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PubMed2005

[Comparision of ovulation synchronization (OVSYNCH) with the selective induction of oestrus using PGF2alpha after rectal palpation in a dairy herd].

The efficacy of a protocol for the synchronization of ovulation followed by timed Al (OVSYNCH) was compared with a reproductive management protocol based on induction of oestrus after rectal palpation using PGF2alpha on a commercial dairy farm in Brandenburg, Germany. Cows in the OVSYNCH group (n = 309) were treated between 62 and 68 days in milk (DIM) with 0.02 mg of buserelin (GnRH analogue) intramuscularly (i.m.). Seven days later 0.75 mg of tiaprost (PGF2alpha-analogue) were administered i.m. to regress the corpus luteum (C.I.). All cows received a second treatment with GnRH another 48 hrs later and were inseminated 16 to 20 hrs after the second GnRH-treatment (72 to 78 DIM). Cows in the PGF group (n = 302) were examined by rectal palpation between 69 and 75 DIM. Cows with a C.I. received 0.75 mg of tiaprost to induce oestrus. Cows were inseminated on observed oestrus. Cows not inseminated within 14 days after treatment were re-examined between 83 and 89 DIM. For both groups, the voluntary waiting period was set at 72 DIM. Service rate was higher (86.4 vs. 63.2 %, P < 0.05), days to first service (77 +/- 11 vs. 86 +/- 19 days, P < 0.001) and days open (102 +/- 34 vs. 109 +/- 35 days, P < 0.05) were shorter in the OVSYNCH group. First service conception rate (40.3 vs. 42.4 %), conception rate to all services (37.9 vs. 40.0 %) and the proportion of pregnant cows at 200 DIM were similar in the OVSYNCH evaluation for the total costs per pregnancy in 71 different cost scenarios showed only slight differences between the groups. Mean costs per pregnancy were euro 235.43 and euro 235.08 for the OVSYNCH and the PGF group, respectively. Quality of oestrus as assessed by the Al technician did not reveal a relationship to conception rate after OVSYNCH and timed Al. However, swelling, hyperaemia and a moist vestibulum were positively associated with conception rate.

Animals↗
PubMed1997

Effects of supplementary treatment with bovine growth hormone on hormonal and ovulatory responses to inhibin immunization in ewes.

The aim of this study was to determine whether supplementary treatment with recombinant bovine growth hormone(rbGH) can enhance the ovulatory response of ewes to inhibin immunization. Crossbred ewes (n = 20) were actively immunized against bovine inhibin a1-29 peptide conjugate while 20 ewes served as controls. Oestrus was synchronized using progestagen sponges and ewes were allocated to four groups: control ewes (n = 10); control ewes given rbGH (n = 10); inhibin-immunized ewes (n = 10) and inhibin-immunized ewes given rbGH (n = 10). A single s.c. dose of rbGH (50 mg) was given 7 days before sponge removal. Blood was collected for measurement of inhibin antibody titre, and concentrations of insulin-like growth factor I (IGF-I), FSH, oestradiol and progesterone. Ovulation, pregnancy and lambing rates were also recorded. All inhibin-immunized ewes produced antibodies that bound 125I-labelled (32 kDa) inhibin. The concentration of FSH in the plasma of the ewes after the second booster inhibin immunization was higher than that in control ewes (P < 0.005). Treatment with rbGH promoted a 2-3-fold increase in plasma concentration of IGF-I (P < 0.001); the response was less (P < 0.01) in immunized compared with control ewes. Treatment with rbGH alone had no significant effect on the concentration of FSH or oestradiol or on ovulation rate or litter size. Overall, inhibin-immunized ewes had higher mean FSH concentrations (P < 0.002), higher preovulatory oestradiol surges (P < 0.05) and higher progesterone concentrations in the luteal phase (P < 0.0001). Treatment with rbGH reduced the effects of immunization on FSH (P < 0.01) and progesterone (P < 0.02) concentrations. Immunized ewes showed a threefold increase in ovulation rate (P < 0.001) and a 1.8-fold increase in litter size (P < 0.05) compared with control ewes. In immunized ewes given rbGH, ovulation rate was increased by a factor of 2.2 and litter size by a factor of 1.8. In conclusion, these data do not support the hypothesis that supplementary treatment of ewes with rbGH to raise plasma IGF-I concentrations (and presumably intraovarian IGF-I) can enhance the ovulatory response to inhibin immunization.

Animals↗
PubMed2000

Immediate and delayed effects of heat stress on follicular development and its association with plasma FSH and inhibin concentration in cows.

The aim of this study was to characterize the immediate effects of heat stress on plasma FSH and inhibin concentrations, and its involvement in follicular dynamics during a complete oestrous cycle, and to examine a possible delayed effect of heat stress on follicular development. Holstein dairy cows were oestrous synchronized and randomly assigned to either cooled (n = 7) or heat-stressed (n = 6) treatment groups. During a complete oestrous cycle, control cows, which were cooled, maintained normothermia, whereas heat-stressed cows, which were exposed to direct solar radiation, developed hyperthermia. At the end of this oestrous cycle (treated cycle), both groups were cooled and maintained normothermia for the first 10 days of the subsequent oestrous cycle. Throughout this period, follicular development was examined by ultrasonography, and plasma samples were collected. During the second follicular wave of the treated oestrous cycle, a significantly larger cohort of medium sized follicles (6-9 mm) was found in heat-stressed cows than in cooled cows (P < 0.05). The enhanced growth of follicles in this wave in heat-stressed cows was associated with a higher plasma FSH increase which lasted 4 more days (days 8-13 of the oestrous cycle; P < 0.05), and coincided with a decrease in the plasma concentration of immunoreactive inhibin (days 5-18 of the oestrous cycle; P < 0.05). During the follicular phase (days 17-20 of the treated cycle), heat-stressed cows showed an increase in the number of large follicles (>/= 10 mm), and the preovulatory plasma FSH surge was significantly higher in heat-stressed cows than in cooled cows (P < 0.01). The effect of heat stress was also observed during the first follicular wave of the subsequent cycle: the postovulatory plasma FSH concentration was higher (P < 0.01), but fewer medium follicles developed, and the first follicular wave decreased at a slower rate in previously heat-stressed cows than in cooled cows (0.40 and 0.71 follicles per day, respectively). This study shows both immediate and delayed effects of heat stress on follicular dynamics, which were associated with high FSH and low inhibin concentrations in plasma. These alterations may have physiological significance that could be associated with low fertility of cattle during the summer and autumn.

Animals↗
PubMed2003

Effect of oestradiol benzoate given after prostaglandin at two stages of follicle wave development on oestrus synchronisation, the LH surge and ovulation in heifers.

Oestrus synchronization following prostaglandin-induced luteolysis is variable and dependent on follicle wave status in cattle. Oestradiol benzoate (ODB) has been used following prostaglandin to reduce the interval to oestrus and ovulation, but the effect of follicle wave status at the time of ODB administration is not clear. The aim of this study was to characterize the endocrine and follicular responses following ODB after luteolysis at different stages of the follicle wave. Prostaglandin was administered at either emergence or dominance of the second follicle wave. Twenty-four hours later animals received either 0.5mg ODB in oil or a control oil injection. Follicular development was monitored daily by ultrasonography, oestrous behavior was determined and blood samples were collected. In animals treated with ODB at emergence, there was a reduction (P<0.05) in the maximum diameter of the ovulatory follicle (11.7+/-1.2 mm versus 13.1+/-0.1 mm) and in the interval from prostaglandin to oestrus (52.0+/-2.3 h versus 88.0+/-9.6h), to the LH surge (53.3+/-3.5 h versus 89.1+/-6.5 h) and to ovulation (96+/-0.0 h versus 129.6+/-9.6h), compared with controls. In animals treated with ODB at dominance, there was a reduction (P<0.05) in the interval from prostaglandin to the LH surge (54.0+/-3.1 h versus 70.9+/-4.8 h), but not in the interval from prostaglandin to oestrus (53.3+/-2.7 h versus 65.7+/-4.5 h; P=0.11), to ovulation (96.0+/-0.0 h versus 110.4+/-4.8 h; P=0.12) or the maximum diameter of the ovulatory follicle (12.7+/-0.3 mm versus 13.6+/-0.4 mm; P=0.12), compared with controls. Treatment did not affect (P>0.05) the length of the subsequent oestrous cycle or corpus luteum size. In conclusion, the use of ODB advanced, but did not alter the temporal relationships among oestrus, the LH surge and ovulation, regardless of stage of follicle development at treatment.

Animals↗
PubMed1982

Synchronization of oestrus in ewes with Provera sponges/PMSG, prostaglandin F2 alpha or the prostaglandin analogue, ICI 80996, and fertility following natural mating or artificial insemination.

Following the synchronization of oestrus with different treatments during the breeding season, the fertility of naturally-mated ewes has been compared to that of ewes artificially inseminated with fresh semen. The following treatments were used: I. sponges impregnated with progestagen (50--60 mg of medroxyprogesterone acetate) left in situ for 14 days and an i.m. injection of 500 IU of PMSG at sponge withdrawal; II. injections of prostaglandin F2 alpha (dose: 15 mg/injection) at intervals varying from 9 to 14 days (see tables); III. injections of the prostaglandin F2 alpha analogue, ICI 80996 (dose: 100 micrograms/injection), with the same time intervals as in treatment II. Lambing rate and prolificacy after natural service at the induced oestrus was 55 and 140 p. 100, 32.5 and 153 p. 100 and 60 and 133 p. 100 for ewes receiving treatments I, II or III, respectively, compared to 62.5 and 120 p. 100 for the untreated controls. Double artificial insemination (AI) 48 and 58 h after the sponge/PMSG treatment resulted in a lambing rate of 25 p. 100 and a prolificacy of 133 p. 100. The lambing rate and prolificacy of ewes inseminated 58 and 68 h after the final injection of PGF2 alpha or ICI 80996 were 27.8 and 149 p. 100 and 45.5 and 165 p. 100, respectively. Single AI 55 h after sponge withdrawal gave a lambing rate of 37.8 p. 100. The lambing rate of ewes inseminated 56 h after the final ICI 80996 injection was higher (54.8 p. 100) than that of ewes inseminated at 60 h (37.5 p. 100) or 66 h (30.8 p. 100). However, two inseminations 56 and 66 h after the final ICI 80996 injection gave an even more elevated lambing rate (61.9 p. 100). These results demonstrate that ICI 80996 can successfully control oestrus in the ewe during the breeding season, thus offering an alternative to sponges/PMSG and that the fertility subsequent to both natural mating and AI is equivalent to that of the controls.

Animals↗
PubMed2005

Effect of presynchronization using prostaglandin F2alpha and a milk-ejection test on pregnancy rate after the timed artificial insemination protocol, Ovsynch.

The objective of this research was to determine if PGF2alpha-induced milk letdown (ML) is an accurate indicator of luteolysis, allowing cows to be synchronized to begin the Ovsynch protocol (GnRH-7d-PGF2alpha-2d-GnRH-24h-AI) at the most beneficial time of the estrous cycle (days 5-9), and determine if this would improve pregnancy rate (PR). Lactating Holstein cows between 55 and 70 days in milk were used to evaluate the ML test and PR after the Ovsynch protocol, when initiated on the basis of the test result (PROSYNCH). PROSYNCH cows (n = 60) had one teat cannulated to test for ML and were treated with 500 microg cloprostenol, PGF2alpha analogue (PG). Cows with ML were started on Ovsynch 10 days later, and those without started 3 days later. Cows in the control group (OVSYNCH, n = 64) were injected with physiological saline and observed for ML. This group was started on Ovsynch 10 days after saline treatment. Milk samples were collected thrice weekly to determine progesterone concentrations. ML indicated luteolysis with a sensitivity of 98% and a specificity of 60%. The positive and negative predictive values were 83 and 92%, respectively. Pregnancy rates were 48% for PROSYNCH and 52% for OVSYNCH (P = 0.72). When data from both groups were combined, PR was greater in cows that started the Ovsynch protocol in stage 2 of the estrous cycle (days 5-9, 67%) than all other stages (stage 1: days 1-4, 35%; stage 3: days 10-16, 45%; stage 4: days 17-21, 42%; P < 0.01). The proportion of animals with ovulation after GnRH#1, luteolysis after PGF2alpha, and ovulation after GnRH#2 were all greater in the PROSYNCH group (77% versus 55%, P < 0.02; 83% versus 66%, P < 0.03; 97% versus 84%, P < 0.03, respectively). Therefore, the ML test indicated luteolysis with sufficient precision to time the initiation of the Ovsynch protocol between days 5 and 9 of the cycle, however, this did not alter PR compared to starting the protocol randomly throughout the cycle. Initiating the Ovsynch protocol between days 5 and 9 of the cycle increased PR, and improved the efficacy of each injection.

Animals↗
PubMed2002

Season affects characteristics of the pre-ovulatory LH surge and embryo viability in superovulated ewes.

The aim of this study was to determine whether there are seasonal shifts in ovulatory response, and in the viability of ova recovered from superovulated ewes. Fifty mature ewes underwent a standard oestrous synchronisation (CIDR), superovulation (oFSH) and artificial insemination procedure during October (peak breeding season) and April (transition to anoestrus). In each month peripheral LH and progesterone concentrations were measured around the time of ovulation and embryos were recovered, graded and cryopreserved on day 6 after insemination. During the subsequent breeding season, grade 1 and 2 morulae and unexpanded blastocysts were thawed and transferred singly to synchronous recipients (October, n = 40; April, n = 40) or cultured in vitro for 18-20 h (October, n = 107; April, n = 98). Following culture, viable embryos were stained to count cell nuclei or assayed to measure their capacity for glucose metabolism ([3H]glucose) and protein synthesis ([35S]methionine). Peak LH concentrations were higher in October than in April (38.2 +/- 3.26 ng ml(-1) versus 25.7 +/- 1.99 ng ml(-1), respectively; P < 0.01) and the pre-ovulatory LH surge was advanced by approximately 3 h (P < 0.05). Progesterone concentrations at CIDR withdrawal were lower in October than in April (3.1 +/- 0.16 ng ml(-1) versus 4.3 +/- 0.19 ng ml(-1), respectively; P < 0.001) but were not different at embryo recovery. Season did not affect the numbers of corpora lutea per ewe or the numbers of ova recovered but the proportion of recovered ova that was unfertilised/degenerate was lower in October than in April (0.43 versus 0.58, respectively; P < 0.001). For embryos containing more than 16 cells, there was no effect of season on the median stage of development or morphological grade. The proportions of October and April embryos that established pregnancy following transfer to recipient ewes were 0.78 and 0.70 (not significantly different), and that were viable after in vitro culture were 0.66 and 0.37 (P < 0.05), respectively. Season did not affect the number of nuclei per viable embryo or the capacity for protein synthesis but the glucose uptake of October embryos was approximately double that of April embryos (3163+/-293.4 dpm versus 1550+/-358.9 dpm, respectively; P < 0.05). Results indicate that during the late compared to peak breeding season, there is an increased incidence of fertilisation failure as a possible consequence of seasonal shifts in LH secretion and (or) associated effects on follicular function. Frozen-thawed embryos produced at contrasting stages of the breeding season are equally viable in vivo but those produced during the late, as opposed to the peak breeding season have lower viability following in vitro culture.

Animals↗
PubMed1991

Relationship of oestrus synchronization method, circulating hormones, luteinizing hormone and prostaglandin F-2 alpha receptors and luteal progesterone concentration to premature luteal regression in superovulated sheep.

Ewes were treated with exogenous follicle-stimulating hormone (FSH) and oestrus was synchronized using either a dual prostaglandin F-2 alpha (PGF-2 alpha) injection regimen or pessaries impregnated with medroxy progesterone acetate (MAP). Natural cycling ewes served as controls. After oestrus or AI (Day 0), corpora lutea (CL) were enucleated surgically from the left and right ovaries on Days 3 and 6, respectively. The incidence of premature luteolysis was related (P less than 0.05) to PGF-2 alpha treatment and occurred in 7 of 8 ewes compared with 0 of 4 controls and 1 of 8 MAP-exposed females. Sheep with regressing CL had lower circulating and intraluteal progesterone concentrations and fewer total and small dissociated luteal cells on Day 3 than gonadotrophin-treated counterparts with normal CL. Progesterone concentration in the serum and luteal tissue was higher (P less than 0.05) in gonadotrophin-treated ewes with normal CL than in the controls; but luteinizing hormone (LH) receptors/cell were not different on Days 3 and 6. There were no apparent differences in the temporal patterns of circulating oestradiol-17 beta, FSH and LH. High progesterone in gonadotrophin-treated ewes with normal CL coincided with an increase in total luteal mass and numbers of cells, which were primarily reflected in more small luteal cells than in control ewes. Gonadotrophin-treated ewes with regressing CL on Day 3 tended (P less than 0.10) to have fewer small luteal cells and fewer (P less than 0.05) low-affinity PGF-2 alpha binding sites than sheep with normal CL. By Day 6, luteal integrity and cell viability was absent in ewes with prematurely regressed CL. These data demonstrate that (i) the incidence of premature luteal regression is highly correlated with the use of PGF-2 alpha; (ii) this abnormal luteal tissue is functionally competent for 2-3 days after ovulation, but deteriorates rapidly thereafter and (iii) luteal-dysfunctioning ewes experience a reduction in numbers of small luteal cells without a significant change in luteal mass by Day 3 and, overall, have fewer low-affinity PGF-2 alpha binding sites.

Animals↗
PubMed1992

Relationship between the onset of oestrus, the preovulatory surge in luteinizing hormone and ovulation following oestrous synchronization and superovulation of farmed red deer (Cervus elaphus).

The timing of ovulation relative to the onset of oestrus and the preovulatory surge in luteinizing hormone (LH) was studied in red deer following treatments to synchronize oestrus and induce either a monovulatory or superovulatory response. Mature hinds (n = 36) were allocated randomly to two mating groups (n = 16 + 20), with respective treatments staggered by 4 weeks during the 1990 rut (March-April). Each hind was treated with an intravaginal controlled internal drug releasing (CIDR)-type S device for 14 days. Treatments to induce a monovulatory response included CIDR device alone (treatment A; n = 4 + 8) and additional injection of 200 iu pregnant mares' serum gonadotrophin (PMSG) at device removal (treatment B; n = 4 + 4). Treatments to induce a superovulatory response included injections of 200 iu PMSG and 0.5 units ovine follicle-stimulating hormone (FSH) at about time of removal of CIDR devices (treatment C; n = 4 + 4) and further treatment with gonadotrophin-releasing hormone (GnRH) analogue 18 h after removal of CIDR devices (treatment D; n = 4 + 4). The hinds were run with crayon-harnessed stags from insertion of CIDR devices (12 March or 9 April) and blood samples were taken every second day to determine plasma progesterone. Further blood samples were collected for determination of plasma LH and progesterone via indwelling jugular cannulae every 2 h for 72 h from removal of CIDR devices. Hinds were allocated randomly to an initial ovarian examination by laparoscopy at either 16 or 20 h (A and B), or 12 or 16 h (C and D) after the onset of oestrus, with laparoscopy repeated at intervals of 8 h until either ovulation was recorded (A and B), or for four successive occasions (C and D). All hinds received cloprostenol injections 15 days after device removal. A total of 28 hinds (78%) exhibited oestrus and a preovulatory LH surge, with mean (+/- SEM) times to onset of oestrus of 44.6 +/- 1.0 h (A; n = 7), 37.4 +/- 2.0 h (B; n = 7), 16.3 +/- 1.7 h (C; n = 6) or 14.0 +/- 1.7 h (D; n = 8). Failure to exhibit oestrus or LH surge was most prevalent among hinds in treatment A early in the rut.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1998

Gonadotrophin storage patterns in the ewe during the oestrous cycle or after long-term treatment with a GnRH agonist.

The storage pattern of gonadotrophins in the ewe pituitary was investigated during the oestrous cycle and after desensitization to GnRH using long-term treatment with a GnRH agonist, buserelin. Oestrous cycles in ewes were synchronized with progestagen sponges. Animals were allocated to two experiments. In the first, ewes were killed 36 h (before the preovulatory surge, n = 4), 48 h (end of the preovulatory surge, n = 5), 72 h (post-ovulation, n = 4) and 240 h (luteal phase, n = 3) after sponge removal. In the second experiment, another progestagen sponge was inserted in ewes 84 h after removal of the first sponge. Four ewes were infused continuously with buserelin (50 micrograms/day) for 15 days before killing. A further four ewes received no buserelin (controls). Pituitaries were collected and processed for immunocytochemistry to detect monohormonal (LH or FSH) and multihormonal (LH/FSH) cells. The percentages of LH or FSH immunoreactive cells in the pituitary were lower at the end of the preovulatory surge (7.4 +/- 0.3% and 1.2 +/- 0.3% respectively) compared with the other stages (11.4 +/- 0.5% and 5.4 +/- 0.7% respectively). Analysis of dual immunostaining showed the existence of monohormonal cells for LH and multihormonal cells (LH/FSH). No monohormonal cell for FSH was detected except at the end of the preovulatory surge when a few monohormonal FSH cells appeared (0.1 +/- 0.01% of pituitary cells). The percentage of monohormonal LH cells in the pituitary gland was similar in all studied stages of the oestrous cycle, whereas the percentage of multihormonal cells was lower at the end of the surge. In agonist-treated ewes, the percentages of LH or FSH immunoreactive cells (5.3 +/- 0.5% and 1.5 +/- 0.8% respectively) were decreased compared with controls (9.4 +/- 1% and 7.5 +/- 1.1% respectively). Analysis of the double immunostaining revealed a few monohormonal FSH cells (0.2 +/- 0.01% of pituitary cells) in agonist-treated ewes but not in controls. The percentage of monohormonal LH cells in the pituitary gland increased from 1.9 +/- 0.2% in controls to 3.8 +/- 0.3% in agonist-treated ewes, whereas multihormonal cells dropped from 7.5 +/- 1.1% to 1.3 +/- 0.7%. Our data suggest, therefore, that multihormonal cells contribute to gonadotrophin secretion, either during the preovulatory surge of the oestrous cycle or during the 'flare-up' effect initially induced by a GnRH agonist. Moreover, the appearance of monohormonal FSH cells in some conditions reflects a differential regulation of LH and FSH.

Animals↗
PubMed2006

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: II. Endometrial gene expression related to maintenance of pregnancy.

The objectives were to examine the effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on expression of key endometrial genes and proteins regulating prostaglandin synthesis in lactating dairy cows. Two diets were fed, at about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with calcium salts of fish oil-enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol and cows were inseminated artificially or not inseminated on d 0 (d 0 = time of synchronized ovulation; 77 +/- 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST, and were slaughtered on d 17 to recover uterine secretions and endometrial tissue. Number of cows in the control diet: 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows and in the FO diet: 4 bST-treated FO-cyclic (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. The FO diet increased progesterone receptor (PR) mRNA, and treatment with bST increased PR mRNA concentration in endometrium of no bST-C, but not in no bST-FO-C or no bST-P cows. Concentrations of estrogen receptor-alpha (ERalpha) mRNA and protein, and oxytocin receptor (OTR) mRNA were decreased in no bST-P cows compared with no bST-C cows. Treatment with bST tended to increase OTR and ERalpha mRNA concentrations in cyclic cows fed control or FO diets. Immunohistochemistry demonstrated effects of bST, FO, and pregnancy on distributions of ERalpha and PR proteins in endometrium. Pregnancy and FO feeding decreased ERalpha abundance in luminal epithelium. Prostaglandin H synthase-2 (PGHS-2) protein was elevated in pregnant cows and localized to the luminal epithelium. Both FO and bST treatments reduced staining intensity of PGHS-2 protein. Concentrations of prostaglandin E synthase mRNA were elevated in either cyclic or pregnant cows in response to bST, whereas bST decreased prostaglandin F synthase mRNA in pregnant cows. Uterine lumen fluids had more PGF2alpha and prostaglandin E2 in pregnant than cyclic cows. Uterine lumen fluids of bST-P cows contained more prostaglandin E2 than those from no bST-P cows. In summary, both pregnancy and bST altered endometrial gene expression, and cyclic cows responded differently to bST than pregnant cows. Feeding FO modulated PR, ERalpha, and PGHS-2 expression and distribution among endometrial cell types in a manner that may favor establishment and maintenance of pregnancy.

Animals↗
PubMed2006

Effect of estradiol benzoate or GnRH treatment prior to superstimulation in CIDR-treated, Korean native cows (Bos taurus).

The objective of this study was to evaluate the effectiveness of superovulatory protocols by synchronizing the emergence of the follicular wave using estradiol benzoate (EB) or GnRH in CIDR-treated, Korean cows. Sixty-six cows were used in the study and these were divided into three groups. The standard group comprised cows that were between days 8 and 12 of their estrous cycle (n=22). The remaining 44 cows, at all other stages of the estrous cycle, received CIDR and were assigned to two treatment groups that received either 2mg EB (EB-CIDR group, n=22) or 100 microg GnRH (GnRH-CIDR group, n=22) 1 day after CIDR insertion. Gonadotropin treatment began between the 8th and 12th days of the estrous cycle in the standard group, 5 days after EB injection in the EB-CIDR group, and 3 days after GnRH injection in the GnRH-CIDR group. All cows were superovulated with porcine FSH (pFSH) twice daily, with the dose (total 28 mg) decreasing gradually over 4 days. On the 5th and 6th injections of pFSH, 25 and 15 mg doses of PGF(2alpha) were administered. CIDR was withdrawn at the 7th pFSH injection and the cows received 200 microg GnRH at 24h after CIDR withdrawal. Cows were artificially inseminated twice at 36 and 48 h post-CIDR withdrawal and embryos were recovered 7 days after the 1st insemination. The numbers of preovulatory follicles (22.9-28.2), ovulated preovulatory follicles (17.6-21.7) and CL (15.9-17.9) detected by ultrasonography did not differ among groups (P>0.05). Similarly, the numbers of total ova (6.7-10.0), transferable embryos (4.0-6.0), degenerate embryos (1.1-1.8) and unfertilized ova (1.3-4.3) did not differ among groups (P>0.05). Progesterone and estradiol concentrations during superovulation treatments and at embryo recovery were also the same in all groups (P>0.05). We conclude that in CIDR-treated Korean native cows, superovulatory treatments that follow administration of either EB or GnRH (at any stage of the estrous cycle) result in both a superovulatory response and embryo yield comparable to conventional superovulation protocols.

Animals↗
PubMed1997

Absence of response to oestrus induction and synchronization treatment is related to lipid mobilization in suckled beef cows.

Energy status, follicular growth, oestradiol and LH secretion were investigated in 17 suckled Charolais cows synchronised 59.0 +/- 3.6 days after calving with a 10 day ear implant containing 3 mg of Norgestomet. The cows received 3 mg of Norgestomet and 5 mg of oestradiol valerate by IM injection at implant insertion (day 0) and 600 IU PMSG at implant removal (day 10). They were artificially inseminated (AI) 48 and 72 h after implant removal. Energy status was assessed by measuring weekly plasma concentrations of non-esterified fatty acids (NEFA), beta hydroxy-butyrate (BHB), glucose and insulin 7 weeks before AI. Progesterone plasma concentrations were measured during the same period to assess the presence of a functional corpus luteum. Follicular growth was followed daily by ultrasonography from day -3 to day 13. Oestradiol secretion was measured on day -3, day 6 and day 10 from five hourly samples. Oestradiol and LH plasma concentrations were measured hourly from 29 to 48 h after implant removal for seven cows. Cows were checked for pregnancy by ultrasonography 45 days after AI. Pregnant cows (P) were compared with non-pregnant cows (NP) for energy status, follicular growth, and oestradiol secretion by split-plot ANOVA. Two cows (11.8%) were cyclic before treatment, seven ovulated after treatment (41.2%) and five were found pregnant 45 days after AI (29.4%). There was no difference in body condition score and body weight between P and NP cows on day 0 (2.5 +/- 0.2 and 685 +/- 24 kg vs 2.5 +/- 0.1 and 670 +/- 13 kg; P > 0.05). Mean plasma NEFA concentrations before treatment were significantly lower in P than in NP cows (218 +/- 29 mu eq/L vs 279 +/- 18 mu eq/L; P < 0.05). No significant differences between P and NP cows were found for BHB, glucose and insulin concentrations. P cows presented more medium sized follicles (5 mm < or = diameter < 10 mm) than NP females during the period of observation (2.65 +/- 0.19 vs 2.50 +/- 0.12; P = 0.05). Plasma oestradiol concentrations were not different between P and NP cows on day -3 (8.4 +/- 0.7 pg/mL vs 7.7 +/- 0.4 pg/mL, P > 0.05), day 6 (10.4 +/- 0.6 pg/mL vs 9.8 +/- 0.4 pg/mL, P > 0.05) but were higher in P than in NP cows on day 10 (10.9 +/- 0.6 pg/mL vs 7.8 +/- 0.4 pg/mL; P < 0.05). After implant removal, oestradiol secretion only increased in P cows and a LH peak occurred whereas no increases in oestradiol (11.0 +/- 0.4 pg/mL vs 6.3 +/- 0.3 pg/mL, P < 0.05) and LH (6.0 +/- 0.5 ng/mL vs 1.2 +/- 0.5 ng/mL, P < 0.05) secretion were observed in NP cows. The conclusion was that follicular growth, oestradiol secretion, ovulation and pregnancy rate after oestrus synchronisation treatment are related to mobilization of energy stores before treatment in suckled beef cows in the same body condition score.

Animals↗
PubMed1998

Effect of acute treatment with progesterone on the timing and synchrony of ovulation in Bos indicus heifers treated with a norgestomet implant for 17 days.

The aim of the present study was to develop a treatment protocol for the precise synchronization of oestrus that would avoid the development of persistent dominant ovarian follicles. Bos indicus heifers, in which oestrous cycles had been presynchronized, were allocated randomly, according to the day of their oestrous cycle, to one of five treatment groups. All heifers received a subcutaneous ear implant containing 3 mg of norgestomet for 17 days starting on day 0 and an injection of an analogue of prostaglandin F2 alpha on days 0 and 4. Heifers in group 1 (control group; n = 7) received no other treatment, while heifers in groups 2 (n = 8), 3 (n = 7), 4 (n = 7), and 5 (n = 7) received a single progesterone-releasing controlled internal drug release device (CIDR) for 24 h on days 10, 12, 14 and 16, respectively. Treatment with a single CIDR delayed the mean time of ovulation and the day of emergence of the ovulatory follicle in heifers treated on days 14 and 16 compared with control heifers (P < 0.05). There was less variation in the interval to ovulation in heifers treated on day 10 compared with other treated heifers (P < 0.05). The variation among heifers in the day of emergence of the ovulatory follicle and the age of the ovulatory follicle at ovulation was less for all groups treated with a CIDR than for the control group (P < 0.05). The duration of dominance and variation in the duration of dominance of the ovulatory follicle was less in heifers treated with a CIDR device on days 10 and 16 than for control heifers (P < 0.05). Mean age (days from emergence to ovulation) of the ovulatory follicle did not differ among treatment groups (P > 0.05). Concentrations of LH and oestradiol decreased coincident with increased concentrations of progesterone on the days of CIDR treatment in treated compared with control heifers (P < 0.02) but increased again after removal of the CIDR. A smaller proportion of follicles in the growing phase of follicular development at the time of CIDR treatment become atretic compared with follicles that had reached a plateau phase of follicular growth (14.3% (1/7) versus 90.5% (19/21), respectively; P < 0.001). It was concluded that acute treatment with progesterone can influence the growth pattern of ovarian follicular development. However, the effect varies with the stage of ovarian follicular development. Short term treatment with progesterone 7 days before the end of a 17 day period of norgestomet treatment resulted in precise synchrony of ovulation without the ovulation of a persistent dominant ovarian follicle.

Animals↗
PubMed2002

Comparison of three approaches for synchronization of ovulation for timed artificial insemination in Bos indicus-influenced cattle managed on the Texas gulf coast.

Our objectives were to compare the relative efficacies of three protocols designed to synchronize ovulation for timed artificial insemination (AI) of predominantly Brahman-influenced cows and heifers. In Exp. 1, 273 Brahman x Hereford (F1) cows at three locations were stratified by BW, body condition score (BCS), age, and days postpartum and assigned randomly to three treatments: 1) Syncro-Mate-B (SMB), 2) norgestomet-prostaglandin (NP), and 3) Ovsynch. The management goal required that cows have a minimum BCS of 5 and be at least 36 d postpartum (PP) at treatment onset. However, final results included 23 cows (8.4%) whose BCS fell below 5. In Exp. 2, 286 pubertal beef heifers were stratified by BW and BCS and allocated randomly to the three treatments. Heifers were predominantly Brahman crossbred (n = 265; Brahman x Hereford, F1; Santa Cruz) or purebred Brahman-influenced (Santa Gertrudis) with a smaller number (n = 21) of Hereford heifers also included. For both experiments, SMB treatment consisted of a 9-d norgestomet ear implant plus an estradiol valerate/norgestomet injection on d 0. Norgestomet-prostaglandin-treated females were implanted with a SMB implant without the estradiol valerate/norgestomet injection at the time of implant insertion and received 25 mg prostaglandin F2alpha (PGF) i.m. 2 d before implant removal. Ovsynch consisted of 100 microg GnRH i.m. on d 1, 25 mg PGF i.m. on d 8, and a second GnRH injection on d 10. Beginning on d 9, calves were removed for 48 h in Exp. 1. Cattle in SMB and NP groups in both experiments were timed-inseminated 48 to 54 h after implant removal and at 12 to 24 h after the second GnRH injection (Ovsynch). Timed AI conception rates did not differ between the SMB (45.1%) and Ovsynch (42.4%) groups; however, conception rate in the NP group tended (P < 0.12) to be lower overall than in the other groups due to a reduced (P < 0.05) conception rate in cows that were < 60 d PP at treatment onset. Conversely, timed-AI conception was greatest (P < 0.056) in NP (54.7%) compared with SMB (40.4%) and Ovsynch (39.1%) for heifers in Exp. 2. We conclude that in mature, suckled beef cows with Brahman genetic influence, SMB and Ovsynch perform similarly when cow eligibility relies primarily on BCS and minimum days PP. The NP treatment results in lower conception in cows < 60 d PP compared with SMB and Ovsynch. However, in nulliparous Brahman-influenced heifers that are confirmed to be pubertal, NP may be superior to the other two treatments for timed AI.

Age Factors↗
PubMed2001

Plasma inhibin A in heifers: relationship with follicle dynamics, gonadotropins, and steroids during the estrous cycle and after treatment with bovine follicular fluid.

The relationship between follicle growth and plasma inhibin A, FSH, LH, estradiol (E), and progesterone was investigated during the normal bovine estrous cycle and after treatment with steroid-free bovine follicular fluid (bFF) to arrest follicle development. In the first study, four heifers were monitored over three prostaglandin (PG)-synchronized cycles. Blood was collected every 2-8 h, and ovaries were examined daily by ultrasonography. Inhibin A was measured using a modified enzyme-linked immunosorbent assay that employed a new monoclonal antibody against the alpha subunit of bovine inhibin. Plasma inhibin A ( approximately 50 pg/ml before luteolysis) rose steadily during the induced follicular phase (P < 0.05) to a peak ( approximately 125 pg/ml) coincident with the preovulatory E/LH/FSH surge. After ovulation, inhibin A fell sharply (P < 0.05) to a nadir ( approximately 55 pg/ml) coincident with the secondary FSH rise. During the next 3 days, inhibin A increased to approximately 90 pg/ml in association with growth of the new dominant follicle (DF). Plasma E also rose twofold during this period, whereas FSH fell by approximately 50%. Inhibin A was negatively correlated with FSH (r = -0.37, P < 0.001) and positively correlated with E (r = 0.49, P < 0.0001). Observations on eight cycles (two cycles/heifer), in which growth of the ovulatory DF was monitored from emergence to ovulation, showed that the first-wave DF (DF1) ovulated in three cycles and the second-wave DF (DF2) in five cycles. After PG, plasma inhibin A and E increased similarly in both groups, with concomitant falls in FSH. In the former group, the restricted ability of DF1 to secrete both inhibin A and E was restored after luteolysis. Results indicate that dynamic changes in the secretion of both E and inhibin A from the DF contribute to the fall in FSH during the follicular phase and to the generation and termination of the secondary FSH surge, both of which play a key role in follicle selection. In the second study, bFF (two dose levels) was administered to heifers (n = 3-4) for 60 h starting from the time of DF1 emergence. Both doses suppressed FSH (P < 0.05) and blocked DF1 growth to the same extent (P < 0.01), although inhibin A levels were only marginally raised by the lower dose (not significant compared to controls). The high bFF dose raised (P < 0.001) inhibin A to supraphysiological levels ( approximately 1 ng/ml). A large "rebound" rise in FSH occurred within 1 day of stopping both treatments, even though the inhibin A level in the high-dose bFF group was still approximately threefold higher than that in controls. This indicates that desensitization of gonadotropes to inhibin negative feedback is a contributory factor, together with reduced ovarian output of E, in generation of the post-bFF rebound in FSH.

Animals↗
PubMed1998

Measurement of inhibin-A (alpha beta A dimer) during the oestrous cycle, after manipulation of ovarian activity and during pregnancy in ewes.

A new two-site ELISA was validated for ovine plasma and used to measure circulating inhibin-A concentrations during a synchronized oestrous cycle in four ewes and throughout pregnancy in six ewes. Inhibin A concentrations were also determined in four ewes during chronic treatment with a GnRH agonist and after subsequent exposure to pregnant mares' serum gonadotrophin (PMSG) to stimulate ovarian follicular development. Concentrations of FSH, LH, oestradiol and progesterone were determined by radioimmunoassay. The detection limit of the inhibin-A ELISA was approximately 50 pg ml-1 and no significant crossreaction was observed with a range of related molecules including activin-A, inhibin-B, activin-B, follistatin and alpha 2-macroglobulin. Inhibin-A concentrations were below the detection limit in plasma from hypophysectomized and ovariectomized ewes. During the oestrous cycle, plasma inhibin-A concentrations (approximately 0.3-0.4 ng ml-1) did not vary during the follicular phase whereas plasma oestradiol increased approximately tenfold. After the preovulatory LH/FSH surge, inhibin-A fell to a nadir (approximately 0.15 ng ml-1) coincident with the peak of the postovulatory FSH rise. During the next 2 days, FSH concentrations fell to basal values as inhibin-A concentrations increased (P < 0.05) to a peak (approximately 0.5 ng ml-1) 3 days after the preovulatory LH/FSH surge. Over the following 3 days, FSH values increased again (P < 0.05) as inhibin-A concentrations fell to approximately 0.25 ng ml-1 (P < 0.05). Chronic GnRH agonist treatment suppressed FSH concentrations by about 50%, while inhibin-A and oestradiol concentrations fell below detection limits. Within 2 days after the PMSG injection, concentrations of inhibin-A (approximately 4.5 ng ml-1) and oestradiol (approximately 20 pg ml-1) had increased to very high values, while FSH concentrations had been reduced by a further 50%. Plasma concentrations of inhibin-A and FSH were similar to those in nonpregnant ewes during the first 60 days of gestation, but inhibin-A values fell markedly (sevenfold; P < 0.01) between days 60 and 90, coincident with a twofold decrease in FSH (P < 0.05). Inhibin A and FSH concentrations remained low for the remainder of gestation and were positively correlated throughout pregnancy (r = 0.48; P < 0.005). These observations support an endocrine feedback role for ovarian inhibin-A and oestradiol in controlling the secondary (postovulatory) FSH surge in ewes, but indicate that an increase in oestradiol is responsible for the characteristic reduction in FSH during the early to mid-follicular phase. The reduced secretion of FSH from mid- to late pregnancy cannot be attributed to increased inhibin-A secretion by the feto-placental unit, but most likely reflects increased steroid secretion from this source.

Analysis of Variance↗
PubMed2005

Restoration patterns for luteinising hormone and ovarian function following treatment with GnRH agonist implants (deslorelin) for 7, 14 or 21 days in cycling dairy cows.

Continuous GnRH agonist treatment of cows results in downregulation of GnRH responsiveness and a state of induced anoestrus. Inducing anoestrus in a precisely controlled manner could have several potential applications in dairy herd management. However, relatively little is known regarding the processes involved in restoring reproductive normality following an induced anoestrus. This study describes an experiment that was conducted to examine patterns of recovery of LH release and follicle growth in non-lactating Holstein cows immediately following cessation of treatment for 7, 14 or 21 days with a deslorelin implant. Oestrus cycles were synchronized at 7 days intervals and a deslorelin implant inserted in every cow 13 days after detected oestrus so that a group had implants for either 21 days (n = 9), 14 days (n = 10) or 7 days (n = 9). On the day of implant removal every ovarian follicle greater than 4 mm in diameter was ablated using ultrasound guided vacuum needle aspiration in an attempt to standardize follicle sizes. Daily ovarian ultrasound examinations were performed on each cow until 35 days after implant removal and again at 45, 59 and 74 days. A subgroup of four cows randomly selected from each treatment group had frequent serial blood samples collected over 8 h at 4 and 10 days after implant removal for LH profiling. There was no significant effect of treatment duration on any LH parameter and results were pooled. Mean LH pulse amplitude increased by 67% between 4 and 10 days after implant removal (0.34 ng/ml versus 0.57 ng/ml; 4 days versus 10 days post-implant, P < 0.001). Mean pulse frequency remained unchanged between the two samplings (5.9 pulses versus 6.9 pulses per 8 h; 4 days versus 10 days post-implant, P > 0.1). Smoothed mean LH concentrations were unaffected by treatment duration or time (0.36 ng/ml versus 0.41 ng/ml; 4 days versus 10 days post-implant, P > 0.1). The pattern of follicle growth and ovulation did not differ significantly between treatment durations and pooled means were used for comparative descriptions. The emergence of a new follicle wave could be detected beginning at 4 days after implant removal (mean 7.9 +/- 0.8 days). After emergence, a period of rapid follicle growth generally ensued with signs of oestrus occurring when the follicle reached 12.3 +/- 0.5 mm and ovulation when mean follicle diameter was 13.1 +/- 0.7 mm at 13.6 +/- 1.5 days after implant removal. Oestrus preceded ovulation in all cases where ovulation subsequently occurred. The mean interovulatory interval after implant removal was similar amongst groups (18.2 +/- 1.3 days). Follicle growth could be categorized into three groups based on the time to emergence and fate of the first wave DF. Spontaneous recovery was characterised by ovulation of the newly emerged DF. Failure to ovulate the first DF was associated with the formation of a persistent follicle by 35 days after implant removal with some 25% (7/28) of cows showing persistent follicles. Delayed emergence (>14 days after implant removal) was detected in 11% (3/28) of cows, but when a follicle did eventually emerge it was seen to ovulate normally. The correlation between number of days to first oestrus expression and LH pulse amplitude on 4 days post-implant removal was significant (R2 = 44%, P < 0.05). A model was then proposed for the restoration of reproductive function following GnRH agonist removal. In conclusion, duration of deslorelin treatment was associated with only small changes in LH and follicle parameters. The formation of persistent follicles delayed the recovery of more cows than delayed emergence of a new follicle wave after implant removal.

Animals↗