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The present study evaluated the effect of progesterone supplementation during a double PGF2α injection synchronization protocol on pregnancy per artificial insemination (P/AI) and on the incidence of twin births. Seven hundred and eighty three dairy cows were synchronized with two injections of PGF2α 14 days apart, starting on day 35 postpartum for their first postpartum insemination. Six days before the second PGF2α injection, cows were treated either with a progesterone-releasing intravaginal device (PRID-Delta) and an intramuscular injection of 500mg of progesterone (n=387) or served as control (n=396) and did not receive progesterone treatment. Cows were inseminated 12h after being detected in estrus. Pregnancy was diagnosed 40-45 days later by transrectal palpation. Progesterone administration improved (P<0.05) the percentage of cows detected in estrus in multiparous [(192/255) 75% vs (161/267) 60%], but not in primiparous cows [93/132 (70%) vs 90/129 (70%)]. Progesterone treatment increased P/AI in multiparous [53/192 (28%) vs 27/161 (17%)] but not in primiparous cows [25/93 (27%) vs 29/90 (32%)]. The incidence of twin births tended to be lower (P=0.09) in cows treated with progesterone [1/74 (1%)] than in the control group [4/53 (7%)]. It is concluded that progesterone administration during estrus synchronization with two PGF2α injections in dairy cows improves estral response and increases P/AI in multiparous, but not in primiparous cows, and tended to decrease the incidence of twin births., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study compared the responses shown by lactating dairy cows to four different P4-based protocols for AI at estrus. Cows with no estrous signs 96 h after progesterone intravaginal device (PRID) removal were subjected to fixed-time AI (FTAI), and their data were also included in the study. In Experiment I, follicular/luteal and endometrial dynamics were assessed every 12 h from the beginning of treatment until AI. The estrous response was examined in Experiment II, and fertility was assessed in both experiments. The protocols consisted of a PRID fitted for five days, along with the administration of different combinations of gonadotropin releasing hormone (GnRH), equine chorionic gonadotropin and a single or double dose (24 h apart) of prostaglandin F2α. In Experiment I (40 cows), animals receiving GnRH at the start of treatment showed a significantly higher ovulation rate during the PRID insertion period while estrus was delayed. In Experiment II (351 cows), according to the odds ratios, cows showing luteal activity at the time of treatment were less likely to show estrus than cows with no signs of luteal activity. Treatment affected the estrous response and the interval from PRID removal to estrus but did not affect conception rates 28-34 days post AI. Primiparous cows displayed a better estrous response than multiparous cows. Our findings reveal acceptable results of 5-day P4-based protocols for AI at estrus in high-producing dairy cows. Time from treatment to estrus emerged as a good guide for FTAI after a 5-day P4-based synchronization protocol.

This study evaluated whether the administration of 50 and 100mg bovine somatotropin (bST) at the start of synchronization and at the time of natural mating in ewes improves the ovulation rate, embryonic development and pregnancy rate of transferred embryos. Forty-eight donors were assigned to three treatments: the bST-100 treatment (n=15) received 100mg bST at the start of synchronization and at natural mating, the bST-50 treatment (n=15) received 50mg bST on the same schedule as the previous group, and the control (n=18) did not receive any bST. Two embryos were transferred to each recipient (n=121): 35 received embryos from bST-100; 50 received embryos from bST-50, and 36 received embryos from the control. The superovulatory rate, percentage of recovered structures, cleavage rate, percentage of transferable embryos, embryo quality and development and pregnancy rate were analyzed using the GENMOD procedure of SAS. The number of corpora lutea and the cell number were analyzed using the GLM procedure of SAS. The insulin and IGF-1 concentrations were analyzed with ANOVA for repeated measures. The bST application did not affect the superovulatory rate, number of corpora lutea and recovered structures (P>0.05). The numbers of transferable embryos and embryos reaching the blastocyst were higher (P≤0.01) in the bST-50 (96.4±3.6% and 69.0±7.8%) than the bST-100 (93.0±4.5% and 27.2±38.9%) and control (87.7±5.4% and 50.4±6.4%) groups. The insulin and IGF-1 concentrations were higher (P<0.05) in the bST-treated groups, but the insulin concentration was higher (P<0.05) in the bST-100 group than in the bST-50 group. The pregnancy rate was similar (P=0.21) in ewes receiving embryos from the two treatments [bST-50, (70.0%); bST-100, (62.5%), and control, (56.6%)]. The administration of 50mg bST at the start of synchronization and at natural mating in superovulated ewes was concluded to enhance the proportion and development of transferable embryos. However, bST did not affect the pregnancy rate of transferred embryos., (Copyright © 2014. Published by Elsevier B.V.)

Plasma estradiol and progesterone (P4) concentrations during the peri-ovulatory period are positively correlated with pregnancy success in cattle. The aims of this study were to assess the effects of estrus occurrence and early diestrus P4 concentrations on pregnancy per timed-embryo transfer (P/TET). A total of 267 crossbred beef heifers [222 with corpus luteum (CL) and 45 without a CL but with a follicle >8mm at beginning of the estrous synchronization protocol) received an intra-vaginal P4 device and intramuscular administration of estradiol benzoate. Progesterone devices were removed 8 days later (Day 0), and heifers received d-cloprostenol, eCG and estradiol cypionate. Estrous behavior was monitored twice daily for 3 days after P4 device removal. Plasma P4 concentration was measured by radioimmunoassay at Day 7 and Day 9. At Day 9, heifers with a CL (n=236; i.e. submission rate of 85.5%; 236/276) undergoing TET received an in vitro-produced embryo. Heifers expressing a standing behavioral estrus had a greater P/TET than heifers that did not express a standing estrus [62.4% (106/170) compared with 47.0% (31/66)]. The probability of pregnancy was positively correlated with plasma P4 concentration at TET. When heifers were grouped by quartiles of P4 concentration at TET (Q1=0.64±0.16, Q2=1.70±0.04, Q3=2.90±0.07 and Q4=5.52±0.27ng/mL) the P/TET were 45.8% (Q1; 27/59)(c), 52.25% (Q2; 31/59)(bc), 66.1% (Q3; 39/59)(ab) and 67.8% (Q4; 40/59)(a). Additionally, heifers that became pregnant had greater P4 concentrations at TET (2.87±0.16ng/mL; n=137) than heifers that did not become pregnant (2.45±0.24ng/mL; n=99). No statistical difference was observed regarding P4 concentrations on Day 7, regardless of standing estrus or pregnancy status. In cattle, manifestation of estrous behavior and plasma P4 concentration at TET increase the probability of pregnancy in in vitro-produced embryo recipients., (Copyright © 2014. Published by Elsevier B.V.)

Anti-Müllerian hormone (AMH) has been correlated with phenotypic indicators of fertility. However, the effects of exogenous hormones used during estrus synchronization on AMH have not been evaluated. Therefore, the objective of this experiment was to determine whether concentrations of AMH at estrus are similar between a synchronized compared with a natural estrous cycle. Nulliparous dairy and beef heifers (n = 68) were synchronized with the Select Synch + controlled internal drug release (CIDR) protocol (GnRH + CIDR-7 d-CIDR removal + PG). Heifers were observed for expression of estrus every 6 h until 84 h after the injection of PG. Visual detection of the subsequent estrus, considered natural estrus, occurred every 6 h from day 16 to 24 after synchronized estrus. At the time of standing estrus, ovarian structures in heifers were evaluated by transrectal ultrasonography. Blood samples were collected at estrus for analysis of concentrations of AMH during the synchronized and natural estrous cycles. The GLM and CORR procedures of SAS were used to analyze data. Concentrations of AMH between natural and synchronized estrus were positively correlated (r = 0.67; P < 0.001). Mean concentration of AMH did not differ (P > 0.05) between the natural (0.0543 ± 0.0076 ng/mL) or synchronized (0.0428 ± 0.0076 ng/mL) estrous cycles. In conclusion, concentrations of AMH were similar between natural and synchronized estrous cycles. Concentrations of AMH in natural and synchronized estrous cycles were highly correlated within individual heifers and varied among heifers with beef heifers having increased (P < 0.05) concentrations of AMH compared with dairy heifers (0.0638 ± 0.01 and 0.0402 ± 0.01 ng/mL, respectively)., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The aim of this study was to demonstrate successful breeding of sows with a single fixed-time insemination following ovulation induction by buserelin, a GnRH analogue. In a first step, the optimal dose of buserelin (6, 10, or 16 μg) injected at 77 hours after weaning was determined in weaned sows (N = 15, 11, and 12, respectively) using its ability to induce an LH surge of similar magnitude as in control sows (N = 15) and induce ovulation. In 29/38 treated sows (76%), ovulation was induced and synchronized between 32 and 44 hours after injection, and the proportion of females ovulating during this time window was similar between groups at 73%, 73%, and 83% (6, 10, or 16 μg, respectively). Interestingly, whereas ovulation of 100% multiparous sows was induced and synchronized in the 32 to 44 hours posttreatment time window, successful induction was achieved in a lower proportion of primiparous sows (50%, 50%, and 67% following 6, 10, or 16 μg, respectively), the dose effect being nonsignificant. The magnitude of the LH surge was similar between control and treated sows, irrespective of the buserelin dose injected. Neither ovulation rate nor the number of good embryos on Day 5 postovulation differed between groups. Interestingly, the frequency of follicular cysts at slaughter was significantly affected by treatment (P < 0.05), being minimal and maximal in sows treated with 10 or 6 μg buserelin, respectively. In a second step, 419 sows from commercial herds in Spain, Germany, and France were randomly allocated to a control or treated group. The control sows were inseminated twice 12 ± 4 hours apart once estrus was detected. Treated sows received 10 μg buserelin at 86 ± 3 hours after weaning and were inseminated once 30 to 33 hours later. Farrowing rate of treated sows (87%, 166/192) was similar to that of control sows (84.5%, 169/200). Litter size was also similar between treated and control sows (13.6 ± 3.8 vs. 13.7 ± 3.2). In multiparous sows, neither duration of lactation nor magnitude of the fat loss during lactation significantly affected treatment effects. It is concluded that ovulation of weaned multiparous sows can be tightly synchronized by buserelin (10 μg) administration at 86 hours postweaning. This allows breeding once at a fixed time following buserelin injection while maintaining reproductive performance at a level similar to that of sows bred twice during estrus., (Copyright © 2013 Elsevier Inc. All rights reserved.)

We evaluated the cortisol response of adult female eland (n=8) that were handled in hydraulic chute daily or 3×/week. Females were divided into two groups and each group (n=4) successively received two estrous cycle synchronization treatments: (1) two injections of prostaglandin (PG-PG) F2α at 11 day intervals and (2) oral administration of altrenogest for 7 days and an injection of PGF2α on day 7 (Alt-PG). Blood samples were collected 3×/week during the synchronization (Synch) and expected luteal phase (Nonintensive) periods, and daily during the expected time of induced (Intensive 1) or natural (Intensive 2) estrus. Overall, mean cortisol levels were highest during Intensive 1, followed by Intensive 2, Synch and Nonintensive periods. Individual eland were the most significant source of variation for cortisol level. The frequency of handling and the synchronization treatment significantly affected cortisol levels in 3/8 and 4/8 females, respectively. In conclusion, in response to increased frequency of handling, eland cortisol levels rose transiently and returned to baseline within few days after more intensive handling. Thus, the eland females were tolerant to and recovered from the effects of repeated daily handling., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Insulin-like peptide 3 (INSL3), a major product of testicular Leydig cells, is also expressed by the ovary, but its functional role remains poorly understood. Here, we quantified expression of INSL3 and its receptor RXFP2 in theca interna cell (TIC) and granulosa cell compartments of developing bovine antral follicles and in corpora lutea (CL). INSL3 and RXFP2 mRNA levels were much higher in TIC than granulosa cell and increased progressively during follicle maturation with INSL3 peaking in large (11-18 mm) estrogen-active follicles and RXFP2 peaking in 9- to 10-mm follicles before declining in larger (11-18 mm) follicles. Expression of both INSL3 and RXFP2 in CL was much lower than in TIC. In situ hybridization and immunohistochemistry confirmed abundant expression of INSL3 mRNA and protein in TIC. These observations indicate follicular TIC rather than CL as the primary site of both INSL3 production and action, implying a predominantly autocrine/paracrine role in TIC. To corroborate the above findings, we showed that in vitro exposure of TIC to a luteinizing concentration of LH greatly attenuated expression of both INSL3 and its receptor while increasing progesterone secretion and expression of STAR and CYP11A1. Moreover, in vivo, a significant cyclic variation in plasma INSL3 was observed during synchronized estrous cycles. INSL3 and estradiol-17β followed a similar pattern, both increasing after luteolysis, before falling sharply after the LH surge. Thus, theca-derived INSL3, likely from the dominant preovulatory follicle, is detectable in peripheral blood of cattle, and expression is down-regulated during luteinization induced by the preovulatory LH surge. Collectively, these findings underscore the likely role of INSL3 as an important intrafollicular modulator of TIC function/steroidogenesis, while raising doubts about its potential contribution to CL function.

Previous studies reported increased fertility using Ovsynch for presynchronization before Ovsynch (Double-Ovsynch), as compared with presynchronization with two prostaglandin F(2α) (PGF(2α)) treatments before Ovsynch (Presynch-Ovsynch). This study compared ovarian follicular dynamics and hormone concentrations during Double-Ovsynch versus Presynch-Ovsynch. Lactating Holstein cows (N = 193) were assigned to one of two treatment groups: (1) Presynch (N = 93), two injections of PGF(2α) 14 days apart, followed by the Ovsynch-timed AI protocol 12 days later; and (2) Double-Ovsynch (N = 100), one injection of GnRH, PGF(2α) 7 days later, and GnRH 3 days later, followed by the Ovsynch-timed AI protocol 7 days later. All cows received the same Ovsynch-timed AI protocol: GnRH (G1) at 68 ± 3 days in milk (mean ± SEM), PGF(2α) 7 days later, and GnRH (G2) 56 hours after PGF(2α). Ultrasonographic evaluations of the ovaries and blood sampling were performed at G1, PGF(2α), G2, and 6 days after the G2 injection of the Ovsynch-timed AI protocol. Double-Ovsynch decreased the percentage of cows with low circulating progesterone (P4) concentrations (<0.50 ng/mL) at G1 (12.0% vs. 30.1%; P = 0.003) and increased the percentage of cows with medium P4 concentrations (0.50 > P4 ≤ 3.0 ng/mL) at G1 (80.0% vs. 57.0%; P < 0.01), and with CL at G1 (94.0% vs. 67.8%; P < 0.01). Double-Ovsynch also increased the percentage of cows with high P4 (>3.0 ng/mL) at PGF(2α) (88.0% vs. 76.3%; P = 0.04) and tended to increase average circulating P4 at PGF(2α) (3.52 ± 0.17 ng/mL vs. 3.09 ± 0.21 ng/mL; P = 0.11). Double-Ovsynch also tended to increase percentage of cows ovulating to G1 (80.0% vs. 69.9%; P = 0.11) and G2 (98.0% vs. 93.5%; P = 0.08). Thus, presynchronization of cows with Double-Ovsynch induced ovulation in noncycling cows and appeared to increase most aspects of synchronization during the Ovsynch protocol., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The study was aimed at induction/synchronization of estrus in postpartum anestrous Kankrej cows of zebu cattle maintained at an organized farm. The study included use of different hormone protocols, viz., Ovsynch, CIDR (controlled internal drug release), Ovsynch plus CIDR, and Heatsynch with estimation of plasma progesterone on days 0, 7, 9/11 (artificial insemination--AI) and on day 20 post-AI following fixed time insemination. Thirty selected anestrous animals were divided into five equal groups (four treatment and one control), and the findings were compared with the normal cyclic control group of six cows. All the protocols were initiated in cows with postpartum anestrous period of more than 4 months, considering the day of first GnRH injection or CIDR insertion as day 0. The animals were bred by fixed time artificial insemination. Pregnancy was confirmed per rectum on day 60 post-AI in non-return cases. The conception rates at induced/first heat in Ovsynch, CIDR, Ovsynch + CIDR, and Heatsynch protocols were 33.33, 66.66, 50.00 and 16.67%, respectively. The corresponding overall conception rates of three cycles post-treatment were 50.00% (3/6), 100.00% (6/6), 66.66% (4/6), and 50.00% (3/6). In normal cyclic and anestrous control groups, the pooled pregnancy rates were 83.33% (5/6) and 16.67% (1/6), respectively. The pooled mean plasma progesterone (nanograms per milliliter) concentrations were significantly (P < 0.05) higher on day 7 in Ovsynch (5.727 ± 1.26), CIDR (4.37 ± 0.66), Ovsynch plus CIDR (3.55 ± 0.34), and Heatsynch (5.92 ± 1.11) protocols as compared with their corresponding values obtained on days 0, 9/11 (AI), and on day 20 post-AI. In anestrous control group, the mean progesterone concentration at the beginning of experiment was 0.67 ± 0.33 ng/ml, which was at par with values of all other groups. The overall plasma progesterone levels on the day of initiating treatment were low in all groups, with smooth small inactive ovaries palpated per rectum twice at 10 days interval, suggesting that most of the animals used in the study were in anestrous phase. Mean (± SE) values of plasma progesterone (nanograms per milliliter) on day 20 post-AI were higher in conceived cows than the non-conceived cows of all the groups, but differed significantly (P < 0.05) only in normal cyclic group. These results suggest that use of different hormone protocols particularly Ovsynch, CIDR, and Ovsynch + CIDR may serve as an excellent tool for induction and synchronization of estrus and improvement of conception rate in postpartum anestrous Kankrej cows.

Our objective was to determine if repeated exposure of lactating dairy cows to human chorionic gonadotropin (hCG) would induce an antibody (Ab) response against hCG. Cows either received an hCG injection (hCG; n = 24, each given 2000 IU im) or no treatment (CON; n = 22) 18 days after a timed AI (TAI) and 7 days before initiation of Ovsynch for resynchronization of ovulation and TAI. A subgroup of cows continued in the experiment to receive a second hCG injection (n = 17) 35 days after the first exposure to hCG, whereas another subgroup served as controls (n = 9). Another subgroup of cows continued in the experiment to receive a third hCG injection (n = 11) 35 days after the second exposure to hCG, whereas cows not receiving hCG served as controls (n = 8). A binding radioimmunoassay was used to detect hCG antibodies in serum samples collected 0, 7, 14, 21, and 28 days after treatment. A positive Ab response (>6.2% bound) was defined as three standard deviations above CON binding. No cows had hCG antibodies at Day 0 before the first exposure to hCG. After the first hCG treatment, there was no difference (P = 0.52) between Ab positive cows in CON (0%) and hCG (4%) treatments. At the second hCG treatment, on Day 0 there was no difference (P = 0.65) between CON (0%) and hCG (6%) cows, whereas, more (P = 0.02) hCG cows (47%) were positive than CON cows (0%) within 28 days of the hCG injection. At the third hCG injection, hCG cows tended (P = 0.09) to have a greater percentage of Ab positive (36%) than CON cows (0%), whereas after the injection, a greater (P < 0.01) percentage of hCG cows were positive (hCG = 73% vs. CON = 0%). After the second and third exposure to hCG, 8 of 17 and 8 of 11 cows within the hCG group had greater percent Ab bound at 7, 14, 21, and 28 days after hCG than cows in CON and those with no Ab response. The greatest percent Ab binding occurred at 14 days after the second and third hCG exposure. We concluded that some but not all lactating dairy cows developed an Ab response after repeated exposure to hCG and that maximum response occurred within 14 days after hCG exposure., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The objectives were to (1) compare blood metabolites and reproductive outcomes in lactating dairy cows not inseminated before (early) and after (late) 100 d in milk (DIM) because of prolonged anovulation or anestrus; and (2) evaluate reproductive responses of cows ≤100 DIM to GnRH+PGF(2α) treatments after a fixed-time artificial insemination (AI; Ovsynch) or after induced estrus (Select Synch). In blood samples collected before initiating hormone-based breeding programs, anovular cows ≤ 100 DIM had the greatest serum total protein and globulin concentrations and the lowest tri-iodothyronine concentrations. Anovular and ovular cows >100 DIM had the greatest serum urea concentrations. Ovaries in cows (n=40) >100 DIM were examined by transrectal ultrasonography, and those without a detectable corpus luteum (CL; anovular) were given GnRH and then PGF(2α) 7 d later (Select Synch), whereas cows with a CL (ovular) were given 2 PGF(2α) injections 12 d apart. Cows were inseminated at observed estrus after the second or only PGF(2α) injection. More ovular (79%; 15/19) than anovular cows (24%; 5/21) were detected in estrus. No differences were detected between ovular and anovular cows in DIM at first AI, conception rate to first AI, cumulative pregnancy rates, number of services per conception, or days open. Cows (n=93) ≤100 DIM were assigned randomly to 3 treatments: (1) control (n=20) AI at estrus; (2) GnRH and then PGF(2α) on 7d (Select Synch; n=42) and monitored for signs of estrus for 5d and AI accordingly; or (3) 2 GnRH injections 9 d apart with PGF(2α) given 48 h before second GnRH injection and AI at 16h after the second GnRH injection (Ovsynch; n=31). Among cows ≤100 DIM, controls had more days to first service (149±16 d) than Select Synch cows (117±7 d). Ovsynch cows had the fewest days to first service (84±10 d) compared with control (149±16 d) and Select Synch (117±7 d) cows. Conception rates in control (25%) and Select Synch (26.2%) cows did not differ from those in Ovsynch cows (29%). Ovsynch cows had greater cumulative pregnancy rates and fewer days open than control (161±20 vs. 258±29 d), but did not differ from Select Synch (233±19d). Timed AI produced comparable fertility and superior cumulative pregnancy rates, fewer days to first service, and fewer days open than AI at observed estrus in cows inseminated ≤100 DIM., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The effect of gonadotropin-releasing hormone analogue (GnRH-A) or follicular aspiration at the onset of progesterone-based timed artificial insemination (TAI) on subsequent follicular growth and synchronization of ovulation was examined in early postpartum Japanese Black cows. A total of 40 (22 in Exp. 1 and 18 in Exp. 2) Japanese Black cows at 20-30 days postpartum were fitted with a progesterone releasing internal device (PRID) for 7 days, injected with a prostaglandin F2α analogue upon removal of the PRID and GnRH-A 48 h later, and inseminated 18 h after GnRH-A injection. In Exp. 1, the animals were divided into three groups (untreated control, GnRH-A injection or follicular aspiration) of different treatments on the first day of PRID insertion (day 0), and the synchronized ovulation rate in the follicular aspiration group (100%; 8/8) tended to be higher (P = 0.077) than that in the control group (42.9%; 3/7). In Exp. 2, follicular growth in the GnRH (n = 9) and follicular aspiration (n = 9) groups was monitored by ultrasonography. Four out of the nine animals in the GnRH group had a corpus luteum on either day 4 or day 7 (OV group), and the other five animals had no induced ovulation (NOV group). The diameter of the ovulatory follicle on day 9 in the OV group (1.44 ± 0.11 cm) tended to be greater (P = 0.078) than that in the NOV group (1.13 ± 0.07 cm). Follicular aspiration at the onset of PRID-based TAI of early postpartum Japanese Black cows, regardless of the resumption of ovarian cyclicity, tended to result in a higher rate of synchronization of ovulation than that of the untreated controls.

Objectives were to develop a timed artificial insemination (TAI) resynchronization program to improve pregnancy per AI and to evaluate responses of circulating progesterone and pregnancy-associated glycoproteins in lactating cows. Cows (n=1,578) were presynchronized with 2 injections of PGF2alpha, given 14 d apart starting on d 45+/-3 postpartum, followed by Ovsynch [2 injections of GnRH 7 d before and 56 h after injection of PGF2alpha, TAI 16 h after second injection (d 0)]. The Resynch-treated cows received an intravaginal progesterone insert from d 18 to 25, GnRH on d 25, and pregnancy diagnosis on d 32, and nonpregnant cows received PGF2alpha., GnRH 56 h later, and TAI 16 h later (d 35). The control cows were diagnosed for pregnancy on d 32 and nonpregnant cows received GnRH, PGF2alpha 39 d after TAI, GnRH 56 h later, and TAI 16 h later (d 42). Pregnancy was reconfirmed on d 60 after AI. Ovarian structures were examined in a subset of cows at the time of GnRH and PGF2alpha injections. Blood samples for analyses of progesterone and pregnancy-associated glycoproteins were collected every 2 d from d 18 to 30 in 100 cows, and collection continued weekly to d 60 for pregnant cows (n=43). Preenrollment pregnancies per AI on d 32 did not differ for cows subsequently treated as Resynch (45.8%, n=814) and control (45.9%, n=764), and pregnancy losses on d 60 were 6.7 and 4.0%, respectively. Resynchronized service pregnancy per AI (36%, n=441; 39.5%, n=412) and pregnancy losses (6.3 and 6.7%) did not differ for Resynch and control treatments, respectively. Days open for pregnant cows after 2 TAI were less for the Resynch treatment than for the control treatment (96.2+/-0.82 vs. 99.5+/-0.83 d). Cows in the Resynch treatment had more large follicles at the time of GnRH. The number of corpora lutea did not differ between treatments at the time of PGF2alpha. Plasma progesterone for pregnant cows was greater for Resynch cows than for control cows (18-60 d; 6.6 vs. 5.3 ng/mL), and plasma concentrations of progesterone on d 18 were greater for pregnant cows than for nonpregnant cows (5.3 vs. 4.3 ng/mL). Plasma pregnancy-associated glycoproteins during pregnancy were lower for cows in the Resynch treatment compared with control cows on d 39 (2.8 vs. 4.1 ng/mL) and 46 (1.3 vs. 3.0 ng/mL). Cows pregnant on d 32 that lost pregnancy by d 60 (n=7) had lower plasma concentrations of pregnancy-associated glycoproteins on d 30 than cows that maintained pregnancy (n=36; 2.9 vs. 5.0 ng/mL). Pregnancy-associated glycoproteins on d 30 (>0.33 ng/mL) were predictive of a positive d 32 pregnancy diagnosis (sensitivity=100%; specificity=90.6%). In conclusion, Resynch and control protocols had comparable pregnancy per AI for first and second TAI services, but pregnancy occurred 3.2 d earlier in the Resynch group because inseminations in the Resynch treatment began 7 d before those in the control treatment. Administration of an intravaginal progesterone insert, or GnRH, or both increased progesterone during pregnancy. Dynamics of pregnancy-associated glycoproteins were indicative of pregnancy status and pregnancy loss., (Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The aims of this study were to determine whether a single treatment of estradiol dipropionate (EDP) could induce pseudopregnancy in gilts and to determine the effectiveness of PGF(2alpha) treatment on estrus synchronization in EDP-induced pseudopregnant gilts. In experiment 1, gilts were treated with 20 mg of EDP (n=11) or vehicle (n=5) on Day 12 (Day 0=onset of estrus). Establishment of pseudopregnancy was defined as a lack of estrus and maintenance of the plasma progesterone concentration above 1 ng/ml between Days 12 and 36. Nine of 11 gilts (82%) treated with EDP became pseudopregnant. The plasma estradiol-17beta level was significantly higher in the EDP-treated gilts than in the control gilts until Day 29. In experiment 2, PGF(2alpha) was administered twice with a 24-h interval from Day 36 in pseudopregnant gilts (n=6) or Day 10 in cyclic gilts (control; n=5). Estrus after PGF(2alpha) treatment was observed in 83% of the pseudopregnant gilts. The interval from the day of the first PGF(2alpha) treatment to the onset of estrus and the peak of the LH surge was significantly shorter in the pseudopregnant gilts than in the control gilts. In experiment 3, six pseudopregnant gilts were bred by artificial insemination at the estrus after PGF(2alpha) treatment. The farrowing rate and average litter size did not differ between the PGF(2alpha)-treated pseudopregnant and cyclic gilts. These results indicate that a single treatment of EDP on Day 12 of the estrous cycle can induce pseudopregnancy in pigs and that a convenient protocol for administering PGF(2alpha) to EDP-induced pseudopregnant pigs is available for estrus synchronization programs in cyclic pigs.

Numerous studies have shown that it is possible to manipulate follicular and luteal dynamics, thereby eliminating the need for oestrus detection in embryo transfer (ET) programmes. Fixed-time ET (FTET) protocols are based on the use of gonadotrophin-releasing hormone (GnRH) and prostaglandin (PG) F or progesterone/progestogen (P4)-releasing devices and oestradiol. The FTET protocols increases the proportion of recipients transferred, and therefore pregnancy rates, compared with the use of PGF followed by ET 7 days after oestrus. Furthermore, the addition of equine chorionic gonadotrophin (eCG) to the P4 and oestradiol-based FTET protocols results in an even higher proportion of recipients transferred, and thus higher pregnancy rates. The beneficial effect of eCG treatment may be related to increased growth of the dominant follicle and increased plasma P4 concentrations during the subsequent luteal phase. In Bos taurus x Bos indicus recipients, pregnancy rates were positively correlated with the diameter of the corpus luteum (CL) and the number of CL at ET. When repeat-breeder Holstein cows were used as recipients, FTET protocols increased number of recipients transferred and pregnancy rates compared with the traditional PGF-based synchronisation protocols. In conclusion, the use of FTET protocols eliminates the need for the detection of oestrus and results in a greater proportion of recipients transferred and satisfactory pregnancy rates. Thus, FTET optimises the use of recipients, reducing labour and animal handling and facilitating the use of ET.

Three experiments were conducted on anestrous ewes of Suffolk, Dorset, and Katahdin breeding to examine the potential value of GnRH to improve ovulation and pregnancy in response to introduction of rams. In Experiment 1, treatment with GnRH 2d after treatment with progesterone (P(4); 25mg i.m.) at introduction of rams was compared to treatment with P(4) alone at the time of introduction of rams. Treatment with GnRH did not increase percentages of ewes with a corpus luteum (CL) 14d after introduction of rams, pregnant 32d after treatment with PGF(2)alpha 14d after introduction of rams, or percent of treated ewes lambing to all services. In Experiment 2, treatments with GnRH on day 2, 7, or both after introduction of rams were compared. Treatments did not differ in mean estrous response, percentages of ewes with a detectable CL or number of CL present on day 11, or mean pregnancy and lambing rates. Therefore, neither one nor two injections of GnRH at these times appeared to be effective to induce anestrous ewes to breed. In Experiment 3, treatments compared included GnRH 4d before introduction of rams, GnRH 4d before and 1d after introduction of rams, ram introduction alone, and treatment with P(4) (25mg i.m.) at the time of introduction of rams. Percentages of ewes with concentrations of P(4) greater than 1ng/mL (indicating formation of CL had occurred) 7d after ram introduction tended to be greater (P<0.07) in ewes treated with GnRH or P(4) than in control ewes treated with ram introduction alone. However, there was no difference in P(4) concentrations between groups by day 11 or 12 after introduction of rams. Estrous response rates and percentages of ewes pregnant 95d after PGF(2)alpha was administered (on day 12 after introduction of rams) tended to be greater (P=0.08 and 0.06, respectively) in ewes treated with GnRH or P(4) than in ewes exposed to rams only. There was no difference in response variables between ewes treated with GnRH 4d before introduction of rams and ewes treated with GnRH 4d before and 1d after introduction of rams. In conclusion, treatment with GnRH 4d before ram introduction showed promise as an alternative to treatment with P(4) to improve the ovulatory response and reproductive performance of ewes introduced to rams during seasonal anestrus.

Synchrony of estrous, and consequently of conception and birth of young, may be of adaptive significance for certain mammals. Among the species in which estrous synchrony has been suspected several times are elephants, but clear evidence is still missing. We determined estrous cycles of African elephants (Loxodonta africana) (n=4) at the Vienna Zoo, Austria, between June 2003 and January 2006 by measuring serum progesterone levels from weekly blood samples. Except for the dominant female when she was intensively lactating, all animals showed clear cycles or progesterone release with a mean period of 105.3+/-15.37 days. For most of the study period, estrous cycles were asynchronous between females. However, after re-occurrence of the progesterone cycle in the dominant female following the first period of lactation, all four females showed high synchrony of progesterone release over the two subsequent cycles. Large changes in individual period lengths indicated that synchronization was due to the adjustment of cycle length in subdominants to that of the dominant female. We used a bootstrap procedure, based on resampling measured times of progesterone peaks, to determine if this apparent synchrony could have been caused by chance alone. This statistical analysis indicated that between-individual variances of the timing of progesterone peaks were much smaller that to be expected by chance (P=0.009). This finding represents the first evidence for estrous synchrony between elephants. We discuss various hypotheses to explain the biological function of cycle synchrony in elephants.

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

The objective was to investigate ovarian follicular development and hormone concentrations in previously inseminated cows with estrous cycles resynchronized with various resynchronization treatments. Lactating dairy cows were treated with a previously used intravaginal progesterone releasing device (IVD) for 7 d (EB+IVD 7+EB, n=15) or 8 d (EB+IVD 8+EB, n=16), starting 13 d (Day 13) after a first estrus (Day 0) and AI. Estradiol benzoate (EB; 1 mg im) was given at device insertion and 24 h after removal. Other cows were given the same treatment as the EB+IVD 8+EB cows, but were not treated with EB at IVD insertion (IVD 8+EB, n=11). There were no differences (P>0.05) between EB+IVD 7+EB and EB+IVD 8+EB treatments for follicle dynamics and plasma progesterone concentrations during treatment. Based on a comparison between the IVD 8+EB treated cows and the pooled results of the EB+IVD 7+EB and EB+IVD 8+EB treated cows, EB at device insertion increased the number of follicular waves between Days 13 and 20 (mean+/-S.E.M.; 2.3+/-0.14 vs 2.7+/-0.10, P=0.033), delayed emergence of follicles that were dominant or emerging on Day 20 (17.2+/-0.36 vs 14.1+/-0.65 d, P<0.001), reduced diameters of dominant or emerging follicles on Day 20 (9.0+/-0.58 vs 12.7+/-0.59, P<0.001), and reduced plasma progesterone concentrations by 0.85+/-0.44 ng/mL (P=0.059) during treatment. Furthermore, comparison of the IVD 8+EB to the EB+IVD 8+EB treated cows demonstrated that treatment with EB at device insertion also reduced the diameter of ovulatory follicles (14.2+/-0.58 vs 19.0+/-0.71 mm, P=0.001), delayed emergence of ovulatory follicles (17.0+/-0.32 vs 13.5+/-1.26, P=0.020), and reduced the interval from emergence to ovulation (7.0+/-0.32 vs 10.5+/-1.26 d, P=0.020). We concluded that administration of EB altered ovarian follicular dynamics and tended to reduce plasma progesterone concentrations during treatment with an IVD that was used to resynchronize estrous cycles. However, use of a 7-d compared to an 8-day treatment with an IVD did not significantly affect follicle dynamics nor plasma progesterone concentrations during treatment.

The objective of this study was to examine the effects of dietary n-3 polyunsaturated fatty acid (n-3 PUFA) supplementation on embryo yield and quality in heifers. Animals were individually offered barley straw and concentrate diets supplemented with either palmitic acid (C16:0; CON) or a partially rumen protected n-3 PUFA-enriched supplement. Following oestrous cycle synchronisation, superovulation was induced using FSH. Blood samples were collected for the measurement of fatty acids, metabolites, insulin and IGF-1. On day 7 post-insemination the number of ovulations was estimated and embryos recovered non-surgically and quality graded. At embryo recovery 50 ml of the uterine flushing was collected from each horn for fatty acid analysis. Grade 1 embryos were isolated, snap frozen in liquid nitrogen and stored at -80 degrees C. mRNA expression for six genes, LIF, BAX, Cx43 and E-CAD associated with embryo development, and PPAR-alpha and -delta, associated with lipid metabolism was analysed. The n-3 PUFA supplementation increased plasma n-3 PUFA concentration (P<0.05) and reduced n-6:n-3 PUFA ratio (P<0.05). Uterine concentration of the n-3 PUFA, eicosapentaenoic acid was increased (P<0.05) and the concentration of arachidonic acid decreased (P<0.05) following n-3 PUFA supplementation. While CON increased triglyceride concentrations, diet did not affect the other plasma metabolites, insulin or IGF-1 (P>0.05). Similarly, there was no effect of diet on superovulation rate, embryo recovery rate, embryo quality (P>0.05) or mRNA expression of the genes examined (P>0.05).

Fertility is often lower in anestrous compared to cyclic ewes, after conventional estrus synchronization. We hypothesized that synchronization of ovarian follicular waves and ovulation could improve fertility at controlled breeding in anestrous ewes. Estradiol-17beta synchronizes follicular waves in cattle. The objectives of the present experiments were to study the effect of an estradiol injection, with or without a 12-d medroxyprogesterone acetate (MAP) sponge treatment, on synchronization of follicular waves and ovulation in anestrous ewes. Twenty ewes received sesame oil (n=8) or estradiol-17beta (350 microg; n=12). Eleven ewes received MAP sponges for 12d and were treated with oil (n=5) or estradiol-17beta (n=6) 6d before sponge removal. Saline (n=6) or eCG (n=6) was subsequently given to separate groups of ewes at sponge removal in the MAP/estradiol-17beta protocol. Estradiol treatment alone produced a peak in serum FSH concentrations (4.73+/-0.53 vs. 2.36+/-0.39 ng/mL for treatment vs. control; mean+/-S.E.M.) after a short-lived (6 h) suppression. Six of twelve ewes given estradiol missed a follicular wave around the time of estradiol injection. Medroxyprogesterone acetate-treated ewes given estradiol had more prolonged suppression of serum FSH concentrations (6-18 h) and a delay in the induced FSH peak (32.3+/-3.3 vs. 17.5+/-0.5 h). Wave emergence was delayed (5.7+/-0.3 vs. 1.4+/-0.7d from the time of estradiol injection), synchronized, and occurred at a predictable time (5-7 vs. 0-4d) compared to ewes given MAP alone. All ewes given eCG ovulated 3-4d after injection; this predictable time of ovulation may be efficacious for AI and embryo transfer.

A total of 799 Holstein cows from 3 herds were randomly assigned at 37 +/- 3 d in milk (DIM) to timed artificial insemination (AI) or insemination at detected estrus. Cows were presynchronized with injections of PGF(2alpha) at 37 and 51 DIM. At 65 DIM, cows received an injection of GnRH, followed 7 d later by PGF(2alpha). Cows in the estrus-detected group were inseminated after being observed in estrus during the 7 d after the last PGF(2alpha). Cows in the timed AI group received an injection of 1 mg of estradiol cypionate (ECP) 24 h after the last PGF(2alpha). If detected in estrus or=1 ng/mL; L = <1 ng/mL), resulting in 8 combinations (LLL, LHL, LLH, LHH, HHH, HHL, HLH, and HLL). Conception rates and pregnancy rates were higher for cows in the timed AI group than in the estrus-detected group at 30, 44, and 58 d (e.g., at 58 d, pregnancy rates were 42.2% for multiparous cows or 34.4% for primiparous cows in the group receiving ECP and timed AI compared with only 20.8 or 18.8% for respective parity subgroups for the treatment group inseminated only at detected estrus). Pregnancy losses were 11.5% from 30 to 58 d and did not differ between treatments. Cyclic cows within both treatments had higher estrous responses, conception rates, and pregnancy rates. Cows that responded to presynchronization and to luteolysis (HHL) had the highest conception and pregnancy rates, followed by cows classified as LHL. Use of 1 mg of ECP to induce ovulation as part of a synchrony regimen improved reproduction at first postpartum insemination in dairy cows.

This study was conducted to identify factors affecting PGF(2alpha) efficacy to synchronize estrus in water buffalo cows. After detection of a corpus luteum (CL) by rectal palpation, cows were treated (im) with dinoprost (12.5, 25 or 50mg) or D(+) cloprostenol (75, 150 or 300 microg) in a total of 66 treatments. Blood samples were collected 0, 24 and 48 h after treatment and ultrasound examinations and observations for estrus were performed daily to the day of ovulation or to 6 days after treatment. No PGF(2alpha) dose-response pattern was observed and overall rates of luteal regression (progesterone <1.0 ng/ml at 48 h), estrus, no detected behavioral estrus with ovulation occurring, and ovulation were 71.2, 36.4, 19.7 and 54.5%, respectively. To analyze plasma progesterone concentrations and ovarian dynamics, cows were divided in three groups according to their response to treatment. Cows that failed to have ovulations from a follicle after treatment (Group A, n = 30) had (P < 0.05) a lower plasma progesterone concentration (2.98 ng/ml) and smaller CL area (CLA; 187.3 mm(2)) before treatment as compared with cows that had an ovulation from a follicle (4.43 ng/ml and 223.7 mm(2), respectively; Groups B and C, n = 36). In cows that failed to ovulate, plasma progesterone concentration decreased in the first 24 h, but did not decline further and was >1.0 ng/ml 48 h after treatment. Moreover, no significant change in CLA after treatment was detected, indicating that treatment induced only partial luteolysis. In cows that ovulated, plasma progesterone concentration and CLA decreased continuously from treatment to ovulation (consistent with complete luteolysis). Threshold values of 2.8 ng/ml for plasma progesterone concentration and 189 mm(2) for CLA were identified as the best predictors of ovulation before treatment (83.3 and 80.6% sensitivity and 58.6 and 65.5% specificity, respectively, with positive and negative predictive values around 71%). When the origin of the ovulatory follicle was investigated, the interval from treatment to ovulation was shorter (91.9 versus 113.3 h; P < 0.05), and the ovulatory follicle had a slower growth rate (1.02 versus 1.55 mm per day; P < 0.005), a lesser increase in diameter from treatment to ovulation (4.7 versus 8.0 mm; P < 0.001), and a greater maximum diameter (13.2 versus 12.1 mm; P < 0.05) in cows that ovulated from the largest follicle present in the ovary before treatment (Group B, n = 27) compared with cows that ovulated from the second largest follicle present in the ovary before treatment (Group C, n = 9). In summary, the efficacy of PGF(2alpha) for causing luteolysis and synchronizing estrus and ovulation in buffalo cows was dependent upon plasma progesterone concentration, CL size and ovarian follicular status before treatment., (Copyright 2002 Elsevier Science B.V.)

Female mice (Mus musculus) frequently are used to study hormonally related differences in susceptibility to infectious organisms or response to pharmaceutical agents. Cyclical variation in hormone levels within a group of mice, however, challenges the experimental design of such studies in that it is often difficult to obtain sufficient numbers of mice in the desired phase of the estrous cycle at the time of treatment. The purpose of this work is to provide investigators with a standardized protocol for inducing estrus in mice through exposure to male urine (Whitten Effect). In addition, we demonstrate how the Whitten Effect can be used to induce susceptibility of mice to Neisseria gonorrhoeae infection. Female BALB/c mice were exposed to male urine via soiled bedding for 0, 24, 48, 72, or 96 h. The effect of exposure on the reproductive cycle was monitored by cytologic examination of vaginal smears and measurement of serum 17-b estradiol levels by using a nonradioactive immunoassay kit. In a separate experiment, mice were exposed to male-urine-soaked bedding for 0, 24, 72, or 96 h prior to intravaginal inoculation with Neisseria gonorrhoeae. Infection was monitored by using vaginal culture for 5 consecutive days. We found that the highest percentage of mice in estrus occurred among mice that were exposed to male-urine-soaked bedding for 96 h. Consistent with this finding was the demonstration that mice were more susceptible to gonococcal infection after exposure to male urine for 3 to 4 days. We conclude that exploitation of this natural murine behavioral response is a simple and inexpensive method by which estrus can be synchronized in a group of mice within a defined period of time. In addition, this protocol can be used to increase mouse susceptibility to experimental gonococcal infection.

The objective of this study was to compare the effects of administration of a single injection of progesterone (P4) and follicle aspiration on Day 7 of the estrous cycle on the timing and synchrony of follicular wave emergence, time of ovulation, and concentrations of P4, estradiol and FSH in Holstein cows. Twenty cows were assigned to 4 groups (n=5 cows per group) in a 2 by 2 factorial arrangement. Cows were treated on Day 7 (Day 0 = estrus) of the estrous cycle with either sham follicular aspiration and an oil vehicle administered intramuscularly (control), aspiration of ovarian follicles (aspiration), 200 mg of P4 im, or aspiration and 200 mg of P4 im (aspiration + P4). On Day 11, PGF(2alpha)(25mg) was administered to all groups. Synchrony of ovulation was less variable in each of the treatment groups compared with the control group (P<0.05), whereas ovulation was delayed in cows in the P4 group (P<0.05). Day of follicular wave emergence was delayed in the cows of the P4 group compared with cows in the aspiration and aspiration + P4 groups (P<0.01), whereas variability in wave emergence was less among both groups of aspirated cows compared with the cows in the control group (P<0.01). More follicles 4 to 7 mm in diameter were detected in the 2 aspiration groups compared with the cows in the control and P4 group (P<0.05). No difference was detected among groups in the maximum concentration of FSH associated with follicular wave emergence. We conclude that both the administration of P4 and the aspiration of follicles on Day 7 of the estrous cycle improves the synchrony of ovulation when luteolysis is induced on Day 11 and results in similar concentrations of FSH at the time of follicular wave emergence, but the timing of wave emergence and the number of follicles post-emergence differ.

This study was conducted to examine the effects of metestrus administration of SyncroMate-B (SMB) on PGF2alpha secretion and corpus luteum (CL) development. In a study replicated over 2 yr, cows were observed for spontaneous estrus in yr 1, and cows received an injection of 25 mg of PGF2alpha and were observed for subsequent estrus in yr 2. At standing estrus (estrus = d 1), cows were randomly allotted to receive either the standard SMB regimen (n = 40) on d 3 of the estrous cycle or no treatment (n = 8). Fifty percent (n = 20) of SMB-treated cows were administered PGF2alpha on d 10 of the estrous cycle 48 h prior to implant removal. Twice-daily blood samples were collected in the morning (AM) and evening (PM) from d 2 AM through d 14 AM of the treated estrous cycle and subsequently analyzed for progesterone (P4) and PGF2alpha metabolite (PGFM). Prior to statistical analysis, SMB- and SMB/PGF2alpha-treated cows were sorted according to P4 concentration at d 10 of the treated estrous cycle to either a CL functional group (P4 > or = 1 ng/mL; n = 20) or a CL nonfunctional group (P4 < 1 ng/mL; n = 17). Following d 10 AM administration of PGF2alpha, functional and nonfunctional groups were further subdivided based on treatment. The groups were as follows: untreated control cows (n = 8); SMB-treated cows retaining a functional CL (SMB-F; n = 8); SMB-treated cows with a nonfunctional CL (SMB-N; n = 11); SMB/PGF2alpha-treated cows retaining a functional CL (SMB/PG-F; n = 12); and SMB/PGF2alpha-treated cows with a nonfunctional CL (SMB/PG-N; n = 6). Of all SMB-treated cows, 54% retained a functional CL through d 10 AM of the treated estrous cycle. Mean serum P4 concentrations increased for cows in all groups until d 7, after which P4 concentrations increased for cows in SMB/PG-F, SMB-F, and control groups and decreased for cows in SMB/PG-N and SMB-N groups. Following PGF2alpha administration on d 10, mean serum P4 concentrations remained < 1 ng/mL for cows in SMB/PG-N and SMB-N groups, decreased to < 1 ng/mL for cows in the SMB/ PG-F group, and remained > 1 ng/mL for cows in SMB-F and control groups. Mean serum PGFM concentrations tended (P = .06) to increase in cows with nonfunctional CL compared with control cows on d 8 AM and were greater (P < .05) in cows with functional CL on d 8 PM through d 9 PM. These results indicate that retention of a functional rather than a nonfunctional CL following metestrus administration of SMB is dependent on a premature release of uterine PGF2alpha.

Estimation of the long-term consequences on reproduction performance of the oestrus synchronisation treatments that are annually applied to ewes was carried out on nine officially controlled dairy flocks in the Roquefort region of France. A hormonal treatment combining the insertion of a vaginal fluoro-gestone acetate (FGA) sponge for 14 days and the injection of about 500 IU of pregnant mare serum gonadotropin (PMSG) at withdrawal was applied to the ewes in seven of the nine flocks. The ewes in the two other flocks were used as controls. Blood samples were taken from each female just before the treatment (to test for the presence of residual antibodies) and 20 days after the PMSG injection. Anti-PMSG antibody binding rates were calculated for each blood sample. The residual binding rate increased with age and induce negative effects on the following years reproduction performances, i.e., they increased the probability that the ewes would not become pregnant.

Two experiments were conducted to determine if corpus luteum regression, formation, and function were associated with the decreased calving rate observed in beef females administered PGF2 alpha 5 days before Syncro-Mate B (SMB) treatment. Experiment 1 included 31 beef heifers 11 to 13 months old and experiment 2 included 31 beef heifers 19 to 21 months old. Heifers were randomly assigned to 1 of 2 groups (control and PGF2 alpha 5 days before SMB treatment). Heifers were bled 10 days before PGF2 alpha treatment, immediately before PGF2 alpha and SMB treatments, at the time of implant removal, and twice weekly after implant removal. Heifers in experiment 2 were observed twice daily for estrus for 5 days after PGF2 alpha treatment and for 3 days after norgestomet implant removal. Based on the blood samples collected before SMB treatment, 15 heifers in experiment 1 and every heifer in experiment 2 were with estrous cycles. All heifers in experiment 1 had progesterone concentrations < 0.5 ng/ml 2 days after implant removal. However, progesterone concentrations during the luteal phase in control heifers with estrous cycles were higher (P < 0.05) than in PGF2 alpha treated heifers with estrous cycles and in heifers previously without estrous cycles. In experiment 2, based on the occurrence of estrus and progesterone concentrations, heifers were also classified as metestrus or diestrus at the time of SMB treatment. The data were analyzed as a 2 x 2 factorial with treatment (control or PGF2 alpha) and stage of the cycle (metestrus and diestrus) as main effects. More metestrus heifers (40%) had progesterone concentrations > 1.0 ng/ml 2 days after implant removal than diestrus heifers (0%). In addition, progesterone concentrations during the luteal phase in metestrus heifers were lower (P < 0.05) than in diestrus heifers. PGF2 alpha treatment had no effect (P > 0.25) on the number of heifers with > 1.0 ng/ml progesterone 2 days after implant removal and progesterone concentrations during the luteal phase. There was no treatment by stage of the estrous cycle interactions. In summary, the administration of PGF2 alpha 5 days before SMB decreased the calving rate by causing more heifers to be metestrus at SMB treatment. Fewer metestrus heifers (than diestrus heifers) were synchronized (with < 1.0 ng/ml of progesterone 2 days after implant removal) to SMB treatment and those synchronized had lower progesterone concentrations during the luteal phase.

Although different FSH preparations and injection regimens are used to superovulate cattle, the optimum treatment regimen and blood concentrations of FSH to induce effective superovulatory responses are currently not known. The current objective was to evaluate the pattern of follicular growth, oestradiol-17 beta(E2) concentrations and yield of embryos in heifers following superovulation with two different pFSH preparations reportedly differing in LH content. In experiment 1, 90 synchronised heifers were superovulated at mid-cycle using a 2 x 2 factorial design comparing Folltropin (Vetrepharm; low LH) with Pluset (Serovet; FSH:LH ratio 1:1) administered either as a single or multiple (8 for Folltropin, MF and 10 for Pluset, MP) injections. Animals were inseminated during oestrus which was induced with prostaglandin F2 alpha analogue and embryos were recovered 7 days later. Overall, Pluset treatments compared with Folltropin resulted in more ovulations and unfertilized or degenerate embryos (P < 0.05). Multiple injections resulted in more (P < 0.05) freezable (MF = 55 +/- 1.2; MP = 3.8 +/- 1.0) and transferable embryos (MF = 2.68 +/- 0.9; MP = 2.71 +/- 0.9) than single injections (SF = 2.2 +/- 0.5 and 1.0 +/- 0.3 respectively; SP = 2.6 +/- 0.8 and 1.3 +/- 0.4 respectively); there was also a higher (P < 0.05) percentage embryo recovery rate. In two subsequent experiments, animals (n = 17) were superovulated with either single or multiple injections of Folltropin or Pluset as described and blood samples were collected and analysed for E2 concentrations. Ovarian scanning was carried out until 72 h after the first FSH injection, to count medium (5-9 mm) and large (> or = 10 mm) follicles. Heifers treated with SP had higher E2 concentrations in comparison with heifers treated with SF at 18, 36-48 and 84-96 h after the FSH injection. There were no differences in E2 concentrations in heifers treated with MF or MP treatments. Heifers treated with SP had greater numbers of follicles compared to SF treated heifers (21.0 +/- 3.1 vs 13.9 +/- 2.2; P = 0.089) on the third day after FSH injection. There were no differences between the numbers of medium and large follicles in heifers treated with MF or MP at any time throughout the experimental period. These data indicate that a single injection of Folltropin or Pluset can result in multiple ovulations and that the E2 profiles are different following single injections of either Folltropin or Pluset.

The effects of a progesterone antagonist (onapristone) on heat synchronization, luteinizing hormone (LH) surge, ovulation, oocyte maturation and fertilization of superovulated ewes were studied. Its effects on in vitro bovine oocyte maturation and fertilization were also studied. Estrus synchronization and superovulation treatments were applied to 39 adult ewes using an intravaginal sponge with fluorgestone acetate for 9 days with injections of prostaglandin F2 alpha and pregnant mare's serum gonadotrophin given 24 h before sponge withdrawal. The animals were randomly assigned to four different groups; T1 receiving only the synchrony treatment (n = 11); T2 ewes received two injections of onapristone (1 mg kg-1, i.v.) 12 h apart from 3 h after sponge withdrawal (n = 10); T3 ewes received two injections of progesterone 12 h apart from sponge withdrawal (n = 10); and, T4 ewes received both onapristone and progesterone as described (n = 8). Ewes were mated by a fertile male during estrus. Progesterone and LH were measured during the superovulation period in plasma samples taken every 4 h. Uterine flushings for ova recovery were performed at 5 days (n = 25), 48 h (n = 5) and 24 h (n = 5). Non-fertilized oocytes collected at 24 and 48 h were checked for meiosis resumption. The effects of two doses of onapristone (D1 and D2) on in vitro bovine oocyte maturation (control = 100, D1 = 100 and D2 = 100) and fertilization (control = 107, D1 = 40 and D2 = 75) were also studied. The percentage of animals showing heat signs was significantly lower in group T3 (50% vs. 100%). The onset of oestrus (27.6, 24.8, 68.8 and 25.5 h, respectively for T1, T2, T3 and T4) and an LH surge (32.3, 28.8, 76.5 and 30.5 h, respectively for T1, T2, T3 and T4) after sponge withdrawal were significantly delayed in group T3. There were no significant differences in the intervals between estrus and LH surge among groups (4.61 +/- 0.75 h). The response and ovulation rates until 40 h after sponge withdrawal (group T3 excluded) were similar among groups, but the fertilization rates were significantly lower in groups T2 and T4 when compared with T1 (2% and 3% vs. 41%, respectively; P < 0.001) due to sperm arrest in the cervix. Ova recovery rate decreased significantly from 24-48 h to 5 days and was not affected by treatments (76.9% vs. 37.1% respectively). Onapristone did not affect the resumption of meiosis. Fertilization of bovine oocytes in vitro decreased significantly only in group D2 when compared to control (48% vs. 62.6%, respectively). In conclusion, onapristone treatment during the preovulatory period did not interfere with normal synchronization of estrus, ovulation and oocyte maturation but severely compromised fertilization by arresting spermatozoa in the cervix.

The variability between animals in the timing of oestrus after administration of a synchronization treatment seems to explain the low rate of fertility in goats inseminated at a predetermined time after progesterone withdrawal. Two experiments were performed during the breeding season to test whether the variation was due to the exogenous hormone regime or to the endogenous physiology of the animals. Twenty-one goats were given a synchronization treatment consisting of a vaginal sponge impregnated with 45 mg of fluorogestone acetate (FGA) for 11 days associated with intramuscular injection of 400 I.U. of equine chorionic gonadotrophin (eCG) and 50 microg of cloprostenol 48 h before sponge removal. Progesterone concentrations were measured during the subsequent cycle and the patterns were modelled to allow precise determination of the onset of luteolysis. Oestrus and the luteinizing hormone (LH) surge began 33.0+/-6.8 h and 76.0+/-33.0 h after sponge withdrawal, v. 43.4+/-5.7 h and 90.0+/-36.0 h after natural luteolysis. For both observations, the between-goat variability was larger during the natural than during the synchronized oestrus (P < 0.05). The duration of the oestrous cycle was independent of the number of corpora lutea (CL), whereas the duration of luteal phase was shorter in goats with 2-3 CL (16.4+/-0.9 day than in those with 1 CL: 17.7+/-1.3 day; P < 0.05). In the second experiment, 20 goats were ovariectomized and given a vaginal sponge as described above. Sixteen h after sponge removal, they were injected with 50 microg of oestradiol benzoate (ODB). This treatment was repeated with the second sponge being inserted 1-2 days after observation of oestrus. Oestrus and LH surge were observed: 32.8+/-6 8 h v. 27.8+/-7.8 h after the first ODB injection, and 36.6+/-7.3 h v. 34.3+/-4.8 h after the second ODB injection. No relationship was observed between data of the two experiments. In both cases, the variability in the occurrence of oestrus and LH surge was of the same order as observed in the first experiment. This study shows that the timing of oestrus and LH surge is less variable after progestagen treatment than during a natural oestrous cycle. Moreover, a significant proportion of variability is inherent in the delays following the oestradiol peak, suggesting that further improvement in the synchronizing capacity of treatment based on progestagen administration is unlikely.

Ovarian follicular development and plasma concentrations of progesterone (P4), estradiol-17 beta (E2), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were compared during a synchronized estrous cycle between heifers and mature cows displaying contrasting superovulatory responses. Six heifers < 2 years old with a history of good responses to superovulatory (SOV) treatment and six cows 9 to 13 years old with poor responses to SOV treatments were used. Follicular development was monitored by daily ultrasonography. Blood samples were collected two to three times daily for P4 and E2 and thrice daily for LH and FSH analysis. Intensive sampling (samples every 15 min for 6 hr) was performed at critical periods of follicular development to analyze the pulsatile secretion of gonadotropins. In both cattle groups, a transient increase (P = 0.0001) in E2 occurred 4 to 5.7 d after the preovulatory LH surge or 2.3 d before the dominant follicle reached its maximum size. FSH concentrations increased (P = 0.006) before the emergence of the second cohort of follicles and then decreased despite no change in the concentration of E2. Contrary to our expectation and despite differences between groups in terms of age, number of previous SOV treatments, and divergent responses to superovulation, follicular development was similar in both groups. However, during the luteal phase, concentrations of E2 and FSH and LH pulse amplitudes were less (P < or = 0.05) in cows than in heifers. Therefore, follicular development monitored by ultrasonography and endocrine profiles during a synchronized estrous cycle are of limited value to predict quality of embryo donors.

The active transport of mineral substances by cell membranes plays an important role in the neurohumoral regulation of reproductive processes in farm animals. A great importance is attributed to lipoproteins as sources of the primary precursor of steroid synthesis in the placenta and ovaries (Knopp et al., 1986; Grummer et al., 1988). The effects of hormones on K metabolism in the blood serum of ewes were studied by Hawk et al. (1961). Krajnicáková et al. (1993) observed the dynamic changes of electrolytes in the course of the reproductive cycle in ewes. Na and K levels and their relation to ovarian hormones were determined in the systemic blood stream of ewes during oestrus synchronization and pregnancy. Ten Slovak Merino ewes aged 3-5 years and weighing 40-50 kg were included in the experiment. The animals were reared conventionally. A feed ration consisted of silage, hay, feeding beet-pulp, straw and in the last two months of pregnancy it was supplemented with roughage. Water and salt were given ad libitum. Blood samples were obtained from the jugular vein prior to synchronization (day 0) and on days 3, 7 and 14 of synchronization with Agelin vaginal implants. After removing the implants the animals received 500 I.U. PMSG (Pregnant Mare Serum Gonadotropin). The animal in oestrus cycle were inseminated with fresh diluted semen. Reinsemination was not performed in this experiment. Sampling was continued on the day of insemination and on days 7, 14 and in month 2, 3, 4 and 5 of pregnancy. Na and K levels were measured by atomic absorbtion spectrophotometry using the Atom Spek device (RANG-HIGLER). Progesterone (P4) and 17 beta-oestradiol (E2) levels were assayed radioimmunologically using the respective kits (RIA-test-PROG and RIA-test-ESTRA) manufactured by URVJ Kosice. During the observation period Na levels fluctuated within 131.50 +/- 1.71 and 153.55 +/- 4.77 nmol/l of serum (Tab. I) and significantly decreased between day 14 of synchronization and the end of the investigation (P < 0.05; P < 0.01; P < 0.001). Potassium levels were observed to significantly decrease from the day of insemination on, throughout the first days of pregnancy and the above-mentioned months of gravidity, the values ranging between 4.57 +/- 0.52 and 3.93 +/- 0.36 nmol/l.(ABSTRACT TRUNCATED AT 400 WORDS)

Our investigations were concerned with dynamic changes in total lipids (CL), cholesterol (CHOL) and progesterone (P4) in blood serum of sheep in the period of oestrus synchronization treatment and during mating and gravidity. Our experiment was carried out using 10 animals housed under the conditions of productive rearing. Blood samples were taken from v. jugularis on day of swab application (day 0) and on days 3 and 7 of the action of Agelin vaginal swabs, on day of insemination, and on days 7, 14, 17 and in the 2nd, 3rd and 4th month of gravidity. Blood serum was used to determine total lipids and cholesterol by means of Bio-Lachema tests, and P4 concentrations employing RIA-test-Prog kits (URVJT, Kosice). A statistically significant decrease in concentrations of total lipids (Fig. 1, Tab. I) in sheep blood serum was recorded on day of insemination (P < 0.05) compared to day 0, with the value 1.59 +/- 0.31 g/l of serum, and in the 3rd month of gravidity (P < 0.01), at concentrations 1.36 +/- 0.38 g/l of serum. The determined decrease in their values in the mentioned period can be modulated by the mutually changing ratio of steroid hormones or by inhibition of synthesis of lipoproteins responsible for changes in total plasma lipids. Changes in cholesterol concentrations (Fig. 2, Tab. I) during the introduction of swabs were insignificant and ranged from 1.60 +/- 0.42 to 1.73 +/- 0.33 mmol/l of serum. An insignificant increase in cholesterol concentrations (P < 0.05), with its highest levels 1.98 +/- 0.43 mmol/l of serum, was recorded in the 3rd month of gravidity.(ABSTRACT TRUNCATED AT 250 WORDS)

Knowledge of pathogenesis of sexual dysfunctions at altered thyroid activity is limited by the knowledge of multiple and ubiquitous action of its hormones throughout the organism. One of the possibilities of modulatory influence of thyroid hormones on sexual functions can be realized through the participation of thyroxine and triiodothyronine in the synthesis and metabolism of primary substrate of steroid synthesis--cholesterol. The presented work is aimed at the study of simultaneous dynamic changes of concentrations of thyroxine (T4), triiodothyronine (T3), 17 beta-estradiol (E2), progesterone (P4) and cholesterol (Chol) during synchronization of the rutting period and gravidity at parallel correlative evaluation of mutual relations of the followed parameters in ten Merino sheep in the seasonal period. Synchronization was achieved by chlorsuperlutin (Agelin--vaginal swabs, Spofa; 20 mg of chlorsuperlutin/swab) and PMSG (500 I. U./animal). Blood was sampled by means of a jugular vein puncture at the time of swab insertion (-13th day) and after three (-10th day) and seven (-7th day) following days, at the removal of swabs and application of PMSG (-3rd day), on the day of insemination (zero day), on the 7th, 14th and 17th day and in the middle of the 2nd, 3rd, 4th and 5th month of gravidity. In the phase of oestrus synchronization a significant increase of E2 concentrations on days -7 and -3 of the experiment (0.47 +/- 0.079 and 0.542 +/- 0.177 nmol.l-1 of serum, P less than 0.001; P less than 0.001) was observed compared to the E2 values on day -13 (0.084 +/- 0.036 nmol.l-1 of serum). Parallel to these observations, marked intermittent changes of T4 (Tab. I, Graph 1) were recorded with the lowest values of this parameter observed on days -10 (41.75 +/- 20.23, P less than 0.05) and -3 (50.22 +/- 18.77, P less than 0.05) and the highest on day -7 (96.77 +/- 17.51 nmol.l-1, P less than 0.01) and day zero (85.40 +/- 19.59 nmol.l-1 of serum, P less than 0.05) in comparison with the -13th day (67.22 +/- 18.29 nmol.l-1 of serum). Concentrations of P4 (Tab. I, Graph 4) declined to the lowest values on day zero observation (0.09 +/- 0.08 nmol.l-1 of serum, P less than 0.05 vs 3.40 +/- 3.61 nmol.l-1 on day -13). No significant changes of concentrations of T3 (Tab. I, Graph 2) and Chol (Tab. I, Graph 5) were observed during oestrus synchronization. During gravidity, concentrations of E2 (Tab. I Graph 3) showed an increasing trend compared to the -13th day.(ABSTRACT TRUNCATED AT 250 WORDS)

The results of four experiments are presented in summary form. The data are considered in relationship to the improvement of the fecundity and fertility of the Australian ewe breeding flock. In the first, three commonly used methods of oestrous synchronization were examined and showed differences that are attributed to the different patterns of hormonal changes associated with the methods demonstrated. The second experiment looked at the use of active immunization against testosterone and concluded that this method can improve fecundity but not fertility. The third experiment, a group of five trials, studied the use of progestagen sponges and PMSG in anoestrous ewes as a means of inducing normal fertility. The extensive data produced in this experiment allowed the relationships between ovulation rate and fertility and between fertility and prolificacy (fecundity) to be examined. Fertility appeared greatest when the mean flock ovulation rate was about 2.5. At this ovulation rate prolificacy was also improved and a high proportion of twins were produced. We concluded that high fertility and low prolificacy (i.e. of 1.00) are an unlikely combination. In the final experiment the effect of post-mating hormonal supplementation on fertility was examined and a number of earlier reports were confirmed by showing that fertility can be improved with supplementary progesterone between days 10 and 25 post-mating. The effect appears to be modified by hormonal and nutritional factors.