (&gt; 5 mm) were not detected at day 110 of pregnancy. In vitro testosterone output by follicles was constant throughout pregnancy. Oestradiol output remained steady until day 80, but decreased markedly at day 110 of pregnancy. This decrease was associated with a reduction in aromatase activity in follicles obtained at this stage. Experiment 2 examined the effect of administration of high concentrations of progesterone between day 100 and day 120 after mating on resumption of follicular growth in ewes that underwent Caesarean section at day 99 of pregnancy. In ewes that underwent Caesarean section, progesterone supplementation was successful in mimicking the profile found in pregnant ewes, but did not prevent re-initiation of follicular growth, as demonstrated by the presence of large follicles (&gt; 5 mm) at day 120 after mating. Experiment 3 examined the effects of PGF(2alpha)-induced regression of the corpus luteum of day 100 of pregnancy on resumption of follicular growth. High concentrations of PGF(2alpha) (0.28 mg kg(-1) body weight) administrated at day 100 of pregnancy were required to initiate regression of the corpus luteum. At day 120 after mating, the mean (+/- SEM) diameter of the largest follicle in PGF(2alpha)-treated ewes (3.40 +/- 0.47 mm) was significantly greater (P &lt; 0.05) than that in control pregnant ewes (2.52 +/- 0.34 mm). Experiment 4 examined the effect of removal of the fetus and of the corpus luteum at day 100 of pregnancy on resumption of ovulation. Removal of the corpus luteum by PGF(2alpha) treatment at the time of removal of the fetus resulted in earlier occurrence of short luteal phases (27.8 versus 40.6 days, PGF(2alpha)-treated versus non-treated) but did not alter the timing of the first normal luteal phases (41 days). In conclusion, the results from these experiments indicate that placental compounds play a major role in inhibiting follicular growth and maturation during late pregnancy in sheep.

Control of ovarian follicular growth and maturation by the corpus luteum and the placenta during pregnancy in sheep

Ovarian follicular growth and maturation and its control throughout pregnancy have not been described fully in sheep. Experiment 1 characterized the size and maturation (steroid production in vitro and aromatase activity) of ovarian follicles obtained at days 20, 50, 80 and 110 of pregnancy compared with those obtained at day 12 of the oestrous cycle. There was no difference in the number of small follicles (< 3 mm in diameter) between cyclic and pregnant ewes, regardless of the stage of pregnancy. There was a marked reduction (P < 0.01) in the number of medium follicles (3-5 mm) starting at day 80 of pregnancy. Large follicles (> 5 mm) were not detected at day 110 of pregnancy. In vitro testosterone output by follicles was constant throughout pregnancy. Oestradiol output remained steady until day 80, but decreased markedly at day 110 of pregnancy. This decrease was associated with a reduction in aromatase activity in follicles obtained at this stage. Experiment 2 examined the effect of administration of high concentrations of progesterone between day 100 and day 120 after mating on resumption of follicular growth in ewes that underwent Caesarean section at day 99 of pregnancy. In ewes that underwent Caesarean section, progesterone supplementation was successful in mimicking the profile found in pregnant ewes, but did not prevent re-initiation of follicular growth, as demonstrated by the presence of large follicles (> 5 mm) at day 120 after mating. Experiment 3 examined the effects of PGF(2alpha)-induced regression of the corpus luteum of day 100 of pregnancy on resumption of follicular growth. High concentrations of PGF(2alpha) (0.28 mg kg(-1) body weight) administrated at day 100 of pregnancy were required to initiate regression of the corpus luteum. At day 120 after mating, the mean (+/- SEM) diameter of the largest follicle in PGF(2alpha)-treated ewes (3.40 +/- 0.47 mm) was significantly greater (P < 0.05) than that in control pregnant ewes (2.52 +/- 0.34 mm). Experiment 4 examined the effect of removal of the fetus and of the corpus luteum at day 100 of pregnancy on resumption of ovulation. Removal of the corpus luteum by PGF(2alpha) treatment at the time of removal of the fetus resulted in earlier occurrence of short luteal phases (27.8 versus 40.6 days, PGF(2alpha)-treated versus non-treated) but did not alter the timing of the first normal luteal phases (41 days). In conclusion, the results from these experiments indicate that placental compounds play a major role in inhibiting follicular growth and maturation during late pregnancy in sheep.

Serum Values of Estradiol (E 2) Adjusted to Serum Volume May Change the Evaluation Outcome of Ovarian Response, Treatment and Results of IVF Patients When Compared to the Absolute Values

Objectives: Evaluation of ovarian response to FSH stimulation in IVF cycles is made by testing hormone levels (per ml) in the blood (serum). However, serum volume is about 5% of body weight (BW), therefore serum volume is dependent on BW of the individual patient. Ovarian secretion, follicle and oocyte size are not dependent on BW; therefore, absolute serum level measurements may not represent the real values and ovarian response. The decision on which day hCG (D-hCG) should be given or cycle cancellation depends mainly upon the daily examination of E2 level. Our objective was to examine the relationship between of absolute E2 (a-E2) and adjusted to E2 (sa-E2) levels on D-hCG and IVF outcome.

Hormonal and ovarian responses to a 5-day progesterone treatment in anoestrous dairy cows in the third week post-partum

Primiparous cows with low body condition at calving have an extended anovulatory period. Induction of ovulation and oestrus is possible with progesterone treatment but the response to this treatment differs between Friesian and Jersey breeds. The objective of this study was to describe changes in pulsatile LH secretion and the synchrony of developing ovarian follicles that occur during a progesterone treatment period of 5 days in primiparous anovulatory cows. The experimental model compared the progesterone treatment with spontaneous post-partum changes as well as a breed comparison in a factorial design. Thirty-six cows (Jersey n=19 and Friesian n=17) were managed to calve with a low body condition score (BCS<4.5). Daily changes in ovarian follicle size were observed with transrectal ultrasonography in each cow from 8 days post-partum. Thirty of these cows were diagnosed to be anovulatory at 12–18 days post-partum (day 0) and allocated to a treatment ( n=16) or a control group ( n=14), balanced for breed. Each treated cow had a progesterone-releasing controlled internal drug-releasing (CIDR) device inserted vaginally for 5 days while control cows were left untreated. Changes in plasma LH concentrations were measured with intensive blood sampling over 8 h on days −1, 1, and 4. Blood samples were also collected daily (06:00 h) for determination of plasma progesterone as well as oestradiol concentrations on days 6 and 8. Treatment with progesterone was associated with a transient initial decrease (day 1) in both LH pulse frequency and mean LH concentrations after device insertion, but both had returned to pre-treatment levels by day 4. Jersey cows had a greater pulse frequency, but there was no breed difference in mean LH concentrations. Patterns of ovarian follicle growth were affected by progesterone treatment with an increase in diameter of the dominant follicle (DF) identified after treatment initiation. This followed an earlier emergence of a new DF after device insertion. Follicular response to progesterone was dependent on the diameter of the DF present at treatment initiation. Those follicles ≥9 mm were replaced by a new DF during treatment such that the DF observed at the time of device removal was large (≥9 mm) and growing in 13/16 cases. Progesterone was not effective for the induction of an LH surge, ovulation and oestrus in anovulatory cows with a low BCS. However, treatment was associated with synchronous development of a DF so that it was large and growing at the end of the treatment period in most cases. This synchronous development may be due to the transient suppression of LH and the presence of an LH-dependent DF.

Pharmacodynamics of the contraceptive patch

Objective: To select a contraceptive patch size that provides ovulation suppression and cycle control equal to or greater than a well accepted oral contraceptive (OC).Methods: Subjects were randomly assigned to 1 of 4 treatment groups (for 4 cycles): 10-cm2 patch (3.0 mg of 17-deacetylnorgestimate [17d-NGM]/0.38 mg of ethinyl estradiol [EE]) (n = 153), 15-cm2 patch (4.5 mg of 17d-NGM/0.56 mg of EE) (n = 157), 20-cm2 patch (6.0 mg of 17d-NGM/0.75 mg of EE) (n = 150), or oral NGM 250 μg/EE 35 μg (n = 150). Regimens: Three 7-day patches (21 days) plus 1 patch-free week; 1 pill daily (21 days) plus 1 pill-free week, with a 1-day late start in cycle 4 for all subjects. Suppression of ovarian activity was determined by serum progesterone levels on days 7, 14, 21 and 28 of cycles 1, 3, and 4, ovarian follicle size by ultrasonography in 109 subjects on days 2 (or 3), 7, 14, 21, and 28 of cycles 1, 3 and 4, and cycle control via diary cards.Results: The proportions of subjects with progesterone values greater than 3 ng/mL in cycles 1 and 3 were 41.9%, 10.2%, 5.5%, and 6.1% for the 10-, 15-, and 20-cm2 patch and OC groups, respectively. Maximum mean follicular diameter in cycles 1, 3, and 4 ranged from 10 to 11.1 mm in the 20-cm2 patch (EVRA) group and from 10.7 to 14 mm for the OC group. The percentages of subjects in the 20-cm2 patch (EVRA) and OC groups with breakthrough bleeding/spotting were similar for cycles 1 and 2 and, in cycle 3, were better for EVRA (10.5% vs 15%).Conclusion: The 20-cm2 contraceptive patch (EVRA) provides high levels of suppression of ovarian activity and effective cycle control compared with a widely accepted OC, while providing more complete suppression of follicular development.

Ultrasonographic study of ovarian function during early pregnancy and after parturition in the ewe

Transrectal ultrasonography of ovaries was performed in 11 ewes from Days 10 to 26 and on Day 30 of pregnancy to record the number, size, and position of ovarian follicles ≥3 mm in diameter and corpora lutea (CL). Transrectal and/or transabdominal ultrasonography of the uterus was performed on Days 10 to 26, 30, 35, 40, 45 and 50 of gestation to ascertain the number and position of the conceptuses. In a second experiment, ultrasonography was conducted in 15 ewes on Days 10, 25, 30, 45 and 50 of pregnancy and from Days 13 to 29 after parturition. Ovarian data were classified into ovaries without CL (Group 1), ovaries with the CL in 1 ovary (Group 2), and ovaries with the CL in both ovaries during pregnancy (Group 3). In early pregnant ewes, the total number of follicles and the diameter of all follicles ≥3 mm were smaller (P<0.05) in the CL-bearing ovaries (both Group 2, n=7 and Group 3, n=8) than in the non-CL-bearing ovaries (Group 1, n=7), while the largest follicle diameter was significantly smaller in Group 3 than in Group 1 or 2 ovaries. The number of 3-mm follicles in Group 2 ovaries was lower (P<0.05) than in Group 1 or 3 ovaries, but the mean number of follicles ≥5 mm in diameter was significantly lower in Group 3 than in Group 1. The total luteal volume per ovary was higher (P<0.001) in Group 2 than in Group 3 ovaries of early pregnant ewes. The total follicle diameter and the number of follicles growing from 3 to ≥5 mm in diameter was lower (P<0.05) for Group 2 ovaries of ewes that carried twins (n=3) compared with Group 2 ovaries from ewes with singletons (n=4). There were no differences in follicular dynamics between Group 3 ovaries and the ovaries of Group 2 in ewes that carried twins. No follicles >3 mm were seen in the ovaries of postpartum ewes that contained CL during gestation, until Days 21 and 25 postpartum for Groups 2 (n=10) and 3 (n=8), respectively, and follicles reaching ≥5 mm in diameter were detected in only 2 ovaries (Group 2), on Days 27 and 28 postpartum, respectively. We conclude that during early pregnancy in ewes there is a suppression of antral follicle growth which appears to be exerted primarily by the developing conceptus but remains confined to CL-bearing ovaries. Residual local inhibition of follicular development extends into the postpartum period.

Effects of altering dietary fatty acid composition on prostaglandin synthesis and fertility

Several studies over the past 20 years have demonstrated that subjects on diets composed of substances with high levels of n-3 polyunsaturated fatty acids (PUFAs) (e.g. fish) have a decreased incidence of heart disease. On this basis, a recent report from the Department of Health has advised UK consumers to decrease the proportion of saturated as opposed to unsaturated fats in their diet and to increase the ratio of n-3 to n-6 PUFAs. This could be achieved by altering the amounts of these constituents in milk and meat. n-3 Fatty acids can most easily be added to animal feed as either fish oil or linseed oil and can be increased in the blood and milk of ruminants following protection to avoid hydrogenation in the rumen. In western countries the ratio of consumption of n-6 to n-3 PUFAs is greater than 10 and current evidence tends to suggest that a ratio nearer 5 would be more desirable and compatible with cardiovascular well being. As fertility in the UK dairy herd is already poor, it is important to establish whether alterations in dietary n-3 and n-6 PUFAs affects herd fertility before widespread changes in animal diets are recommended. Therefore, this review considers the role played by PUFAs and eicosanoids in fertility, with particular reference to the implications for farm livestock production.The evidence reviewed shows that alteration of the concentration and ratio of n-6 and n-3 PUFAs in feeds can influence prostaglandin synthesis/metabolism in a number of mammalian systems. The changed patterns of prostaglandin synthesis can as a consequence, affect the diverse functions (e.g. hormone secretion) that are normally mediated via prostaglandins. Similarly, changes in prostaglandin synthesis effected through manipulation of PUFAs has a major bearing on fertility (as PGs affect many reproductive parameters, e.g. ovulation). Several studies in cattle and other mammals, show that feeding or infusing different types of fat with varying PUFA content to females can alter: the number and size of ovarian follicles, the ovulation rate, progesterone production by the corpus luteum, the timing of luteolysis and gestational length. In the male most recent work has focussed on sperm production and experiments in fowl have demonstrated clear effects of dietary PUFAs on both the sperm membrane phospholipid composition and on fertilizing ability.

Post partumSubclinical Hypocalcaemia and Effects on Ovarian Function and Uterine Involution in a Dairy Herd

Episodes of subclinical hypocalcaemia occurred in the first 6 weeks after calving in 6 out of 12 multiparous Friesian cows calving in the winter period and 7 out of 23 calving in the summer in a subtropical environment. In the winter calving group, there were significant (P< 0.05) decreases in mean plasma calcium concentrations (PTCa) on days 6, 27 and 36 after calving in those animals unable to maintain PTCa >2.0 mmol/L in the first 21 days. These cows had a significantly higher mean body weight and higher mean milk production than normocalcaemic cows. In the summer calving group, the seven cows with episodes of subclinical hypocalcaemia had significantly lower mean PTCa over periods of 21 and 60 days. Intervals until involution of the uterus were negatively (P< 0.05) correlated with mean PTCa over the first 21 and 30 days after calving in the winter and summer calving groups (r= –0.62 and –0.42), respectively. The mean size of the first ovulated follicles was significantly smaller in cows showing episodes of subclinical hypocalcaemia in the winter group and the mean number of ovulatory size follicles was less in these cows at 15 days (P< 0.001) (1.2 ± 0.17vs2.3 ± 0.21) and at 30 days (P< 0.03) (3.5 ± 0.43vs5.2 ± 0.54). During the first three dioestrus periods, mean plasma progesterone concentration as a function of corpus luteum area was significantly lower (P< 0.05) in the cows with episodes of subclinical hypocalcaemia in the winter group.

Ovulation and estrus characteristics in crossbred Brahman heifers treated with an intravaginal progesterone-releasing insert in combination with prostaglandin F2alpha and estradiol benzoate.

Crossbred Brahman heifers (n = 60) were studied to determine the effect of a 7-d intravaginal progesterone-releasing insert (INSERT) in combination with PG (Lutalyse; 25 mg i.m.) and estradiol benzoate (EB; .5 mg i.m.) on time of ovulation and estrous behavior. In Phase I, heifers at unknown stages of the estrous cycle were assigned by BW and body condition score to one of the three treatments on d 0: 1) INSERT for 7 d and PG on d 7 (CONTROL; n = 10); 2) INSERT for 7 d, PG on d 7, and EB 24 h after INSERT removal (EB24; n = 10); or 3) INSERT for 7 d, PG on d 7, and EB 48 h after INSERT removal (EB48; n = 10). Blood samples were collected every 8 h after INSERT removal. Also, blood sampling and ultrasonography began 8 h after the onset of estrus, determined with HeatWatch devices, and every 4 h thereafter to detect ovulation. In Phase II, Phase-I treatments (n = 10/treatments) were replicated, but only behavioral estrus data were collected to minimize handling of heifers. Frequent handling of heifers did not influence (P > .1) the interval from INSERT removal to the onset of HeatWatch and visual estrus and duration of estrus, so behavioral estrus data were combined for Phases I and II. Interval from INSERT removal to HeatWatch estrus was decreased (P < .05) in EB24 (45.5 h) vs EB48 (55.9 h) and CONTROL (59.2 h). Interval from INSERT removal to ovulation differed (P < .04) between CONTROL, EB24, and EB48 (93.5, 74.5, and 78.9 h, respectively). Ovulatory follicle size was similar (P > .1) between CONTROL, EB24, and EB48 (14.4, 12.5, and 14.1 mm, respectively). Duration of estrus was similar for CONTROL, EB24, and EB48 (14.0, 15.1, and 17.6 h, respectively). No difference (P > . 1) was observed in number of mounts received between CONTROL, EB24, and EB48 (28.0, 25.7, and 39.4, respectively), but number of mounts received increased in Phase II vs Phase I (40.0 and 22.2, respectively; P < .05). In conclusion, EB hastened the interval from INSERT removal to ovulation without altering duration of estrus or number of mounts received. Frequent handling of heifers did not affect interval to first mount received after INSERT removal or duration of estrus, but it decreased the total number of mounts received.

Follicular activity and ovulation regulated by exogenous progestagen and PMSG in anestrous ewes

Follicular dynamics and ovulation were compared in 3 groups of anestrous ewes: those treated with medroxyprogesterone acetate (MAP) sponges for 12 d, then with 750 IU PMSG at the time of sponge removal (P4+PMSG, n=6), or PMSG alone (n=6) and untreated controls (n=6). Waves of follicular activity were observed in all the animals. In the P4+PMSG treatment group, MAP priming permitted more ovulatory follicles (P<0.001) to be recruited without changing follicle growth rate; MAP priming also delayed the time of ovulation (P < 0.001) and the time of the LH surge (P<0.01), which allowed for an increase in the size of ovulatory follicles (P<0.05). Ovulation also resulted in normal luteal function after P4+PMSG (P<0.01) but not after PMSG alone, since premature luteal regression occurred in 80% of the cases and was related to the presence of follicles >4 mm when P4 levels were < 1 ng/mL on the day following ovulation. The results showed that MAP priming increased the ovulation rate by increasing the number of follicles that responded to PMSG.

Effects of level of food intake on ovarian follicle number, size and steroidogenic capacity in the ewe.

The effect of high (H) or low (L) levels of food intake, during the preceding 4 weeks, on ovarian follicle numbers and steroidogenic capacity were investigated in groups of 12 adult Scottish Blackface ewes. Ewes of the two treatments had similar levels of body condition at the time of study but there was a twofold difference in levels of food intake. Ovaries were surgically removed on day 11 or 12 of the oestrous cycle (luteal phase; n = 6 per nutritional treatment) or at 30 h after injection (i.m.) of prostaglandin F2alpha analogue on day 11 or 12 of the cycle (follicular phase; n = 6 per nutritional treatment). Ovarian follicles > 1 mm diameter were dissected out and incubated individually for 2 h at 37 degrees C, in 1 ml of medium 199 which was then assayed to determine concentrations of oestradiol and testosterone. There were significantly more small follicles (1-2.5 mm diameter) in (H) than (L) ewes (P < 0.05) but no treatment difference in the numbers of large follicles (> 2.5 mm diameter) during either phase of the cycle and no difference in the mean diameters (mm) of the two largest follicles in each animal. However, although there were higher rates of synthesis of both oestrogen (P < 0.05) and testosterone (P < 0.01) in the large follicles of (L) ewes as compared with (H) ewes, there was a lower oestrogen/testosterone ratio in (L) than (H) follicles which may indicate a lower level of aromatase activity in (L) follicles. It is concluded that the effects of level of food intake on ovulation rate are expressed through differences in late stages of follicle development, probably through effects on the intrafollicular steroid milieux.

Follicle size-dependent induction of prostaglandin G/H synthase-2 during superovulation in cattle.

Under physiological conditions, prostaglandin G/H synthase-2 (PGHS-2) is induced in bovine preovulatory follicles by the endogenous surge of gonadotropins. To characterize the pattern of follicular PGHS-2 expression during superovulation in cattle, heifers were treated with exogenous FSH and ovulation was induced with hCG. Animals were ovariectomized 0, 18, and 24 h post-hCG, and extracts of follicles > or = 6 mm were analyzed by Western blotting. Follicular fluid concentrations of prostaglandin (PG) E2, PGF2alpha, progesterone, and estradiol-17beta were determined by RIAs, and the morphology of the cumulus oocyte complex was examined. Results showed that PGHS-2 protein was absent in all follicles isolated at 0 h post-hCG (n = 119) and in small follicles (6 to < 8 mm) isolated between 0 and 24 h post-hCG (n = 27 follicles). In contrast, 12.3% of medium (8 to < 10 mm) and 43.7% of large (> or = 10 mm) follicles were PGHS-2-positive at 18 h post-hCG, and these percentages rose at 24 h to 45.9% and 91.0% in medium and large follicles, respectively (p < 0.05). Follicular fluid concentrations of PGE2 and PGF2alpha were low in follicles isolated at 0 h and increased only in PGHS-2-positive follicles isolated 24 h post-hCG (p < 0.05). Concentrations of progesterone and estradiol-17beta at 0 h were 28.2 +/- 5.8 and 291.8 +/- 13.0 ng/ml, respectively, and a shift from estradiol-17beta to progesterone dominance (luteinization) occurred at 24 h post-hCG only in PGHS-2-positive follicles. Also, expansion of the cumulus oocyte complex was detected at 24 h post-hCG only in PGHS-2-positive follicles. Lack of PGHS-2 induction in follicles of ovulatory size (> 8 mm) was associated with an apparent failure to respond to hCG (absence of luteinization and cumulus expansion). Collectively, these results demonstrate the presence of a time- and follicle size-dependent induction of PGHS-2 in bovine follicles during superovulatory treatment and suggest that PGHS-2 expression can be used as a marker for follicular commitment to ovulation during ovarian hyperstimulation protocols.