Fertility-associated antigen on Nelore bull sperm and reproductive outcomes following first-service fixed-time AI of Nelore cows and heifers

The objective was to determine whether the presence of fertility-associated antigen (FAA) on sperm collected from Nelore (Bos indicus) bulls can be used to assess potential fertility of sperm for use at first-service fixed-time AI (TAI). Six Nelore bulls were selected based on FAA status (FAA-negative: N = 3; FAA-positive: N = 3) and the ability to produce neat semen with ≥ 70% morphologically normal sperm and 60% estimated progressive motility before cryopreservation. In Experiment 1, suckled multiparous Nelore cows (N = 835) were evaluated for body condition score (BCS) and received an intravaginal progesterone device (CIDR) and 2.0 mg of estradiol benzoate (Day 0). On Day 9 the CIDR was removed, 12.5 mg of PGF_2α and 0.5 mg of estradiol cypionate were administered, and calves were removed for 48 h. All cows received TAI on Day 11 (48 h after CIDR removal). Pregnancy per TAI (P/TAI) was not different between FAA-positive and FAA-negative bulls (41.5% vs. 39.3%, respectively). There was an effect of AI technician on P/TAI (36.0% vs. 43.9%; P < 0.05) and BCS tended to affect P/TAI (P = 0.09), as cows with BCS ≥ 2.75 were 1.4 times more likely to become pregnant compared with cows with BCS < 2.75. In Experiment 2, nulliparous Nelore heifers (N = 617) were evaluated for BCS and received a CIDR and estradiol benzoate (2.0 mg) on Day 0. On Day 7, all heifers received PGF_2α (12.5 mg). On Day 9, CIDR inserts were removed and all heifers received estradiol cypionate (0.6 mg) and 200 IU eCG. All heifers received TAI on Day 11 (48 h after CIDR removal). Pregnancy/TAI was different (P = 0.04) between FAA-positive and FAA-negative bulls (33.7% vs. 40.7%, respectively). Presence of FAA on sperm was unsuccessful in assessing the potential fertility of sperm for use in TAI.     

Efficiency of superstimulatory protocol P-36 associated with the administration of eCG and LH in Nelore cows

Recent work with P-36 demonstrates that the replacement of the last two doses of Follicle-Stimulating Hormone (FSH) with equine chorionic gonadotropin (eCG) increases embryo yields. However, it is unclear if the positive effect of eCG is related to its FSH-like activity, LH-like activity, or both. This study aimed to verify the replacement of eCG with pLH on the last day of superstimulatory treatment. Twenty-five Nelore cows were allocated to four groups: P-36 (control), P-36/eCG, P-36/LH2, and P-36/LH4. All animals underwent four treatments in a crossover design. The control group cows were superstimulated with decreasing doses of porcine Follicle-Stimulating Hormone (pFSH, 133 mg, im). In the P-36/eCG, P-36/LH2, and P-36/LH4 groups, the last two doses of pFSH were replaced in the former group by two doses of eCG (200 IU each dose, im) and in the latter two groups by two doses of pLH (1 and 2 mg each dose, im), respectively. Donors received fixed-time artificial insemination 12 and 24 hours after pLH. Embryo flushing was performed on D16. Data were analyzed by ANOVA (Proc Mixed, SAS). There was a trend of decreasing ovulation rate when comparing groups LH2 and eCG (P = 0.06). However, there was no significant difference in the mean number of viable embryos among groups P-36 (3.3 ± 0.7), P-36/eCG (4.5 ± 0.5), P-36/LH2 (3.7 ± 0.8), and P-36/LH4 (4.2 ± 1.0). It is concluded that the replacement of eCG by pLH on the last day of superstimulatory treatment can be performed with no significant variation in the production of viable embryos.     

Ovum pick up, in vitro embryo production, and pregnancy rates from a large-scale commercial program using Nelore cattle (Bos indicus) donors

The objective was to clarify in vitro production of bovine embryos in Brazil. Data from 656 ovum pick-up/in vitro production (OPU/IVP) procedures, performed on 317 Nelore (Bos indicus) donors, without hormone stimulation or control of ovarian follicular waves, were analysed. Donors were subjected to OPU from one to nine times (no specific schedule), with < 15 d between consecutive procedures. There were 20,848 oocytes, of which 15,747 (75.53%) were considered viable, 5,446 embryos were obtained, 5,398 embryos were immediately transferred, resulting in 1,974 pregnancies (36.57%) at Day 30 and 1,788 (33.12%) pregnancies at Day 60. The average number of total and viable oocytes produced per OPU session was (mean ± SEM) 30.84 ± 0.88 and 23.35 ± 0.7 (average of 8.1 ± 0.3 embryos and 3.0 ± 0.1 pregnancies per OPU-IVP procedure). Since oocyte production varied widely among donor, they were designated as very high, high, intermediate, and low, with 58.94 ± 2.04, 32.61 ± 0.50, 22.13 ± 0.50, and 10.26 ± 0.57 oocytes, respectively, produced by 78, 80, 79, and 80 donors. The number of viable oocytes recovered ranged from 0 to 128; since donors with numerous viable oocytes produced many viable embryos and pregnancies, oocyte production was useful for donor selection. However, there was no significant effect of the number of OPU sessions per donor on mean numbers of oocytes produced. In conclusion, we confirmed field reports of high oocyte production by some Nelore donors and demonstrated individual variation in oocyte yield, which was associated with embryo production and pregnancy rates.     

The effect of prostaglandin F2 alpha treatments in superovulated cattle on estrus response and embryo production

Approximately 10% of cows in a commercial embryo transfer center that were superovulated for embryo production did not show estrus at the right time and therefore did not produce embryos. This problem was investigated by studying the effects of prostaglandin F2 alpha (PGF) treatment regime and dose rate on the superovulatory process. The cows in estrus following superovulation (96% vs. 86.6%), the % cleaved (62% vs. 51%) and the transferable embryo production (5.4 vs. 3.8) was increased when 50 mg. PGF was administered in three divided doses rather than in two doses. In a second experiment doses of 15 mg., 30 mg. and 45 mg. (each administered as three divided doses 6 hours apart) all produced the same estrus response (95.6, 97.9 and 95%) and production of transferable embryos (4.9, 3.6 and 4.6). Three-times-a-day PGF reduced the time interval from treatment to the onset of estrus, but the time from PGF to estrus was not correlated with embryo production.     

Day 0 protocol: superstimulatory treatment initiated in the absence of a large follicle improves ovarian response and embryo yield in goats

A new superstimulatory protocol (Day 0 Protocol) to initiate FSH treatment in the absence of a large follicle was compared to a traditional protocol in goats. The Day 0 Protocol (n=44) consisted of pre-treatment with progesterone and eCG to synchronize ovulation and the emergence of Wave 1, with FSH starting 84 h after the end of progesterone exposure (i.e., soon after ovulation). The traditional protocol (n=46) consisted of 11 d of progesterone exposure, with FSH treatment beginning 2 d before the end of progesterone exposure. Treatment with FSH was initiated in the absence of a large follicle in 37/44 and in 6/46 goats in the Day 0 Protocol and traditional protocol, respectively (P<0.01). There was more CL in the Day 0 Protocol than in the traditional protocol (breeding season: 9.6+/-0.6 and 6.3+/-0.8, P<0.05; non-breeding season: 14.3+/-1.5 and 10.7+/-1.5; P<0.05). More Grades 1 and 2 embryos were recovered in the Day 0 Protocol than in the traditional protocol (breeding season: 4.8+/-0.7 and 1.8+/-0.5, P<0.05; non-breeding season: 5.6+/-1.1 and 3.5+/-0.7, P=0.07). Similarly, the proportion of embryos that were Grades 1 and 2 was higher for the Day 0 Protocol than for the traditional protocol (breeding season: 81/114, 71%, versus 16/43, 37%, P<0.05; non-breeding season: 118/203, 58% versus 95/205, 46%, P<0.05). In summary, the Day 0 Protocol, was effective in initiating superstimulatory treatment in the absence of a large follicle, and compared to the traditional protocol, induced a higher ovulation rate and better embryo yield in goats.     

The effect of gamma irradiation dose on cucumber (Cucumis sativus L.) haploid embryo production

Pollination with irradiated pollen was the only effective way for the induction of haploid embryo in cucurbits. The possibility of using lower doses of γ rays (Co⁶⁰ source) was studied. The effect of 0.2 and 0.3 kGy of rays was tested on five cucumber lines and three hybrids in the first experiment. It was found that there was hardly any difference between the total number of embryos produced by all studied forms. The highest number of isolated embryos was obtained from hybrids Gy-3 M and BxOg (111 and 188 respectively). The plant regeneration was estimated at 3.3 %. The best two lines and one hybrid were used in the second experiment to find the lowest possible dose of irradiation. The dose 0.05 kGy produced only diploid embryos and was rejected as too low. Other three doses (0.1; 0.2; and 0.3 kGy) effected embryo development in relation to the irradiation applied with the highest number obtained at the lowest dose. However the number of plants regenerated from each combination was similar. The plant regeneration in this experiment was 7.7 %. The effect of 0.1 and 0.3 kGy was tested during the next two years on one highly vigorous variety. It was confirmed that 0.1 kGy stimulated the development of higher number of haploid embryos.     

The influence of ovulation number and ovarian dimensions on embryo production in superovulated cattle

Twenty-one cycling Angus heifers and five Holstein cows received a subcutaneous (SC) injection of 50 mg of progesterone (P) in oil for 14 consecutive days. On day 6 of (P) treatment, animals were injected intramuscularly (IM) with 6 mg of estradiol valerate, and on day 13, received an IM injection of 2,000 IU of Pregnant Mare Serum Gonadotropin. Three additional Angus heifers were used as non-hormone treated controls. Seventeen of 21 heifers and 4 of 5 cows (81%) exhibited estrus within 48 to 132 hr following P treatment. Two of the five animals in which estrus was not observed were palpated as pregnant and discarded from the study. Treatment animals showing estrus were randomly assigned either to Group I, animals bred by natural service, or Group II, animals artificially inseminated with two straws of frozen semen at 12-hr intervals for a total of four breedings. Twenty-one animals were slaughtered 2 to 6 days after the onset of estrus, and those animals in which estrus was not detected were slaughtered 10 days after the last P injection. Two of the 24 treated animals had no ovulations. A total of 397 ovulation points ($\text{397}\text{22}$) were counted for a mean ovulation rate of 18 ovulations per animal. One hundred and fifty-six ova were recovered ($\text{156}\text{397}$) for a collection rate of 39%. Group I animals had 44 of 66 (67%) of their ova fertilized while 23 of 71 (32%) of the ova in Group II were fertilized. Nineteen unfertilized eggs were collected from the three animals not observed in estrus. No differences in fertilization rates between the Group I and Group II animals were found. Mean ovarian width, length and weight in the treated animals was measured and found to be 3.5 ± 1.1 cm, 4.8 ± 1.4 cm, and 21.7 ± 21.2 gm, respectively. Ovarian width, length and weight were all positively correlated with the number of ovulations per ovary r=.74, r=.74, and r=.55, respectively. No significant correlation existed between ovarian width (r=.16), lenght (r=.21), or weight (r=.13) when compared to ova recovery rate. This result suggests that ovarian size or weight may not be the limiting factor involved in embryo recovery.     

Fixed-time AI protocols replacing eCG with a single dose of FSH were less effective in stimulating follicular growth, ovulation, and fertility in suckled-anestrus Nelore beef cows

The aim of the present study was to evaluate the effects of a single treatment with FSH on diameter of the largest follicle and on conception rates of suckled Bos indicus beef cows submitted to timed artificial insemination (TAI). Four hundred fifty-six suckled anestrous Nelore beef cows at 30-60 days postpartum were assigned to treatments. At the first day of the estrous synchronization protocol (Day 0), all cows received a progesterone-releasing intravaginal device plus 2mg of estradiol benzoate. On Day 8, cows were assigned to blocks according to the diameter of the largest follicle and then allocated to one of three treatment groups (Control, FSH, or eCG) within each block. Simultaneously to progesterone device withdrawal on Day 8, cows in the eCG treatment group (n=150) received 300 IU of eCG and cows in FSH treatment group (n=153) received 10mg of FSH, and Control cows (n=153) did not receive any additional treatment. Additional treatments with 150 μg of cloprostenol and 1mg of estradiol cypionate (EC) were also administered concurrently to progesterone device removal in all cows on Day 8. Two days later (D10), TAI and ovarian ultrasonic examinations to evaluate follicle size were performed in all cows. On Day 12, a subset of cows (n=389) were submitted a second ultrasonic exam to confirm ovulation. Final follicular growth (mm/day) was less (P=0.006) in both Control (0.95±0.11) and in FSH-treated cows (0.90±0.10) than in eCG-treated cows (1.40±0.13). Interestingly, there was a treatment-by-BCS interaction in ovulation results (P=0.03), in which, eCG treatment increased percentage of cows having ovulations with a lesser BCS. Similarly, there was a treatment-by-BCS interaction for conception (P=0.04), where the eCG treatment increased fertility in cows with a lesser BCS. In conclusion, FSH failed to stimulate final follicular growth, ovulation, and conception rate in sucked-anestrous beef cows submitted to TAI as effectively as eCG. However, physiological effects of eCG seem to be more evident in cows with a lesser BCS.     

Effects of superovulation on endogenous LH secretion in cattle, and consequences for embryo production

Stimulation of follicular growth during superovulation is achieved by the injection of FSH or compounds with high FSH-bioactivities. However, some LH-activity is required for follicle maturation. It is of relevance to evaluate, therefore, the effect of superovulatory treatments on endogenous LH secretion. Luteinizing hormone is secreted in discrete pulses, and the pattern of pulsatile LH secretion during superovulation is reviewed. Four of five published studies have shown that LH pulse frequency is significantly reduced by injection of eCG or FSH preparations. This suppression appears within 8 h of treatment Effects of superovulation on LH pulse amplitude are less consistent. The reasons for the decrease in pulse frequency have been investigated, and although the answer is not definitive, it would seem that increased follicular estradiol, acting perhaps in synergism with progesterone, may play a role. Changes in plasma progesterone concentrations are not related to changes in LH pulse frequency. What is the significance of decreased LH pulse frequency? We attempted to investigate this by inducing LH pulses during superovulation, but the result was a major reduction in ovulation rate. More research is required to determine if modification of endogenous LH secretion can improve superovulatory responses.     

Effect of a CIDR insert and flunixin meglumine, administered at the time of embryo transfer, on pregnancy rate and resynchronization of estrus in beef cattle

The objectives of this study were to evaluate the effects of flunixin meglumine (FM), an inhibitor of PGF(2alpha) synthesis, and insertion of an intravaginal progesterone-releasing device (CIDR), on pregnancy rates in beef cattle embryo transfer (ET) recipients, and to examine the effect of a CIDR after embryo transfer on the synchrony of the return to estrus in non-pregnant recipients. Cows (n=622) and heifers (n=90) at three locations were assigned randomly to one of four groups in a 2x2 factorial arrangement of treatments with FM administration (500 mg i.m.) 2-12 min prior to ET, and insertion of a CIDR (1.38 g progesterone) immediately following ET as main effects. Fresh or frozen embryos (Stage=4 or 5; Grade=1 or 2) were transferred on Days 6-9 of the estrous cycle and CIDR devices were removed 13 days after ET. Recipients at Location 2 only were observed for signs of return to estrus. Recipients that returned to estrus at Location 2 were either bred by AI or received an embryo 7 days after estrus. Following the initial ET, there was an FMxlocation interaction on pregnancy rate (P<0.01; Location 1, 89% versus 57%; Location 2, 69% versus 64%; Location 3, 64% versus 67% for FM versus no FM, respectively). Pregnancy rates of embryo recipients were not affected by CIDR administration (P>0.05; 65% with CIDR, 70% without CIDR), however, the timing of the return to estrus was more synchronous (P<0.01) for recipients given a CIDR. Pregnancy rate of recipients bred following a return to estrus did not differ between cows receiving or not receiving a CIDR for resynchronization (P>0.13). Effects of FM on pregnancy rate were location dependent and CIDR insertion at ET improved synchrony of the return to estrus.     

Overall efficiency of in vitro embryo production and vitrification in cattle

In 5 replicates a total of 719 immature oocytes recovered from 94 slaughterhouse-derived bovine ovaries were matured and fertilized in vitro, then cultured for 7 to 9 d on a granulosa cell monolayer in TCM 199 supplemented with calf serum. Of 338 blastocysts (47% of oocytes cultured), 301 were vitrified in Hepes/bicarbonate buffered TCM-199 medium, 20% calf serum and dimethylsulfoxide and ethylene glycol as the cryoprotectants. After thawing in 1 M sucrose and subsequent culture in vitro, 237 (79%) of the blastocysts re-expanded and 177 (59%) hatched. Re-expansion and hatching rates differed between the blastocysts vitrified on Day 7 and Day 8 (84 and 69% vs 70 and 41%, respectively). We conclude that the applied methods are relatively simple and inexpensive to use, with an overall efficiency of the in vitro production/vitrification procedure being 1.9 hatched blastocyst/ovary. Therefore, this system seems suitable for large-scale production of cryopreserved bovine embryos for various purposes.     

Fertility, production and culling following cesarean section in dairy cattle

A population-based historical cohort study design was used to investigate the effects of a cesarean section on calf mortality, fertility, milk production and culling on 35 Dutch dairy farms. The data were collected during a routine herd health and production control program. Cows undergoing a cesarean section had a higher calf mortality rate, a longer interval from first service to conception and a smaller risk for retained placenta than control cows, those with spontaneous deliveries and those with dystocias. Cumulative milk production at 100 days was lower and the risk of being culled was higher for cows with cesarean sections than for the control cows.     

Influence of CIDR treatment during superovulation on embryo production and hormonal patterns in cattle

One of the major sources of success in embryo transfer is timing of AI relative to the LH surge and ovulation. The aim of this study was to compare the embryo production following superovulation during a PGF2alpha (control cycle) or a CIDR-B synchronized cycle (CIDR-B cycle). CIDR-B (CIDR-B ND, Virbac, Carros, France) was inserted on Day 11 of a previously synchronized cycle and left for 5 days. A total dose of 350 microg FSH was administered (eight injections i.m. for 4 days; first on Day 13, decreasing doses) and PGFalpha analog (750 microg i.m.: Uniandine ND, Schering-Plough, Levallois-Perret, France) injected at the time of third FSH injection. Artificial inseminations were performed 12 and 24 h after standing estrus (Day 0). Embryos were collected on Day 7. Luteinizing hormone was measured by EIA (Reprokit Sanofi, Libourne, France) from blood samples collected every 3 h for 36 h, starting 24 h after PGF2alpha (control cycle) or 12 h after CIDR-B removal (CIDR-B cycle). The effects of treatment group and interval between the LH peak and AI (two classes, < 10 and > or = 10 h) on embryo production and quality were analyzed by ANOVA. No effect of treatment was observed on embryo production variables. The intervals between the end of treatment and onset of estrus and between end of treatment and LH surge were greater in heifers treated during a control than a CIDR-B cycle, respectively (45.5 +/- 1.4 versus 31.9 +/- 0.7; 42.0 +/- 1.6 versus 31.0 +/- 1.5; P < 0.05), but maximal LH and estradiol concentrations, at the preovulatory surge were similar in control and CIDR-B synchronized heifers. The numbers of viable and Grade I embryos were significantly increased (P < 0.01) when animals had an interval from LH peak to first AI > or = 10 h (7.2 +/- 0.9 and 3.5 +/- 0.6) when compared to shorter intervals (4.2 +/- 1.1 and 2.0 +/- 0.7) whereas total number of embryos was unchanged (11.8 +/- 1.4 versus 10.3 +/- 1.8). It is concluded that late occurrence of LH peaks in relation to estrous behavior is associated with a lower embryo quality when first AIs are performed systematically 12 h after standing estrus. Further studies are needed to know if results may be improved when making AI at a later time after standing estrus or if LH assays are useful to better monitor AI time.     

Potential use of ovum pick-up for embryo production and breeding in cattle

The efficacy of transvaginal ultrasound-guided puncturing of ovarian follicles for collecting immature oocytes in cattle was studied. Three experiments were conducted to examine the effects of puncturing on follicle recruitment and on the number of oocytes collected. Puncture sessions were executed twice weekly at regular intervals of 3 and 4 d respectively. The oocytes were matured, fertilized and allowed to develop in vitro and the number of transferable embryos was recorded. The health of the cows was checked daily. In Experiment 1, dairy cows (n=10) were punctured over a period of 5 mo, and the collected oocytes were fertilized with the semen of 1 bull. In Experiment 2, oocytes were collected from one 12 year old high pedigree dairy cow and an one month pregnant cow were punctured. The oocytes of the old cow were fertilized with semen of 8 different bulls. In Experiment 3, beef cows (n=6) were punctured over a 2 mo period and the semen of 2 different bulls of the same breed was used to fertilize the oocytes from 3 of these cows. In Experiment 1, 14.5 +/- 0.4 (mean +/- SEM) follicles were punctured per session, and 8.0 +/- 0.3 (mean +/- SEM) oocytes were recovered. A mean of 16% of the oocytes developed into transferable embryos with a pregnancy rate of 40%. The results did not differ between the months of the experiments, indicating that the transvaginal puncturing method can be used successfully over a 5 mo period. No detrimental effects were observed after clinical and post mortem examinations, nor did breed, age or reproductive status appear to affect the results. However, large differences were observed between individual cows and between cow/bull combinations.     

Production and breeding of transgenic cattle using in vitro embryo production technology

Transgenic technology permits major modifications of phenotype by introducing subtle changes in genotype. For domestic farm species, genetic modification may be used to enhance agricultural production or to generate novel genotypes capable of producing heterologous proteins for biomedical applications. The advent of in vitro embryo production techniques has facilitated the large-scale, commercial use of transgenic technology in cattle. Accordingly, we employed in vitro-produced zygotes and embryos in an effort to generate transgenic cattle. Overall, pronuclei in 36,530 in vitro matured and fertilized zygotes were microinjected with a construct designed to express human alpha-lactalbumin in the mammary gland. Of these, 1,472 developed and were transferred to recipients, including 148 twin transfers. Initial pregnancy rate on Day 30 of gestation was 28% (374/1,324). Subsequent calving rate was 17% (226/1,324). Eighteen calves (8%) were transgenic. In vitro produced embryos were used to facilitate breeding of transgenic bulls. Frequency of transgene transmission varied from 3 to 54% between bulls, indicating varying degrees mosaicism. Embryos produced in vitro by these bulls were biopsied and screened for transgenesis prior to transfer to recipients; so far all (6/6) calves born from screened, transgenic embryos were themselves transgenic.     

Rat embryo quality and production efficiency are dependent on gonadotrophin dose in superovulatory treatments

The present study was performed to optimize a superovulation protocol in rats in order to produce a large number of good-quality embryos suitable to develop rat embryonic stem (rES) cells. We first evaluated the ovulation kinetics of three rat strains: Wistar, Fisher and ACI/N. Animals (n=30 per strain) were treated with 50 IU of pregnant mare serum gonadotrophin (PMSG), and ovulation was induced with 50 IU of human chorionic gonadotrophin (hCG) 50 h apart. Next, we evaluated the dose-response curves of PMSG and hCG in Wistar rats in order to obtain the highest number of embryos. The parameters evaluated for superovulation efficiency were: percentage of mated females, percentage of pregnant females and the average number of embryos collected per female. The results of these experiments suggested that the best dose combination was 50 IU for each hormone. Subsequent experiments, again with Wistar rats, were designed to test which of four hormonal combination treatments (30/30, 30/50, 50/30, and 50/50 IU of PMSG/hCG) will produce the largest numbers of good-quality embryos. Embryo quality was evaluated by embryo development uniformity, embryo morphology, embryo survival in an in vitro culture and embryo ability to generate rES-like cells. Results from these experiments showed that 30/50 IU of PMSG/hCG was the treatment that induced the best embryo quality. In conclusion, our results indicated that, in Wistar rats, the most appropriate hormonal combination dose for superovulation protocols with high number of good-quality embryos was 30 IU of PMSG and 50 IU of hCG given 50 h apart. We are performing further studies with rES-like cells produced with the present methodology to evaluate if they are able to participate in the production of germ-line chimeras.     

The effect of a progesterone releasing intravaginal device (PRID) on pregnancy risk to fixed-time insemination following diagnosis of non-pregnancy in dairy cows

The objective was to compare the probability of pregnancy after fixed-time insemination in cows diagnosed as non-pregnant and re-inseminated following the Ovsynch protocol, with or without exogenous progesterone. Cows (n=415) used in this study originated from 25 farms. Upon diagnosis of non-pregnancy between 30 and 60 days after AI, cows were randomly assigned to receive either a progesterone releasing intravaginal device (PRID; n=208) or a placebo intravaginal device (PID; n=207). All cows received GnRH at enrollment (Day 0), PGF(2alpha) concurrent with intravaginal device removal 7 days later, GnRH on Day 9 and fixed-time insemination 16h later (Day 10). Cows observed in estrus prior to Day 7, had the device removed and were inseminated. Ovaries were examined by transrectal palpation at the time of enrollment and the prominent structures were assessed and recorded. Body condition score, lameness status, interval from previous insemination, and times bred at enrollment were recorded. At intravaginal device removal, the occurrence and intensity of vaginitis was determined according to the amount of debris on the device. Overall, the intravaginal device retention rate was 91%. A total of 5.2% of PID-treated cows and 2.9% of PRID-treated cows were detected in estrus within the 7 days treatment period. Pregnancy status was diagnosed between 30 and 56 days after insemination and all cows were followed for a minimum of 150 days after enrollment. Approximately 28% of cows had evidence of mild vaginitis in response to the intravaginal device, whereas 6% of cows had copious debris associated with the intravaginal device at removal. The probability of pregnancy after fixed-time insemination was 43.8% versus 34.9% in PRID-treated versus PID-treated animals. Exogenous progesterone provided through an intravaginal device to non-pregnant cows that had not displayed estrus improved the probability of pregnancy after fixed-time AI.     

Exogenous progesterone enhances ova and embryo quality following superstimulation of the first follicular wave in Nelore (Bos indicus) donors

Three experiments were conducted to evaluate the effects of exogenous progesterone on superovulatory response and ova/embryo quality in Bos indicus donors superstimulated during the first follicular wave (FFW). We hypothesized that exogenous progesterone during gonadotropin treatments would improve ova and embryo quality. In Experiment 1, 18 Nelore cows were randomly allocated to three groups: (1) FFW; (2) FFW plus a progesterone-releasing device (FFW+P4); and (3) control (E2+P4). Cows in the FFW groups were superstimulated beginning at synchronized ovulation, whereas cows in the control group were superstimulated after synchronization of follicular wave emergence with estradiol plus progesterone (E2+P4). There were no differences in mean (± SD) numbers of transferable embryos between FFW+P4 (8.0 ± 4.5) and control (6.7 ± 4.8) groups, but both were higher (P = 0.006) than the FFW group (0.2 ± 0.4). In Experiment 2, FFW and FFW+P4 were compared in 20 Nelore donors; exogenous progesterone increased the number of transferable embryos (3.9 ± 3.4 vs. 1.3 ± 4.1, P = 0.003). In Experiment 3, FFW and FFW+P4 were compared in 10 Nelore donors except that cows were slaughtered 12 h after pLH (Lutropin-V^®, Bioniche Animal Health, Belleville, ON, Canada) treatment. More mature cumulus oocyte complex (COC) (expanded cumulus cell layers) were collected in the FFW+P4 group than in the FFW group (21.8 ± 13.1 vs. 10.8±14.7; P = 0.003). In summary, superovulatory response was satisfactory when FSH (Folltropin-V^®, Bioniche Animal Health) treatment was initiated at emergence of the first follicular wave in Nelore (Bos indicus) donors, and the hypothesis that administration of exogenous progesterone during the treatment will improve oocyte and embryo quality was supported.