Endocrine alterations associated with extended time interval between estrus and ovulation in high-yield dairy cows

Short fertile half-lives of the male and female gametes in the female tract necessitate accurate timing of artificial insemination. We examined the possible association between extension of the estrus to ovulation (E-O) interval and alterations in concentrations of estradiol, progesterone, and the preovulatory LH surge before estrus and ovulation. High-yielding Holstein cows (n = 74 from a total of 106) were synchronized and were examined around the time of the subsequent estrus. They were observed continuously for estrual behavior. Blood samples were collected before and after estrus, and ultrasound checks for ovulation were made every 4 h. About three-quarters of the cows exhibited short (but normal) E-O intervals of 22 to 25 h (25%) or normal intervals of 25 to 30 h (47%); 17% of them displayed a long (but normal) E-O interval of 31 to 35 h, and about 10% exhibited a very long E-O interval of 35 to 50 h. Extended E-O interval comprised estrus-to-LH surge and LH surge-to-ovulation intervals that were both longer than normal. Pronounced changes in hormonal concentrations were noted before ovulation in the very long E-O interval group of cows: progesterone and estradiol concentrations were reduced, and the preovulatory LH peak surge was markedly less than in the other 3 groups. Postovulation progesterone concentrations during the midluteal phase were lesser in the very long and the long E-O interval groups compared with those in the short and normal interval groups. Season, parity, milk yield, and body condition did not affect the estrus to LH surge, LH surge to ovulation, and E-O intervals. The results indicate an association between preovulatory-reduced estradiol concentrations and a small preovulatory LH surge, on the one hand, and an extended E-O interval, on the other hand. Delayed ovulation could cause nonoptimal timing of AI, a less than normal preovulatory LH surge that may be associated with suboptimal maturation of the oocyte before ovulation, or reduced progesterone concentrations before and after ovulation. All may be factors associated with poor fertility in cows with a very long E-O interval.     

Effects of GnRH administered to cows at the onset of estrus on timing of ovulation,endocrine responses,and conception

Two experiments examined effects of GnRH administered within 3 h after onset of estrus (OE) on ovulation and conception in dairy cows. In experiment 1, 46 cows received either saline, 250 microg of GnRH, or 10 microg of the GnRH analogue, Buserelin. Cows were observed for estrus, blood samples were collected, and ovulations were monitored by ultrasound. In controls, 76% of cows had intervals from estrus to ovulation of < or = 30 h and 24% had intervals > 30 h. Treatment with either GnRH or GnRH analogue (data combined) increased magnitude of LH surges and decreased intervals from estrus to LH surge or to ovulation. Treated cows all ovulated < or = 30 h after OE. Among control cows, plasma estradiol concentrations before estrus correlated positively with amplitudes of LH surges. Higher plasma progesterone was observed in the subsequent estrous cycle in GnRH-treated cows compared to control cows with delayed ovulations. Experiment 2 included 152 primiparous and 211 multiparous cows in summer and winter. Injection of GnRH analogue at OE increased conception rates (CR) from 41.3 to 55.5% across seasons. In summer, GnRH treatment increased CR from 35.1 to 51.6%. Across seasons, GnRH increased CR from 36.0 to 61.5% in cows with lower body condition at insemination and GnRH increased CR (63.2 vs. 42.2%) in primiparous cows compared to controls. Use of GnRH eliminated differences in CR for cows inseminated early or late relative to OE and increased CR in cows having postpartum reproductive disorders. In conclusion, GnRH at onset of estrus increased LH surges, prevented delayed ovulation, and may increase subsequent progesterone concentrations. Treatments with GnRH increased conception in primiparous cows, during summer, and in cows with lower body condition.     

Naturally occurring mastitis effects on timing of ovulation, steroid and gonadotrophic hormone concentrations, and follicular and luteal growth in cows

The effects of naturally occurring subclinical chronic or clinical short-term mastitis on time of ovulation, plasma steroid and gonadotropin concentrations, and follicular and luteal dynamics were examined in 73 lactating Holstein cows. Cows were sorted by milk somatic cell count and bacteriological examination into an uninfected group (n = 22), a clinical mastitis group (n = 9; events occurring 20 ± 7 d before the study), and a subclinical chronic mastitis group (n = 42). In addition, uninfected and mastitic cows were further sorted by their estrus to ovulation (E-O) interval. About 30% of mastitic cows (mainly subclinical) manifested an extended E-O interval of 56 ± 9.2 h compared with 28 ± 0.8 h in uninfected cows and 29 ± 0.5 h in the other 70% of mastitic cows. In mastitic cows with extended E-O interval, the concentration of plasma estradiol at onset of estrus was lower than that of uninfected cows or mastitic cows that exhibited normal E-O intervals (3.1 ± 0.4, 5.8 ± 0.5, and 5.5 ± 0.5 pg/mL, respectively). The disruptive effect of mastitis on follicular estradiol probably does not involve alterations in gonadotropin secretion because any depressive effects of mastitis on pulsatile LH concentrations were not detected. Cortisol concentrations did not differ among groups. The preovulatory LH surge in mastitic cows with delayed ovulation varied among individuals, being lower, delayed, or with no surge noted compared with the normal LH surge exhibited by uninfected cows or mastitic cows with normal E-O interval (6.8 ± 0.7 ng/mL). The diameter of the second-wave dominant follicle was larger and the number of medium follicles was smaller in uninfected and subclinical cows with normal intervals compared with subclinical cows with extended intervals (13.4 ± 0.5 vs. 10.9 ± 0.9 mm, and 3.8 ± 0.2 vs. 6.7 ± 0.14 follicles, respectively). Mid-luteal progesterone concentrations were similar in uninfected and mastitic cows. These results indicate for the first time that around 30% of cows with subclinical chronic mastitis exhibit delayed ovulation that is associated with low plasma concentrations of estradiol and a low or delayed preovulatory LH surge.     

Use of estradiol cypionate as a substitute for GnRH in protocols for synchronizing ovulation in dairy cattle

Our purpose was to determine whether estradiol cypionate (ECP) could be substituted for the second GnRH injection of the standard Ovsynch protocol (injection of GnRH given 7 d before and 48 h after PGF(2alpha), with timed AI [TAI] 12 to 20 h after the second GnRH injection). Lactating dairy cows ranging from 61 to 82 d in milk at TAI were studied in 14 replicates. Main effects were hormone (ECP vs. GnRH) to induce ovulation and exposure to progesterone (P4) or not during the week preceding PGF(2alpha)-induced luteolysis. Four treatments were: 1) 100 microg of GnRH at 48 h after PGF(2alpha) (Ovsynch; n = 27); 2) same as Ovsynch, plus a P4-releasing intravaginal insert (CIDR) placed for 7 d beginning at the first GnRH injection (Ovsynch + CIDR, n = 20); 3) same as Ovsynch, but substituting 1 mg of ECP for GnRH, and injecting ECP at 24 h after PGF(2alpha) (Heatsynch; n = 33); or 4) Heatsynch + CIDR (n = 26). The largest follicle was identified by ultrasonography 24 h after PGF(2alpha) and was monitored every 6 h until ovulation. Incidence of estrus was less after GnRH (54%) than after ECP (87%), but more GnRH-treated cows had LH surges detected (95 vs. 65%) and ovulated (100 vs. 86%). Duration of LH surges, but not peak concentrations, was less after GnRH than after ECP (6.1 +/- 0.7 vs. 12.2 +/- 0.9 h). Pre-treatment with P4 reduced the incidence of LH surges but had no effects on incidence of estrus or ovulation. Intervals to the LH surge and ovulation were less after GnRH than after ECP, but intervals between onset of the LH surge and ovulation did not differ (26 +/- 2 vs. 30 +/- 3 h). We concluded that substituting ECP for GnRH resulted in more cows in estrus and slightly fewer ovulating.     

Relationships between time of day, estrous behavior, and the preovulatory luteinizing hormone surge in Holstein cows after treatment with cloprostenol

Using continuously observed primiparous cows, comparisons were made between cloprostenol-induced and preceding or succeeding spontaneous estruses in a crossover design. There were no significant differences between spontaneous and induced estruses in duration or intensity. In a second experiment, estrus was controlled by administration of two cloprostenol injections, 12 h apart, initiated at 0800 or 2000 h during the midluteal phase. Continuous observation and frequent blood sampling were used to determine relationships between behavior and plasma concentrations of LH and estradiol-17 beta. Relationships between plasma concentrations of estradiol-17 beta and the intensity or duration of estrous behavior were not detected. The intervals from treatment to the onset of estrus and to the LH surge were not affected by time of treatment but were highly correlated. The onset of the LH surge always followed the onset of standing to be mounted by 1 to 2 h. Initiating treatment with cloprostenol at 2000 h resulted in an increased proportion of animals exhibiting onset of estrus and increased mounting behavior at night relative to treatment at 0800 h. For efficient detection of estrus after cloprostenol treatment it is suggested that cloprostenol be administered early in the working day.     

Impaired final follicular maturation in heifers after superovulation with recombinant human FSH

The aim of this study was to investigate whether human FSH without contaminating LH can exert a normal superovulation response in cows. One group of heifers (n = 9) was stimulated with recombinant human FSH (rhFSH), an FSH source without any LH activity, and another group (n = 9) was treated with equine chorionic gonadotrophin (eCG), an FSH source with high LH activity. Daily transrectal ultrasonography showed that eCG- and rhFSH-stimulated heifers (n = 9 per group) had the same follicular growth characteristics and equal numbers of follicles > 8 mm in diameter after 3 days of stimulation. The treatment groups differed considerably in steroid production: rhFSH-treated heifers produced much lower oestradiol concentrations than did eCG-stimulated heifers during the first days of stimulation and much lower progesterone concentrations in the period after the LH surge. During the 27-35 h after prostaglandin injection, rhFSH-treated heifers had fewer LH pulses than did eCG-treated heifers (0.3 versus 3.0 per heifer, respectively; n = 3 per group). All rhFSH-treated heifers (n = 6) underwent a preovulatory LH surge, but this occurred significantly later than in the eCG-treated heifers (n = 4; 39.4 +/- 1.9 h versus 47.1 +/- 1.5 h in rhFSH- and eCG-treated heifers, respectively). Multiple ovulations occurred in only three of six rhFSH-treated heifers, but in all four eCG-treated heifers with an LH surge. At 24 h after the LH surge, the percentage of metaphase II stage oocytes with cortical granules distributed close to the oolemma was significantly lower in the rhFSH group (7.3%) than in the eCG group (55.9%). In conclusion, final follicular maturation is impaired in heifers treated with rhFSH, which might be due to the combination of a lack of LH activity in the gonadotrophin preparation and the severe suppression of LH pulsatility.     

Relationships between FSH patterns and follicular dynamics and the temporal associations among hormones in natural and GnRH-induced gonadotropin surges in heifers

Preovulatory LH and FSH surges and the subsequent periovulatory FSH surge were studied in heifers treated with a single injection of GnRH (100 microg, n = 6) or saline (n = 7). Blood samples were collected every hour from 6 h before treatment until 12 h after the largest follicle reached > or =8.5 mm (expected beginning of follicular deviation). The GnRH-induced preovulatory LH and FSH surges were higher at the peak and shorter in duration than in controls, but the area under the curve was not different between groups. The profiles of the preovulatory LH and FSH surges were similar within each treatment group, suggesting that the two surges involved a common GnRH-dependent mechanism. Concentrations of FSH in controls at the nadir before the preovulatory surge and at the beginning and end of the periovulatory surge were not significantly different among the three nadirs. A relationship between variability in the periovulatory FSH surge and number of 5.0 mm follicles was shown by lower FSH concentrations during 12-48 h after the beginning of the surge in heifers with more follicles (11.0 +/- 1.0 follicles (mean+/-s.e.m.) n = 7) than in heifers with fewer follicles (5.7 +/- 0.4, n = 6). This result was attributed to increased FSH suppression from increased numbers of follicles reaching 5.0 mm. Grouping of heifers into those with longer vs shorter intervals from a 4.5 mm to an 8.5 mm largest follicle did not disclose any relationship between length of the interval and FSH characteristics (e.g. profile of surge, area under curve, FSH concentrations at specific events). The hypothesis of a relationship between variation in the periovulatory FSH surge and variation in follicular dynamics was supported for the number of 5.0 mm follicles but not for the hour the largest follicle reached 8.5 mm. Thus, the expected time of follicle deviation was not altered by the extensive variation in the wave-stimulating FSH surge.     

Anogenital distance is associated with postpartum estrous activity,intensity of estrous expression,ovulation, and progesterone concentrations in lactating Holstein cows

The objectives of this retrospective observational study were to determine the associations of anogenital distance (AGD) with (a) postpartum estrous activity, (b) diameter of the preovulatory follicle, (c) intensity of estrous expression, (d) postestrus ovulation, (e) corpus luteum (CL) size, and (f) concentrations of progesterone at estrus and on d 7 after estrus. Lactating Holstein cows (n = 178; 55 primiparous, 123 multiparous) were enrolled into the study during the first postpartum week. All cows were continuously monitored by a pedometer-based automated activity monitoring (AAM) system for estrus. Postpartum estrous activity was assessed using the AAM estrus alerts, in which cows with at least one true estrus alert (i.e., a relative increase in steps from each cow's baseline detected by the AAM and the presence of at least one follicle >15 mm, a CL <20 mm, or no CL detected by ultrasound) by the first 50 d in milk (DIM) were considered to have commenced estrous activity. At the estrus alert >60 DIM, ovulation was determined by ultrasound at 24 h, 48 h, and 7 d after estrus, and blood samples were collected at estrus alert and on d 7 after estrus for progesterone analysis. The AGD was measured from the center of the anus to the base of the clitoris and classified as either short- or long-AGD using 2 cut-points of 148 mm (predictive of the probability of pregnancy to first insemination; short-AGD, n = 115; long-AGD, n = 63) and 142 mm (the median AGD; short-AGD, n = 90; long-AGD, n = 88). Regardless of the cut-point used, early postpartum estrous activity by 50 DIM (67 vs. 54%), duration of estrus (11.6 vs. 9.7 h), and preovulatory follicle diameter (20 vs. 19 mm) were greater in short-AGD than in long-AGD cows. Increased peak of activity at estrus in short-AGD cows (354 vs. 258% mean relative increase) was affected by an interaction between AGD and parity in which multiparous long-AGD cows had lesser relative increase in activity than primiparous cows (217 vs. 386%, respectively). Mean progesterone concentration at estrus was lesser in short-AGD (0.47 vs. 0.61 ng/mL) than in long-AGD cows. The ovulatory response at 24 h did not differ, but at 48 h (91 vs. 78%) and on d 7 after estrus (97 vs. 84%) it was greater in short-AGD cows. Although CL diameter on d 7 after estrus did not differ, short-AGD cows had greater progesterone concentration 7 d after estrus than long-AGD cows (4.1 vs. 3.2 ng/mL, respectively). In conclusion, greater proportions of short-AGD cows commenced estrous activity by 50 DIM, had larger preovulatory follicles, exhibited greater duration of estrus, had reduced progesterone concentration at estrus, had greater ovulation rates and progesterone concentration 7 d after estrus compared with long-AGD cows, with no difference in CL size between AGD groups. Because all the differences in physiological characteristics of short-AGD cows reported herein favor improved reproductive outcomes, we infer that these are factors contributing to improved fertility reported in short-AGD cows compared with long-AGD cows.     

Preovulatory follicle characteristics and oocyte competence in repeat breeder dairy cows

The varied and elusive etiology of repeat breeding (RB) in dairy cows necessitates evaluation of oocytes and follicles, which have not previously been assessed together. Accordingly, we evaluated characteristics of preovulatory follicles and the competence of oocytes in control (CTL) and RB cows. The estrous cycles of 35 cows (18 CTL and 17 RB) were synchronized using PGF_2α and estrus detection. Cows with a corpus luteum were treated with PGF_2α and, 14 to 15 d after a visible behavioral estrus, they were administered a second PGF_2α, followed 48 h later by follicular fluid (FF) aspiration of the preovulatory follicles. Estradiol (E₂)-active preovulatory follicles did not differ in diameter between the 2 groups of cows. However, FF of RB cows had higher E₂ concentrations than that of CTL cows: 1,854.9 and 1,073.6 ng/mL, respectively, but similar androstenedione and progesterone concentrations. In the second part of the study, 14 consecutive ovum pick-ups (OPU) were performed in 5 CTL and 5 RB cows, at 3- to 4-d intervals. The RB and CTL cows did not differ in average numbers of follicles available per cow per session (7.1 and 7.3, respectively), oocyte recovery rates (42.2 and 44.1%, respectively), or cleavage rates (57.6 and 63.4%, respectively), but blastocyst production was markedly less in RB than in CTL cows (12.5 and 29.2%, respectively). We conclude that part of the RB cows' etiology occurs at an earlier phase of folliculogenesis, thereby impairing oocyte competence, and subsequently reducing the probability of normal fertilization, which diminish embryo vitality and development.     

Effects of progesterone intravaginal devices on synchronization of estrus in postpartum dairy cows

Effects of two intravaginal treatments of progesterone on the synchronization of behavioral estrus were compared in three groups of lactating dairy cows at 7 to 8 wk postpartum. Group 1 had progesterone-releasing intravaginal devices with attached estradiol benzoate capsules inserted for 12 d. Groups 2 and 3 had controlled internal drug release devices, containing progesterone alone, inserted for 9 or 12 d. Behavioral estrus was detected as early as 24, 32, and 36 h after removal of the devices in groups 1, 2, and 3, respectively. Estrus was better synchronized and the proportion of cows showing estrus after device removal was greater in group 1 than in groups 2 and 3. After removal of devices, mean milk progesterone of all cows that did not show estrus was higher than that of cows that showed estrus. In groups 1, 2, and 3, 1 of 7, 7 of 12, and 4 of 12 cows not showing estrus had high milk progesterone after removal of the devices, indicating the presence of functional corpora lutea. In cows not displaying estrus milk progesterone concentrations decreased, indicating these cows were cyclic, although behavioral estrus was absent.     

Luteinization of porcine preovulatory follicles leads to systematic changes in follicular gene expression

The LH surge initiates the luteinization of preovulatory follicles and causes hormonal and structural changes that ultimately lead to ovulation and the formation of corpora lutea. The objective of the study was to examine gene expression in ovarian follicles (n = 11) collected from pigs (Sus scrofa domestica) approaching estrus (estrogenic preovulatory follicle; n = 6 follicles from two sows) and in ovarian follicles collected from pigs on the second day of estrus (preovulatory follicles that were luteinized but had not ovulated; n = 5 follicles from two sows). The follicular status within each follicle was confirmed by follicular fluid analyses of estradiol and progesterone ratios. Microarrays were made from expressed sequence tags that were isolated from cDNA libraries of porcine ovary. Gene expression was measured by hybridization of fluorescently labeled cDNA (preovulatory estrogenic or -luteinized) to the microarray. Microarray analyses detected 107 and 43 genes whose expression was decreased or increased (respectively) during the transition from preovulatory estrogenic to -luteinized (P<0.01). Cells within preovulatory estrogenic follicles had a gene-expression profile of proliferative and metabolically active cells that were responding to oxidative stress. Cells within preovulatory luteinized follicles had a gene-expression profile of nonproliferative and migratory cells with angiogenic properties. Approximately, 40% of the discovered genes had unknown function.     

Incorporation of dietary n-3 fatty acids into ovarian compartments in dairy cows and the effects on hormonal and behavioral patterns around estrus

The objective of this study was to examine the incorporation of dietary n-3 fatty acids (FAs) into ovarian compartments and the effects on hormonal and behavioral patterns around estrus. Multiparous 256-day pregnant cows were fed either a standard diet both prepartum and postpartum (PP) (control; n=22) or supplemented with extruded flaxseed (E-FLAX) providing C18:3n-3 at 172.2 and 402.5 g/day per cow prepartum and PP respectively (n=22). The estrous cycle was synchronized, and at day 7 of the cycle, the cows were injected with prostaglandin F(2)(α) (PGF(2)(α)) and then subjected to 5 days of intensive examination. Compared with those in the control, in the E-FLAX group, the interval from PGF(2)(α) injection to behavioral estrus peak tended to be longer (3.6 h; P<0.1), that to estradiol (E(2)) peak was 6.5 h longer (P<0.03), and that to LH peak tended to be longer (5.3 h; P<0.07). The durations of behavioral estrus and E(2) surge were longer, and the area under the E(2) curve was greater in the E-FLAX cows. Afterward, 7-8 days following behavioral estrus, follicular fluids (FFs) from >7 mm follicles were aspirated. The proportions of n-3 FA increased in plasma, FF, and granulosa cells in the E-FLAX group. The concentrations of PGE(2) in the E(2)-active follicles tended to be lower in the E-FLAX cows (P<0.06). In conclusion, several modifications in hormonal and behavioral estrus patterns were demonstrated in cows fed n-3 FA, which might be attributed to alterations in membrane FA composition and partly mediated by lower PGE(2) synthesis.     

Testosterone antagonist (flutamide) blocks ovulation and preovulatory surges of progesterone, luteinizing hormone and oestradiol in laying hens

The preovulatory release of luteinizing hormone (LH) in the domestic hen occurs after the initiation of a preovulatory surge of testosterone. The objective of this study was to determine whether this testosterone surge has functional significance in the endocrine control of ovulation. Groups of laying hens (n = 10-22) were treated with the androgen receptor antagonist, flutamide, at 8 h intervals for 24 h at doses of 0, 31.25, 62.5, 125 and 250 mg. All doses reduced egg laying (P < 0.001), with the highest dose being the most effective. In a second study, laying hens (n = 9) were treated with 250 mg flutamide at 8 h intervals for 24 h with a control group being given placebo (n = 10). Blood samples were taken for hormone measurements at 2 h intervals for 18 h starting 4 h before the onset of darkness. The percentage of hens laying per day did not differ between groups before treatment (control, 88% vs flutamide, 86%). Ovulation was blocked in all hens treated with flutamide within 2 days while the control hens continued to lay at the pretreatment rate (80%). Preovulatory surges of plasma testosterone, progesterone, oestradiol and LH were observed in control hens but with the exception of testosterone, flutamide treatment blocked the progesterone, oestradiol and LH surges. LH concentrations declined progressively with time in the flutamide-treated hens. It is concluded that inhibition of testosterone action blocks egg laying and the preovulatory surges of progesterone, luteinizing hormone and oestradiol demonstrating a key role for the preovulatory release of testosterone in the endocrine control of ovulation in the domestic hen.     

Ovarian follicular responses to high doses of pulsatile luteinizing hormone in lactating dairy cattle

Two experiments in lactating dairy cows examined ovarian follicular responses to high, frequent doses of exogenous LH pulses at levels associated with follicular cysts. In Experiment 1, estrus was synchronized in 12 cyclic lactating cows >40 d postpartum. Emergence of the second follicular wave (d 0) was determined by ultrasonography. Starting on d 1, cows received LH (40 microg/h; n = 7) or saline (2 mL/h; n = 5) in hourly pulses for up to 5 (n = 5) or 7 (n = 7) d. On d 2, all cows received two injections of PGF2alpha, 12 h apart. In experiment 2, 14 lactating cows (7 to 12 d postpartum) received LH (40 microg/h; n = 7) or saline (1 mL/h; n = 7) in hourly pulses for 7 d, beginning 24 h after start of the first follicular wave. Daily samples were used to determine serum concentrations of progesterone (P4), estradiol-17beta (E2), LH, and FSH. Profiles of LH were determined from blood samples collected at 12-min intervals for 8 h on d 3. During infusion of LH, serum P4 and FSH were similar across treatments in both experiments. Serum E2 concentrations were similar in experiment 1, but serum E2 was greater on d 2, 3, and 5 in LH-treated cows in experiment 2. Infusion increased LH pulse frequency and amplitude in both experiments. Formation of cysts did not differ between LH- and saline-treated cows in either experiment (1 of 7 vs. 0 of 5 and 1 of 6 vs. 0 of 7, respectively). Cows that ovulated had similar intervals to ovulation in experiment 1 [6.0 +/- 0.1 d (LH) vs. 6.4 +/- 0.2 d (saline)], but in experiment 2, ovulation was 14 d earlier in LH-treated cows (5.6 +/- 1.8 d vs 19.9 +/- 1.5 d). In conclusion, high concentrations of LH are not solely responsible for formation of cysts in lactating dairy cows. Pulsatile infusion of LH stimulated follicular growth and steroidogenesis and decreased time to first ovulation in anestrous postpartum cows.     

The absence of corpus luteum formation alters the endocrine profile and affects follicular development during the first follicular wave in cattle

We previously established a bovine experimental model showing that the corpus luteum (CL) does not appear following aspiration of the preovulatory follicle before the onset of LH surge. Using this model, the present study aimed to determine the profile of follicular development and the endocrinological environment in the absence of CL with variable nadir circulating progesterone (P(4)) concentrations during the oestrous cycle in cattle. Luteolysis was induced in heifers and cows and they were assigned either to have the dominant follicle aspirated (CL-absent) or ovulation induced (CL-present). Ultrasound scanning to observe the diameter of each follicle and blood collection was performed from the day of follicular aspiration or ovulation and continued for 6 days. The CL-absent cattle maintained nadir circulating P(4) throughout the experimental period and showed a similar diameter between the largest and second largest follicle, resulting in co-dominant follicles. Oestradiol (E(2)) concentrations were greater in the CL-absent cows than in the CL-present cows at day -1, day 1 and day 2 from follicular deviation. The CL-absent cows had a higher basal concentration, area under the curve (AUC), pulse amplitude and pulse frequency of LH than the CL-present cows. After follicular deviation, the CL-absent cows showed a greater basal concentration, AUC and pulse amplitude of growth hormone (GH) than the CL-present cows. These results suggest that the absence of CL accompanying nadir circulating P(4) induces an enhancement of LH pulses, which involves the growth of the co-dominant follicles. Our results also suggest that circulating levels of P(4) and E(2) affect pulsatile GH secretion in cattle.     

Dietary unsaturated fatty acids influence preovulatory follicle characteristics in dairy cows

Dietary unsaturated fatty acids (UFAs) have been implicated in several reproductive processes in dairy cows through a variety of mechanisms. This study examined the effects of periparturient supplementation of rumen bypass fats low or high in proportion of UFAs (oleic and linoleic) on preovulatory follicle characteristics. Forty-two 256-day pregnant dairy cows were divided into three groups and were fed a control diet (n=14) or supplemented with fats either low (LUFA; n=14) or high (HUFA; n=14) in UFAs. At 14-15 days following behavior estrus, the cows received a prostaglandin F(2)(alpha) injection and 48 h later >7 mm follicles were aspirated. Progesterone (P(4)), androstenedione (A(4)), and estradiol (E(2)) were determined in the follicular fluid. Out of 75 follicles, 37 follicles that were aspirated between 55 and 70 days post partum were regarded as E(2)-active follicles (E(2)/P(4) ratio >1) and subjected for further analysis. The diameter of preovulatory follicles was greater in cows fed HUFA than in those fed control or LUFA. The concentrations and content of A(4) and E(2) in follicles and E(2)/P(4) ratio were higher in the HUFA group than in the other two groups. The P450 aromatase mRNA expression in granulosa cells that were collected from the aspirated preovulatory follicles was also higher in the HUFA cows than in the other groups. A significant correlation was observed between E(2) concentrations in preovulatory follicles and E(2) concentrations in plasma at aspiration. In conclusion, dietary UFA increased the size of and elevated steroid hormones in preovulatory follicles, which may be beneficial to consequent ovarian function.     

Enhanced precision of estrus and luteinizing hormone after progesterone and prostaglandin in heifers

Objectives were to determine if low progesterone and increased basal luteinizing hormone for 3 d after injecting prostaglandin F2 alpha would affect precision of preovulatory surges of luteinizing hormone and onset of estrus in Holstein heifers. In experiments 1 and 2, heifers received progesterone devices for 10 d and prostaglandin F2 alpha on the 7th d after inserting progesterone device. For controls, progesterone devices were removed when prostaglandin F2 alpha was injected, and control devices were installed for 3 d. In experiment 1, jugular blood was sampled every 2 or 4 h from injection of prostaglandin F2 alpha until 108 h after removing progesterone to determine basal and peak secretion of luteinizing hormone. In treatment heifers, basal luteinizing hormone tended to increase, and intervals to peak secretion of luteinizing hormone were shorter and more precise than for controls. In experiment 2, heifers were observed for estrus for 30 min every 4 h for 96 h after removing progesterone devices. Intervals to onset of estrus for experimental heifers were shorter and more precise than for controls. After exogenous prostaglandin F2 alpha, 1 to 2 ng of progesterone per milliliter of serum for 2 to 3 d allows increased basal luteinizing hormone and is associated with increased precision of preovulatory secretion of luteinizing hormone and onset of estrus. This may be an acceptable method to control ovulation precisely for fixed time insemination of cattle.     

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 a synchronized estrous cycle (d 0 = estrus), cows were treated with a Syncromate-B implant and two injections of PGF2, (25 mg, 0700 h; 15 mg, 1900 h, 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 32 mg 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 22 h after the onset of estrus. Cows in experiment 3 received a used controlled intravaginal drug releasing (CIDR) device and two injections of PGF2alpha (25 mg, 0700 h; 15 mg, 1900 h; 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 (20 mg 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 10.7 mm on d 5 to 15.4 mm on d 10 (experiments 1 and 2), and from 9 mm on d 5 to 20.4 mm on d 11 (experiment 3). From d 11 to 14, the number of class 1 (2 to 5 mm) follicles was lower in the aspirated group than in the control group; the number of class 2 (6 to 9 mm) follicles was higher on d 12 and 13 for the aspirated group (experiments 1 and 2). The number of class 3 (> or =10 mm) 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.     

A GnRH/LH surge without subsequent progesterone exposure can induce development of follicular cysts

Our hypothesis was that follicular cysts would develop if cows experienced an estradiol-induced GnRH LH surge in the absence of an ovulatory follicle. Further, we hypothesized that estradiol would fail to induce a subsequent GnRH/LH surge in these cows until they were treated with progesterone. In experiment 1, seven cows were synchronized with a controlled internal drug releasing device (CIDR) for 9 d and each received 500 microg of cloprostenol on d 7. All follicles (> or = 5 mm in diameter) were aspirated at the time of CIDR removal using transvaginal follicular aspiration. Two days after aspiration, cows were treated with 5 mg of estradiol benzoate (EB) to induce a GnRH/LH surge in the absence of an ovulatory-sized follicle. All cows had an LH surge following the estradiol treatment and three of seven developed an anovulatory condition that resembled follicular cysts. The four cows that did not develop follicular cysts luteinized remaining cells from one aspirated follicle each. Thus, all cows with a progesterone elevation after the estradiol/GnRH/LH surge had subsequent ovulatory cycles, whereas the absence of progesterone was followed by follicular cysts. After 49 d, the anovulatory cows were induced back to normal cyclicity by insertion of a CIDR for 7 d. In two subsequent experiments, nine of 26 cows were induced to have follicular cysts by follicular aspiration followed by 5 mg of EB. After 26 d of observation, all cystic cows received a second treatment with 5 mg of EB and none of the cows showed an LH surge or ovulation. Cystic cows were untreated (n = 4 controls) or treated for 7 d with a CIDR (n = 5). All cystic cows were subsequently treated for a third time with 5 mg of EB. All CIDR-treated cows had an LH surge and ovulated, whereas none of the control cows had an LH surge or ovulation after the estradiol treatment. Thus, a large follicle anovulatory condition, similar to follicular cysts, can be induced by estradiol induction of a GnRH/LH surge in the absence of subsequent luteinization, and this condition prevents a GnRH/LH surge in response to high doses of estradiol. Progesterone eliminates this condition by reinitiation of GnRH/LH surges in response to estradiol.     

Effect of adrenocorticotropin and cortisol on luteinizing hormone surge and estrous behavior of cows

In six cows, twice daily administration of 100 IU corticotropin for 3.5 days during the follicular phase delayed the preovulatory luteinizing hormone surge and onset of behavioral estrus. Corticotropin increased progesterone and decreased estradiol and basal luteinizing hormone concentrations of blood. Following corticotropin withdrawal, a shortened period of behavioral estrus (50% of control) was accompanied by an apparent luteinizing hormone surge and ovulation. To ascertain if these effects were caused by the elevated corticosteroid concentrations induced by corticotropin, we infused intravenously four heifers with cortisol sodium succinate. During this infusion, luteinizing hormone surge and estrous behavior were inhibited; however, estradiol and basal luteinizing hormone concentrations were not affected. Furthermore, there was no luteinizing hormone surge, ovulation, or behavioral estrus after cessation of cortisol treatment. These results are consistent with a role for corticosteroids in mediating inhibitory effects on reproduction produced by corticotropin administration.