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A normal, live 35-kg water buffalo bull calf was born 300 days after it was nonsurgically collected as a 7-day blastocyst from a water buffalo donor and transferred nonsurgically to an unrelated water buffalo recipient. The development of estrus synchronization, superovulation and estrus detection methods in water buffalo are described.
This study was conducted to determine whether acute fasting in Holstein heifers enhances the suppressive effect of an intravenous injection of ethanol on pulsatile LH release (LH pulse) and, additionally, to establish whether or not the mechanism is estrogen-dependent. After estrus synchronization (Day 0 = estrus), 29 heifers were either fasted (fasting group; n = 14) or fully fed as a control (control group; n = 15) from Days 1 to 4. On Day 4, blood samples were taken at 10-min intervals for 4 h before (pre-injection period) and after (post-injection period) an intravenous injection of 1.5 mL of saline, 1.5 mL of ethanol , or 35 mg of tamoxifen dissolved in 1.5 mL of ethanol . We analyzed the mean LH level, the number of LH peaks per 4 h, and the amplitude of LH peaks. No differences were observed in the LH pulse in the pre-injection period between the control and the fasting group. However, in the post-injection period, compared with the saline injected control heifers, ethanol suppression of the LH pulse was observed in the number of LH peaks of the ethanol injected control heifers and in all pulse parameters of the ethanol injected fasting heifers. Furthermore, tamoxifen inhibited suppression of ethanol on LH pulse was observed in the control and fasting heifers injected with tamoxifen dissolved in ethanol. It was concluded that acute fasting in Holstein heifers has an enhancing effect on ethanol inhibition of the LH pulse and that the mechanism may be estrogen-dependent.
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Two experiments were conducted to determine the conception rates of heifers time-inseminated following melengestrol acetate/prostaglandin F(2alpha) (MGA/PG) estrous synchronization treatment. In Experiment 1, timed insemination of heifers at 72 h after the PG injection, without regard for behavioral estrus, tended to improve (P < 0.15) the percentage of heifers pregnant to artificial insemination (AI) compared with that of synchronized heifers bred 12 h after they were first detected in estrus. In the timed-insemination treatment, heifers exhibiting behavioral estrus 48 to 72 h after PG tended to have an increased (P < 0.15) conception rate to AI compared with heifers exhibiting estrus within 48 h of PG administration. In Experiment 2, the number of heifers conceiving to AI following the MGA/PG estrous synchronization regimen was increased by mass insemination of all heifers not exhibiting estrus by 72 h after PG. The pregnancy rate to AI was higher in heifers with serum progesterone (P(4)) concentrations higher than 1 ng/ml compared with that of heifers with concentrations lower than 1 ng/ml. Of heifers with serum P(4) greater than 1 ng/ml, the pregnancy rate to AI tended to be higher when concentrations exceeded 2 ng/ml than when concentrations were 1 to 2 ng/ml. In cyclic heifers, timed insemination can increase the percentage of heifers pregnant after being synchronized with MGA/PG.
The objective of this experiment was to determine the effect of prepartum lipid supplementation on the number and quality of embryos recovered following ovarian super-ovulation in postpartum suckled beef cows. Mature cows (n = 40) were assigned to one of two treatments (lipid versus. no lipid) and supplemented for approximately 40 days prior to calving. Supplements provided to cows were isocaloric and isonitrogenous. The treatment group was fed 1.6 kg hd(-1) per day of whole soybeans (WSB; 19.8% ether extract, and 41.8% crude protein) and the control group received a supplement consisting of 1.8 kg hd(-1) day of a soybean meal and soy-hull combination (SBS; 2.15% EE and 36.81% CP). Cows were synchronized using a GnRH [Cystorelin((R)) 100 microg im]-GnRH-PGF(2alpha) [Lutalyse 25 mg im] protocol. Cows were administered two injections of GnRH seven days apart and PG seven days after the second GnRH injection. Twenty-eight cows (WSB, n = 15; SBS, n = 13) responded to estrus synchronization and were superstimulated. Super-ovulation was initiated on day 8-10 of the synchronized cycle by twice-daily injections of pFSH (Pluset) over four days in decreasing doses using a total of 608.4 IU per cow. Prostaglandin F(2alpha) was administered 96 and 108 h after super-stimulation was initiated with FSH. Days postpartum (WSB = 59 days; SBS = 57 days) at initiation of FSH treatments were similar (P > 0.10) for both treatments. Cows were monitored for estrus activity by the HeatWatch Estrus Detection System. Twenty-seven cows (WSB, n = 15; SBS, n = 12) exhibited estrus after FSH and inseminated at 0, 12, and 24 h after the onset of estrus with 1, 2, and 1 units of semen, respectively. Embryos were recovered and evaluated 7-8 days later. Only cows that responded to FSH and that were inseminated were used for statistical analysis. Data were analyzed using the General Linear Models Procedure of SAS. Body condition scores did not differ (P > 0.10) between treatments when cows were evaluated at the initiation of the experiment, two weeks prior to calving, and at initiation of superovulation with FSH. Estrous cyclicity prior to the initiation of estrus synchronization did not differ (P > 0.10) between treatments. There was no difference (P > 0.10) between treatments in recovery of total embryos (WSB, 14.7 +/- 3.5; SBS, 17.5 +/- 3.0), transferable embryos (WSB, 10.3 +/- 2.5; SBS, 13.6 +/- 2.6), degenerate embryos (WSB, 3.3 +/- 1.1; SBS, 1.6 +/- 1.7) or unfertilized ova (WSB, 1.1 +/- 0.5; SBS, 2.3 +/- 1.2). Cows that were supplemented with whole soybeans prior to parturition failed to produce an increased total number of ova or transferable embryos following super-ovulation.
Current study assessed differences in the response of sheep to estrus synchronization either by the administration of two doses of prostaglandin or by the insertion of an intravaginal progestagen sponge. The preovulatory follicular dynamics and estradiol secretion, the ovulatory response and progesterone secretion and the number and quality of embryos were studied in 27 ewes treated with two doses of 100 microg of cloprostenol, 10 days apart, and in 29 sheep treated with progestagen sponges for 14 days. Percentage of sheep responding to the synchronization treatments with signs of estrus behaviour was similar between both groups (81.5% versus 72.4%, respectively). The use of progestagen resulted in a higher diameter of the largest follicle (6.6+/-0.2 versus 5.9+/-0.2, P<0.05), and a lower number of small (6.7+/-0.3 versus 9.6+/-0.4, P<0.005) and total follicles (10.3+/-0.3 versus 12.9+/-0.4, P<0.005). However, mean plasma estradiol concentration during the follicular phase was higher in cloprostenol treated sheep (P<0.005). The mean ovulation rate was similar in both treatments (1.7+/-0.2 versus 1.7+/-0.3), but progesterone concentration during the early luteal phase was again higher in sheep treated with cloprostenol (P<0.05). The mean number of retrieved oocytes/embryos was very similar in both treatments (1.2+/-0.2 versus 1.4+/-0.2) and showed similar fertilization rates (70.6% versus 66.7%), but, although differences did not reach statistical significance, final viability rate was higher in cloprostenol than in progestagen treated ewes (58.9% versus 46.1%, P=0.07). Current results give new evidences supporting the negative effects of progestagens on the functionality of ovulatory follicles and support the development of new protocols for assisted reproduction including the use of prostaglandin analogues.
Eighty-four nulliparous ewes were used to examine the effect of short-term insulin treatment on feed intake and reproductive performance. Following estrus synchronization, ewes were observed for estrus (= Day 0) and were penned individually beginning on Day 7. Ewes were fed twice daily and feed intake was recorded. On Days 9 through 13, ewes were treated s.c. with 1 IU/kg BW insulin (n = 44) or an equivalent volume of saline (n = 40). On Day 14, ewes were placed with fertile rams and number of ewes in estrus (bred) was recorded. Thirty days post-breeding, ewes were checked for pregnancy via ultrasonography. Feed intake and percentage of ewes in estrus did not differ between saline- and insulin-treated ewes. Similarly, neither pregnancy rate (69 +/- 8.7% vs. 80 +/- 8.1%, respectively) nor lambing rate (61 +/- 8.9% vs. 78 +/- 8.4%, respectively) differed between treatments. The number of lambs born per ewe was, however influenced by a breed-group effect (P < 0.0002). Romanov ewes had more (P < 0.001) lambs than the other breed groups in the study. Therefore, treating well-nourished, nulliparous ewe lambs with insulin did not increase reproductive efficiency, possibly because the ewes were already at a maximal nutritional and/or reproductive state.
The experimental objective was to compare synthesis of oviductal secretory proteins of dairy cows bearing a persistent dominant follicle (PDF) versus a fresh dominant follicle (FDF) at estrus. On Day 7 after synchronized estrus (Day 0), cows received an intravaginal progesterone device and injection of prostaglandin F2alpha (PGF2alpha). On Day 9, cows received an injection of a GnRH agonist (FDF group; n = 3) or received no injection (PDF group, n = 3). On Day 16, all cows received PGF2alpha, and progesterone devices were removed. At slaughter on Day 18 or Day 19, oviducts ipsilateral and contralateral to the dominant follicle were divided into infundibulum, ampulla, and isthmus regions. Explants from oviductal regions were cultured in minimal essential medium supplemented with [3H]leucine for 24 h. Two-dimensional fluorographs of proteins in conditioned media were analyzed by densitometry. Rate of incorporation of [3H]leucine into macromolecules was greater in the infundibulum, ampulla, and isthmus of FDF cows (p < 0.01). Overall, intensities of radiolabeled secretory protein (P) 2 and P13 were greater for FDF than for PDF. In the ampulla, P14 was more intense for FDF while P7 was more intense for PDF. Abundance of P1 in the isthmus was greater for PDF cows. Across regions, P5, P6, P8, P9, and P11 were more intense for PDF than for FDF in the ipsilateral side. In the contralateral side, P19 was more intense for PDF than for FDF, whereas P6, P8, P9, and P11 were more intense for FDF. Differences in biosynthetic activity and in secreted oviductal proteins from cows bearing a PDF may contribute to the decrease in fertility associated with a PDF.
A synchronization treatment was initiated when each of 1227 heifers (four trials) was tailpainted. The tailpaint was sprayed with an aerosol raddle at the end of the treatment period. The heifers were in herds of 20 to 279 animals. Each herd was observed for estrus at selected post treatment intervals. A heifer was considered to be (or to have been) in estrus when the raddle was rubbed off. In three of the trials, animals which had the raddle removed were inseminated at 48 h following the end of the synchronization treatment. The tailpaint of an inseminated animal was scored from 0 (less than 10% of the paint remained) to 5 (more than 90% of the paint remained) and was then reraddled with a second color. The detection-insemination sequence was always repeated at 72 and 96 h, and sometimes at 120 h. Animals which had been previously inseminated, but then had paint scores reduced by at least 2 units were reinseminated 24 h later. Over the four trials, 94.5% of the heifers were detected in estrus through the use of the tailpaint and raddle system. The remaining 67 animals included only 10 (0.8%) which had ovulated without being detected in estrus. The reinsemination rate on consecutive days was 11.3% and was highest among animals that had a tailpaint score of 4 or 5 at 48 h. The proportion of animals detected in estrus at selected posttreatment intervals varied with the different synchronization treatments used within one herd, or with the same treatment used in different herds. The combination of tailpaint, raddling, tailpaint scoring and reraddling is a simple sequence which can be effectively used to detect estrus among heifers synchronized in research or commercial herds.
The Controlled Internal Drug Releasing (CIDR) device is an intravaginal pessary containing progesterone (P4) designed for synchronizing estrus in ruminants. To date, there has been little information available on the timing, duration, and quality of the follicular phase after CIDR removal and how those characteristics compare with natural periovulatory endocrine events. The present communication relates the results of methods we used to characterize the endocrine events that followed CIDR synchronization. Breeding-season ewes were given an injection (10 mg) of Lutalyse (PGF2 alpha), and then studied during three consecutive estrous cycles, beginning in the luteal phase after the estrus induced by PGF2 alpha. Cycle 1 estrus was synchronized with 1 CIDR (Type G) inserted for 8 d beginning 10 d after PGF2 alpha. Cycles 2 and 3 were synchronized with two CIDRs for 8 d beginning 10 d after previous CIDR removal. Cycle 1 estrous behavior and serum gonadotropins showed a follicular phase (the interval from CIDR withdrawal to gonadotropin surge [surge] peak) of 38.2 +/- 1.5 hr. Two CIDRs lengthened the interval to 46.2 +/- 1.5 hr (P < 0.0001). At CIDR removal, circulating P4 concentrations were higher in ewes treated with two CIDRs (5.1 +/- 0.3 and 6.4 +/- 0.4 ng/mL in Cycles 2 and 3 vs. 2.7 +/- 0.3 ng/mL in Cycle 1), whereas estradiol concentrations were higher in the 1 CIDR cycle (3.3 +/- 0.5 pg/mL in Cycle 1 vs. 0.5 +/- 0.1, and 0.7 +/- 0.2 pg/mL in Cycles 2 and 3), suggesting that the lower levels of P4 achieved with one CIDR was not sufficient to arrest follicular development. There were no differences in any other endocrine variable. Both one and two CIDR synchronization concentrated surges within a 24-hr period in 92% of the ewes in Cycles 1 and 2. Cycles 3 ewes were euthanized at estimated luteal, early follicular, late follicular, LH surge, and secondary FSH rise timepoints. Endocrine data and ovaries showed that 88% of the ewes synchronized with two CIDRs were in the predicted stage of the estrous cycle. These data demonstrate that the CIDR device applied during the luteal phase effectively synchronizes estrus and results in a CIDR removal-to-surge interval of similar length to a natural follicular phase.
An experiment was conducted to study an estrous synchronization regimen that involved the use of prostaglandin F(2alpha) (PGF(2alpha)) alone or in combination with estradiol benzoate (EB) and appointment breeding. Fifty-three registered Angus yearling heifers and 167 registered Angus cows (3 to 9 yr of age) were given two injections of PGF(2alpha) 11 d apart. Forty-eight hours after the second injection of PGF(2alpha') a random sample consisting of 117 cows and heifers was injected with EB in sesame seed oil. All females in the herd were artificially inseminated 80 h after the second injection of PGF(2alpha). Nearly equal percentages (25.1 vs 25.6%; P = 0.93) of treated (EB) and control (no EB) females conceived at the appointment breeding. Use of EB tended to reduce (P = 0.06) natural service conception rate (83.4 vs 93.1% for EB and control groups, respectively). Estrous synchronization treatment did not affect interval from Day 1 of the breeding season to calving.
In 664 primiparous gilts inseminated twice after oestrus synchronization, the timing of insemination had an important effect on conception rate and litter size. Best results (586 liver piglets per 100 pregnancies) were achieved by a first insemination on the afternoon of the 6th day. Another trial, with sexually mature gilts which were subsequently slaughtered, invloved artificial insemination at a planned time after synchronization of oestus with TRUISYNCHRON plus injection of Prolosan. Pregnancy rate and embryo count on the 29th day after synchronization were good when the first insemination was made on the evening of the 5th day and the second on the evening of the 6th day. Insemination on the morning and evening of the 6th day gave better results than on the evening of the 5th and morning of the 6th day.
The present study was designed to establish whether factors such as previous estrus synchronization, corpus luteum and embryo number at the time of pregnancy diagnosis, changes in body condition score, milk production, clinical disease (mastitis or lameness) and the inseminating bull affect pregnancy loss from 38 to 90 days of gestation. We derived data from 601 pregnant lactating dairy cows from a single herd. Pregnancy diagnosis was performed by ultrasonography between Day 38 and 44 following insemination. We also recorded corpus luteum and embryo number at this time. Pregnancy loss was defined as a negative pregnancy diagnosis on the second palpation per rectum undertaken between 90 and 96 days after insemination. Data were analyzed using multiple logistic regression methods. Cows that had an additional corpus luteum were eight times less likely to miscarry. The risk of pregnancy loss was 3.1 times higher in cows bearing twins. A one unit reduction in body condition score from previous partum to 30 days postpartum resulted in a 2.4-fold increase in pregnancy loss. We noted a higher incidence of pregnancy loss in cows inseminated using semen from one of the six bulls used. This particular bull led to a 3.4-fold increase in the rate of pregnancy loss. Logistic regression analysis showed no significant effects of previous estrus synchronization, milk production, clinical disease, body condition at previous partum or at pregnancy diagnosis, or body condition change between previous partum and pregnancy diagnosis. Our findings indicate a positive relationship between the presence of an additional corpus luteum and the maintenance of gestation. Risk factors for pregnancy loss were twin pregnancy, reduced body condition after previous parturition and the inseminating bull.
One hundred and sixty-four beef heifers representing Angus, Brahman and their crosses were subjected to estrus synchronization treatment following evaluation of weight, body condition score and reproductive tract. Heifers were assigned to 1 of 2 methods of estrus detection, either visual observation for signs of standing estrus or a rump-mounted pressure-sensitive detection device. All heifers were artificially inseminated during a 25d insemination period and then bred by a bull. The effectiveness of estrus detection and timely insemination were evaluated by the type of detection method, breed and breeding event resulting in a pregnancy. Although there was not a significant difference in first service conception for method of detection, at the end of a 25-d insemination period, 60.5% of the visually observed heifers were pregnant, while only 45.8% of the heifers detected by the mount detection device were pregnant (P = 0.05). The reduced 25-d conception rate in the pressure-sensitive detection group suggested that insemination of detected heifers may not have been optimal for pregnancy. The mean time to estrus after implant removal and the time of insemination were evaluated. Heifers pregnant at the first service had a shorter time to estrus (32.11 +/- 1.6 h, P=0.13), a longer mean interval from the start of estrus to insemination (12.10 +/- 1.2 h, P = 0.16) and a positive mean interval between the end of standing estrus and the time of insemination (3.17 +/-1.7 h, P=0.19) than heifers not conceiving at the synchronized estrus (38.5 +/- 2.1, 7.42 +/- 1.9 h and -2.04 +/- 2.1 h, respectively). Breed differences were observed in estrus durations (Angus 8.52 +/- 1.2 h, Brahman 6.65 +/- 1.2 h, crossbred 11.90 +/- 1.2 h; P = 0.03), number of mounts (19 +/- 3.6, 25 +/- 5.4, 37 +/- 5.5, respectively; P=0.02) and gestation length (281 +/- 1.2, 291 +/- 1.8, 286 +/- 1.1 d, respectively; P = 0.001).
A series of experiments was conducted to determine why ovariectomized heifers exhibit estrus after they are treated with the estrus synchronization product, Syncro-Mate B(*) (SMB). In Experiment 1, 23 of 40 (58%) ovariectomized heifers exhibited estrus after treatment with SMB. The mean concentration of estradiol-17beta (E(2)) in serum was lower (P < 0.001) before treatment than after implant removal in ovariectomized heifers treated with SMB. Six of 10 heifers from which serum was collected to determine concentrations of LH exhibited estrus and 5 of 6 had a surge of LH in serum after implant removal. In Experiment 2, when no estradiol valerate (EV) was given or when the norgestomet implant period was extended from 9 to 18 d, no heifer exhibited estrus after implant removal. The mean concentration of E(2) for 3 d after implant removal was lower (P < 0.001) in ovariectomized heifers with implants for 18 d versus those with implants for 9 d and was also lower (P < 0.001) in ovariectomized heifers treated only with norgestomet compared with those receiving the standard SMB treatment. When estradiol-17beta was substituted for EV in the SMB treatment, serum E(2) was lower (P < 0.001) after implant removal than in heifers receiving the standard SMB treatment. Experiment 3 demonstrated that combining a norgestomet implant or implant plus a 3-mg injection of norgestomet with EV did not alter concentrations of E(2) in serum on the days when synchronized estrus would be expected following SMB treatment. The results indicate that the SMB-induced estrus in ovariectomized heifers is dependent upon EV in the SMB treatment. Apparently, EV elevates the concentration of E(2) in serum, and the E(2) remains sufficiently high to induce estrus after implant removal.
Variability of the endometrium thickness and surface epithelium in sheep after the effect of 20 mg of chlorsuperlutin (vaginal swabs soaked in this preparation) was studied. The results obtained demonstrate that at the estrus synchronization by means of the above mentioned preparation similar changes of the endometrium can be observed as those at the estrus without controlled sexual activity. It was found out that the variability of the endometrium thickness, both in cows and heifers, depended on the follicle-stimulating hormone evoking mucosal edematization in view of the phase of the cycle when the ovulation-inhibiting factor came into play. The endometrium thickness of 1499.5 mu on an average was found in sheep with synchronized estrus, and the epithelium thickness was, on an average, 24.7 mu. We observed, according to the results of the experiment, that the endometrium thickness varied even in a given animal; the changes were the same on the endometrium of both horns of uterus independent of the ovary showing cyclical changes. The endometrium gets thinner towards to oviduct, its thickness not being uniform in individual parts of uterus. The results can be distorted due to some artificial factors, e.g. the way of sampling, fixating media, etc. Nevertheless, an assessment of the variability of the endometrium thickness belongs to objective methods to examine the functional activity of ovaries and to find out the state of uterine mucosa.
Treatment with GnRH and PGF2 alpha is a practical method for controlling ovarian follicular and luteal functions and increasing the precision of estrus synchronization in cyclic and acyclic postpartum cows and heifers. This method reduces considerably the period of time needed for estrus detection; it synchronizes the estrous cycle of 70 to 80% of the cyclic cows to within a 4-d interval without any detrimental effect on the fertility rate (65 to 85%). Moreover, resumption of ovarian activity and normal fertility in acyclic cows in favored. Administration of GnRH eliminates the large follicles by ovulation or atresia and induces emergence of a new follicular wave within 3 to 4 d after treatment at any stage of the estrous cycle, but it limits further growth of these emerging follicles by increasing atresia. The precision of estrus and the unaltered fertility rate is due to the synchronized selection of a new larger growing follicle, which becomes the ovulatory follicle after PGF(2 alpha)-induced luteolysis 6 d after GnRH treatment. Also, fixed-time AI programs without the need for estrus detection may be possible using a second injection of GnRH in a GnRH-PGF(2 alpha)-GnRH protocol to ovulate the selected follicle at a precise time. We describe a physiological model to explain how the precision of estrus is improved following PGF(2 alpha)-induced luteolysis, via the effect of pretreatment with GnRH on follicular development and luteal functions in cattle. Application of this model to the development of reliable methods of fixed-time insemination is also explored.
Forty-four crossbred postpubertal bovine females were used to study how mating with a bull affected estradiol-17beta (E(2)) secretion and timing of the preovulatory LH surge. Estrous cycles were synchronized with two injections of prostaglandin-F(2alpha) (PGF(2alpha)) 11 d apart. Females were either isolated from males (NE) or exposed to epididectomized bulls (BE) after the second PGF(2alpha) injection. Females exposed to bulls were allowed to mate once and then were separated from the bull. Blood samples were collected at 2-h intervals from the second PGF(2alpha) injection until 12-h post injection to monitor progesterone (P(4)) and luteinizing hormone (LH) concentrations and at hourly intervals from 12 h to 60 h post-injection to monitor LH secretion and timing of the preovulatory LH surge. Samples were also collected at 4-h intervals until 60 h post-injection to monitor estrogen (E(2)) secretion. LH surges were detected in 16 and 14 of 22 females from the BE and NE groups, respectively, during the 60-h period after PGF(2alpha) injection Mean P(4) concentrations and time of P(4) decline to <1 ng/ml were not different between the two treatment groups (P>0.30). Mean E(2) concentration during the 60-h sampling period was different (P<0.003) between BE and NE groups, and a significant treatment effect (P<0.002) occurred 48 h, 52 h and 60 h after the second PGF(2alpha) injection. However, mean LH concentration before the LH surge, duration of the LH surge and peak LH concentration during the surge were not different between the BE and NE groups (P>0.40). Mean time for the second PGF(2alpha) injection to the beginning of the LH surge was 51.6 +/- 1.5 h (X +/- S E) for the females not exposed to bulls and 48.5 +/- 1.4 h for females exposed to bulls (P>0.14). In this study, the presence of and/or mating by a bull did not affect LH secretion or timing of the preovulatory LH surge after PGF(2alpha) administration.