Synchronization of ovulation in yaks (Poephagus grunniens L.) using PGF(2alpha) and GnRH

The objective of this study was to test the efficacy of estrus synchronization in yaks using the Ovsynch protocol. To eight non-lactating cycling yaks were administered GnRH analogue followed by PGF(2alpha) analogue treatment 7 days later and further injected with a second injection of same GnRH analogue 2 days after the PGF(2alpha) analogue administration. Ovulation was detected by rectal palpation at 2 h intervals from the initial signs of estrus till ovulation. For LH and progesterone the blood samples were collected at 15 min intervals starting from 1 h prior to the second injection of GnRH analogue until 6 h later and further at 2 h intervals till 2 h after the ovulation. Ovulation was detected in seven out of eight yaks after Ovsynch treatment. The mean time interval from the second GnRH injection to ovulation was 24.8+/-1.95 h with a range of 20-34 h and the mean interval from the LH peak and ovulation was 19.96+/-1.91 h with a range of 14-29 h. The high degree of ovulation synchronization could be attributed to the highly synchronized LH peaks in the treated animals. It was concluded that this estrus synchronization protocol could be applied for fixed time AI in yak.     

Synchronisation of oestrus in dairy cows using prostaglandin F2alpha,gonadotrophin-releasing hormone,and oestradiol cypionate

An oestrous synchronisation protocol was developed for use in lactating dairy cows using PGF(2alpha), GnRH, and oestradiol cypionate (ECP). In experiment 1, lactating dairy cows received two injections of PGF(2alpha) (on days 0 and 11) (PP; n=10) or two injections of PGF(2alpha) (days 0 and 11) and 100 microg of GnRH on day 3 (PGP; n=10). In experiment 2, cows were treated with PGP (n=7), or PGP and 1 mg of ECP at the same time (PGPE(0); n=7) or 1 day after the second PGF(2alpha) injection (PGPE(1); n=7). In experiment 3, 101 lactating dairy cows in a commercial herd were assigned to one of three treatments; PP, PGP, or PGPE(1). Follicular growth was measured by ultrasound in experiments 1 and 2. Every cow (experiments 1, 2, and 3) was blood sampled at selected intervals for progesterone and oestradiol assays and inseminated at oestrus. In experiment 1, a higher percentage of GnRH-treated cows ovulated after the first PGF(2alpha) injection (90% versus 50%; P<0.05). The GnRH-treated cows tended to have a larger dominant follicle present at the time of the second PGF(2alpha) injection (16.5+/-0.5 mm versus 15.0+/-0.7 mm; P<0.10). The percentage of cows that ovulated after the second PGF(2alpha) injection was similar (60%). In experiment 2, cows treated with ECP had higher peak preovulatory concentrations of oestradiol in plasma (6.99+/-0.63 versus 3.63+/-0.63; P<0.01) following the second PGF(2alpha) injection and a higher percentage ovulated (86% versus 43%; P<0.05). A higher percentage of PGPE(1)-treated cows in experiment 3 were observed in standing oestrus and ovulated after the second PGF(2alpha) injection (standing oestrus, 26.4, 34.3, and 62.6%, P<0.01; ovulated, 56, 63, and 78%, P<0.05; PP, PGP, and PGPE(1), respectively). In conclusion, the PGP protocol increased the number of cows that ovulated after the first PGF(2alpha) injection and produced a more mature dominant follicle at the time of the second PGF(2alpha) injection. Adding ECP to PGP (PGPE(1)) enhanced the expression of oestrus and increased ovulation percentage. The combination of PGP and ECP is potentially a new method to routinely synchronise oestrus and ovulation in dairy cows.     

Estrous behavior and the estrus-to-ovulation interval in Nelore cattle (Bos indicus) with natural estrus or estrus induced with prostaglandin F2 alpha or norgestomet and estradiol valerate

Estrous behavior and the estrus-to-ovulation interval are essential for estimating the best time to artificially inseminate cattle. Because these parameters are not well characterized in the Nelore breed (Bos indicus), the main purpose of the this study was to determine the estrus-to-ovulation interval in Nelore heifers and cows with natural estrus or with estrus induced by treatments with PGF2 alpha or norgestomet and estradiol valerate (NEV). The cows and heifers were observed continuously (24 h a day) to determine the onset of estrus and to study estrous behavior in the cows. Ten hours after the start of estrus the ovaries were scanned every 2 h by ultrasonography to monitor the dominant follicle until ovulation. Blood samples were collected periodically to determine progesterone levels by RIA. Administration of PGF2 alpha (2 injections, 11 days apart) did not induce estrus in most Nelore females in spite of the presence of functional CL, indicated by progesterone concentrations above 6.0 ng/ml in 25 of 28 animals. Treatment with NEV induced high sexual receptivity in cows (10/11), but only 66% ovulated. Cows with natural or induced estrus exhibited behavioral estrus of 10.9 +/- 1.4 h, and ovulation occurred 26.6 +/- 0.44 h (n = 26) after the onset of estrus. In most of the cows (53.8%) estrus began at night (between 1801 and 600 h), and 34.6% it started and finished during the night. It is concluded that in Nelore females ovulation occurs approximately 26 h after the onset of estrus. Additionally, estrous behavior is shorter than in European breeds, and there is a high incidence of estrus at night, which makes it difficult to detect and, consequently, impairs Al in Nelore cattle. The observation that a high percentage of Nelore females with an active CL did not respond to usual dosages of PGF2 alpha warrants further investigation.     

Ovarian and endocrine responses to prostaglandin F2alpha (PGF2alpha) given at the expected time of the endogenous FSH peak at mid-cycle in ewes

In the ewe, a rise in circulating concentrations of FSH preceding follicular wave emergence begins in the presence of growing follicles from a previous wave. We hypothesized that prostaglandin F(2alpha) (PGF(2alpha)) given at the time of an endogenous FSH peak in cyclic ewes would result in synchronous ovulation of follicles from two consecutive waves, increasing ovulation rate. Twelve Western White Face (WWF) ewes received a single i.m. injection of PGF(2alpha) (15 mg/ewe) at the expected time of a peak in FSH secretion, from Days 9 to 12 after ovulation. The mean ovulation rate after PGF(2alpha) treatment (2.3+/-0.3) did not differ (P>0.05) from the pre-treatment ovulation rate (1.7+/-0.1). Five ewes ovulated follicles from follicular waves emerging before and after PGF(2alpha) injection (3.0+/-0.6 ovulations/ewe) and seven ewes ovulated follicles only from a wave(s) emerging before PGF(2alpha) treatment (2.0+/-0.3 ovulations/ewe; P>0.05). The mean interval from PGF(2alpha) to emergence of the next follicular wave (1.0+/-0.4 and 4.0+/-0.0 d, respectively; P<0.001) and the interval from PGF(2alpha) treatment to the next FSH peak (0 and 3.5+/-0.4d, respectively; P<0.05) differed between the two groups. Six ewes ovulated after the onset of behavioral estrus, with a mean ovulation rate of 1.7+/-0.2, and six ewes ovulated both before and after the onset of estrus (3.0+/-0.5 ovulations/ewe; P<0.05). None of the ovulations that occurred before estrus resulted in corpora lutea (CL) with a full life span. At 24h before ovulation, follicles ovulating before or after the onset of estrus differed in size (4.1+/-0.3 or 5.5+/-0.4mm, respectively; P<0.05) and had distinctive echotextural characteristics. In conclusion, the administration of PGF(2alpha) at the expected time of an FSH peak at mid-cycle in ewes may alter the endogenous rhythm of FSH secretion and was not consistently followed by ovulation of follicles from two follicular waves. In non-prolific WWF ewes, PGF(2alpha)-induced luteolysis disrupted the normal distribution of the source of ovulatory follicles and may be associated with untimely follicular rupture and luteal inadequacy.     

Efficacy of PGF(2alpha) to synchronize estrus in water buffalo cows (Bubalus bubalis) is dependent upon plasma progesterone concentration,corpus luteum size and ovarian follicular status before treatment

This study was conducted to identify factors affecting PGF(2alpha) efficacy to synchronize estrus in water buffalo cows. After detection of a corpus luteum (CL) by rectal palpation, cows were treated (im) with dinoprost (12.5, 25 or 50mg) or D(+) cloprostenol (75, 150 or 300 microg) in a total of 66 treatments. Blood samples were collected 0, 24 and 48 h after treatment and ultrasound examinations and observations for estrus were performed daily to the day of ovulation or to 6 days after treatment. No PGF(2alpha) dose-response pattern was observed and overall rates of luteal regression (progesterone <1.0 ng/ml at 48 h), estrus, no detected behavioral estrus with ovulation occurring, and ovulation were 71.2, 36.4, 19.7 and 54.5%, respectively. To analyze plasma progesterone concentrations and ovarian dynamics, cows were divided in three groups according to their response to treatment. Cows that failed to have ovulations from a follicle after treatment (Group A, n = 30) had (P < 0.05) a lower plasma progesterone concentration (2.98 ng/ml) and smaller CL area (CLA; 187.3 mm(2)) before treatment as compared with cows that had an ovulation from a follicle (4.43 ng/ml and 223.7 mm(2), respectively; Groups B and C, n = 36). In cows that failed to ovulate, plasma progesterone concentration decreased in the first 24 h, but did not decline further and was >1.0 ng/ml 48 h after treatment. Moreover, no significant change in CLA after treatment was detected, indicating that treatment induced only partial luteolysis. In cows that ovulated, plasma progesterone concentration and CLA decreased continuously from treatment to ovulation (consistent with complete luteolysis). Threshold values of 2.8 ng/ml for plasma progesterone concentration and 189 mm(2) for CLA were identified as the best predictors of ovulation before treatment (83.3 and 80.6% sensitivity and 58.6 and 65.5% specificity, respectively, with positive and negative predictive values around 71%). When the origin of the ovulatory follicle was investigated, the interval from treatment to ovulation was shorter (91.9 versus 113.3 h; P < 0.05), and the ovulatory follicle had a slower growth rate (1.02 versus 1.55 mm per day; P < 0.005), a lesser increase in diameter from treatment to ovulation (4.7 versus 8.0 mm; P < 0.001), and a greater maximum diameter (13.2 versus 12.1 mm; P < 0.05) in cows that ovulated from the largest follicle present in the ovary before treatment (Group B, n = 27) compared with cows that ovulated from the second largest follicle present in the ovary before treatment (Group C, n = 9). In summary, the efficacy of PGF(2alpha) for causing luteolysis and synchronizing estrus and ovulation in buffalo cows was dependent upon plasma progesterone concentration, CL size and ovarian follicular status before treatment.     

The variability in the interval between estrus and ovulation in cattle and its determinants

Fertility of Holstein cows has been decreasing for years and, to a lesser extent, the fertility of heifers too but more recently. A hypothesis to explain this phenomenon may be that the chronology of events leading to ovulation is different for those animals bred nowadays when compared to what was reported previously; this would result in an inappropriate time of insemination. Therefore, two experiments were designed to investigate the relationships among estrus behavior, follicular growth, hormonal events and time of ovulation in Holstein cows and heifers. In the first experiment, the onset of estrus, follicular growth, patterns of estradiol-17beta, progesterone and LH, and the time of ovulation were studied in 12 cyclic Holstein heifers that had their estrus synchronized using the Crestar method; this was done twice, 3 weeks apart. The intervals between estrus and ovulation, estrus and the LH peak, and between the LH peak and ovulation were, respectively, 38.5 h +/-3.0, 9.1 +/- 2.0 and 29.4 h +/-1.5 (mean+/- S.E.M). The variation in the interval between estrus and the LH peak explained 80.6% of the variation in the interval between estrus and ovulation. The intervals between estrus and the LH peak, and estrus and ovulation were correlated with estradiol-17beta peak value (r=-0.423, P <0.04 and r=-0.467, P<0.02, respectively). Positive correlation coefficients for the number of follicle larger than 5 mm, and negative correlation coefficients for the size of the preovulatory follicle with the intervals between estrus and LH peak, LH peak and ovulation, and estrus and ovulation suggest an ovarian control of these intervals. In respect to its role to explain the variation in the interval between estrus and ovulation, the variation in the interval between estrus and the LH peak was evaluated further in a second set of experiments utilizing 12 pubertal Holstein heifers and 35 Holstein cows. The duration of the interval between the beginning of estrus and the LH peak was longer in heifers than in cows (4.15 h versus -1.0 h; P <0.002); the variation for this interval was higher in cows than in heifers (S.E.M.= 1.2 h versus 0.8 h; P=0.01). According to the results of these studies it can be proposed that estradiol and other product(s) of ovarian origin regulate not only the duration of intervals between the onset of estrus and the LH surge but also between the LH surge and ovulation. From the results obtained in the first experiment, it may be postulated that differences observed between cows and heifers for the duration of the interval between onset of estrus and the LH surge as well as for the variation of this interval would be observed also for the interval between the onset of estrus and ovulation. Therefore, on a practical point of view, the long interval between the onset of estrus and ovulation and the high variation of this interval, especially in cows, may be a source of low fertility and should be considered when analysing reproductive disorders.     

Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.     

LH surge in Nelore cows (Bos indicus), after induced estrus or after ovarian superestimulation

Considering that there is limited information about the preovulatory LH surge in Zebu cattle (Bos indicus), the purpose of the present work was to assess the LH surge in Nelore cows during the estrous cycle and after ovarian superestimulation of ovarian follicular development with FSH. This information is particularly important to improve superovulatory protocols associated with fixed-time artificial insemination. Nelore cows (n=12) had their estrus synchronized with an intravaginal device containing progesterone (CIDR-B) associated with estradiol benzoate administration (EB, 2.5 mg, i.m., Day 0). Eight days later all animals were treated with PGF2alpha (Day 8) in the morning (8:00 h) and at night, when CIDR devices were removed (20:00 h). Starting 38h after the first PGF2alpha injection, blood sampling and ovarian ultrasonography took place every 4h, during 37 consecutive hours. Frequent handling may have resulted in a stress-induced suppression of LH secretion resulting in only 3 of 12 cows having ovulations at 46.7+/-4.9 and 72.3+/-3.8 h, respectively, after removal of CIDR-B. Thirty days later, the same animals received the described hormonal treatment associated with FSH (Folltropin), total dose=200 mg) administered twice a day, during 4 consecutive days, starting on Day 5. Thirty-six hours after the first injection of PGF2alpha, to minimize stress, only seven blood samples were collected at 4h interval each, and ultrasonography was performed every 12 h until ovulation. In 11 of 12 cows (92%) the LH surge and ovulation were observed 34.6+/-1.6 and 59.5+/-1.9 h, respectively, after removal of progesterone source. The maximum values for LH in those animals were 19.0+/-2.6 ng/ml (mean+/-S.E.M.). It is concluded that, in Nelore cows submitted to a ovarian superstimulation protocol, the LH surge occurs approximately 35 h after removal of intravaginal device containing progesterone, and approximately 12h before the LH surge observed after an induced estrus without ovarian superstimulation.     

Ovulation rate after GnRH or PGF2alpha administration in early postpartum dairy cows

The objectives of this study evaluating induction of ovulation in early postpartum dairy cows were to: compare two methods of GnRH (100 mcg) administration (i.m. route and s.c. implant), and determine if prostaglandin F(2alpha) (PGF) causes release of LH or ovulation similar to that reported for GnRH. In trial #1, serum LH peaked at 2h after i.m. administration of GnRH and was declining at 4h. The s.c. GnRH implant also caused an elevation in serum LH at 2 and 4h after treatment, with LH declining at 6h. Serum LH was unchanged in control cows. Experimental treatment caused ovulation in 4 of 14 GnRH i.m. treated cows, 4 of 12 GnRH implanted cows and 0 of 13 control cows. Parity had no effect on LH response but did affect resulting ovulation rate as multiparous cows were more likely to ovulate than were primiparous cows in response to either GnRH treatment. All cows that ovulated had a follicle larger than 12 mm at the time of treatment. In trial #2, serum LH increased as before after i.m. administration of GnRH, however, serum LH was unchanged in cows treated with PGF or saline. Gonadotropin releasing hormone caused more cows to ovulate than did PGF or saline treatments, and GnRH shortened the interval from treatment to the onset of CL function over the PGF treatment; 13.9+/-2.6, 28.2+/-4.1 and 22.3+/-4.1 days for GnRH, PGF and saline, respectively. In summary, there was no difference in the ability of s.c. implantation and i.m. administration of GnRH to cause ovulation. Prostaglandin F(2alpha) did not cause release of LH or ovulation. In 22 early postpartum dairy cows treated with 100 mcg GnRH i.m. in these two trials, nearly all cows (95%) responded with a release of LH but only 45% (10/22) responded with an ovulation and subsequent formation of a CL.     

Exposure to endotoxin during estrus alters the timing of ovulation and hormonal concentrations in cows

The effect of intramammary (IMM) or intravenous (IV) administration of E. coli endotoxin (LPS), at the onset of estrus, at the time of ovulation was examined. Steroid and gonadotropin concentrations around ovulation were also determined. Lactating Holstein cows (n=33) were assigned to saline-controls (n=12) and treated with LPS-IV (0.5mug/kg; n=13) or LPS-IMM (10mug; n=8). Synchronized cows were observed continuously for estrus. LPS (or saline) was injected within 30min from the onset of standing estrus, at peak estradiol concentrations. The typical rise of body temperature, somatic cell count, cortisol, and NAGase activity was noted. One-third of both LPS-IV- and LPS-IMM-treated cows were manifested by an extended estrus to ovulation (E-O) interval of around 75h or did not ovulate, compared with about 30h in the other 2/3 of LPS cows and all controls. Estradiol concentrations 24h before and after LPS did not differ between groups. However, LPS-IV cows with extended intervals exhibited another estrus and an additional rise of estradiol followed by delayed ovulation. LPS-treated cows with a delayed E-O interval had low or delayed LH surge; two LPS-treated cows did not exhibit LH surge and did not ovulate. All control cows exhibited normal hormone levels. Delayed ovulation was associated with a delayed rise of luteal progesterone. The results indicated that exposing cows to endotoxin during estrus induced a decreased and delayed LH surge in one-third of the cows. This was associated with delayed ovulation, which reduces the chances of successful fertilization.     

Environmental,genetic and social factors affecting the expression of estrus in beef cows

Genetic, social and environmental factors affecting behavioral estrus were evaluated in Angus (n = 10), Brahman (n = 10) and Senepol (n = 10) cows during a PGF2alpha synchronized estrus and subsequent spontaneous estrus. Cows were equally stratified by breed to two groups of 15. Both groups were pre-synchronized with a modified two-injection PGF2alpha protocol. At the start of the experiment, cows were treated with 25 mg PGF2alpha followed by a second and third administration of 12.5 mg PGF2alpha, 11 and 12 days later to induce synchronized estrus. The subsequent estrus was designated as spontaneous estrus. Behavioral estrus data including the onset and end of estrus, estrous duration and the total number of mounts received for the synchronized and spontaneous estruses were collected using HeatWatch". Interval from the third PGF2alpha, treatment to the onset of a HeatWatch" estrus occurred earlier (P < 0.05) in Angus (31 +/- 5 h) than Brahman (53 +/- 7 h) or Senepol (53 +/- 4 h) cows, with dominant Senepol and Brahman cows taking longer to exhibit estrus after PGF2alpha than subordinate cows. The duration of the synchronized estrus tended to be shorter (P < 0.06) in Senepol (12 +/- 3 h) than in Angus (19 +/- 2 h) or Brahman (17 +/- 2 h) cows. Behavioral estrus data between the two periods were confounded by greater temperature-humidity index (THI) values during spontaneous estrus. The THI during spontaneous estrus appeared (P = 0.09) to affect the duration of estrus (9 +/- 1 h versus 16 +/- 1 h) and did affect (P < 0.0001) the total number of mounts received (8 +/- 4 mounts versus 34 +/- 4 mounts) during spontaneous estrus compared to synchronized estrus. Breed had no effect (P > 0.10) on the duration and total number of mounts received during synchronized and spontaneous estruses. In conclusion, type of estrus (synchronized or spontaneous), THI, social dominance and breed exerted significant effects on characteristics associated with behavioral estrus in beef cattle in subtropical environments.     

The effect of a GnRH analogue on the dynamics of follicular development and synchronization of estrus in lactating cyclic dairy cows

A GnRH analogue was used to synchronize ovarian follicular development prior to an injection of PGF(2alpha) for the synchronization of estrus in lactating Holstein cows. On Day 12 (estrus = Day 0) of the experimental cycle, cows (n = 8) were injected with 8 mug Buserelin (BUS group), followed by 25 mg PGF(2alpha) 7 d later (Day 19). Control cows (n = 7) received PGF(2alpha) on Day 12 (PGF group). Ovaries were scanned daily via ultrasonography, and plasma progesterone and estradiol concentrations were determined. Sizes of all visible follicles were recorded. Follicles were classified as small (3 to 5 mm), medium (6 to 9 mm), or large (>/= 10 mm). Between Days 12 and 16 of the cycle, the number of large follicles in PGF cows remained unchanged (1.2), whereas in the BUS group, the number of large follicles decreased from 1.3 on Day 12 to 0.5 on Day 15. Only 4 of 7 PGF cows ovulated a second-wave dominant follicle. In the BUS group, 7 of 8 cows ovulated a GnRH analogue induced dominant follicle that was first identified on Day 15. During the follicular phase (last 5 d prior to estrus), plasma progesterone declined in association with CL regression in both groups, and estradiol concentrations increased, reaching higher (P<.0.05) preovulatory peak concentration in BUS cows than in PGF cows (14.0 +/- 1.0 vs 10.4 +/- 1.1 pg/ml). The number of medium-size follicles was smaller and the number of small-size follicles tended to be higher in BUS cows than in the PGF-treated group. On the day of estrus, the size of the ovulatory follicle (16.1 vs 13.3 mm) and the size difference between the ovulatory and second largest follicle (11.4 vs 6.2 mm) were both larger in BUS cows than in PGF-treated cows, suggesting a more potent dominance effect of the ovulatory follicle in the BUS cows. This study suggests that a GnRH analogue can alter follicular development prior to synchronization of estrus with an injection of PGF(2alpha) in lactating dairy cows.     

Determination of ovulation time in bitches based on teasing, vaginal cytology, and elisa for progesterone

The estrous cycle of 16 mature mongrel female dogs was monitored to evaluate the accuracy of teasing, vaginal cytology and quantitative ELISA progesterone assay to determine ovulation. The dogs were presented to male, and blood samples and vaginal swabs were taken daily during proestrus and estrus. Selected serum samples collected during estrus were assayed for endogenous LH by radioimmunoassay (RIA). Plasma samples collected during proestrus and estrus were assayed for progesterone with a commercially avialable ELISA kit. Ovulation was considered to take place 48 h after the preovulatory LH peak. Vaginal cytology smears were stained with Wright's stain and evaluated for the percentage of superficial squamous cells. Day 1 of diestrus (Day 1) was defined as a drop of 20% or more in the total number of superficial cells. Two standard curves (linear and best fitted curves) commonly used with ELISA were compared together and with the RIA progesterone assay. Ovulation was estimated to occur when progesterone concentration was 4.9 +/- 1.0ng/ml (mean +/- SD, n = 15), with a range of 3.4 to 6.6 ng/ml. Based on vaginal cytology, ovulation took place 6.9 +/- 1.6 d (n = 15) after 80% of the squamous cells were superficial and 6.8 +/- 1.4 d (n = 16) before Day 1. Ovulation took place 2.1 +/- 3.9 d (n=11) after the first day of standing estrus and 8.8 +/- 1.5 d (n = 10) before the last day of receptivity. The two standard curves were found parallel to each other and to the RIA progesterone assay. Based on the results of the present study, ELISA progesterone assay and determination of the first day of estrus by vaginal cytology are reliable methods for predicting ovulation, whereas the last day of receptivity as determined by teasing and Day 1 as determined by vaginal cytology are reliable methods to retrospectively estimate ovulation time.     

Timing of ovulation in relation to onset of estrus and LH peak in yak (Poephagus grunniens L.)

The objective of the study was to determine the timing of ovulation in relation to onset of estrus and the preovulatory LH peak in yaks. For this purpose, a sensitive LH enzymeimmunoassay previously established in buffaloes was successfully validated for measuring the hormone in yak plasma. Plasma LH and progesterone were estimated from blood samples collected from eight non-lactating cycling yaks at 2 h intervals after estrus onset until 6 h after ovulation (ovulation was confirmed by palpation of ovaries per rectum). The mean+/-S.E.M. preovulatory plasma LH peak was 10.11+/-0.35 ng/ml with the values ranging from 8.75 to 11.51 ng/ml in individual yaks. The mean+/-S.E.M. duration of the LH surge was 7.25+/-0.55 h with a range of 6-10 h. Onset of LH surge (mean+/-S.E.M.) occurred 3.0+/-0.65 h after the onset of estrus. Mean plasma progesterone stayed low (<0.25 ng/ml) during the entire duration of sampling. Ovulation occurred 30.5+/-0.82 h (range, 28-34 h) after the onset of estrus and 20.25+/-1.03 h after the end of LH surge. The occurrence of the LH peaks within a narrow time frame of 4-8h post estrus onset in yaks could have contributed to the animals ovulating within a narrow time interval.     

Synchronization of estrus and ovulation and associated endocrine changes in Bos indicus cows

The effects of 4 estrus synchronization treatments on intervals to and synchrony of estrus and ovulation, on timing of the preovulatory LH surge and associated changes in plasma progesterone, LH, FSH, and 17beta-estradiol (E(2)) were investigated in 48 Bos indicus cows. Treatment 1 consisted of 2 injections of PGF(2alpha) 14 d apart (n = 12); Treatment 2 of a subcutaneous 3-mg norgestomet implant and an intramuscular injection of 3 mg of norgestomet and 5 mg estradiol valerate, with the implant removed 10 d later (n = 12; norgestomet-estradiol); Treatment 3 of norgestomet-estradiol, with a subcutaneous injection of PMSG given at time of implant removal (Day 10; n = 12); and Treatment 4 of norgestomet implant (as for Treatments 2 and 3) inserted for 10 d, with an intramuscular injection of PGF(2alpha) given at the time of implant removal (n = 12). The experiment was conducted in 2 replicates (24 cows/replicate, 6 cows/group). Estrus, ovulation and timing of the preovulatory surge of LH varied less in cows treated with norgestomet-estradiol and PMSG than in cows in Treatments 1 and 4 (P < 0.008). Treatment with PMSG reduced variation in ovulation times and timing of the LH surge in cows treated with norgestomet-estradiol (P < 0.02). Concentrations of E(2) were higher in cows in Treatments 2 and 3 on the final day of treatment and at about 6 h post ovulation compared with cows in Treatments 1 and 4 (P < 0.05). Different methods for synchronizing estrus did not alter sequential endocrine and behavioral changes in relation to the timing of the LH peak, and the results were consistent with current recommendations for insemination times in Bos taurus cattle.     

Efficacy of Heatsynch protocol for induction of estrus, synchronization of ovulation and timed artificial insemination in yaks (Poephagus grunniens L.)

The objective of this study was to test the efficacy of induction of estrus, synchronization of ovulation and timed artificial insemination in anestrous yaks using the Heatsynch protocol. In Experiment 1, 10 anestrous yaks were administered an analogue of gonadotropin releasing hormone (GnRH) followed by prostaglandin (PG)F2alpha 7 days later and then estradiol cyponate (ECP) 24 h after that. Ovulation was detected by rectal palpation at 2h intervals beginning at the initial signs of estrus. Blood samples were collected at 2h intervals beginning at the time of ECP injection up to 2h after the occurrence of ovulation for the determination of LH and progesterone. All the animals responded to the Heatsynch protocol with expression of estrus and synchronization of ovulation. The mean time interval from the ECP injection to ovulation was 59.4+/-2.62 h (range 50-72 h). The interval from the LH peak to ovulation was 30.2+/-2.3 h. The high degree of synchrony in ovulation could be attributed to the synchrony in the timing of LH peaks. In Experiment 2, 10 anestrous yaks were treated with the Heatsynch protocol (as in Experiment 1) and TAI was performed at 48 and 60 h after the ECP treatment. Concurrently, 16 cycling yaks were inseminated approximately 12 h after detection of spontaneous estrus. Pregnancy rates were similar in both groups, 40% for TAI and 43.75% for yaks inseminated following spontaneous estrus (p>0.05). From this study, two conclusions can be drawn. First, the Heatsynch protocol can be successfully used to induce and synchronize estrus in anestrous yaks and, second, ovulation following the Heatsynch protocol is synchronized adequately to permit the use of fixed time AI in this species.     

Progesterone and follicular changes in postpartum noncyclic dairy cows after treatment with progesterone and estradiol or with progesterone, GnRH, PGF2alpha, and estradiol

A previous study showed that noncyclic dairy cows treated with 10 microg of GnRH and a progesterone-releasing CIDR insert on Day 0, 25 mg of PGF2alpha and CIDR removal on Day 7, followed by 1 mg estradiol benzoate on Day 9 for those cows that still had not shown estrus (CGPE program) had higher conception rate (47% vs. 29%) than cows treated only with CIDR and estradiol benzoate as above (CE program). This study was to investigate the mechanisms by which the CGPE program improved conception rate compared with the CE program. Sixteen noncyclic Holstein-Friesian cows were randomly assigned to 2 groups balanced for the size and growth pattern of the dominant follicles, which were determined by ultrasonography over a 3-d period. One group received the above CGPE treatment, and the other group received the CE treatment. Follicular and luteal development were monitored by daily ultrasonography. Blood samples were collected daily from Day -2 to Day 11, and thereafter milk samples were collected thrice weekly for a further 24 d. Blood and milk samples were analyzed for progesterone. The GnRH treatment induced ovulation in 7 of 8 cows, resulting in elevated (P<0.05) progesterone concentrations between Days 4 and 7 for cows in the CGPE group. All induced CL underwent luteolysis by 24 h after PGF2alpha treatment. Within 5 d of CIDR removal, 7 of 8 cows in both the CE and CGPE groups ovulated. The interval from emergence of the ovulatory follicle to ovulation was similar (P=0.32) but less (P<0.05) variable for the CGPE group (9.0+/-0.3 d) compared with the CE group (10.3+/-1.2 d). Progesterone concentration in milk samples was similar between the two groups up to 10 d after ovulation. In summary, the GnRH treatment induced ovulation or turnover of dominant follicles, induced a synchronized initiation of a new follicular wave, and increased the progesterone concentration from 4 d after treatment. These could be the reasons for the increased conception rate of cows treated with the CGPE program.     

Estrus synchronization and artificial insemination of hair sheep ewes in the tropics

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2alpha (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2alpha injection or at CIDR or sponge removal, sterile rams were placed with the ewes. Jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at -20 degrees C until LH, and progesterone concentrations were determined by RIA. There was no difference (P>0.10) in time to estrus among the CIDR-, PGF2alpha- or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2alpha-treated ewes showed signs of estrus by this time (P<0.06). The time from ram introduction to ovulation was not different (P>0.10) among the CIDR-, PGF2alpha- or sponge-treated ewes. The time to the preovulatory LH surge was similar (P>0.10) among CIDR, PGF2alpha and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P>0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2alpha and bred by TAI using frozen-thawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9% (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2alpha. Even though there were no significant differences in the timing of ovulation or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technician.     

Buserelin in a superovulatory regimen for Holstein cows. I. Pituitary and ovarian hormone response in an experimental herd

Two experiments were conducted with frequent blood sampling in standard superovulatory regimens using follicle stimulating hormone (FSH) and prostaglandin F(2) alpha (PGF) to study the effects of the gonadotropin releasing hormone analog, Buserelin, on changes in FSH, luteinizing hormone (LH), progesterone (P(4)) and estradiol (E(2)). In Experiment I, Buserelin (20 mug) was administered to a total of 28 dry Holsteins. One group was treated with Buserelin 36 and 60 h after PGF administration, a second group was treated 60 h after PGF, and a third group served as the controls. In Experiment II, 30 dry Holsteins received Buserelin (10 mug). One group was treated 48 h after PGF, a second group at 54 h after PGF, a third group 24 h after estrus was first observed and a fourth group was a control. The general pattern of a decrease in P(4) following PGF, an increase in E(2), the onset of estrus, an LH peak, and finally, an increase in P(4) in superovulated cows was observed. Buserelin consistently produced a sharp LH peak at 36 h when given 36 h after PGF. At later intervals, it produced either a major or minor peak depending upon whether a spontaneous LH peak had already occurred. There was too much individual cow variation in the interval from PGF to a spontaneous LH peak to consistently induce a uniform LH peak, except when Buserelin was given 36 h after PGF, which may be early for normal oocyte maturation. There was no treatment effect on FSH, and embryo recovery rate was unaffected by treatment (P>0.05).     

Behavioral estrous signs can predict the time of ovulation in mithun (Bos frontalis)

The objective of this study was to investigate the relationship of different behavioral estrous signs and time of ovulation to identify if behavioral estrous sign(s) can be used as predictor of time of ovulation in mithuns. Data were collected for 54 ovulations from 16 mithuns. The animals were monitored for onset of estrus by observing different behavioral estrous signs at 2 h interval and bull parading thrice a day for 30 min and were further confirmed by plasma progesterone profile. All animals were also observed for any of the estrous signs at every 2 h interval for 30 min and mounting behavior was studied by bull parading at every 2 h for 30 min after onset of estrus. Time of ovulation was detected by rectal palpation at 2 h interval from onset of estrus till ovulation. Behavioral signs of estrus was more intense in primiparous than multiparous mithuns. Ovulation occurred at 26.1+/-1.1 h (ranging between 20 and 31 h) after the onset of estrus. As the method used to determine the onset of estrus is time consuming, labor intensive and no device is yet available to detect onset of estrus automatically, so this cannot be used practically as a predictor of time of ovulation. The mithun cow at estrus to be mounted by bull was recorded in all cases (100%). Ovulation occurred 23.5+/-1.5 h (ranging between 19 and 27 h) after first mounting. Although promising, mounting cannot be assessed automatically, which limits its practical use as a predictor of ovulation. Standing heat was recorded in 98.1% of total estrus studied in mithun cows and ovulation occurred 21.8+/-1.3 h (ranging between 19 and 25 h) after first observed standing heat. Standing heat can be detected automatically using mounting detectors. Hence, standing heat can be used practically as ovulation predictor in mithuns. In conclusion, cow to be mounted by mithun bull is the best predictor of ovulation, but non-availability of devices to detect it automatically restricts its practical application. Standing heat that recorded 98.1% estrus cases in mithun cows, can also be detected automatically using mounting detector, therefore be used widely as an ovulation predictor in field condition for mithun cows.