Fertility in the mare after repeated transvaginal ultrasound-guided aspirations

Ovum pick-up (OPU) by transvaginal ultrasound guided aspiration (TUGA) is a procedure applied in equine-assisted reproduction programs such as oocyte transfer and in vitro embryo production. Despite a large number of studies reporting that it is a repeatable and safe technique, little information is available about the effect of repeated punctures on fertility of mares. Moreover, even if flushing follicles improves the oocyte recovery rate, to our knowledge the efficiency of flushing estrous and diestrous follicles has not been evaluated. The aims of the present study were (1) evaluate if repeated TUGAs negatively effects fertility and (2) investigate the influence of flushing the follicular cavity (as compared to aspiration only-unflushed) on the recovery rate from follicles of different sizes and in different stages of the estrous cycle. Seventy-six TUGAs were carried out on 20 mares during the breeding season; 153 follicles were aspirated and 31 oocytes were recovered (20.3% per follicle; 40.8% per TUGA attempt). Of the 76 aspirations, 52 were carried out during estrus and 24 in diestrus. Flushing the follicular cavity significantly increased (P < 0.01) the oocyte recovery rate from estrous follicles (13/28, 46.4% flushed versus 3/24, 12.5% aspirated only) but not (P > 0.05) from diestrous follicles of different diameters (3/30, 10% flushed versus 2/36, 5.6% aspirated only for follicles <2 cm in diameter; 6/20, 30% flushed versus 4/15, 26.7% aspirated only for follicles > or =2 cm in diameter). Mares underwent ultrasonic examinations after every aspiration and no alteration was found with the exception of two mares in which the corpus luteum (CL) did not form following aspiration of estrous follicle. Of the 20 mares involved in this study, 10 were artificially inseminated with fresh semen from a single fertile stallion at the first spontaneous heat following the previous aspiration. Of the 10 inseminated mares, 7 were found to be pregnant 16, 30 and 50 days after artificial insemination (AI), indicating that repeated TUGAs did not adversely affect fertility.     

Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows

The aim of the present study was to confirm earlier findings, obtained with a small number of animals, that gonadotropin-releasing hormone (GnRH) can shorten corpus luteum functional life when it is administered 24 h after cloprostenol (PG) treatments given 7-9 days after estrus. In addition, the effects of two treatments, PG alone or PG + GnRH given before mid-diestrus, on signs of estrus were studied. Sixty cows in farm conditions were used in the experiment. Eight days after natural estrus, they were given an intramuscularly (i.m.) treatment of cloprostenol (0.5 mg). The animals were then divided into two groups. One group (n = 25) received an i.m. treatment of gonadorelin (0.1 mg) 24 h after the PG treatment (PG + GnRH group), while another group (n = 35) served as controls without any further treatment (PG group). Estrous signs were recorded. Progesterone concentrations were measured from samples of whole milk. No short cycles were observed in the PG group, whereas 33% of the cows in the PG + GnRH group exhibited premature luteal regression (P < 0.05). Cloprostenol treatment on Day 8 had no effect on the intensity of the estrous signs. Instead, GnRH treatment 24 h after PG treatment weakened the estrous signs significantly (P < 0.01). It is concluded that GnRH administration 24 h after a PG treatment given 8 days after estrus can cause short estrous cycles in some cows on an individual basis.     

Effects of norgestomet, altrenogest, and/or estradiol on follicular and hormonal characteristics of late transitional mares

Two experiments were conducted to investigate the effect of norgestomet and altrenogest, alone or in combination with estradiol, on late transitional mares. In the first experiment, 32 mares were assigned to four treatment groups: controls (C), those treated with 1.5 mg of norgestomet (N1), 3.0 mg norgestomet (N2) or 26 mg altrenogest (AT). Treatments were initiated during the months of April and May and given daily for 15 d. During treatment, altrenogest suppressed estrous behavior and diameter of the largest follicle, whereas norgestomet had no effect at either dose. The rise in serum luteinizing hormone (LH) levels following the withdrawal of altrenogest treatment was significantly greater than that for the other three groups. In the second experiment, 24 late transitional mares were assigned to three treatments: controls (C), those receiving 26 mg altrenogest (AT) daily, or 26 mg altrenogest plus 10 mg estradiol (AE) daily for 16 d. Both altrenogest treatments suppressed estrous behavior and follicular growth compared with controls. However, suppression of follicular activity was significantly greater for the combined steroid treatment. Following treatment, the interval to ovulation and estrus was longer for the combined steroid group. We concluded that: 1) norgestomet at a dose up to 3.0 mg per day had no effect on follicular activity, estrous behavior or serum LH levels in late transitional mares, 2) estradiol combined with altrenogest had greater suppressive activity on follicular growth than altrenogest alone, and 3) the greater suppression by the combined steroid treatment had no advantage over altrenogest alone on induction of estrus and ovulation in late transitional mares.     

Effect of phenylbutazone (PBZ) on luteolysis in the mare induced by uterine biopsy

Uterine biopsy in the mare on day 4 post-ovulation causes an acute inflammatory reaction which results in premature luteolysis. In this study, seven mares (4 to 6 years of age) were used in a switchback experimental design to test the hypothesis that in the mare parenterally administered PBZ will block luteolysis induced by uterine biopsy on day 4 post-ovulation. All mares were allowed two normal estrous cycles (range 18 to 24 days). On the first day of estrus of the third estrous cycle each mare was intravenously given 2 grams PBZ (treatment) or 10 ml 0.9% saline (control) daily until signs of estrus were exhibited. The day of ovulation (day 0) was determined by rectal palpation and subsequently verified by peripheral plasma progesterone concentrations. On day 4 following ovulation all mares were subjected to uterine biopsy, and subsequent estrus detection was performed daily using an andro-genized gelding. A total of 19 estrous cycles (ten for PBZ treatment and nine for controls) were evaluated. Mean number of days (+/-SE) from uterine biopsy to induced estrus was 5.00+/-0.16 for control cycles and was significantly different (P<0.025) when compared with 9.20+/-0.34 days for treatment cycles. Results of this study suggest that PBZ can block luteolysis in the mare induced by uterine biopsy on day 4 post-ovulation, possibly as a result of accumulating PBZ in acutely inflamed uterine tissue and inhibiting prostaglandin synthesis.     

Effect of progesterone impregnated vaginal sponges and PMSG administration on estrus synchronization in mares

Estrus synchronization trials with mares were carried out using progesterone impregnated vaginal sponges and pregnant mare serum gonadotropin (PMSG) injections. In Phase 1, 10 non-pregnant, non-lactating mares were administered 1 g progesterone via vaginal sponges (5 x 6 cm) without regard to stage of estrous cycle. Sponges were replaced on day 7 of trial for an additional seven days. On day 12, PMSG (1000 IU, IM) was administered to five mares (Group A); five control mares (Group B) received no injections. There was no difference (P>.05) in estrus synchronization between Group A and Group B. Total sponge retention was 75%. In Phase 2, 11 non-pregnant, non-lactating mares were administered 2 g progesterone via vaginal sponges (10 x 6 cm) without regard to stage of estrous cycle. Sponges were replaced on day 7 of trial for an additional seven days. Estrus behavior was exhibited in 54.5% of mares by day 19. Total sponge retention was 95.4%. There was no difference (P>.05) in estrus synchronization or sponge retention between Phase 1 and Phase 2. The larger Phase 2 sponges showed less (P<.01) posterior movement within the vagina than the smaller Phase 1 sponges.     

The effects of dose and duration of administration of pFSH during the first follicular wave on the ovulation rate of beef heifers

Four experiments were carried out to examine the effects of administration of pFSH (Vetrepharm) from Day 3 of the estrous cycle in conjunction with PG on Day 5 on follicular populations and ovulation rate in heifers. In Experiment 1, 47 heifers were allocated to 1 of 4 treatment groups (n = 11 to 12 per group): a) control, b) 1.5 mg pFSH, c) 2.0 mg pFSH or d) 2.5 mg pFSH until estrus. Heifers assigned to the 3 treatments had a higher ovulation rate than the controls (P < 0.05). In Experiment 2, 45 heifers were allocated to 1 of 5 treatment groups (n = 8 to 10 per group): a) control, b) 1.0 mg pFSH until PG, c) 1.0 mg pFSH until estrus, d) 1.5 mg pFSH until PG or e) 1.5 mg pFSH until estrus. From Day 5, heifers assigned to pFSH treatments had more large follicles than the controls (P < 0.05). There was no effect of treatment on the incidence of twin ovulations. In Experiment 3, 43 heifers were assigned to 1 of 3 groups (n = 11 to 16 per group): a) control, b) 1.0 mg pFSH until estrus or c) 1.5 mg pFSH until estrus. At slaughter, 14 d after administration of PG, the incidence of twin ovulations was 0/11, 7/16 and 8/16 for Groups a, b and c, respectively (P = 0.011). In Experiment 4, pFSH (1.5 mg) was administered to 3 groups during the development of the first dominant follicle: a) growth phase (n = 19); b) static phase (n = 17); and c) decline phase (n = 17). All pFSH-treated heifers had a higher ovulation rate than the controls (P < 0.05); heifers assigned to Group c had a higher ovulation rate than those in Groups a or b (P < 0.05). More heifers assigned to Group c (7/17) superovulated than in the other 2 groups (P < 0.05). In conclusion, administration of 1.0 or 1.5 mg pFSH twice daily beginning at Day 3 of the estrous cycle in association with the induction of luteolysis increased the ovulation rate significantly when pFSH treatment was continued to onset of estrus. The ovulation rate and the occurrence of multiple ovulations were significantly higher when pFSH was administered at the time that the first dominant follicle was in decline.     

Influence of reproductive stage at PRID insertion on synchronization of estrus and ovulation in mares

In the present study, we investigated the effects of reproductive status, size of follicles and plasma progesterone concentrations of mares at PRID insertion on the efficacy of the treatment, estrous cycle patterns, plasma concentrations of progesterone and LH. The progesterone-releasing device (PRID) was administered intravaginally to 28 Haflinger mares for 11 days at different reproductive stages: anestrus (n=6), estrus (n=11) and diestrus (n=11). Plasma concentrations of progesterone at insertion (Day 1) of PRID differed among treatment groups (anestrus: 0.2-0.6 ng mL(-1), estrus: 0.2-0.5 and diestrus: 1.6-10.8 ng mL(-1); P<0.001). Total secretion of progesterone (area under curve (AUC)) during treatment period revealed highest values in diestrus (38.2+/-3.1 ng mL(-1)h(-1)) followed by estrus (25.1+/-2.7) and anestrus (21.0+/-0.4 ng mL(-1)h(-1); P<0.05). Progesterone area under curve (AUC) was positively correlated with initial progesterone concentrations (R=0.5; P<0.05), but it did not correlate with the interval from PRID removal to ovulation. Plasma concentrations of LH during treatment period, were significantly lower in anestrous mares (184.6+/-28.6 ng mL(-1)h(-1)) when compared to estrous and diestrous mares (349.7+/-53.3 and 370.5+/-40.3 ng mL(-1)h(-1); P<0.05). Follicular size at PRID insertion had no effects on the intervals from PRID removal to subsequent estrus and ovulation. Follicle diameters at removal of PRID were significantly correlated with the interval from coil removal to estrus (R=-0.55, P<0.05) and ovulation (R=-0.72, P<0.0004) in cyclic mares. In anestrus 0 of 6 (0%) mares, in estrus 5 of 11 (45.5%) and in diestrus 6 of 11 (54.5%) mares ovulated within a defined interval of 1 day before to 1 day after mean interval from PRID removal to ovulation. In cyclic mares, response to treatment was significantly higher when compared to anestrous mares: almost all mares responded with estrus and ovulation independent from the stage of the estrous cycle at the start of treatment. However, accuracy of synchronization was still unsatisfactory. In cyclic mares, the plasma progesterone concentrations at insertion of PRID seem to be more important for the efficacy of the treatment than the assignment to estrous cycle stages.     

Steroid priming shortens prostaglandin-based estrus synchronization program from 14 to 7 days in cattle

Single injection of estrogen and progesterone before prostaglandin (steroid priming) was used to shorten the prostaglandin-based estrus synchronization program. Sixty-five cyclic Sistani cattle, with parity ranging from 1 to 4 and postpartum period of >80 days were selected at unknown stages of the estrous cycle and assigned to 2 groups according to their age, weight and parity. Females in the control group (n=33; 58.4 +/- 4.3 months; 277 +/- 8 kg LW) received two consecutive injections of prostaglandin F2alpha analogue (500 microg; Cloprostenol, PG) 14 days apart (Day 0 = First PG injection). On Day 7, treated females (n=32; 60 +/- 4.8 months; 292 +/- 9 kg LW) were given an intramuscular injection of 100 mg progesterone and 2 mg estradiol benzoate followed by prostaglandin 7 days later, concurrent with the second PG injection of the control group. Estrus detection was carried out every 6 hours for 7 days, commencing from 24 hours after the last PG injection. Females that allowed to be mounted were identified (standing estrus) and inseminated with frozen semen 12 hours later. Pregnancy was diagnosed on Day 50 after AI through palpation per rectum. Data were analyzed using Chi-squared and t-test. The tightness of estrus synchrony (%), the interval from the end of treatment to estrus (h) and conception rates (%) were similar (P > 0.05) between control (69.6%, 77.7 +/- 5.96 h and 56.5%) and treatment (68.2%, 82.6 +/- 7.64 h and 54.5%) groups. In conclusion, steroid priming is an efficient way to shorten the prostaglandin-based estrus synchronization program from 14 to 7 days without compromising estrous response and fertility.     

Sexual behavior in ovariectomized and seasonally anovulatory pony mares (Equus caballus)

Ten ovariectomized (OVEX) and ten intact, but seasonally anovulatory (ANOV), pony mares were observed for sexual activity with five stallions, using a “harem group” social testing paradigm (two OVEX and two ANOV mares plus one stallion per group) for 15 consecutive daily tests lasting 20 min each. All mares in both conditions showed proceptive behavior in at least one test, all mares but one were mounted, and 14 of 20 mares received ejaculations. No statistical differences were found between the two conditions for any measure of proceptivity, copulatory activity, or days in estrus. The quality of estrus was judged to be equivalent to that displayed by periovulatory mares during their initial and terminal days of estrus, but less intense than that seen near ovulation. Mares in both groups were in estrus during approximately 60–70% of the tests and only 3 of the 20 mares were sexually refractory for more than five consecutive tests. Thus, the typical 2-week phase of sexual refractoriness seen in intact diestrous mares was absent in OVEX and ANOV mares, suggesting that the ovary plays a major role in actively suppressing estrous responses during the luteal phase of the cycle.     

Efficacy of a prostaglandin analogue in reproduction in the cycling mare.

A prostaglandin F analogue caused luteolysis in normal cycling non-lactating mares, and lactating mares (treated after the foal estrus). Effective doses ranged from 1.0 to 4.0mg given as a single subcutaneous injection 8–10 days after ovulation. A dose of 0.5mg was ineffective, hence the dose-response relationship was steep, indicative of a quantal type of response. Mares usually returned to estrus within 2–4 days and ovulated by 7 days after treatment. Mares bred naturally or by artificial insemination at the induced estrus and ovulation were fertile. The compound was without side-effects, and hence should be of value in manipulating the estrous cycle of the mare.     

Regulation of estrus and ovulation in mares with progesterone or progesterone and estradiol biodegradable microspheres with or without PGF2alpha

A single injection of a microsphere preparation, designed to deliver 1.25 gm progesterone and 100 mg estradiol-17beta at a controlled rate, for a duration of 12 to 14 days, produces accurate control of estrus and fertile ovulations in mares. Theatment is followed by PGF(2)alpha injection 14 days after steroid injection. The objectives of the present study were to determine whether estradiol added to the progesterone treatment or PGF(2)alpha administered at the end of the steroid treatment regimen, would improve synchronization of estrus and ovulation. A total of 45 cyclic horse mares was randomly assigned to 1 of 5 treatment groups as follows: Group 1 (control, n=9) sterile microsphere vehicle + sterile PGF(2)alpha vehicle 14 days after treatment with microsphere vehicle; Group 2 (n=9) progesterone and estradiol microspheres + PGF(2)alpha 14 days after treatment with microspheres; Group 3 (n=9) progesterone and estradiol microspheres + PGF(2)alpha vehicle 14 days after treatment with microspheres; Group 4 (n=9) progesterone + PGF(2)alpha 14 days after treatment with microspheres; and Group 5 (n=9) progesterone + PGF(2)alpha vehicle 14 days after treatment with microspheres. Addition of estradiol (P<0.05) or PGF(2)alpha (P<0.05) to the treatment regimen increased synchronization efficary by reducing variation in days to ovulation. All treatments significantly reduced variation in days to estrus compared with that of the controls; however, mares in the progesterone groups had an increased incidence of silent or shortened estrous behavior (<- 2 days) following treatment. Estradiol added to the treatment regimen increased (P<0.05) the number of mares with post treatment estrus > 2 days in duration compared with mares treated with progesterone (78 vs 33%, respectively). Therefore, estradiol and PGF(2)alpha each appear to reduce variation in days to ovulation while estradiol seems to promote better expression of posttreatment estrous behavior.     

Effect of prior FSH treatment on the estrus and ovulation responses to eCG in prepubertal gilts

The objective of this study was to determine the effect of pre-treatment of prepubertal gilts with FSH on the estrus and ovulatory responses to eCG injection at two ages. A total of 149 prepubertal Hypor gilts were selected at 150 days (n=76) or 180 days (n=73) of age and assigned to injection of 400 IU eCG plus 200 IU hCG (PG600), 600IU eCG alone (Folligon), pre-treatment with 72 mg FSH (Folltropin) administered as 6 x 12 mg injections at 12 h intervals with 600 IU Folligon 12h after last FSH injection, or non-injected controls. To facilitate detection of estrus, gilts were exposed to a mature boar for 15 min daily for 7 days. To determine ovulatory responses, blood samples were obtained on the day of injection and 10 days later and assayed for progesterone content. Following treatment at 150 days, one control gilt (5.3%) was deemed estrus but ovulation did not occur. Compared to treatment with Folligon alone, PG600 injection tended (P=0.1) to increase the estrus response (52.6% compared with. 26.3%) and increased (P<0.01) the ovulatory response (89.5% compared with. 47.4%). The estrous response in gilts pretreated with Folltropin was intermediate (42.1%) but the ovulatory response (47.4%) was the same as for Folligon alone. Following treatment at 180 days, two control gilts (10.5%) were deemed estrus and ovulation did occur in these gilts. There was no difference between hormone-treated groups for estrus or ovulatory responses, although the ovulatory response of PG600-treated gilts tended (P=0.1) to be greater than for the Folligon-treated group (89.5% compared with 66.7%), with Folltropin-pretreated gilts being intermediate (76.5%). These data demonstrate that the estrus and ovulatory responses of gilts were greater for PG600 than for Folligon and that while responses to PG600 were not affected by gilt age, for the combined Folligon groups, estrous response (P<0.02) and ovulatory response (P<0.05) improved with increased gilt age.     

Estrous cycle characteristics, luteal function, secretion of oxytocin (OT) and plasma concentrations of 15-keto-13,14-dihydro-PGF2alpha (PGF2alpha-metabolite) after administration of low doses of prostaglandin F2alpha (PGF2alpha) in pony mares

In the present study, the kinetics of the prostaglandin F2alpha (PGF2alpha)-metabolite 15-keto-13,14-dihydro-PGF2alpha after a single intramuscular application of various doses of the natural PGF2alpha dinoprost at Day 7 of the cycle in the mare were investigated. Effects of low doses on estrous cycle length and life span of corpus luteum were examined, because release of PGF2alpha is still under discussion to have detrimental influence on success rates of transcervical transfer of equine embryos. Eight Shetland pony mares were each randomly assigned to each of four treatments: (a) 0.8 mg/100 kg (group T1), (b) 0.4 mg/100 kg (group T2), (c) 0.2 mg/100 kg BM dinoprost i.m. (group T3), and (d) 1 ml physiological saline i.m. (group CO). Treatments were administered as single doses on Day 7 of the estrous cycle. Administration of dinoprost caused dose-dependent rises of plasma concentrations of PGF2alpha-metabolite, although values of individual mares showed great variation within groups. Prostaglandin treatments resulted in a distinct decrease of plasma progesterone concentrations to values between 1.6 and 7.9 ng/ml within 24 h. Treatment groups had significantly lower progesterone area under the curve (AUC: T1 942.8+/-175.9, T2 1050+/-181.2 and T3 1117+/-179.8 ng/ml/h) when compared with controls (CO 1601.9+/-227.6; t-test, P<0.05 ). There was a small, but significant negative correlation between AUC of progesterone and of PGF2alpha-metabolite ( R=-0.4; P=0.05 ). Administration of PGF2alpha caused secretion of oxytocin in three (T1, T2) and two (T3) mares out of eight ranging from 19.3 to 63.1 pg/ml. The AUC of oxytocin was positively correlated with AUC of PGF2alpha-metabolite ( R=0.4, P<0.05) and negatively correlated with AUC of progesterone ( R=-0.4, P<0.05). Administration of dinoprost yielded significantly shorter intervals from treatment to estrus and ovulation (values in parentheses), respectively, when compared with controls: T1 3.9+/-0.7 days ( 12.1+/-0.7 days), T2 4.5+/-0.6 ( 12.3+/-0.6 ), T3 4.9+/-0.5 ( 12.3+/-0.6 ), and CO 8.9+/-0.6 days ( 16.5+/-0.8 days) (t-test, P<0.01 ) (Fig. 2). Different doses of PGF2alpha caused similar effects. Data suggest that progesterone concentrations at applications influence efficacy of treatments more than doses administered, as demonstrated by their high correlation with estrous cycle patterns. It is important to note that differences we achieved are gradual and that all mares responded to treatment by luteolysis and premature estrus, regardless of doses applied.     

Progesterone and estradiol in biodegradable microspheres for control of estrus and ovulation in mares

Progesterone and estradiol 17-beta in poly (DL-lactide) microspheres were used to control estrus and ovulation in mares after luteolysis was induced by prostaglandin F(2)infinity. Mares were given a single intramuscular injection of biodegradable poly (DL-lactide) microspheres, 1 day following prostaglandin treatment, containing no hormones (control), 0.625 g progesterone and 50 mg estradiol (low dose), 1.25 g progesterone and 100 mg estradiol (medium dose), or 1.875 g progesterone and 150 mg estradiol (high dose; n=15 mares per group). Mares treated with the low dose had significantly longer intervals (P<0.05) to estrus and ovulation than the control mares; however, low dose mares had shorter intervals (P<0.05) to estrus than high dose mares and shorter intervals to ovulation than medium and high dose mares. Regression analysis indicated that the medium dose was sufficient for maximizing interval to ovulation while the high dose maximized interval to estrus. All groups of mares exhibited similar (P>0.05) post-treatment estrus lengths. A clinical response scoring system based on synchrony of both estrus and ovulation within a treatment group was also used to measure the effectiveness of treatments on control of estrus and ovulation. Clinical response scores did not differ (P>0.05) among treatment groups. Mares were randomly assigned for insemination at the beginning of the first post-treatment estrus. Rates for embryo recovery performed by uterine lavage 7 days post-ovulation did not differ (P>0.05) among groups. Concentrations of serum progesterone increased in mares receiving progesterone and estradiol microspheres. At 10 to 14 days post-injection of microspheres, progesterone concentrations were higher (P<0.05) and remained above 1 ng/ml in the mares receiving the high dose. Progesterone concentrations were also higher (P<0.05) on Days -3 to -1 (Day 0 = day of post-treatment ovulation) in mares receiving the high dose when compared to control mares. Gonadotropin concentrations were suppressed (P<0.05) in the medium and high dose groups.     

Intrauterine infusion of prostaglandin E2 and subsequent luteal function in cattle.

The objective was to evaluate the effect of intrauterine infusion of prostaglandin E2 (PGE2) on luteal function in cattle. Heifers and cows were randomly assigned after two normal estrous cycles to either PGE2 or control treatment groups. Females in Treatment A were infused with 1 mg of PGE2 once daily into the uterine horn ipsilateral to the corpus luteum between days 7-10 of the estrous cycle with a 0.25 ml plastic semen straw and an artificial insemination pipette. Females in Treatment B were similarly infused with 1 mg of PGE2 once daily in 20 ml of a carrier vehicle via a catheter on days 10 and 11 of the estrous cycle. Control animals were infused with the carrier vehicle using either a semen straw (Treatment C) or via a catheter (Treatment D) on the same days of the estrous cycle. Blood samples were collected daily to monitor plasma progesterone concentrations during the treatment period. Females infused with PGE2 on days 7-10 of the estrous cycle returned to estrus in a mean of 23.5 days (range 22-25 days) and were similar (P > 0.05) to those infused on days 10 and 11 which returned to estrus in 23.5 days (range 22-25 days). Animals similarly infused with carrier vehicle on the same days of the estrous cycle returned to standing estrus in 20.2 days (range 17-23 days). Plasma progesterone concentrations indicated an extended period of elevated progesterone concentrations in PGE2-treated animals compared with control animals. These results indicate that short term administration of PGE2 early in the estrous cycle may result in extended luteal maintenance.     

Study of sexual behaviours with different types of estrus synchronization protocols in Boer goats

There is still a lack of information on estrus synchronization in goats. Understanding the estrus synchronization protocols and the subsequent effects is important to improve the efficiency of assisted reproductive technologies (ARTs) and subsequently would improve the breeding procedures. This study will help in determining the most suitable estrus synchronization protocol and understand better the effect on the sexual behaviour and hormonal effects in goats. A total of 127 Boer does were used and divided into three groups with different duration of CIDR insertion intravaginally either for 14 (two groups) or 9 days (one group). Approximately 0.5 ml Estrumate^® (PG) was administered intramuscularly to all groups at CIDR removal, and only groups PMSG14 and PMSG9 were administered with 200IU of Pregnant Mare Serum Gonadotropin (PMSG) intramuscularly. Estrus signs were observed at 4 h intervals and blood samples were collected for progesterone and luteinizing hormone determination. The percentage of does in estrus within 24 to 72 h post CIDR removal was significantly higher (P<0.05) in groups with PMSG compared to the group without the PMSG. The numbers of does display estrus signs within 24 to 28 h post CIDR removal were significantly higher (P<0.05) in group shorter period (9 days) compared to groups with 14 days CIDR. The P4 concentrations at 24 hours post CIDR removals and LH concentration was not significantly different (P>0.05) in all groups. The time of the LH peak in the group without the PMSG was significantly delayed (P<0.05) when compared to group 9 days CIDR and administered with PMSG. It is recommended to use the treatment for 9 days CIDR since the estrous cycle can be shortened.     

Management of reproduction in dairy heifers based on the synchronization of estrous cycles

Israeli-Holstein breed dairy heifers (n = 571), 13 to 15 mo old, were utilized in two experiments. In Experiment 1, the reproductive performance of synchronized heifers was compared with that of untreated controls. The heifers in both groups were inseminated following the detection of estrus. In Experiment 2, all heifers were synchronized and inseminated following the detection of estrus. Half of the animals in this experiment also received one or two fixed-time inseminations 72 and 96 h after the last synchronization treatment. Synchronization of estrous cycles was performed by two prostaglandin F(2alpha) (PG) injections given 12 d apart. In the control group of Experiment 1, observation of estrous behavior and insemination of heifers detected in estrus were carried out daily throughout the experiment. In the synchronized groups of Experiments 1 and in 2, the management of reproduction consisted of estrus detection followed by the insemination of heifers in estrus carried out only during 6 d of every 3 wk. Five days following the second PG injection, 86% of the heifers were detected in estrus, 71% of them at 49 to 96 h after treatment. In Experiment 1, age at first insemination, age at conception, and conception rate were, respectively, 425 d, 446 d and 54% in the control group vs 432 d (P<0.02), 449 d and 62% in the PG-treated group. In Experiment 2, the respective figures were 436 d, 462 d and 59% in the group inseminated following the detection of estrus vs 427 d (P<0.002), 464 d and 51% (P<0.05) in the group in which heifers were inseminated at estrus and also received one or two fixed-time inseminations.     

The efficacy of recombinant equine follicle stimulating hormone (reFSH) to promote follicular growth in mares using a follicular suppression model

The efficacy of a recently engineered single chain recombinant equine follicle stimulating hormone (reFSH) was investigated in estrous cycling mares whose gonadotropins and follicular activity had been suppressed by concurrent treatment with progesterone and estradiol (P&E). Time of estrus was synchronized in 15 estrous cycling mares during the breeding season with prostaglandins F_2α (PGF_2α). The day after ovulation, mares were treated once daily with P&E for 14 days. Mares received a second injection of PGF_2α on day 6 of the synchronized estrous cycle to induce luteolysis. On day 8 post-ovulation mares were randomly assigned to three groups: small dose reFSH-treatment group (0.5 mg reFSH IV, twice daily); large dose reFSH-treatment group (0.85 mg reFSH IV twice daily); control group (saline IV, twice daily). reFSH treatment occurred concurrently with the last week of P&E treatment. After a follicle or cohort of follicles reached 35 mm in diameter, mares were injected with 0.75 mg of recombinant equine luteinizing hormone (reLH) to induce ovulation. Post-treatment ovulation was assessed. Daily blood samples were collected for analysis of FSH, LH, estradiol, progesterone, and inhibin by radioimmunoassay (RIA). On the first day of reFSH/saline treatment, blood samples were collected periodically from 1 h prior to treatment to 6 h post-injection via an indwelling jugular catheter to determine acute changes in FSH concentrations. Monitoring of follicular activity, estrus, and ovulation was performed daily by utilizing a stallion and transrectal ultrasonography.A difference (p ≤ 0.05) between the largest diameter follicle in the reFSH-treatment groups compared to controls occurred on day 14 post-ovulation, the day treatments ended, and the difference continued until day 21 post-ovulation. reFSH-treatment groups had larger (p ≤ 0.05) numbers of 20–29 mm follicles (days 13–18), 30–34 mm follicles (days 15–20) and ≥35 mm follicles (days 16–21) than controls. Mares treated with reFSH, at either dose, took less time (average: 2.95 ± 0.42 days) to develop 2–3 times more pre-ovulatory follicles than control mares (7.8 ± 0.51 days) (p ≤ 0.05). The number of ovulations between treated mares and controls were similar due to a greater incidence of ovulation failure in reFSH-treated mares. During reFSH treatment, concentrations of plasma FSH, inhibin and estradiol were greater (p ≤ 0.05) compared to control concentrations. Plasma LH concentrations in reFSH-treated mares were suppressed and did not exhibit the ovulatory surge of controls (p ≤ 0.05). Plasma progesterone concentrations were not different across groups.These findings demonstrate the specific effects of reFSH to increase number of total follicles including pre-ovulatory follicles in mares with endogenous pituitary gonadotropins and follicular growth suppressed by a regimen of P&E.     

Superovulatory response of river bufalo (Bubalus bubalis )

The ovarian response of 25 buffalo-cows was visually assessed, and their oviducts and uteri separately flushed 3 to 6 d post superovulatory estrus at slaughter. Ten buffalo-cows slaughtered on Days 5 and 6 were examined per rectum for corpora lutea (CL) and follicles > 8 mm prior to slaughter, and the estimate was compared later with the actual ovarian response. Five out of the ten buffalo-cows were nonsurgically flushed in vivo on Day 5 of the estrous cycle, a day before slaughtering, and as a result, six ova/embryos were recovered. After the flushing of the reproductive tract at slaughter, one more ovum was recovered from the uterus of each of the three buffalo-cows. As a result of treatment of three groups of five buffalo with 3000 IU pregnant mare serum gonadotrophin (PMSG) on Days 6, 10 or 14 of the estrous cycle, 3.8, 6.2 and 3.4 CL on the average were recovered, respectively (Experiment I). A mean number of 8.8 and 9.0 CL, respectively, was obtained in two groups of five buffalo each, after treatment with 40 mg of follicle stimulating hormone (FSH) on Day 10 of the stage of the estrous cycle (Experiment II) and 3000 IU PMSG regardless of the stage of cycle (Experiment III). The percentage of ova/embryos recovered in the three experiments was 32.8, 20.4 and 22.2, respectively.     

Ovarian response and endocrine changes in buffalo superovulated at midluteal and late luteal stage of the estrous cycle: a preliminary report

A study was designed to determine whether superovulatory and endocrine responses in buffalo differ when gonadotropin treatment is initiated at midluteal and late luteal stages of the estrous cycle. Twenty-eight buffalo were randomized into 4 groups (A, B, C and D). Buffalo in Groups A and B (n = 8 each) were superovulated with Folltropin (total dose 25 mg) and Lutalyse. Treatments in Group A were initiated between Days 8 to 10 (midluteal group) and in Group B between Days 13 to 15 (late luteal group) of the estrous cycle. Buffalo in Groups C and D (n = 6 each) were not superovulated and served as controls. Blood samples from all groups of buffalo were collected daily for plasma progesterone and estradiol determinations. The number of corpora lutea (CL) and unovulated follicles was recorded (following per rectum palpations) 5 or 6 d post-estrus. Buffalo in Groups A and B exhibited estrus in larger proportions and earlier (49.33 +/- 3.82 h and 46.67 +/- 2.46 h, respectively) than the control Groups C and D (77.33 +/- 5.33 h and 78.0 +/- 3.83 h, respectively). Mean number of CL was higher in Group B (3.38 +/- 0.46) than in Group A (2.25 +/- 0.75), however,the difference was not significant (P > 0.05). Plasma progesterone concentrations on the day of treatment were higher in late luteal superovulated and control groups than in midluteal superovulated and control groups. In both Groups A and B progesterone levels were significantly related (r = 0.78,0.76; P < 0.05) to the number of CL palpated after the superovulatory estrus. Progesterone levels on the day of estimation of ovarian response were approximately 4 times higher in Groups A and B than in Groups C and D. Peak estradiol concentrations were approximately twice as high in superovulated groups as in control groups.