Induction of short-term pseudopregnancy in gilts by the administration of estradiol benzoate or estradiol dipropionate to achieve ovulatory synchronization for embryo collection

A study was conducted to investigate whether ovulation in gilts could be synchronized for embryo collection by the administration of estradiol benzoate (EB) or estradiol dipropionate (EDP) to induce pseudopregnancy, followed by the treatment with prostaglandin F_2α (PGF_2α) on 10 days after. Ten gilts each received a total of 20 mg of EB or EDP on Day 10 or EB on Day 10 and 14 to induce pseudopregnancy (Day 0 = onset of estrus). Donors received PGF_2α 10 or 15 days (as a control) after the first administration of estrogens and subsequently eCG and hCG, and were then inseminated artificially. The embryos were collected 7 days after the administration of hCG, and assessed for embryo yield and their developmental stages. All protocols resulted in good embryo yield (9.8–13.2 embryos in average), and the embryos showed average ability to develop to the expanded blastocyst stage (3.29–4.03 as developmental scores) without any significant differences among the protocols. These results suggest that the administration of PGF_2α 10 days after the treatment of gilts with EB or EDP would allow synchronization of ovulation and embryo collection, as well as shortening the period from estrus detection to embryo collection, thus improving embryo collection efficiency.     

Characterization of blood flow and the effects of exogenous estradiol benzoate on residual follicles formed after ultrasound-guided transvaginal follicle aspiration in cattle

Background: Removal of the follicular content by ultrasound-guided transvaginal follicle aspiration(TVFA) may fail to induce immediate atresia and loss of function, resulting in the occurrence of residual follicles(RF). The aims of this study were to characterize the blood flow in RF and to determine the effects of the treatment with estradiol benzoate on RF fate. Lactating, cyclic Holstein-Gir crossbred cows were used. In Experiment 1, follicular wave emergence(D0) was synchronized in cows(n = 10) and follicular growth was then monitored by transrectal ultrasonography from D0 to D8, followed by TVFA of the largest follicle present on the ovaries 24 h later. Color Doppler ultrasound imaging was used to examine blood flow on the follicular wall, which was recorded immediately before and every 12 h after TVFA, up to 72 h. In experiment 2, cows(n = 22) were randomly allocated to receive either 2 m L of saline i.m.(Control group, n = 11) or 2 m L estradiol benzoate i.m.(EB group, n = 11)immediately after TVFA. Ovaries were scanned every 12 h to confirm the presence and to measure the diameter of RF. The contents of the RF, if present, were collected 72 h after the first TVFA, using the same aspiration procedures.Follicular fluid from original follicles and RF were stored at-20 °C until hormonal assays.Results: In Experiment 1, there was no reduction(P 0.05) of blood flow in the remaining follicle walls after TVFA and maximum blood flow values were observed at 49.5 ± 19.7 h post-TVFA. In Experiment 2, formation of RF after TVFA was proportionally similar between Controls(5/9) and EB(5/10) cows. Also, RF diameter did not differ between groups(P 0.05). Nonetheless, the content of RF from cows in the EB group had lower(P = 0.0004)estradiol(E2) concentration and lower(P = 0.0005) E2:P4 ratio compared with Controls.Conclusions: In conclusion, 1) the persistence of vascularization in the remaining follicle wall may contribute to the formation of RF after follicle aspiration, and 2) the treatment with estradiol benzoate does not prevent formation of RF,but does reduce their estradiol production.     

Genetic partitioning of variation in ovulatory follicle size and probability of pregnancy in beef cattle

The objectives of this research were to partition variation in ovulatory follicle size into genetic and nongenetic components and to assess the utility of ovulatory follicle size as an indicator trait associated with reproductive success in beef cattle. Data were collected during the years 2002 to 2005 from 780 beef females that ranged in age from 1 to 12 yr (mean of 2.4 observations per female). Data were analyzed with a multiple trait Gibbs sampler for animal models to make Bayesian inferences from flat priors. A chain of 500,000 Gibbs samples was thinned to every 200th sample to produce a posterior distribution composed of 2,500 samples. Heritability estimates (posterior mean +/- SD) were 0.16 +/- 0.03 for follicle size and 0.07 +/- 0.02 and 0.02 +/- 0.01 for pregnancy rate as a trait of the female and service sire, respectively. Posterior means of genetic correlations were all <0.10, with 0.00 contained within the respective 90% probability density posterior intervals. Results indicate that whereas follicle size is of greater heritability than pregnancy rate, its usefulness to improve reproductive rate is greatest as an ancillary phenotype in multiple trait selection.     

The effects of administering estradiol benzoate plus progesterone during the growth or static phases of the dominant follicle in CIDR-treated lactating dairy cows

We studied the effects of administering estradiol benzoate (EB) plus progesterone (P4) as part of a CIDR-based protocol during the growth or static phases of dominant follicle development on follicular wave emergence, follicular growth, synchrony of ovulation and pregnancy rate following CIDR withdrawal, treatment with PGF(2alpha) and GnRH, and fixed-time artificial insemination (TAI). Forty-one previously synchronized lactating Holstein dairy cows were randomly allocated to three treatment groups. The control group (n=14) received a CIDR on the third day after ovulation only (Day 0). The two treatment groups were administered CIDRs comprising 2 mg EB and 50 mg P4 either on the third (T1, n=14) or eighth day (T2, n=13) after ovulation (Day 0). All cows received PGF(2alpha) after CIDR removal on Day 7, GnRH on Day 9, and TAI 16 h after GnRH treatment. The proportion of cows with follicular wave emergence within 8 days of treatment differed (P<0.01) among the control (14.3%), T1 (85.7%), and T2 groups (92.9%). However, the mean intervals between treatment and wave emergence were not significantly different. There were significant differences in the diameters of the dominant follicles on Day 7 (P<0.01) and in preovulatory follicles on Day 9 (P<0.01), with the largest follicles observed in the control group and the smallest follicles observed in the T2 group. In contrast, the numbers of cows showing synchronous ovulation after GnRH treatment (92.9 to 100.0%) and pregnancy following TAI (46.2 to 50.0%) were similar between the treatment groups. The results showed that, irrespective of the phase (growth or static) of the dominant follicle, administration of 2 mg EB plus 50 mg P4 to CIDR-treated lactating dairy cows induced consistent follicular wave emergence and development, synchronous ovulation after GnRH administration, and similar pregnancy rates following TAI.     

The influence of exogenous pituitary growth hormone on the ovulatory response to PMSG and hCG and the LH response to estradiol benzoate in prepubertal gilts

Two experiments were performed to examine the influence of exogenous growth hormone on the reproductive axis in gilts. Experiment one employed 26 Yorkshire × Landrace prepubertal gilts, which were selected at 150 d and 86.5 ± 1.5 kg bodyweight (BW) and assigned equally to two treatments. Gilts received injections of either porcine growth hormone at 90 μg/kg BW, or vehicle buffer, from 150 to 159 d. At 154 d gilts received 500 IU PMSG, followed 96 hr later by 250 IU hCG. Gilts were slaughtered at 163 days and their ovaries recovered to determine ovulatory status. In each treatment, gilts failed to show any ovarian response to PMSG/hCG. All remaining control gilts ovulated and their ovaries appeared morphologically normal. In gilts receiving exogenous growth hormone, fewer ovaries (4/11, P<.01) appeared morphologically normal. The ovaries of all other growth hormone injected gilts had very large (12–25 mm) non-luteinized follicles. In experiment two, 20 prepubertal Yorkshire × Landrace gilts were selected at 138 days and 85 kg BW. These gilts received injections of growth hormone at 90 μg/kg BW (n=9) or vehicle (n=11) from 138 to 147 days. At 143 days, all gilts were given an injection of estradiol benzoate (EB) at 15 μg/kg BW. Blood samples were taken at the time of EB injection, at 24 and 36 hr and then at 6 hr intervals until 78 hr. All samples were assayed for serum LH concentrations. The EB induced LH peak height was lower (P<.04) in gilts receiving exogenous growth hormone than in controls. The results presented indicate that the daily injection of growth hormone at 90 μg/kg BW reduced the estradiol-induced release of LH in addition to reducing the number of corpora lutea in gonadotrophin stimulated gilts.     

Concentration of receptors for estradiol and progesterone in canine endometrium during estrus and diestrus

Receptors for estrogen and progesterone were measured in cytosols prepared from specimens of canine endometrium obtained at late proestrus, day 4 of estrus, day 2 of diestrus, and at 10 day intervals from days 10 through 80 of diestrus. Twenty nine adult bitches were used, with 2 to 4 dogs used at each time point. Concentrations of estradiol receptors measured in endometrial cytosols from late proestrus through day 10 of diestrus were similar (mean +/- SEM: 9.9 +/- 2.2, 10.5 +/- 1.2, 16.3 +/- 1.6, and 16.2 +/- 2.9 pmol/g of tissue at proestrus, day 4 of estrus, days 2 and 10 of diestrus, respectively). As serum concentrations of progesterone increased during early diestrus, the concentration of estradiol receptors decreased and were significantly (P less than 0.05) lower on days 30 (4.9 +/- 1.3 pmol/g of tissue) and 40 (3.7 +/- 0.6 pmol/g of tissue) of diestrus. After day 40 of diestrus, when serum concentrations of progesterone were approaching basal concentrations, the concentration of estradiol receptors increased and remained significantly (P less than 0.05) higher from days 60 to 80 of diestrus (day 60, 13.4 +/- 2.9; day 70, 15.7 +/- 1.7; day 80, 19.8 +/- 2.4 pmol/g of tissue). As observed for estrogen receptors, the concentration of endometrial receptors for progesterone also gradually increased from late proestrus (4.9 +/- 1.3 pmol/g of tissue) to day 2 of diestrus (6.4 +/- 0.3 pmol/g of tissue).(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicle-stimulating hormone isoforms and plasma concentrations of estradiol and inhibin A in dairy cows with ovulatory and non-ovulatory follicles during the first postpartum follicle wave

Following parturition, all cows display a wave of ovarian follicular growth, but a large proportion fail to generate a preovulatory rise in estradiol, and hence fail to ovulate. Follicle-stimulating hormone (FSH) exists as multiple isoforms in the circulation depending on the type and extent of glycosylation, and this has pronounced effects on its biological properties. This study examined differences in plasma FSH, estradiol, and inhibin A concentrations, and the distribution of FSH isoforms in cows with ovulatory or atretic dominant follicles during the first postpartum follicle wave. Plasma FSH isoform distribution was examined in both groups during the period of final development of the dominant follicle by liquid phase isoelectric focusing. Cows with an ovulatory follicle had higher circulating estradiol and inhibin A concentrations, and lower plasma FSH concentrations. The distribution of FSH isoforms displayed a marked shift toward the less acidic isoforms in cows with ovulatory follicles. A higher proportion of the FSH isoforms had a pI>5.0 in cows with ovulatory follicles compared to those with atretic follicles. In addition, cows with ovulatory follicles had greater dry matter intake, superior energy balance, elevated circulating concentrations of insulin and insulin-like growth factor-I, and lower plasma nonesterified fatty acids. The shift in FSH isoforms toward a greater abundance of the less acidic isoforms appears to be a key component in determining the capability for producing a preovulatory rise in estradiol, and this shift in FSH isoforms was associated with more favorable bioenergetic and metabolic status.     

Manipulation of follicle development to ensure optimal oocyte quality and conception rates in cattle

Over the last several decades, a number of therapies have been developed that manipulate ovarian follicle growth to improve oocyte quality and conception rates in cattle. Various strategies have been proposed to improve the responses to reproductive biotechnologies following timed artificial insemination (TAI), superovulation (SOV) or ovum pickup (OPU) programmes. During TAI protocols, final follicular growth and size of the ovulatory follicle are key factors that may significantly influence oocyte quality, ovulation, the uterine environment and consequently pregnancy outcomes. Progesterone concentrations during SOV protocols influence follicular growth, oocyte quality and embryo quality; therefore, several adjustments to SOV protocols have been proposed depending on the animal category and breed. In addition, the success of in vitro embryo production is directly related to the number and quality of cumulus oocyte complexes harvested by OPU. Control of follicle development has a significant impact on the OPU outcome. This article discusses a number of key points related to the manipulation of ovarian follicular growth to maximize oocyte quality and improve conception rates following TAI and embryo transfer of in vivo- and in vitro-derived embryos in cattle.     

Experimental use of prostaglandin F2 alpha to synchronize estrus in cattle]

Experiments were conducted for the use of prostaglandin to synchronise oestrus in dairy cows. One single dose of 5 mg F2-Alpha prostaglandin was administered via the intrauterine route. The reference values of the control groups were matched by first insemination following induced oestrus among the best of the test animals which were inseminated twice. These values reached 72 and 96 hours following application accounted for 46.7 and 50.0 per cent. The yellow body phase, a prerequisite for oestrus production by prostaglandins, had been induced in the test animals by cycle blocking, using gestagen. Possible use of the method in a bio-engineering context for large-scale oestrus synchronisation in cows is discussed.     

Influence of the length of progestagen treatment and the time of oestradiol benzoate application on the ovulatory follicle size and ovulation time in anoestrous and cyclic beef cows

Previous research from our laboratory in beef cattle suggests that the pre-ovulatory follicle size, maturity and subsequent susceptibility to gonadotropin are influenced by the length of progestagen treatment in artificial insemination programme in beef cows. To test this hypothesis, two experiments were conducted. In experiment 1, 35 anoestrous beef cows received an intravaginal sponge containing 200 mg of medroxyprogesterone acetate. The treatment lasted for 7 (n = 12), 8 (n = 11) or 9 (n = 12) days. Half of the animals in each group were injected with 0.7 mg of oestradiol benzoate (EB) at device removal (0 h) and the other half 24 h later. In experiment 2, 38 cycling beef cows were treated with the same protocols as in experiment 1. Ultrasound examinations were performed to determine the follicular diameter at device removal (dominant follicle), interval to ovulation and ovulatory follicle diameter. The dominant follicle of anoestrous cows with progestagen for 7 days (8.4 ± 1.6 mm) resulted smaller (p < 0.05) than the cows treated for 8 (10.5 ± 1.6 mm) and 9 days (10.6 ± 1.2 mm). However, regardless of the length of the treatments, ovulation time after device removal was longer (p < 0.05) when EB was injected 24 h after withdrawal than at 0 h in anoestrous cows (EB0 = 52.7 ± 4.0 h; EB24 = 70.8 ± 6.2 h) and in cyclic cows (EB0 = 50.0 ± 21.0 h; EB24 = 73.0 ± 20.0 h). In anoestrous cows, the treatment with progestagen for 9 days and EB at 24 h increased the diameter of the ovarian follicle (p = 0.033) but did not affect the diameter of the ovulatory follicle in cyclic cows. In conclusion, increasing the length of progestagen treatment for 8 or 9 days compared to 7 days increased the diameter of the dominant follicle, in anoestrous and cyclic beef cows. Oestradiol benzoate administered at device removal resulted in a shorter interval from device removal to ovulation compared with EB injection 24 h after the end of a progestagen treatment.     

Relationship between size of the ovulatory follicle and pregnancy success in beef heifers

Previous research indicated that the size of the ovulatory follicle at the time of insemination significantly influenced pregnancy rates and embryonic/fetal mortality after fixed-timed AI in postpartum cows, but no effect on pregnancy rates was detected when cows ovulated spontaneously. Our objective was to evaluate relationships of fertility and embryonic/fetal mortality with preovulatory follicle size and circulating concentrations of estradiol after induced or spontaneous ovulation in beef heifers. Heifers were inseminated in 1 of 2 breeding groups: (1) timed insemination after an estrous synchronization and induced ovulation protocol (TAI n = 98); or (2) AI approximately 12 h after detection in standing estrus by electronic mount detectors during a 23-d breeding season (spontaneous ovulation; n = 110). Ovulatory follicle size at time of AI and pregnancy status 27, 41, 55, and 68 d after timed AI (d 0) were determined by transrectal ultrasonography. Only 6 heifers experienced late embryonic or early fetal mortality. Interactions between breeding groups and follicle size did not affect pregnancy rate (P = 0.13). Pooled across breeding groups, logistic regression of pregnancy rate on follicle size was curvilinear (P < 0.01) and indicated a predicted maximum pregnancy rate of 68.0 +/- 4.9% at a follicle size of 12.8 mm. Ovulation of follicles < 10.7 mm or > 15.7 mm was less likely (P < 0.05) to support pregnancy than follicles that were 12.8 mm. Ovulatory follicles < 10.7 mm were more prevalent (28% of heifers) than ovulatory follicles > 15.7 mm (4%). Heifers exhibiting standing estrus within 24 h of timed AI had greater (P < 0.01) follicle diameter (12.2 +/- 0.2 mm vs. 11.1 +/- 0.3 mm) and concentrations of estradiol (9.9 +/- 0.6 vs. 6.6 +/- 0.7) and pregnancy rates (63% vs. 20%) than contemporaries that did not exhibit behavioral estrus. However, when differences in ovulatory follicle size were accounted for, pregnancy rates were independent of expression of behavioral estrus or circulating concentration of estradiol. Therefore, the effects of serum concentrations of estradiol and behavioral estrus on pregnancy rate appear to be mediated through ovulatory follicle size, and management practices that optimize ovulatory follicle size may improve fertility.     

Incorporation of estradiol benzoate to CIDR protocol improves the reproductive responses in crossbred dairy heifers

The present study was designed to determine the effect of estradiol benzoate (EB) on reproductive response following a controlled internal drug release (CIDR) protocol in crossbred (Sahiwal × Friesian) dairy heifers. In the first trial, a total of 100 crossbred dairy heifers were treated with CIDR protocol for 7 days and injected with the PGF_2α on day 6. After 24 h of CIDR removal, one group (EB?=?50) was injected with estradiol benzoate whereas the other (control?=?50) remained untreated. Estrus intensity and response were recorded visually and ovulation rate was recorded by ultrasonography. All heifers were artificially inseminated at 48 and 60 h following CIDR removal. Heifers were scanned for pregnancy within days 30–40 of artificial insemination (AI). In the second trial, two subgroups of heifers were included to observe the estrus and ovulatory events. The results of the first trial revealed that estrus response was achieved 100% in both the treatment groups. Estrus intensity (2.9?±?0.1 vs. 2.0?±?0.7) and ovulation rate (100 vs. 88%) differed significantly (P?<?0.05) between the EB and control groups. However, a tendency for higher pregnancy per AI was observed (54 vs. 36%; P?=?0.07) in EB than that in control groups. The results of the second trial revealed that a significantly (P?<?0.05) shorter estrus and earlier ovulatory events were observed in EB-treated heifers. It is concluded that the incorporation of estradiol benzoate to the CIDR protocol is helpful to improve the estrus signs and enhance the ovulation and the pregnancy per AI in crossbred dairy heifers.     

A comparison of effects of zearalenone and estradiol benzoate on reproductive function during the estrous cycle in gilts

Effects of estradiol benzoate (EB) and zearalenone (Z) on luteal maintenance and plasma hormone concentrations were studied in 45 gilts. Gilts were allocated to receive either 20 mg Z, 2 mg EB or no treatment (C) on d 1 to 5 (T1), 6 to 10 (T2) or 11 to 15 (T3) of an estrous cycle (five per treatment). Onset of estrus was designated as d 0 of the estrous cycle. Zearalenone was added to the daily ration and EB was administered via an intramuscular injection. Blood samples were collected every 10 min over a 4-h period on the first 2 d prior to onset of treatment; the first, third and fifth days of treatment; and the first two and the fifth day after the end of the treatment periods. Gilts receiving EB and Z during T2 and T3 had longer (P less than .05) inter-estrous intervals than C gilts. The range in inter-estrous intervals for Z and EB treatments was 28 to 74 and 27 to 63 d, respectively. Mean plasma progesterone concentrations were elevated (P less than .05) during T2 and T3 in EB and Z-treated gilts when compared with C females. Estradiol benzoate treatment during T2 and T3 reduced (P less than .05) mean plasma luteinizing hormone (LH) concentrations more than C or Z treatments. Mean plasma concentrations of 13, 14-dihydro-, 15-keto-prostaglandin F2 alpha (PGFM) during T3 were higher (P less than .05) in C and Z gilts on d 13 and 15 post-estrus when compared with EB gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

A progestin-based treatment with a high dose of estradiol benzoate normalizes cyclic changes in endometrial EGF concentrations and restores fertility in repeat breeder cows

The objective of the present study was to examine the efficacy of a progestin-based treatment with a high dose of estradiol benzoate (EB) to normalize the epidermal growth factor (EGF) profile in the uterine endometrium and restore fertility in repeat breeder cows. Repeat breeder cows without peaks in their endometrial EGF concentrations on Days 3 and 14 of the estrous cycle were used throughout the study. The effect of 1 (standard dose), 2.5 and 5 mg of EB in a progestin-based treatment protocol (EB1, EB2.5 and EB5 treatments, respectively; n=5 for each group) on endometrial EGF concentrations was first examined. The EB1 and EB2.5 treatments in the repeat breeder cows produced a suppressed response in endometrial EGF compared with EB1 treatment in the fertile controls (n=5) and failed to restore the normal EGF profile during the next estrous cycle. However, EB5 treatment produced an increase in EGF concentrations similar to the fertile controls and normalized the endometrial EGF profile. The effects of the EB1 and EB5 treatments (n=30 for each group) on the endometrial EGF profile and fertility were then examined in the repeat breeder cows. The proportion of cows, with an EGF profile normalized by the treatments was higher in the EB5 group (66.7%) than in the EB1 (30.0%) and untreated control (13.3%; n=30) groups (P<0.01). The pregnancy rates of the cows having a normal EGF profile after treatment in the EB1 and EB5 groups were similar (88.9 and 85.0%, respectively) and higher than those of the cows having an abnormal profile within the same groups (19.0 and 30.0%, respectively, P<0.01). In summary, the endometrial response to EB in terms of the EGF concentration was suppressed in repeat breeder cows. A high dose (5 mg) of EB in a progestin-based treatment was found to be effective for restoration of a normal EGF profile and fertility in repeat breeder cows having lesser endometrial EGF concentrations on Days 3 and 14.     

Influence of metabolic hormones and nutrition on ovarian follicle development in cattle: practical implications

Nutrition has long been known to have a profound influence on reproductive performance of female cattle, but the underlying mechanism remains poorly understood. Whilst early investigations focused on the modulation of nutrition on hypothalamic-pituitary axis, more recent studies have tested the hypothesis that metabolic hormones as nutritional signals exert a direct effect at the ovarian level. In cattle, treatment with recombinant bovine somatotrophin (rGH) significantly increases the population of small ovarian follicles. This is associated with increases in circulating concentrations of insulin and insulin-like growth factor-I (IGF-I). Subsequent studies, both in vitro and in vivo, have highlighted the importance of IGF-I and/or insulin acting in synergy with FSH and LH. More recently, we demonstrated that feeding heifers with 200% maintenance requirements for a short period significantly increases circulating insulin concentrations and population of small ovarian follicles. Based on these findings, our recent work has aimed at addressing some practical problems in cattle production. Firstly, we showed that both rGH pretreatment and increased dietary intake significantly enhance the response to standard superovulatory regimes. Secondly, we have demonstrated that feeding a diet to increase circulating insulin concentrations during the early lactation can advance the first ovulation postpartum and increase conception rate to the first service in dairy cows. In summary, nutrition influences ovarian follicle development in cattle possibly through changes in metabolic hormones. These interactions can be manipulated to improve reproductive performance.     

The use of estradiol benzoate to manage lambing period in ewes bred at synchronized estrus

The induction and management of lambing was studied in a commercial flock of ewes. These ewes were bred at synchronized estrus and managed to lamb three times in 2 yr, with lambings occurring in the sequence of February, September and June. Ewes treated with 2 mg of estradiol benzoate in oil (i.m.) at approximately 17 d prior to term did not lamb in response to treatment. When 2 mg estradiol benzoate was given at 0800 on d 142 of gestation, lambing occurred an average of 37 +/- 5 h later in ewes treated in June or September and at 70 +/- 10 h later in ewes treated in February. The comparable figures for oil-treated ewes (control) were 70 +/- 5 h and 116 +/- 9 h, respectively. Altering the time of day of injection or using two injections 24 h apart did not alter the time for injection to lambing. Increasing the dose of estradiol benzoate to 15 mg decreased the time from injection to lambing (43 +/- 7 h, ewes treated in February) but also increased the incidence of dystocia (50% incidence). The incidence of dystocia averaged 8% for all other treatments and controls. Death loss of lambs to 1 wk of age was 12% and was not affected by treatment. The length of the lambing period was reduced from 9.0 +/- 3.3 d in control ewes to 3.6 +/- 1.1 d in induced ewes. A single injection of 2 mg estradiol benzoate given at 0800 on d 142 of gestation or a double treatment, with injections at 2000 on d 142 and 143, resulted in 53% of ewes lambing between 0800 and 1600 on each day of the lambing period, compared with 33% in control ewes.