Trials were performed to examine the effectiveness of 250 micrograms (1 ml) of cloprostenol (Oestrophan Spofa) implantation under the mucous membrane of the vaginal vestibule of 128 cows with a clinically pronounced corpus luteum on ovaries. Within 72 hours from administration, oestrus was observed in 112 animals (85.5%). Out of the 97 cows inseminated, 63 cows (64.94%) got in calf. The effectiveness of the luteolytic action was examined on the basis of progesterone check in milk in 56 treated cows. The submucous implantation of cloprostenol rapidly degraded the function of the corpus luteum since from the original level of 14.78 ng/ml progesterone decreased to 0.87 ng/ml within 72 hours. However, luteolysis did not affect all the corpora lutea. Hence the submucous administration of cloprostenol was found to be effective, and at the same time, highly economical, owing to a substantial reduction in the costs of reproduction control as well as the costs of production.

Forty-two cycling, multiparous beef cows (percentage-Brahman) were injected twice at 11-d intervals with 500mug Cloprostenol (a prostaglandin F(2alpha) analog) to induce luteolysis. Cows were randomly assigned for ovariectomy at 12 hr intervals from 0 to 72 hr post-injection. Corpora lutea were excised and one-half of the corpus luteum was stored in phosphate-buffered formalin until mounting and staining with hematoxylin and eosin. The other half of the CL was snap-frozen for determination of progesterone content and concentration. Luteal cell density increased following Cloprostenol injection and was significantly correlated with a shift from predominantly healthy Type I cells to predominantly degenerating Types III and V cells. Cell mitosis tended to decrease by 12 hr and was lower by 24 hr post-injection. Cell pyknosis increased by 24 hr post-injection and was correlated with the decrease in percentage of healthy luteal cells. No pattern was detectable in cell karyorrhexis. Histological regression of the CL was inversely correlated with CL progesterone content. Therefore, we conclude that a reduction in cell mitosis is the earliest morphological sign of degeneration of the CL and that the CL follows a well-defined sequence of regression which is accompanied by a decrease in progesterone content.

A short calving to conception interval is of main importance to achieve high economic efficiency in dairy cow industry. In order to reduce this interval, several hormonal treatments have been put on the market, in which cloprostenol, a synthetic analogue of prostaglandin F2alpha (PGF2alpha). The aim of this study was to compare fertility of cloprostenol-induced oestrus to that of spontaneous oestrus in dairy cows. In a group of 525 cows, 280 (treated group) were administered 0.5 mg cloprostenol i.m. after transrectal corpus luteum (CL) detection, and inseminated at detected oestrus during the following week. The other 245 cows (control group) were inseminated during spontaneous oestrus. Whey progesterone concentrations were checked at treatment and at insemination in order to remove from the study cows whose P4 levels indicate a non-functional CL, or a lack of luteolysis respectively. Moreover, cows that were not inseminated due to genital problems were also excluded from this study. Conception (59% vs 54.5%) and calving rates (93.7% vs 93%) were not significantly different between the two groups.

The aim of this study was to determine if initiation of superovulation in heifers during the time of development of the first dominant follicle (Days 2 to 6) would give equivalent ovulation and embryo production rates as treatment initiated at mid-cycle. Estrus was synchronized in 60 beef heifers using luprostiol (PG) and they were randomly allocated to treatment with 4.5, 3.5, 2.5 and 1.5 mg of porcine follicle stimulating hormone (FSH) administered twice daily, either on Days 2, 4, 5 and 6 (Day-2 group), respectively, or with similar doses at four consecutive days during mid-cycle (Day-10 group, initiation on Day 9 to 11). All heifers received 500 mug cloprostenol at the fifth FSH injection and 250 mug at the sixth injection. Blood samples for progesterone determination were collected at the time of FSH injections. Heifers were slaughtered 7 d post estrus, and the number of ovulations and large follicles >>/=10mm) were determined on visual inspection of the ovary. Following flushing of the uterine horns the quality of embryos and the fertilization rate were determined. Significant differences between treatments were determined using a two-sided t-test, and frequency distributions were compared using Chi-square tests. The mean number (+/-SEM) of ovulations for heifers in the Day-10 group was 12.9+/-1.0, and 8.5+/-0.9 embryos were recovered. Both the number of ovulations (6.7+/-0.8) and embryos recovered (4.1+/-0.6) were lower (P=0.0001) in heifers in the Day-2 group. Following grading based on a morphological basis, a higher number (P=0.002) of embryos was categorized as Grades 1 and 2 (4.1+/-0.6) and Grade 3 (2.1+/-0.4) in Day-10 heifers than in the Day-2 group (Grade 1 and 2, 1.9+/-0.3; Grade 3, 0.7+/-0.2). The number of Grade 4 and 5 embryos (Day 10, 1.6+/-0.2; Day 2, 1.4+/-0.2) and the number of unfertilized ova (Day 10, 0.7+/-0.4; Day 2, 0.2+/-0.1) did not differ between treatments. Progesterone concentrations were lower (P=0.0001) in Day-2 heifers at FSH treatment prior to prostaglandin, and the decline was more rapid following prostaglandin injection at Day 5 (P=0.02). Results of this study indicate that the number of ovulations and embryos recovered was lower in heifers when FSH treatment was initiated on Day 2 compared with Day 10 of the estrous cycle.

Simultaneous injections of PGF and FSH or saline were given to 32 Holstein cows to test their combined ability to improve estrous and ovulation synchrony beyond that of PGF alone. All the cows were randomly assigned to receive PGF on either Day 8 or Day 10 of the estrous cycle (estrus = Day 0), and all the cows in each group were further assigned to simultaneous injection of either FSH or saline. Regression of the corpus luteum (CL), return to estrus and follicular activity were monitored by plasma progesterone assay, twice-daily estrous detection and ultrasonographic examination, respectively. Plasma progesterone concentrations declined to <1.0 ng/ml at 24 hours after PGF treatment in all the cows and FSH did not affect this decline. Return to estrus was not affected by FSH treatment in cows treated on Day 8 or Day 10; however, FSH disrupted normal follicular activity and either delayed normal ovulation following estrus or induced premature ovulation or cyst formation in 4 of 8 PGF/FSH (Day 8) cows and 5 of 8 PGF/FSH (Day 10) cows. These data indicate that exogenous FSH administered simultaneously with a luteolytic does of PGF does not maintain viability of large, dominant follicles and, therefore, is not an effective method for the synchronization of estrus and ovulation.

The aim of this study was to determine whether sodium cloprostenol administered at a continuous low dosage induced luteolysis and polydipsia in early dioestrous bitches. Sodium cloprostenol was administered subcutaneously to greyhounds at doses of 4.04-5.19 microg/kg/day (treated group, n=5) or 0 microg/kg/day (control group, n=5) delivered by mini-osmotic pumps for 7 days. The treated bitches and two of the control bitches were in early dioestrus (Days 5-14, and 6 and 10, respectively) when the mini-osmotic pump was inserted (Day 0). Concentrations of plasmatic progesterone were measured in dioestrous bitches each day from Day -2 to 7, and then weekly until Day 90. Daily intake of water was ascertained in all bitches from Day -2 until Day 10, and their weight was measured on Days -2, 6 and 13. Biochemical analyses on plasma for concentrations of urea and glucose, and urinalyses were performed on all bitches before (Day -1), during (Day 4) and after treatment (Day 10). Concentrations of plasmatic progesterone declined dramatically and rapidly in treated bitches after Day 0 to <2.9 ng/ml but were not similarly affected in the dioestrous control bitches. However, in three of five treated bitches, concentrations of plasmatic progesterone increased to >1 ng/ml in the period from Day 10 to 90 indicating that luteolysis was incomplete. All treated bitches were polydipsic (intake of water >100 ml/kg/day) for 2-6 days during the period of treatment, and for 0-2 days immediately after treatment (Days 7 and 8). One control bitch was polydipsic on Days -2, -1 and 0. The treated bitches were also polyuric since they were hyposthenuric (<1.007, n=4) or isothenuric (1.010, n=1) on Day 4, their weight did not increase and no gastrointestinal or respiratory effects were observed. The control bitches were always hypersthenuric when measured during and after treatment (>1.021). Biochemical analyses of plasma and other data obtained from urinalyses did not reveal any differences between groups. This study indicated that sodium cloprostenol administered at a continuous low dosage induced polydipsia and suppressed luteal function in early dioestrous bitches.

Six Holstein-Friesian cows were immunized against pregnant mare serum gonadotrophin (PMSG) using Freunds' adjuvant during the mid-luteal phase of the estrous cycle. Antibody response was maintained by five booster immunizations at 2- to 3-wk intervals. Four cows were treated with a single intramuscular injection of PMSG (2350 I U) 107 d after primary immunization. Cloprostenol (500 ug) was administered at 56 h and 72 h after the treatment with PMSG; the cows were inseminated three times at 12-h intervals starting 56 h after cloprostenol treatment. Five days after insemination, the animals were slaughtered and their reproductive organs were recovered to quantify the population of corpora lutea and unovulated follicles (>10 mm dia). Antibody titres and progesterone concentrations were determined from blood samples collected either on alternate days or twice a week. Initially, progesterone concentrations were measured in milk samples. All cows produced antibodies, and titres were elevated within 6 to 9 d following each booster immunization. After each boost, however, the antibody titres declined rapidly. Progesterone concentrations declined to below 1 ng/ml after two weeks of initial immunization and remained low throughout the study, except in one cow that ovulated on Day 75. All animals were observed to have large follicular cysts during this period. Treatment with PMSG induced a single ovulation in one cow. Ovulations were neither induced by PMSG nor observed in any of the other animals. In PMSG-treated animals, the mean number of large follicles (5.0) was greater than in those which were not treated (2.0). The results of this study suggest that low titres of antibodies against PMSG are sufficient to disturb ovarian activity, result in follicular cysts and block multiple ovulations in response to exogenous PMSG.

One hundred and twenty crossbred Angus heifers, after exhibiting a 17- to 23-d estrous cycle, were placed into six groups of 20 heifers each and administered 2 ml i.m. propylene glycol containing either 0 (controls), 3.75, 7.5, 15.0 or 30.0 mg of luprostiol, or saline containing 0.5 mg cloprostenol (Groups 1 through 6, respectively). Heifers were observed for estrus every 6 h and all treatments were given 6.5 to 8.0 d after heifers were observed in standing estrus. Blood samples were collected after treatments from 10 heifers in each groups. Blood serum was assayed for progesterone. The synchronization period was considered to be 120 h after administration of luprostiol or cloprostenol. There were 0, 16, 17, 18, 20 and 18 heifers observed in estrus during the synchronization period in Groups 1 through 6, respectively. Progesterone concentrations in blood serum dropped below 1 ng/ml in 0, 8, 9, 10, 10 and 10 of the heifers from which blood samples had been taken in the six groups. All heifers observed in estrus were artificially inseminated. During the synchronization period, 0, 12, 14, 15, 16 and 10 heifers conceived in Groups 1 through 6, respectively. The interval from injection to estrus for the 89 heifers that exhibited estrus in the synchronization period averaged 49.0 h and was not different among the luprostiol and cloprostenol treated groups. Control heifers returned to estrus an average of 13.2 d after the treatment. The number of heifers that conceived at first insemination, regardless of when estrus occurred, was 16, 15, 16, 16, 16 and 12, and the total number that conceived at the first and second inseminations was 18, 18, 17, 19, 19 and 16 for Groups 1 through 6, respectively. Based on serum progesterone concentration and/or interval from treatment to estrus, 15 and 30 mg of luprostiol effectively regressed corpora lutea (100%) when administered between 6.5 and 8.0 d after estrus, and the estrous response and conception rate for these two groups equalled or exceeded that of the control and cloprostenol groups.

A trial was conducted to evaluate the ability of a prostaglandin analog, Luprostiol (LP), to synchronize estrus in Brahman cows and heifers. Animals were injected with either 0, 3.75, 7.5, 15 or 30 mg LP or 500 micrograms cloprostenol (CLP) on d 8 or 9 after estrus (d 0). All concentrations of LP (greater than 0 mg) and CLP caused luteolysis in cows and heifers, as indicated by a decline (P less than .01) in serum progesterone concentration after injection. Animals receiving 0 or 3.75 mg LP had a longer (P less than .04) interval to estrus after injection than did animals in other treatment groups. The proportion of animals exhibiting estrus by 120 h after injection was influenced by dose of LP (P less than .0001; 0, 3.75 mg less than 7.5, 15 and 30 mg and CLP) but not by age. Cows had a lower (P less than .01) progesterone concentration than heifers on d 10, 11 and 12 after LP-induced estrus. Progesterone concentration was lowest (P less than .01) on d 10, 11 and 12 after LP-induced estrus in cows given 15 mg LP or CLP. First-service conception rate was similar between cows and heifers, but it was lower (P less than .01) in animals given 15 or 30 mg LP. Both estrogen and LH concentrations were decreased (P less than .01) at the time of estrus by the 15 and 30 mg of LP. Luprostiol can cause luteolysis and estrous synchrony in Brahman cattle.(ABSTRACT TRUNCATED AT 250 WORDS)

Twelve Friesian cows, 76 to 110 days calved, with blood cell counts and Compton metabolic profile values in the normal range throughout, were housed and fed a marginal diet for varying periods before being injected twice, 11 days apart, with cloprostenol. Artificial insemination was carried out 72 and 96 hours after the second injection. Plasma concentrations of progesterone, oestradiol 17 beta, luteinising hormone, follicle stimulating hormone, and beta-carotene were monitored during this regime as were uterine and ovarian changes. Progesterone profiles were followed for a further 21 days by assay of milk samples. Subsequent rectal examinations showed that five of the 12 cows conceived at controlled oestrus and another four within 52 days of this time. No correlation was observed between time of conception and condition score, metabolic profiles or haematological parameters; there was a correlation between time of conception and plasma beta-carotene concentrations and cows with lower beta-carotene values showed cyclic irregularities or appeared to have depressed steroid hormone production.

One aim of this study was to compare the reproductive performance of cows and heifers when resynchronizing returns to estrus for a second insemination by treating with an intravaginal progesterone-releasing device (IVD) for 7 or 8d when estradiol benzoate (EB) was administered at the start of treatment and again 24h after device removal. An additional aim was to document the pattern of onset and characteristics of estrus with each resynchrony treatment. Lactating cows in three herds were synchronized for a first estrus and AI by treatment with an IVD for 8d, starting on Day 0, cloprostenol (0.5 mg im) at device removal and EB at device insertion (2.0 mg im) and 24h after removal (1.0 mg im). Cows were resynchronized for a second estrus starting on Day 23 by reinsertion of IVDs for 7 (IVD-7-EB; n=449) or 8d (IVD-8-EB; n=445) with EB (1.0 mg im) administered at device insertion and 24h after removal. Cows were resynchronized for a third estrus by administration of EB (1.0 mg im) on Day 46, but subsequent treatments (no further treatment, reinsertion of CIDR or administration of EB on Day 55) varied among herds as part of separate studies. Maiden heifers (7-Day, n=68; 8-Day, n=69) were similarly treated as cows in a separate herd, but doses of EB were always 1.0 mg im at device insertion and 0.75 mg im 24h after removal. Heifers were not resynchronized for a third estrus. Cattle were inseminated on detection of estrus at each synchronized estrus. Cumulative pregnancy rates 4 week (66.0%, 276/418 versus 59.1%, 247/418) and 7 week (72.7%, 304/418 versus 67.7%, 283/418) after the start of AI were greater (P<0.05) in the IVD-7-EB cows compared to the IVD-8-EB cows, respectively; this was associated with a 9% increase in conception rates at the second estrus (P=0.051) in the IVD-7-EB cows. Treatment did not significantly affect reproductive performance in heifers. Characteristics of estrus measured with radiotelemetry did not differ significantly between the two treatment groups, but more cows were detected in estrus 36 h after removal of IVDs in the IVD-8-EB cows compared to the IVD-7-EB cows (P<0.05). We concluded that reproductive performance in resynchronized dairy cows but not heifers was greater following resynchronization of estrous cycles after AI with an IVD for 7 compared to 8d when EB was injected at the start of treatment and 24h after device removal.

The time to resumption of ovarian cycles was determined in 27 post partum beef suckler cows by twice weekly milk progesterone analysis and was 24.5 +/ 1.6 (SEM) days. Treatment of 87 cows with a double injection of 500 micrograms cloprostenol at an interval of 11 days resulted in 55 cows (63.2 per cent) responding by a fall in progesterone levels and ovulation. Thirty-eight of these cows (43.7 per cent of total) subsequently conceived to a timed double insemination. Eight (9.2 per cent) had high progesterone levels at the second injection had failed to respond; three (3.4 per cent) had a prolonged period of low progesterone levels after the first injection which were still low when the second was given. A further 21 cows (24.1 per cent) had low progesterone levels at the time of the second injection but could not be categorised further with certainty because of lack of previous samples. These cows were treated at a significantly earlier stage post partum than the rest and in fact were treated earlier than the recommended day 42. However, four of the 21 cows conceived to the timed insemination resulting in a total conception rate of 48.3 per cent.

Reproductive management programmes based on strategic use of prostaglandin F2 alpha (PGF2 alpha) to induce and synchronize oestrus in post-partum dairy cows are widespread. Repeated shortening of the oestrous cycle during early lactation in high-yielding dairy cows, however, could impair corpus luteum function and thus decrease fertility. The objective of this study was to analyse the effect of repeated treatments with the prostaglandin F2 alpha analogue D (+) cloprostenol sodium on progesterone concentrations indicative of a functional corpus luteum in post-partum dairy cows. Furthermore, the influence of milk production, parity and endometritis on progesterone concentrations under these circumstances were studied. Eighty-four cows of a commercial dairy operation were treated three to four times with D (+) cloprostenol sodium (Preloban; Hoechst Roussel Vet, Wiesbaden, Germany) at 14-day intervals, starting 22-28 days post-partum. Blood samples were collected prior to treatment 1 (sample 1) and 14 days after treatments 1, 2 and 3 (samples 2-4) and serum progesterone (P4) levels were determined. The percentage of cows with P4 levels < 1 ng/ml decreased from 51% in sample 1 to 23% in samples 3 and 4. More primiparous cows had low P4 levels 14 days after the second treatment than older cows (P < 0.05). Cows with low progesterone levels in sample 3 or 4 had lower protein contents in milk on the second milk test day post-partum and in their cumulative milk yield of the first 100 days of lactation. Clinical endometritis at post-partum examination did not influence progesterone levels after treatment with PGF2 alpha. Repeated application of PGF2 alpha (more than twice) in the post-partum period does not influence serum progesterone levels 14 days after treatment. Failure to develop luteal tissue after treatment contributed to the conception failures after first service.

Different therapeutic strategies have been used with the objective of improve luteal function to reduce embryonic losses. The objective of this work was to study the effect of the administration of GnRH or hCG at Day 4 post fixed time artificial insemination (FTAI) on reproductive efficiency in Merino sheep during the breeding season in North Patagonia. Estrus of multiparous Merino ewes (n = 288) was synchronized by two injections of prostaglandins (PG; 125 μg, Cloprostenol), 14 days apart. Cervical FTAI was performed 53-56 h after the second PG with a dose of fresh semen (100 × 10⁶ spermatozoa) from five Merino rams. In all ewes body condition score (BCS) was determined at FTAI. At 4 days post FTAI ewes were randomly assigned into three experimental groups: GnRH group (4 μg, Buserelin; n = 99), hCG group (300 IU, hCG; n = 92) and Control group (1 ml, saline solution; n = 97). Pregnancy and pregnancy losses were evaluated by ultrasonography on Days 33 and 90 post FTAI. Additionally, embryo crown-rump length (CRL) was measured by ultrasonography (n = 12 single-pregnant ewes by experimental group) at the first ultrasound. Date of birth, litter size and lamb weight were recorded (n = 111 pregnant ewes). Pregnancy rate on Days 33 and 90 post FTAI did not differ among treatment groups (P > 0.05). Pregnancy losses at Day 33 post FTAI were lower in the hCG group compared to the GnRH and Control groups (0, 3, 7.2%, respectively; P < 0.05). Pregnancy losses between Days 33 and 90 after FTAI were negligible (P > 0.05). The embryo CRL at Day 33 post FTAI was not increased by the hormonal treatments (P > 0.05). Moreover, it was lower in GnRH group compared to Control group (P < 0.05). Litter size tended to be greater in the GnRH group compared to the hCG and Control groups (P < 0.1). The birth weight of twin lambs tended to be higher in the GnRH group compared to the Control group (P < 0.1). The birth weight of single lambs was not affected by treatments (P > 0.05). Ram fertility and BCS of ewes at FTAI influenced the effect of hormonal treatments on reproductive parameters. In conclusion, administration of hCG or GnRH at Day 4 post FTAI does not improve pregnancy rate but treatment with hCG reduces pregnancy loss on Day 33 post FTAI. GnRH treatment improves litter size and twin lambs birth weight.

The aim of this study was to assess the need of using eCG on short-term estrus synchronization protocol in nulliparous (NUL) and multiparous (MULT) dairy goats during the breeding season. Alpine (n = 20), Nubian (n = 20), and Saanen (n = 16) goats received 60 mg medroxyprogesterone acetate intravaginal sponges for 6 days plus 30 μg d-cloprostenol and 200 IU eCG (G-eCG, n = 28) or saline (G-Control, n = 28) 24 h before sponge removal. The NUL and MULT goats of each breed were equally assigned into the two treatments. Transrectal ultrasonography was used to evaluate ovulatory parameters, and teaser goats were used for estrus detection every 12 h from sponge removal to ovulation. eCG did not affect (P > 0.05) estrus response (~86%), diameter of ovulatory follicles (~6.8 mm), and number of ovulations (~1.6). Nevertheless, eCG led to earlier (P < 0.05) ovulation (G-eCG = 65.1 and G-Control = 73.2 h) and increased (P < 0.05) the ovulation rate (G-eCG = 96.4% and G-Control = 67.9%). In the absence of eCG, no differences regarding reproductive parameters (P > 0.05) were found between parity orders. Alpine MULT goats underwent a superior (P < 0.05) number of ovulations (2.2) in comparison to NUL goats (1.3). In conclusion, the exclusion of eCG from short-term estrus synchronization protocol did not interfere with estrus response but decreased the ovulation rate.

Keywords: Does; Gonadotropin; Hormone; Multiparous; Nulliparous; Reproduction.

In the present study, we investigated the effect of the synthetic analogue of prostaglandin F2α (PGF2α)-cloprostenol-on cultured steroidogenic luteal cells of selected felid species over a two day culture period. The changes induced by cloprostenol were measured based on progesterone concentration and mRNA expression analysis of selected genes. Cloprostenol significantly reduced concentration of progesterone in cell culture medium of small luteal cells isolated from domestic cat corpora lutea (CL) at the development/maintenance stage (p < 0.05), but did not influence progesterone production in cultured cells from the regression stage. A decrease or complete silencing of progesterone production was also measured in cultured luteal cells of African lion (formation stage) and Javan leopard (development/maintenance stage). Gene expression analysis by real-time PCR revealed that treatment with cloprostenol did not have an influence on expression of selected genes coding for enzymes of steroidogenesis (StAR, HSD3B, CYP11A1) or prostaglandin synthesis (PTGS2, PGES), nor did it effect hormone receptors (AR, ESR1, PGR, PTGER2), an anti-oxidative enzyme (SOD1) or factors of cell apoptosis (FAS, CASP3, TNFRSF1B, BCL2) over the studied period. Significant changes were measured only for expressions of luteinizing hormone (P < 0.05), prolactin (P < 0.05) and PGF2α receptors (P < 0.005) (LHCGR, PRLR and PTGFR). The obtained results confirm that PGF2α/cloprostenol is a luteolytic agent in CL of felids and its impact on progesterone production depends on the developmental stage of the CL. Cloprostenol short term treatment on luteal cells was associated only with functional but not structural changes related to luteal regression.

Keywords: cell cultures; cloprostenol; felids; hormone receptors; luteal cells; luteal regression; prostaglandin F2α.

The aim was to develop a program for resynchronization of ovulation (ReBreed21) that allowed reinsemination of non-pregnant Bos indicus heifers every 21 d using timed AI (TAI) without the need for detection of estrus. The Rebreed21 program begins 12 d after previous TAI (Day 0) by inserting an intravaginal P4 implant (Day 12) that is removed 7 d later (Day 19) combined with treatment with 300 IU of eCG. On Day 21, early pregnancy diagnosis (Doppler PD) is performed based on CL vascularity. Non-pregnant (NP) heifers immediately received AI combined with 100 μg of GnRH. The program is replicated 12 d after second TAI to produce a breeding season (BS) of 42 d with 3 potential TAIs. Two experiments were conducted as a proof of concept for this rapid rebreeding program. In Experiment 1, 76 heifers were enrolled in ReBreed21, as explained above. In Experiment 2, 300 Nellore heifers were synchronized for 1st TAI and randomly assigned to one of two groups: ReBreed21 (n = 147) or another early resynchronization procedure, Resynch14 (n = 153) with P4 implant inserted 14 d after previous TAI plus 50 mg of long-acting injectable P4; 8 d later P4 implant removed (Day 22) and early Doppler PD performed; NP heifers received 150 μg of cloprostenol, 0.5 mg of ECP, and 300 IU of eCG with TAI on Day 24. In both experiments, the largest follicle (LF) was measured at each Resynch TAI. Ultrasound was later used to confirm the early Doppler PD and to determine ovulation (OV) to Resynch at 12 d after TAI in ReBreed21 (Day 33 of pregnancy) and 14 d after TAI in Resynch14 (Day 38 of pregnancy). Final PD was performed 40 d after 3rd TAI. Results for Experiment 1 were: diameter of LF 11.8 ± 0.23 mm; 88.9% OV; 20.5% false positives; 38.1% P/AI at 1st TAI; 44.4% overall P/AI for ReBreed21 TAIs; 72.3% total pregnant at end of BS. In experiment 2, Rebreed21 vs. Resynch14 were different for: diameter of LF (10.9 ± 0.17 vs. 10.0 ± 0.17 mm, P = 0.0003), heifers with LF < 8.5 mm (10.2 vs. 26.4%, P = 0.04), or LF ≥ 11 mm (50.0 vs. 37.2%, P = 0.001), and P/AI at first TAI (29.3% [43/147] vs. 20.3% [31/153], P = 0.074) but similar for OV (overall 86.8% [239/275], P = 0.82), false positives (P = 0.52) overall P/AI for Resynch TAIs (33.6 vs. 28.8%, P = 0.4), and total pregnant at end of BS (58.5% [86/147] vs. 55.6% [85/153], P = 0.64). In addition, median time to pregnancy was 9 d earlier (P = 0.0007) for ReBreed21 than Resynch14. Thus, ReBreed21 is a novel protocol that allows earlier reinseminations than Resynch14 but with similar fertility.

Keywords: Doppler ultrasound; Reproductive efficiency; Resynchronization; TAI.

Fifty-six cows received a norgestomet implant and an injection of norgestomet and estradiol valerate; half (n = 28) received 500 IU equine chorionic gonadotrophin (eCG) at implant removal, 9 d later. A third group (n = 25) received 2 doses of cloprostenol (500 micrograms) 11 d apart. Estrous rate was higher (P < 0.05) for cows given norgestomet and estradiol plus 500 IU eCG (75.0%) than for those receiving cloprostenol (44.0%); for those receiving norgestomet and estradiol alone, it was intermediate (67.8%). Pregnancy rates to artificial insemination (after estrus or timed) were higher (P < 0.05) for cows given norgestomet and estradiol than for those given cloprostenol (23 of 28, 82.1% vs 13 of 25, 52.0%), and intermediate (67.8%) for those given norgestomet and estradiol plus eCG. In a second experiment, for heifers treated with norgestomet and estradiol plus eCG (n = 15) or with 2 doses of cloprostenol (n = 16), estrous rates were 66.7% vs 56.2% (P>> 0.5), ovulation rates were 100.0% vs 81.2% (P = 0.08), intervals from implant removal or cloprostenol treatment to estrus were 48.0 +/- 4.4 hours vs 61.3 +/- 7.0 hours (P = 0.12) and to ovulation were 70.4 +/- 4.4 hours vs 93.2 +/- 7.5 hours (P < 0.01), respectively; pregnancy rates were 41.7 and 35.7%, respectively (P>> 0.5). Norgestomet and estradiol were as good as (heifers) or superior to (cows) a 2-dose cloprostenol regimen. In cows given norgestomet and estradiol, injecting eCG at implant removal did not significantly improve estrous or pregnancy rates.

Five experiments were conducted with the objective of developing a method to induce superovulation in ewes with a single i.m. injection of FSH. This was achieved by injection of 10 mg FSH-P in propylene glycol at the same time as luteolysis was induced by cloprostenol on day 13 of the oestrous cycle (day 0 = oestrus). Experiments 1, 2, 3 and 5 were conducted in a single flock of Manchega ewes in Spain during the breeding season. Ovulation rates were determined at laparoscopy. In Expt 1, FSH-P was diluted in saline, and neither 5 mg FSH on day 1 nor 5 or 10 mg FSH-P on day 13 changed the ovulation rate after cloprostenol treatment on day 13. In Expt 2, FSH-P was diluted in propylene glycol and data were collected over 2 years. Ten milligrams FSH-P, on day 13 only, increased (P < 0.01) the mean number of corpora lutea to 5.5 compared with a control value of 1.5. Five milligrams FSH-P on day 13 only had no effect; however, 5 mg FSH-P on day 1 reduced the mean number of corpora lutea formed in ewes receiving 10 mg FSH-P on day 13 to 2.6 (P < 0.01). Saline and propylene glycol, as vehicles for 10 mg FSH-P, were compared directly at two times of injection in Expt 3. FSH-P increased the mean number of corpora lutea when injected on day 13 in propylene glycol (4.7) but not in saline (2.5; P < 0.5).(ABSTRACT TRUNCATED AT 250 WORDS)

Thirty heifers were given 11 subcutaneous injections of 5 mg estradiol benzoate and 200 mg progesterone every 3 d to develop their mammary glands. Three days later groups of animals were treated with 1) 20 mg dexamethasone twice, 2) 500 micrograms cloprostenol thrice, 3) dexamethasone and cloprostenol, 4) oxytocin 4 IU six times, or 5) no further injections. Two further groups of six heifers each (6 and 7) were treated in a manner similar to groups 1 and 3 except the dose of estrogen to develop their mammary glands was doubled to 10 mg/3 d. Six lactating first calf heifers were controls (8). The proportion of animals lactating, combined milk yield for each group (kg), and mean days lactated were 1) 5/6, 3831, 142; 2) 1/6, 912, 195; 3) 6/6, 4898, 194; 4) 3/6, 1066, 128; 5) 1/6, 293, 154; 6) 6/6, 6109, 130; 7) 6/6, 6265, 130; and 8) 6/6, 19, 190, 251. The lactogenic response to dexamethasone and oxytocin is similar to that in sheep, but the response to cloprostenol indicates a species difference. Intensive blood sampling before and after injection of hormones, intended to trigger lactogenesis, showed that plasma prolactin rose to peaks above 210 ng/ml in cows of groups 2, 3, and 4 and were unchanged from the base below 40 ng/ml in groups 1 and 5. Monitoring of steroids after induction treatment showed estradiol-17 beta ranged between 35 and 400 pg/ml and 20 to 80 pg/ml in mammary secretion and plasma and progesterone concentrations were less than at diestrus.(ABSTRACT TRUNCATED AT 250 WORDS)

A total of 480 female camels with a history of conception failure were examined through transrectal palpation, ultrasonography, and vaginal exploration. Animals were categorized according to parity (nulliparous n=200 vs. multiparous n=280), and type of uterine infection (endometritis n=360 vs. metritis n=120). They were randomly assigned to receive one of three intrauterine treatments: (i) 100mL acriflavin 0.1% (group 1, n=170), (ii) 100mL lotagen 4% (group 2, n=200), or (iii) 300mg/100mL gentamicin sulphate (group 3, n=110). All groups received 500microg cloprostenol IM at infusion. Animals were exposed for breeding 7 d later and received 5000 IU hCG im at mating. The criteria for efficacy of treatment were 90 days non-return rate (90 d NRR) and calving rate (CR). The results showed that the 90 d NRR and CR were significantly influenced by parity, type of uterine infection, regime of treatment, and their interactions, P<0.05. Treatment regimes were approximately equally efficient in treating females with endometritis (90 d NRR were 64%, 53.1% and 53.3% and CR were 58.9%, 49.3%, and 42.5% for groups 1, 2, and 3, respectively, P>0.05). In contrast, regimes differed in treating those with metritis (90 d NRR were 55.6%, 75%, and 28.6% and CR were 31.6%, 54.8%, and 12.5% for groups 1, 2, and 3, respectively, P<0.05). In conclusion, a regime consisted of intrauterine lotagen infusion and administration of PGF(2)alpha at infusion and hCG at mating was more efficient for treating female camels with metritis.

Conception rates after timed artificial insemination (TAI) are of paramount importance for the success of protocols based on synchronization of ovulation. Stage of lactation and milk production level are known factors that influence dairy cow fertility. It was the objective of this study to analyse the effect of stage of lactation and milk production level on conception rates and pregnancy rates by 200 days in milk (DIM) in dairy cows synchronized with the Ovsynch protocol (Day -10, Day -1: 0.1 mg of D-Phe6-gonadorelin, Day -3: 0.5 mg of cloprostenol, Day 0: AI). A total of 1,288 dairy cows were assigned to two groups and classified in three production levels (high, average, low). Cows of all milk production levels in Group 1 (Simultaneous Ovsynch, SO) were synchronized with the Ovsynch protocol simultaneously for TAI between 73 and 81 DIM. In Group 2 cows with average milk production were synchronized at the same time as Group 1, while low producing cows were synchronized 3 weeks earlier and high producing cows were synchronized 3 weeks later than Group 1, respectively. First service conception rates (FSCRs) were lower (P<0.05) in cows synchronized earlier than in cows of the same production level synchronized later (low production: 14.4% (22/153) versus 34.5% (51/148); high production: 28.2% (40/142) versus 41.4% (53/128)). Milk production level had no significant impact on conception rates after TAI in cows synchronized at the same stage of lactation. At 200 DIM fewer cows with high production level were pregnant than cows with average or low production (P<0.05). This effect was independent of the stage of lactation at the initiation of Ovsynch. Endometritis at a postpartum examination did not influence conception rates after TAI. In conclusion, stage of lactation, but not milk production level, has a major influence on conception rates after TAI. Early AI after Ovsynch is less efficient and therefore its return on investment should be evaluated carefully.

The objective of this study was to investigate the effect of time of first postpartum ovulation after calving on uterine involution in dairy cows with and without uterine puerperal disease. Transvaginal follicular puncture (FP) of follicles >6 mm suppressed ovulation and development of a CL until Day 42 after calving. Fifty-three lactating Holstein Friesian cows (3.4 ± 1.2 years old, parity 2.5 ± 1.0 [median ± mean absolute deviation]) were divided into groups on the basis of the presence (UD+) or absence (UD-) of uterine disease and whether FP was carried out (FP+) or not (FP-). Uterine disease was defined as the occurrence of retained fetal membranes and/or metritis. This resulted in the following groups: UD-FP- (n = 15), UD-FP+ (n = 13), UD+FP- (n = 13), and UD+FP+ (n = 12). A general examination, vaginoscopy, transrectal palpation, and transrectal B-mode sonography of the reproductive organs were conducted on Days 8, 11, 18, and 25 and then every 10 days until Day 65 after calving. After hormonal synchronization of ovulation (cloprostenol between Days 55 and 60 postpartum and GnRH 2 days later), cows were inseminated in the next spontaneous estrus. On average, the cows ovulated on Day 21.0 ± 6.0 (UD-FP-), 50.0 ± 4.0 (UD-FP+), 16.0 ± 3.0 (UD+FP-), and 48.0 ± 2.0 (UD+FP+) postpartum. Calving-to-conception interval and first-service conception rates were not affected by FP (P>> 0.05). Healthy cows with FP had smaller (P < 0.05) uterine horn and cervical diameters assessed sonographically than cows without FP. FP reduced the prevalence of purulent vaginal discharge and uterine size assessed transrectally in UD+ cows (P < 0.05). The results showed that suppression of an early ovulation by transvaginal FP improved uterine involution in cows with and without uterine disease.

OBJECTIVE

To determine whether there are differences in postpartum gonadotrophic activity between strains of Holstein-Friesian dairy cows genetically selected on mature liveweight that might explain differences between the strains in fertility, and the interval between calving and the resumption of ovarian follicular activity.

METHODS

Mixed-age Holstein-Friesian cows fed generous allowances of ryegrass/white clover pasture, and genetically selected for heavy (H) or light (L) mature liveweight, were given 10 microg buserelin on Days 21, 28, 35 and 42 (Experiment 1a; n=8/group), or Days 7, 14, 21 and 28 (Experiment 1b; n=8/group) postpartum. The same dose of buserelin was also given to first-calved heifers from each strain (Experiment 1c; n=6/group) on Days 7, 14, 21, and 28 postpartum. Luteinising hormone (LH)concentrations were measured in serial blood samples that were taken for up to 240 min after administration of buserelin. In Experiment 2, serial blood samples were taken at 15-min intervals from H and L cows (n=7/group) over 8 h on Days 14, 21, 28 and 35 postpartum, to examine the endogenous secretion patterns of LH and follicle stimulating hormone (FSH). The time-course of the restoration of positive feedback between oestradiol and LH was examined by giving 1 mg oestradiol benzoate(ODB) 48 h after administration of 500 microg cloprostenol to mixed-age cows from each strain on Days 7 and 21 (n=8/group), or 14 and 28 (n=8/group) after calving (Experiment 3). Relationships between nutrition and the restoration of positive feedback were studied by giving 0.75 mg ODB/500 kg liveweight on Day 17 or 18 after calving to pure-bred Holstein (OSH) and New Zealand Friesian (NZF) cows that were fed either pasture (n=12 OSH, 12 NZF) or a total mixed ration (TMR; n=13 OSH,12 NZF) (Experiment 4). Plasma LH and FSH concentrations were measured in samples collected for 42 h (Experiment 3) or 48 h (Experiment 4) after treatment with ODB. Milk progesterone concentrations were measured 3x weekly to define the reproductive status of animals in each experiment. Conception rates were recorded for animals in all of the experiments.

RESULTS

First-service conception rates were lower (p<0.05) in H than L cows (46% vs 59%). In Experiments 1b and 1c, LH response to buserelin increased between Days 7 and 28 postpartum (both p<0.001), but did not differ between strains (p=0.77 and p=0.19, respectively). In Experiment 1a, LH responses to buserelin did not change between Days 21 and 42 postpartum, but overall mean peak concentrations were significantly(p<0.001) greater in L than H cows. In Experiment 2, anoestrous H cows had higher mean (p=0.004) and episodic (p=0.001) concentrations of LH than did L cows, but in cows that had active corpora lutea there were no such differences. There were no differences in FSH concentrations between strains. LH secretion in response to exogenous oestradiol (Experiment 3) increased between Days 7 and 28 postpartum (p<0.001), but there were no differences between strains. Responses were also similar in OSH and NZF cows on Day 17 or 18 postpartum, although there was a significant effect of ration upon the proportion of cows that exhibited an LH surge (20/24 cows on grass vs 12/25 on a TMR; p=0.005).

CONCLUSIONS

These results confirm that H cows have poorer first-service conception rates than L cows, but do not support an hypothesis that there are major differences between these strains of Holstein-Friesian dairy cows in the rate of restoration in the hypothalamo-pituitary axis. However, in anoestrous cows, differences between strains in the endogenous release of LH maybe related to an earlier onset of oestrous cycles in H animals.