Timing of follicular phase events and the postovulatory progesterone rise following synchronisation of oestrus in cows

In cows the timing of both ovulation and the subsequent postovulatory progesterone rise are critical to successful fertilisation and early embryo development. The aim of this study was to determine the degree of variability in the timing of ovulation relative to other follicular phase events and to determine how variations in the timing of follicular phase events contribute to the timing of the postovulatory progesterone rise. Plasma concentrations of progesterone, oestradiol and luteinising hormone (LH) and the timing of oestrus and ovulation were determined following induction of luteolysis were determined in 18 mature, non-lactating Holstein-Friesian cows. Four cows were excluded on the basis of abnormal reproductive function. In the remaining 14 cows oestrus occurred at 57.4+/-4.3h and the LH surge at 54.6+/-4.0h following luteolysis (progesterone <1ngmL(-1)) followed by a fall in circulating oestradiol concentration at 64.6+/-4.4h. Cows ovulated at 88.0+/-4.7h with the postovulatory progesterone rise (to >1ngmL(-1)) occurring 159+/-7.2h after luteolysis. There was considerable variation in the timing of ovulation following luteolysis (range 64-136h) onset of oestrus (range 24-40h) and onset of the LH surge (range 24-44h). Cows were then split on the basis of interval from progesterone fall to progesterone rise giving groups (n=7 per group) with intervals of 180.6+/-6.7 and 138.3+/-5.7h (P<0.001). Between groups, both the intervals from luteolysis to ovulation (98.3+/-6.9 vs 77.7+/-3.4h; P<0.05) and ovulation to progesterone rise (82.3+/-4.2 vs. 60.6+/-5.5h; P<0.01) were longer in late rise cows. There was no difference between groups in the interval from oestrus or LH surge to ovulation. In conclusion the results of this study further highlight the high variability that exists in the timing and interrelationships of follicular phase events in the modern dairy cow, reemphasising the challenges that exist in optimising mating strategies. However, the data do suggest that in cows with poor post ovulatory progesterone secretion, the key problem appears to be poor post ovulatory development rather than a delay in ovulation.     

Timing of preovulatory LH surge and ovulation in superovulated sheep are affected by follicular status at start of the FSH treatment.

The aims of this study were to evaluate the chronology of periovulatory events (oestrus behaviour, LH surge and ovulation) in 16 superovulated Manchega sheep and to determine whether follicular status at start of the FSH supply might affect their occurrence. Mean timing for onset of oestrus behaviour was detected at 28.1 +/- 0.7 h after sponge withdrawal; the preovulatory LH surge and ovulation started at 37.2 +/- 0.7 h and 65.4 +/- 0.7 h after progestagen withdrawal, respectively. The intervals between oestrus, LH surge and ovulation were affected by a high individual variability, which might be the cause for reported decreased efficiency in embryo production. Current results also addressed the role of follicular status at start of the superovulatory treatment on the preovulatory LH surge and the ovulation. The interval LH surge-ovulation was increased in ewes with a growing dominant follicle at starting the FSH treatment (32.3 +/- 0.9 vs 28.6 +/- 0.5 h, p < 0.05). The developmental stage of the largest follicle at starting the superovulatory treatment also affected occurrence of LH surge and ovulation; follicles in growing phase advanced the occurrence of the LH surge and ovulation when compared to decreasing follicles (33.0 +/- 1.0 vs 43.5 +/- 1.1 h, p < 0.05, for LH peak and 60.7 +/- 1.1 vs 72.8 +/- 1.2 h, p < 0.05, for ovulation). Thus, only ewes with growing follicles ovulated prior to 55 h after sponge withdrawal; conversely, no sheep with decreasing follicles ovulated earlier than 67 h, when an 85.7% of the ewes bearing growing follicles has ovulated at 63 h.     

Comparison of oestrus synchronisation programmes in dairy cattle using oestradiol benzoate, short-acting progesterone and cloprostenol, or buserelin and cloprostenol

AIM: To evaluate the efficacy of a programme using oestradiol benzoate, progesterone and the prostaglandin-F2 (PG) analogue, cloprostenol, to synchronise oestrus and ovulation in dairy cows, compared with a programme using a gonadotropinreleasing hormone (GnRH) agonist, buserelin, and cloprostenol. METHODS: Twenty non-lactating dairy cows, at random stages of the oestrus cycle, were randomly assigned to 1 of 2 treatments. In Treatment 1 ( OPPG; n=10), cows were injected with 2 mg oestradiol benzoate intramuscularly (IM) plus 200 mg progesterone subcutaneously (SC) on Day 0, followed by 500 microg cloprostenol IM on Day 9 and 1 mg oestradiol benzoate on Day 10. In Treatment 2 (GPG; n=10), cows were injected with 10 microg buserelin IM on Day 0, 500 microg cloprostenol IM on Day 7 and 10 microg buserelin on Day 9. The ovaries of all cows were examined by ultrasonography, using an 8 MHz probe, from 5 days before the initial treatment until ovulation. Cows were observed for oestrus 3 times daily for 7 days after cloprostenol treatment. Blood samples were collected daily for determination of progesterone, and 6-hourly for 36 h after the second oestradiol or buserelin injection for the determination of follicle stimulating hormone (FSH) and luteinising hormone (LH) concentrations. RESULTS: The percentage of cows observed in oestrus was higher in the OPPG group than in the GPG group (100% vs 55.6%, p=0.018). Treatment with either short-acting progesterone plus oestradiol benzoate or buserelin was followed by atresia or ovulation of the dominant follicle. Emergence of a new follicular wave occurred earlier (p>0.001) in the GPG group (2.2+/-0.2 days) than in the OPPG group (3.6+/-0.2 days). There was no significant difference between treatment groups in the variation of time of follicular wave emergence or size of the largest follicles at either the time of initial treatment (10.8+/-1.4 mm vs 11.1+/-0.8 mm), cloprostenol treatment (13.8+/-0.7 mm vs 14.0+/-1.3 mm) or of ovulation (15.4+/-0.7 mm vs 17.6+/-1.1 mm; p=0.10). The LH surge occurred sooner after the second injection of buserelin (4.0+/-1.0 h) than after the second injection of oestradiol benzoate (22.8+/-1.2 h; p>0.001). The interval between the second injection of oestradiol benzoate or buserelin and ovulation did not differ significantly between treatment groups (1.7+/-0.3 days vs 1.6+/-0.2 days; p=0.69). CONCLUSIONS: The use of short-term progesterone treatment, combined with oestradiol benzoate for follicular wave synchronisation, and cloprostenol to cause lysis of residual luteal tissue, is a promising alternative to established methods of oestrus synchronisation in cows.     

Oestrous Behaviour and Timing of Ovulation in Relation to Onset of Oestrus and LH Peak in Mithun (Bos frontalis) Cows

The objectives of the study were to evaluate the oestrus behaviour and to determine the timing of ovulation in relation to onset of oestrus and the pre-ovulatory LH surge in mithun (Bos frontalis). For this purpose, the blood samples collected at 15-min intervals for 9 h period following onset of oestrus and thereafter, at an interval of 2 h till 4 h post-ovulation for three consecutive cycles from 12 mithun cows were assayed for plasma LH and progesterone. Ovulation was confirmed by palpation of ovaries per rectum at hourly intervals. Various signs of behavioural oestrus were also recorded. The common signs of oestrus and their frequency of occurrence in mithuns were following and mounting by male mithuns (100%), standing to be mounted (100%), frequent urination (62.33%), raising of tail (65.23%), swelling of vulva (54.26%) and congestion of vulvar mucous membrane (69.87%). The pre-ovulatory LH surges consisted of several pulses (2.92 +/- 0.26 pulses/animal; range, 1-4). The mean (+/-SEM) peak level of LH for individual mithun varied from 6.99 +/- 0.44 to 12.69 +/- 2.10 ng/ml and the mean pooled LH peak concentration was 9.10 +/- 0.60 ng/ml. The highest peak (highest amplitude of LH during LH surge) was 10.83 +/- 0.76 ng/ml (range, 8.07-16.49 ng/ml). The duration of LH surge was 6.98 +/- 0.22 h (6-8 h). Onset of LH surge was at 1.23 +/- 0.17 h post-oestrus onset (range, 0.25-2.25 h). Mean plasma progesterone stayed low (<0.24 ng/ml) during the entire duration of sampling. Ovulation occurred at 26.92 +/- 0.31 (range, 26-29 h) after the onset of oestrus and 18.63 +/- 0.35 h (range, 17-20.75 h) after the end of LH surge. The occurrence of the highest LH peaks within a narrow time frame of 2- to 5-h post-oestrus onset in mithuns could have contributed to the animals ovulating within a narrow time interval. These results are very promising from a practical standpoint of potential success when AI program in this species is implemented in a big way. Furthermore, the results of the occurrence of LH pulses during pre-ovulatory LH surges, which are required for ovulation in this species of animals, is unique and species specific.     

Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.     

The effect of timing of administration of oestradiol benzoate on characteristics of oestrus,timing of ovulation and fertility in Bos indicus heifers synchronised with a progesterone releasing intravaginal insert

OBJECTIVE: To compare the timing of onset of oestrus and ovulation, characteristics of oestrus, and fertility in Bos indicus heifers synchronised with a progesterone releasing intravaginal insert (IVP4) and administration of oestradiol benzoate (ODB) either at the time of removal of the insert or 24 h later. Design: Cohort study. PROCEDURE: Bos indicus and Bos indicus cross heifers were treated on two farms (Farm A, n = 273; Farm B, n = 47) with an IVP4 for 8 days with 1.0 mg of ODB administered at the time of device insertion and 250 mg of cloprostenol at the time of device removal. Heifers in the ODB-0 group were administered 0.75 mg of ODB at the time of device removal while heifers in the ODB-24 group were administered the same dose of ODB 24 h after device removal. Heifers were inseminated once daily after detection of oestrus. Heifers not detected in oestrus by 72 h after removal of inserts were inseminated at that time. Oestrus was detected in heifers on Farm A using heatmount detectors while on Farm B oestrus in heifers was monitored using radiotelemetry of mounting pressure. Ovarian follicular development was monitored daily in 30 heifers on Farm B from the time of administration of inserts until ovulation to a maximum of 96 h after removal of inserts, and again 11 days after removal of inserts (Day 19). A blood sample was collected from all heifers on Farm B on Day 19 and analysed for plasma concentration of progesterone. Pregnancy was diagnosed 6 to 8 weeks after insemination. RESULTS: Administration of ODB at the time of removal of inserts shortened the time interval to oestrus and ovulation (P < 0.001), increased the number of mounts recorded during oestrus (P = 0.04) and reduced the odds of pregnancy (P = 0.03). The proportion of heifers ovulating on Farm B was 67% and was not affected by treatment group (P = 0.61). The mean diameter of the largest follicle measured in ovaries was greater at the time of removal of inserts (9.1 +/- 0.6 vs 10.7 +/- 0.4; P = 0.03) and at the expected time of the LH surge (8.1 +/- 0.4 vs 11.5 +/- 0.3 mm; P < 0.001) in heifers that ovulated compared to heifers that failed to ovulate, respectively. Emergence of a new follicular wave was not detected during the synchronisation treatment in heifers that failed to ovulate. Concentrations of progesterone in plasma on Day 19 were less in non-pregnant heifers (P = 0.05) compared to heifers subsequently diagnosed as pregnant to insemination and were affected by the diameter of the ovulatory follicle (P = 0.01). CONCLUSION: Administration of ODB at the time of removal of inserts can shorten the time interval to oestrus and ovulation and can reduce fertility when insemination is carried out once daily. Further work is needed to determine if prolonged suppression of follicular development, anovulatory oestrus and premature ovulation occuring in some heifers is associated with administration of ODB.     

Formation of follicular cysts in cattle and therapeutic effects of controlled internal drug release

Follicular cysts in cattle result from excessive growth of the dominant follicle without ovulation and still constitute a major reproductive disorder in this species. One key hormonal characteristic of cows with follicular cysts is the lack of an LH surge, although they have increased plasma estradiol concentrations. Another is a relatively high level of pulsatile secretion of LH that promotes continued growth of the dominant follicle. These LH characteristics seem to result from a functional abnormality in the feedback regulation of LH secretion by estradiol. Treatment with controlled internal drug release devices that increase circulating progesterone levels is effective in resolving follicular cystic conditions by 1) lowering pulsatile LH secretion and 2) restoring the ability of the hypothalamo-pituitary axis to generate an LH surge in response to an increase in circulating estradiol.     

Effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given GnRH

The effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given 100 microg of GnRH im were determined in three experiments. In Experiment 1, heifers were given GnRH 3, 6 or 9 days after ovulation; 8/9, 5/9 and 2/9 ovulated (P<0.02). Mean plasma concentrations of progesterone were lowest (P<0.01) and of LH were highest (P<0.03) in heifers treated 3 days after ovulation. In Experiment 2, heifers received no treatment (Control) or one or two previously used CIDR inserts (Low-P4 and High-P4 groups, respectively) on Day 4 (estrus=Day 0). On Day 5, the Low-P4 group received prostaglandin F(2alpha) (PGF) twice, 12 h apart and on Day 6, all heifers received GnRH. Compared to heifers in the Control and Low-P4 groups, heifers in the High-P4 group had higher (P<0.01) plasma progesterone concentrations on Day 6 (3.0+/-0.3, 3.0+/-0.3 and 5.7+/-0.4 ng/ml, respectively; mean+/-S.E.M.) and a lower (P<0.01) incidence of GnRH-induced ovulation (10/10, 9/10 and 3/10). In Experiment 3, 4-6 days after ovulation, 20 beef heifers and 20 suckled beef cows were given a once-used CIDR, the two largest follicles were ablated, and the cattle were allocated to receive either PGF (repeated 12h later) or no additional treatment (Low-P4 and High-P4, respectively). All cattle received GnRH 6-8 days after follicular ablation. There was no difference between heifers and cows for ovulatory response (77.7 and 78.9%, P<0.9) or the GnRH-induced LH surge (P<0.3). However, the Low-P4 group had a higher (P<0.01) ovulatory response (94.7% versus 61.1%) and a greater LH surge of longer duration (P<0.001). In conclusion, although high plasma progesterone concentrations reduced both GnRH-induced increases in plasma LH concentrations and ovulatory responses in beef cattle, the hypothesis that heifers were more sensitive than cows to the suppressive effects of progesterone was not supported.     

Effects of estradiol on gonadotrophin release, estrus and ovulation in CIDR-treated beef cattle

The effects of estradiol-17beta (E-17beta) or estradiol benzoate (EB) on gonadotrophin release, estrus and ovulation in beef cattle were evaluated in two experiments. In experiment 1, 16 ovariectomized cows received a previously used CIDR insert from days 0 to 7 and 1mg of EB on day 8; they also received 5mg of E-17beta on days 0 or 1, or 5mg of E-17beta+100mg of progesterone on day 0. There was only an effect of time (P<0.0001) on plasma concentrations of progesterone, estradiol, FSH, and LH. Following treatment with E-17beta, plasma FSH concentrations were suppressed for approximately 36 h, whereas plasma LH concentrations were reduced (P<0.05) for 6 h, but surged within 24 h. Injecting 1mg of EB 24 h after CIDR removal decreased (P<0.02) plasma LH concentrations for 6h, followed by an LH surge at 18 h. In experiment 2, ovary-intact heifers (n=40) received a used CIDR and 5mg of E-17beta+100mg of progesterone on day 0. On day 7, CIDR were removed, PGF given, and heifers received nothing (control) or 1mg of EB 12, 24, or 36 h later. In these groups, plasma LH peaked (mean+/-SEM) 78.0+/-23.0, 37.8+/-8.5, 44.4+/-10.3, and 51.0+/-5.1 h after CIDR removal (means, P<0.001; variances, P<0.001) and intervals from CIDR removal to ovulation were 102.0+/-6.7, 63.6+/-3.6, 81.6+/-3.5, and 78.0+/-4.1h (P<0.05). The interval from CIDR removal to ovulation was shorter and less variable in EB-treated groups; the interval from EB to ovulation was shortest (P<0.05) in the 12-h group. In summary, E-17beta or EB decreased both FSH and LH, but LH increased after 6h (despite elevated progesterone concentrations). Following CIDR removal, 1mg of EB effectively synchronized LH release, and ovulation (in intact cattle), but the interval from CIDR removal to EB treatment affected the time of ovulation.     

Anterior pituitary concentrations of gonadotropins, GnRH-receptors and ovarian characteristics following early weaning in beef cows

Endocrine changes in the hypophyseal-ovarian axis associated with early calf removal were investigated in anestrous beef cows. Tissues were collected and analyzed from multiparous beef cows slaughtered at O (n = 8), 36 (n = 8) or 72 h (n = 8) after calf removal during the fifth week after calving. Cows that exhibited estrus; had postmortem signs of a recent ovulation or had serum concentrations of luteinizing hormone (LH) indicative of an ovulatory surge, were excluded from the analysis. Five control cows that were not slaughtered exhibited estrus from 30 to 84 h after calf removal. Seven additional cows were continuously kept with their calves and did not exhibit estrus until 72 +/- 9 d after calving. Serum concentrations of estradiol-17 beta (estradiol) averaged 8.2 +/- 1.7, 7.5 +/- 2.0 and 9.1 +/- 1.5 pg/ml at 0, 36 and 72 h, respectively, but they averaged 22.8 +/- 4.7 pg/ml prior to estrus in control cows. Therefore, observations were assumed to represent events that occur prior to the rise in serum concentrations of estradiol that occurs during proestrus. Volume of fluid from the largest ovarian follicle tended to be greater (P less than .10) at 72 h (1.5 +/- .2 ml) than at 0 h (1.1 +/- .1 ml) or 36 h (1.0 +/- .1 ml). Follicular-fluid concentrations of estradiol, but not progesterone, were positively correlated (P less than .01) with follicular volume. However, numbers of small (less than 100 microliters), medium (100 to 400 microliters) and large follicles (greater than 400 microliters) as based on fluid volume, as well as follicular-fluid concentrations of estradiol and progesterone, did not differ among treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of ovine follicles destined to form subfunctional corpora lutea

A study was done to test whether ovulatory follicles destined to form subfunctional corpora lutea differed from normal ovulatory follicles in steroidogenic function. Twenty-five ewes were treated with prostaglandin F2 alpha on d 11 of the estrous cycle, then unilaterally ovariectomized before (n = 13) or after (n = 12) the surge of luteinizing hormone (LH) at the induced estrus to collect "control" follicles, which would have produced normal corpora lutea. In 15 ewes, the second ovary was removed 63 to 84 h later to collect "treated" follicles before (n = 7) or after (n = 8) the second expected surge of LH. Five ewes (control) were allowed to ovulate from the remaining ovary at first estrus and another five (treated) at the second estrus (3 to 4 d later). Treated ewes had lower serum progesterone than control ewes during the ensuing cycle (P less than .05). Treated follicles contained less estradiol in the theca (4.4 +/- .6 vs 10.0 +/- 2.5 ng; P less than .05), less androstenedione (.1 +/- .1 vs 1.0 +/- .2 ng) and estradiol (.5 +/- .1 vs 2.9 +/- 2.2 ng) in the granulosa (P less than .05) and less progesterone in the follicular fluid (.8 +/- .4 vs 3.3 +/- .8 ng; P less than .05) than control follicles, when removed before the surge of LH. Follicles removed after the surge of LH did not differ. In conclusion, ovulatory follicles with low steroidogenic function became corpora lutea that secreted lower-than-normal quantities of progesterone.     

Endocrine changes and ultrasonography of ovaries in suckled beef cows during resumption of postpartum estrous cycles

Changes in follicular and luteal structures were assessed and concentrations of estradiol and progesterone were measured in 13 Hereford X Angus suckled beef cows during resumption of estrous cycles. Transrectal ultrasonography was used to monitor follicular size, ovulation, and formation and regression of the corpus luteum (CL). The interval from parturition to first postpartum ovulation (FO) was 82 +/- 4.7 d. Serum progesterone remained low before FO. One cow exhibited standing estrus, two cows showed other signs of estrus, and 10 displayed no signs of behavioral estrus preceding FO. All cows exhibited standing estrus before the second postpartum ovulation (SO). All cows had a short luteal phase after FO, with an average interval of 8.5 +/- .2 d between FO and SO. Concentrations of estradiol in serum during the 8 d preceding ovulation were similar before FO and SO. Maximal diameter of the preovulatory follicle was similar before FO and SO. However, the ovulatory follicle was larger in diameter at 2 d (P = .02) and 3 to 8 d (P less than .005) before FO than before SO. The time from detection until ovulation was less (P = .005) for the ovulatory follicle preceding SO than for the follicle associated with FO (8.5 vs 10.2 d, respectively, SE = .4). The second-largest follicle was larger (P less than .005) in diameter during the 8 d preceding the FO than before the SO. The difference in size between the ovulatory follicle and the second-largest follicle on the day before ovulation was greater (P less than .005) preceding SO than preceding FO (8.7 vs 6.6 mm, respectively, SE = .4).(ABSTRACT TRUNCATED AT 250 WORDS)     

Assessment of the Intensity of Behavioural Traits and Ovulation between Synchronized and Non-synchronized Cows

Thirty cyclic, non-suckled Brahman cows were divided into three groups, all of which were synchronized sequentially with CIDR-B and observed continuously for 100 h to determine different behavioural oestrus signs. Twenty-four hours after implant withdrawal, all synchronized cows in the group, together with all other cows displaying oestrus, were subjected to intensive ultrasonographic observations (every 6 h for 120 h) to pinpoint the moment of ovulation. In the first group, oestrus and ovulation response was 60% (6/10), in the second 44% (4/9) showed oestrus and six ovulated, and in the third group oestrus and ovulation were 80% (8/10). Significant differences were observed between the second and third groups (p < 0.05). No differences were observed in the duration of oestrus, time when oestrus was displayed after implant withdrawal, time of ovulation and onset of oestrus, end of oestrus to ovulation, and intensity of oestrus on a point scale. The relationship between duration of oestrus and time of ovulation was r(2) = 0.16. Ovulation, on average, was 32.1 +/- 14.5 h after the onset of oestrus, 22.3 +/- 16.5 h after the end of oestrus, and 91.8 +/- 16.7 after implant withdrawal, although no significant differences were observed. One non-synchronized animal showed oestrous activity in the second group but failed to ovulate. In the third group, 8 animals showed oestrus, 4 with high concentrations of progesterone. Of the other four one ovulated. In conclusion, oestrous behaviour is not necessarily the best marker to predict the time when ovulation takes place due to variation in the length of the oestrous period and the possible integration of non-ovulatory animals into sexually active groups.     

Peri-oestrus hormone profiles and follicle growth in lactating sows with oestrus induced by intermittent suckling.

This study describes follicle dynamics, endocrine profiles in multiparous sows with lactational oestrus compared with conventionally weaned sows (C). Lactational oestrus was induced by Intermittent Suckling (IS) with separation of sows and piglets for either 12 consecutive hours per day (IS12, n = 14) or twice per day for 6 h per occasion (IS6, n = 13) from day 14 of lactation onwards. Control sows (n = 23) were weaned at day 21 of lactation. Pre-ovulatory follicles (> or =6 mm) were observed in 100% of IS12, 92% of IS6 and 26% of C sows before day 21 of lactation and in the remaining 74% C sows within 7 days after weaning. All sows with pre-ovulatory follicles showed oestrus, but not all sows showed ovulation. Four IS6 sows and one IS12 sow developed cystic follicles of which two IS6 sows partially ovulated. Follicle growth, ovulation rate and time of ovulation were similar. E(2) levels tended to be higher in IS sows (p = 0.06), the pre-ovulatory LH surge tended to be lower in IS12 (5.1 +/- 1.7 ng/ml) than in C sows (8.4 +/- 5.0 ng/ml; p = 0.08) and P(4) levels were lower in IS12 and IS6 than in C sows (at 75 h after ovulation: 8.8 +/- 2.4 ng/ml vs 7.0 +/- 1.4 ng/ml vs 17.1 +/- 4.4 ng/ml; p < 0.01). In conclusion, sows with lactational oestrus induced by IS are similar to weaned sows in the timing of oestrus, early follicle development and ovulation rates, but the pre-ovulatory LH surge and post-ovulatory P(4) increase are lower.     

In vivo evidence that local cortisol production increases in the preovulatory follicle of the cow

The aim of the present in vivo study was to monitor real-time fluctuations of cortisol (Cr) in the wall of preovulatory follicles using a microdialysis system (MDS) implanted in the theca layer as well as changes in ovarian venous plasma (OVP) and jugular venous plasma (JVP). Seven cows were superovulated using FSH and prostaglandin F2alpha injections. Dialysis capillary membranes were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system. Fractions of the perfusates were collected from Day -1 (Day 0=LH surge) to Day 3. No difference in the concentrations of Cr between JVP and OVP was detected throughout the experiment. Circulating concentrations of Cr ranged from 20 to 35 ng/ml 8 h after surgery in ovulatory and anovulatory cows. In five ovulatory cows, the Cr concentration decreased to basal levels (<10 ng/ml) between 12 and 24 h after surgery, however, two anovulatory cows retained high Cr levels (>10 ng/ml) up to 42 h after surgery. There was a clear increase in the local concentration of Cr from 13.3+/-2.1 pg/ml at -24 h to 27.5+/-1.7 pg/ml at 0 h (peak of the LH surge) within the wall of ovulatory follicles. This increase was not detected in anovulatory follicles. This transient increase in Cr occurred only in the follicle wall, but not in the OVP or JVP, indicating that the presence of a local regulatory mechanism for Cr production/conversion in ovulatory follicles, and this mechanism may modulate the inflammatory-like reaction induced by LH surge in the follicle wall. The present results demonstrate that the glucocorticoid environment in the follicular wall adjusts at the local level in bovine ovulatory follicles. This mechanism may protect follicles from the adverse effects of glucocorticoid, and it may prevent excess inflammatory reactions associated with ovulation by temporarily increasing local concentrations of glucocorticoid, thus forming an integral part of the regulatory mechanism in ovarian physiology.     

GnRH treatment at CIDR insertion influences ovarian follicular dynamics in Japanese black cows

Ovarian follicular dynamics and estrous synchronization after Gonadotropin-releasing hormone (GnRH) treatment at Controlled Internal Drug Releasing device (CIDR) insertion were investigated in Japanese Black cows. CIDR was inserted for eight cows at 7 days after estrus. Cows were allocated to either Group A: 8-day CIDR insertion with GnRH treatment on d 0 (n=4, d 0=CIDR insertion) or Group B: 8-day CIDR insertion (n=4). Both groups were injected with prostaglandin F2alpha (PGF2alpha) on d 7. Ultrasonography and blood sampling were performed twice daily. Intensive sampling was performed every 15 min for 8 hr to determine the pulsatile release of LH on d -1, d 5 and d 10. Three of four cows showed intermediate ovulation within 2 days after GnRH treatment during CIDR insertion in Group A, whereas no ovulation was found in Group B. Three of four cows in Group A and all four cows in Group B ovulated after CIDR removal. Plasma progesterone concentrations from d 3 to d 7 in three intermediate ovulatory cows in Group A (8.4 +/- 1.6 ng/ml) was significantly higher than those in Group B (4.1 +/- 1.2 ng/ml; 4 cows) during CIDR insertion (P<0.01). Interval to estrus and ovulation after CIDR removal was observed at 60.0 +/- 12.0 hr and 76.0 +/- 6.9 hr in three cows in Group A, and 75.0 +/- 15.1 hr and 93.0 +/- 20.5 hr in Group B, respectively. There was a significant increase in LH pulse frequency on d 10 compared on d -1 or d 5 in both groups (P<0.05), in addition those on d 10 in Group A tended to be higher than in Group B. As a result, GnRH treatment at CIDR insertion at 7 days after estrus induced intermediate ovulation with formation of corpus luteum (CL) and rather synchronized emergence of ovulatory follicle during CIDR insertion. These induced CL increased plasma progesterone concentrations and contributed to precise synchronization.     

Clinical relevance of pre‐ovulatory follicular temperature in heat‐stressed lactating dairy cows

Temperature gradients in female reproductive tissues seem to influence the success of key processes such as ovulation and fertilization. The objective of this study was to investigate whether pre‐ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures in lactating dairy cows under heat stress conditions. Temperatures within the pre‐ovulatory follicle, on the uterine adjacent surface and 20 cm deep within rectum, were measured using fine thermistor probes within 45 min after sunrise (dawn). Cows were selected from synchronized groups for fixed‐time insemination during the warm period of the year. Five cows under direct sun radiation and 11 cows in the shade were included in the study. None of the cows in the sun area ovulated within 24 hr, whereas 10 of the 11 cows in the sun area ovulated. Four of the 10 ovulating cows became pregnant. In the ovulating cows, follicular temperatures were 0.74 and 1.54°C significantly cooler than uterine surface and rectal temperatures, respectively, whereas temperatures in the uterine area were 0.80°C significantly cooler than rectal temperatures. No significant differences among temperatures were found in non‐ovulating cows. Follicular size was similar for ovulating and non‐ovulating cows. Environmental temperatures in the shade area were 6.4°C significantly lower than those in the sun area. Results of this study indicate that pre‐ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures and those temperature gradients were not found in cows suffering ovulation failure.     

Follicular development,oocyte viability and recovery in relation to follicular steroids,prolactin and glycosaminoglycans throughout the estrous period in superovulated heifers with a normal LH surge,no detectable LH surge,and progestin inhibition of LH surge

Estrous cycles of heifers (n = 137) were synchronized with prostaglandin (PGF_2α) and follicular development stimulated with follicle stimulating hormone. Twenty-eight animals were administered Norgestomet implants 12 hr prior to the initial PGF2α injection to suppress the LH surge that initiates ovulation. Animals were ovariectomized every 12 hr after the initial PGF2α (7–9/time, 12–108 hr and at 192 and 240 hr post PGF2α) and divided into three treatment groups to consist of: 1) animals exhibiting a normal luteinizing hormone (LH) surge (n = 86), 2) animals in which no LH surge was detected (n = 23), and 3) suppression of the LH surge via Norgestomet implants (72–108 hr, n = 28). Follicular diameter was measured and follicular fluid was collected for analysis of prolactin, estradiol, progesterone and glycosaminoglycan concentrations. Progesterone concentrations were increased in animals exhibiting an LH surge as compared to animals in which no LH surge was detected; primarily in large follicles (> 8 mm diameter) after the LH surge. Animals not exhibiting an LH surge also had increased follicular progesterone concentrations compared to Norgestomet-implanted animals (242.3 ± 36.3 vs 86.7 ± 6.4 ng/ml, respectively, P < .01), indicating some LH stimulation. Follicular estradiol in animals exhibiting an LH surge increased up to the time of LH surge detection and then declined whereas animals with no LH surge detected had follicular estradiol concentrations that declined after the PGF_2α injection. No differences were noted between those that did not exhibit an LH surge or in which the LH surge was suppressed with Norgestomet in relation to follicular estradiol concentrations. Follicular estradiol concentrations increased with follicular size in all treatment groups (P < .01). Follicular concentrations of prolactin were increased in small follicles (P < .05; ≤ 4 mm diameter) and follicular prolactin increased from 12 to 36 hr post PGF2α injection, then declined after the LH surge. Follicular glycosaminoglycan concentrations decreased with increases in follicular size (P < .01) and were higher in animals that did not exhibit an LH surge (P < .01). No differences in follicular glycosaminoglycans were noted between Norgestomet-implanted animals and those not exhibiting an LH surge. In the animals representing days 4 and 6 of the subsequent estrous cycle (192 and 240 hr post PGF2α), numbers of small-sized follicles were increased. Follicular progesterone and estradiol concentrations were related to atretic large follicles unovulated from the prior estrus and a wave of growth in small and medium follicles. Follicular prolactin and glycosaminoglycans increased with time of the new estrous cycle and were increased in smaller follicles (P < .01). Suppression of LH with progestin implants (Norgestomet) may relate to early effects of progesterone, which may not be totally eliminated at target tissues and subsequently alters the LH surge, steroidogenesis of the follicle, and ovulation. Oocytes were predominantly found in the follicular fluid from animals in which an LH surge was detected and in the buffer wash of follicles in which no LH surge was detected. Oocyte viability was higher in animals exhibiting an LH surge (75% viable) whereas the oocytes of Norgestomet-implanted animals were 75% degenerate.     

Supra-basal progesterone concentrations during the follicular phase are associated with development of cystic follicles in dairy cows

The objective of this study was to test the hypothesis that supra-basal concentrations of progesterone during the follicular phase are associated with the development of follicular cysts. Twenty-five non-lactating dairy cows were used in the study, which was performed over five identical replicate trials. Luteolysis was induced during the mid-luteal phase. Transrectal ultrasonography was performed daily to determine the occurrence/timing of ovulation. Plasma samples were collected for progesterone, oestradiol and luteinizing hormone (LH) analysis. Three cows failed to ovulate (cystic anovulatory) but did ovulate in a subsequent replicate (cystic ovulatory). Eight cows from the appropriate replicates were used as control cows (normal group). Follicular growth patterns and plasma oestradiol concentrations were similar between the three groups. However, the plasma progesterone concentrations during the follicular phase were twofold higher in the cystic anovulatory group (P < 0.01). Furthermore, no LH surge was detected in these animals. While LH pulse amplitude was similar between groups, LH pulse frequency in the cystic anovulatory group was attenuated (P < 0.05). In conclusion, the formation of follicular cysts were preceded by elevated plasma progesterone concentrations and the suppression of the LH surge.     

Dynamics of the Equine Preovulatory Follicle and Periovulatory Hormones: What's New?

Recent studies (2005–2008) on the interrelationships among the preovulatory follicle and periovulatory circulating hormones are reviewed. Close temporal and mechanistic relationships occur between estradiol/inhibin and follicle-stimulating hormone (FSH), between estradiol and luteinizing hormone (LH), and between progesterone and LH. Estradiol from the dominant follicle forms a surge that reaches a peak 2 days before ovulation. Estradiol, as well as inhibin, has a negative effect on FSH, and estradiol has a negative effect on LH. When estradiol decreases, the negative effect diminishes and accounts for the beginning of an FSH increase and a transition from a slow to rapid increase in LH on the day of the estradiol peak. The decrease in estradiol and the reduction or cessation in the growth of the preovulatory follicle beginning 2 days before ovulation are attributable to the development of a reciprocal negative effect of LH on follicle estradiol production when LH reaches a critical concentration. The LH decrease after the peak of the LH surge on the day after ovulation is related to a negative effect of a postovulatory increase in progesterone. Measurable repeatability within mares between consecutive estrous cycles occurs during the preovulatory period in diameter of the ovulatory follicle and concentrations of LH and FSH. Hormone-laden follicular fluid passes into the peritoneal cavity at ovulation and transiently alters the circulating concentrations of LH and FSH. Double ovulations are associated with greater estradiol concentrations and reduced concentrations of FSH.