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Two experiments determined whether the addition of GnRH at the beginning of an estradiol (E2)/progesterone (P4)- based synchronization protocol and/or a second dose of prostaglandin F2α (PGF2α) the day before P4 device removal improves pregnancy rate in lactating dairy cows. On Day 0, all cows received a CIDR-B device and 2 mg i.m. estradiol benzoate, and half received 200 μg i.m. gonadorelin acetate (GnRH). On Day 7, cows were further subdivided to receive PGF2α (500 μg i.m. cloprostenol) or no PGF2α treatment. On Day 8, CIDR-B were removed, and all cows received PGF2α, 1 mg estradiol cypionate and 400 IU eCG i.m., and an estrus detection aid. Experiment 1 was designed to evaluate the effect of treatments on follicular development from P4 device removal to ovulation, expression of estrus, time of ovulation and serum P4 concentrations. Cows (n = 76) were examined by ultrasonography and bled for serum P4 determinations every 12 h from the time of P4 device removal but were not inseminated. In Experiment 2, all cows (n = 1036) were inseminated based on estrus detection using tail-paint. Cows with50% of the paint rubbed-off by 48 h after P4 device removal were inseminated at that time, whereas those not in estrus received 100 μg i.m. of GnRH and were inseminated 12 h later. In Experiment 1, the interval from P4 device removal to ovulation was 71.7 ± 1.5 h and did not differ among groups. However, cows that received 2 injections of PGF2α had a greater (P 0.01) estrus rate and lower (P 0.01) P4 concentrations at 48 h after P4 device removal than those that received 1 PGF2α (estrus rate: 86.8% vs 68.4% and P4 concentration: 0.12 ± 0.01 vs 0.36 ± 0.07, for 2 and 1 PGF2α, respectively). In Experiment 2, estrus rate was also influenced by the number of PGF2α treatments, regardless of whether cows received or did not receive GnRH on Day 0 (2 PGF2α: 84.7%, 438/517 vs 1 PGF2α: 65.7%, 341/519; P 0.01). Furthermore, there was a GnRH treatment by number of PGF2α treatments interaction (P 0.05) on P/AI that was attributed to greater (P 0.05) P/AI in cows that received GnRH on Day 0 and 2 PGF2α than in the other three treatment groups (EB+1 PGF2α: 45.2%, 119/263; EB+2 PGF2α: 45.8%, 119/260; EB + GnRH + 1 PGF2α: 45.7%, 117/256 and EB + GnRH + 2 PGF2α: 57.2%, 147/257). It was concluded that the addition of GnRH on Day 0 and a second dose of PGF2α the day before P4 device removal improves P/AI in lactating dairy cows synchronized with an estradiol/P4-based protocol.

A research career is not only built on ideas and publishable results; it is more often the product of determination, hard work, collegiality and collaboration. It is through our collaborators, family and friends that we really become better persons, and scientists. It is also a matter of being at the right place at the right time. My work in bovine reproduction has progressed from an interest in superovulation and embryo transfer before I became a veterinarian, to the development and application of this technology and fixed-time artificial insemination in beef and dairy herds. Everything that I have done has been possible because of the people that I have worked with over the years. This manuscript combines some of the very exciting things that I have learned about bovine reproduction over the last 30 years and personal stories behind the projects and ideas that we have pursued during that time.

The objective of ovarian superstimulatory treatments in cattle is to obtain the maximum number of viable embryos by stimulating growth of antral follicles and ovulation of competent oocytes. While factors inherent to the donor animal are critical, an increased knowledge of ovarian physiology, gonadotropin biochemistry and the ability to manipulate ovarian function have provided alternatives for the design of simple and successful protocols for superovulation in cattle. Recent protocols have also been made more user-friendly and allowed for the grouping of donors for successful superovulation. Although the number of reports associating FSH profiles with superovulatory response is limited, studies designed to reduce the number of FSH treatments necessary to induce superstimulation may provide guidance for the development of optimized gonadotropin treatment protocols. Although high peak levels of circulating FSH following a single administration of Folltropin-V have been shown to be associated with a reduced superstimulatory response, the ideal treatment protocol would seem to be to increase circulating FSH levels to values comparable to those required for the induction of follicle wave emergence, and to maintain these levels for at least 72 h (or 36 h for superstimulation prior to ovum pick-up) to allow follicles to reach an ovulatory size and acquire the capacity to ovulate.

The objective of the present study was to evaluate the effect of equine chorionic gonadotropin (eCG) administration associated to different proestrus lengths for Fixed-time AI (FTAI) in beef heifers. In Experiment 1, pre-pubertal heifers (n = 46) received a 6-day estradiol/progesterone-based treatment (J-Synch protocol), and were then allocated into four experimental groups in a 2 × 2 factorial design, to receive or not receive eCG (300 IU) at the time of intravaginal progesterone device removal, and to receive GnRH at 48 h or 72 h after device removal (to induce shortened and prolonged proestrus length, respectively). Endometrial samples were obtained 6 d after ovulation from the cranial portion of the uterine horn. The eCG administration induced greater serum estradiol-17β concentrations before ovulation (P 0.05) and greater proportion of heifers bearing a competent corpus luteum after ovulation (P = 0.054). Delaying GnRH administration from 48 h to 72 h induced a longer interval from device removal to ovulation (i.e., prolonged proestrus; P 0.05), larger diameter of the ovulatory follicle, and greater progesterone concentrations on Day 10-11 after ovulation. Heifers in eCG + GnRH72h group had more uterine receptors in luminal epithelium than those in eCG + GnRH48h group (PR and ERα), and than those in No eCG + GnRH72h group (PR) (P 0.05). No effect of eCG or GnRH treatments was found in endometrial gene expression of progesterone and estrogen receptors. In Experiment 2, a total of 2,598 heifers received the J-Synch protocol associated or not with eCG administration at device removal, followed by FTAI/GnRH at 60 or 72 h after device removal (i.e., prolonged proestrus protocol). Heifers that received eCG had greater P/AI than those not receiving eCG (P 0.05) and there was an interaction between eCG treatment and time of FTAI. The lowest P/AI was found in those heifers that received FTAI/GnRH at 72 h without eCG treatment at device removal (P 0.05), and no differences were found between the other experimental groups. In conclusion, prolonging the length of proestrus in J-Synch protocol improves ovulatory follicular diameter and luteal function; and the administration of eCG at device removal improves preovulatory estradiol concentrations and luteal function. Finally, P/AI was enhanced by eCG treatment and the improvement was more evident when FTAI/GnRH was performed at 72 h after device removal.

Three experiments evaluated the effects of insemination based on estrus expression or treatment with GnRH, on the time of ovulation and pregnancy rates to AI (P/AI) in dairy cows. All cows were treated with a P4 device for 8 d, estradiol benzoate at device insertion and prostaglandin F

The main objective of implementing embryo transfer in beef operations is to accelerate the rate of genetic progress in the herd. Among the main factors that affect the use of these technologies are related to nutrition, management and estrus synchronization. As a result of research conducted over the last 20 years, recipient utilization has increased by applying protocols that synchronize ovulation and allow for embryo transfer without the need for estrus detection, usually referred to as fixed-time embryo transfer (FTET). Although these protocols have performed adequately for several years, recent attention has been directed to the effect of estrus expression and estradiol concentrations during growth of the preovulatory follicle on embryo growth and pregnancy. The experiments reviewed herein demonstrate that estrus expression is associated positively with high pregnancy rates and reduced pregnancy losses in recipients receiving in vitro-produced and in vivo -derived frozen/thawed bovine embryos.

Equine chorionic gonadotropin (eCG) is a heterodimeric glycoprotein hormone produced by pregnant mares that has been used to improve reproductive performance in different domestic species. Several strategies to produce the hormone in a recombinant way have been reported; nevertheless, no approach has been able to produce a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial purposes. For this reason, the only current product available on the market consists of partially purified preparations from serum of pregnant mares (PMSG). Herein, we describe a highly efficient process based on third-generation lentiviral vectors as delivery method for the production of reCG in suspension CHO-K1 cells, with productivities above 20 IU 106 cell−1.d−1 and 70% purification yields after one purification step. Importantly, reCG demonstrated biological activity in cattle, since around 30 μg of reCG were needed to exert the same biologic effect of 400 IU of PMSG in an ovulation synchronization protocol. The results obtained demonstrate that the developed strategy represents an attractive option for the production of reCG and constitutes an auspicious alternative for the replacement of animals as a source of PMSG.

Knowledge of follicular wave dynamics obtained through the use of real-time ultrasonography and the development of the means by which follicular wave dynamics can be controlled have provided practical approaches for the in vivo and in vitro production and transfer of embryos in cattle. The elective control of follicular wave emergence and ovulation has had a great impact on the application of on-farm embryo transfer, especially when large groups of donors need to be superstimulated at the same time. Although estradiol and progestins have been used for many years, practitioners in countries where estradiol cannot be used have turned to alternative treatments, such as mechanical follicle ablation or the administration of GnRH for the synchronization of follicle wave emergence. In vitro embryo production also benefits from the synchronization of follicle wave emergence prior to Cumulus Oocyte Complexes (COCs) recovery. As Bos indicus cattle have high antral follicle population, large numbers of oocytes can be obtained by ovum pick-up (OPU) without superstimulation. However, synchronization of follicular wave emergence and superstimulation is necessary to obtain high numbers of COCs by OPU and blastocysts following in vitro fertilization in Bos taurus donors. Finally, embryos can now be transferred in commercial beef or dairy herds using efficacious synchronization and re-synchronization protocols that are easily implemented by farm personnel. These technologies can also be used to resolve reproductive problems such as the reduced fertility observed during summer heat stress and/or in repeat-breeder cows in commercial dairy herds.

Knowledge of follicular wave dynamics obtained through the use of real-time ultrasonography and the development of the means by which follicular wave dynamics can be controlled have provided practical approaches for the in vivo and in vitro production and transfer of embryos in cattle. The elective control of follicular wave emergence and ovulation has had a great impact on the application of on-farm embryo transfer, especially when large groups of donors need to be superstimulated at the same time. Although estradiol and progestins have been used for many years, practitioners in countries where estradiol cannot be used have turned to alternative treatments, such as mechanical follicle ablation or the administration of GnRH for the synchronization of follicle wave emergence. In vitro embryo production also benefits from the synchronization of follicle wave emergence prior to Cumulus Oocyte Complexes (COCs) recovery. As Bos indicus cattle have high antral follicle population, large numbers of oocytes can be obtained by ovum pick-up (OPU) without superstimulation. However, synchronization of follicular wave emergence and superstimulation is necessary to obtain high numbers of COCs by OPU and blastocysts following in vitro fertilization in Bos taurus donors. Finally, embryos can now be transferred in commercial beef or dairy herds using efficacious synchronization and re-synchronization protocols that are easily implemented by farm personnel. These technologies can also be used to resolve reproductive problems such as the reduced fertility observed during summer heat stress and/or in repeat-breeder cows in commercial dairy herds.

A single dose protocol of FSH for superstimulation in cattle may improve compliance and superovulatory response. A single subcutaneous (sc) administration of pFSH was efficacious, but response depended on body condition and injection site; the adipose tissue pad behind the shoulder was most efficacious. Inconsistent results in Holsteins were partially overcome by sc administration of 75% of the total pFSH dose behind the shoulder on the first day followed by 25% 48 h later. An alternative would be to combine FSH with polymers that cause it to be released slowly over several days. Hyaluronan is found normally in most animal tissues and is nonreactive when administered parentally. A single intramuscular (im) administration of pFSH in a 2.0% hyaluronan induced a superovulatory response that did not differ from twice daily im administration over 4 d. However, 2.0% hyaluronan was viscous and difficult to mix with FSH. Although solutions of 1.0 and 0.5% hyaluronan were less viscous, they lacked efficacy as a single im administration. However, superovulatory response was high when either 1.0 or 0.5% hyaluronan was used in a two-dose im protocol; two-thirds on the first day and one-third 48 h later. A single im administration of FSH in 0.5% hyaluronan effectively induced superstimulation for OPU in cattle. Successful superovulation in the cow was associated with circulating FSH levels that were similar to endogenous FSH levels prior to follicular wave emergence; however, levels must be maintained above baseline for at least 72 h, or 36 h for OPU. Circulating FSH levels following a single sc administration of 400 mg NIH-FSH-P1 behind the shoulder in beef cows increased to 1.0 or 1.2 ng/mL at 12 h and were back near baseline in approximately 60 h, while FSH levels following im administration of 200 mg NIH-FSH-P1 in 0.5% hyaluronan into Holstein donors reached 1.5 ng/mL at 12 h and returned to baseline in approximately 36 h.

Two experiments were conducted with the aim of determining the effect of equine chorionic gonadotropin (eCG) administration on day 14 after insemination on ovarian response and pregnancy establishment in postpartum anestrous beef cows. In both experiments, cows were subjected to a progesterone- and estradiol-based treatment for fixed-time artificial insemination (FTAI) and were randomly allocated into 4 groups to receive or not receive eCG (400 IU) at the time of device removal and/or at 14 d after FTAI. In experiment 1, from day 14 to 22, daily ultrasonographic determinations were performed to monitor ovarian dynamics, and blood was collected to determine hormone concentrations in 60 cows. In experiment 2, confirmation of pregnancy was performed at 30 and 60 d after FTAI in 1,060 anestrous cows assigned to the same experimental design. Cows that received eCG on day 14 after FTAI showed increases in corpus luteum area (P < 0.01), follicle diameter (P < 0.05), serum progesterone concentrations (P < 0.01), and estradiol-17β concentrations (P < 0.01), compared with cows that did not receive eCG on day 14. Pregnancy rate on day 30 was greater in those cows that received both eCG treatments (ie, at device removal and 14 d after insemination) than in those that did not receive eCG treatment (P < 0.05). In conclusion, eCG administered on day 14 after FTAI increases serum progesterone concentrations during the critical period of pregnancy in anestrous cows, and this second eCG treatment seems to have a positive effect on achieving pregnancy.

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of

This study was conducted to evaluate oocyte recovery and in-vitro blastocyst production of donor cows superstimulated for ovarian follicular development with FSH administered as twice-daily injections in saline or a single injection diluted in 0.5 % hyaluronan before oocyte aspiration. In Experiment 1, cows were treated with 160 mg of Folltropin-Vdiluted in saline, administered in four twice-daily i.m. injections for 2 days (Multiple FSH group); 160 mg of Folltropin-V diluted in hyaluronan and administered in a single i.m. injection (Single FSH group); or no FSH treatment (Control). In Experiment 2, donor cows were treated with either a single FSH i.m. injection or there was no treatment (Control) before ovum pick up (OPU) was performed. In both experiments, COCs collected using OPU were classified, matured, fertilized and cultured at 38.8 °C in a humidified atmosphere for 7 days. In Experiment 1, the number of follicles aspirated and COCs recovered were greater (P < 0.05) in cows treated with multiple and single doses of FSH. Number of blastocysts produced, however, did not differ among groups. In Experiment 2, mean number of follicles aspirated and COCs recovered were also greater (P < 0.05) in FSH-treated cows. Nevertheless, number of blastocysts produced did not differ. In summary, single and multiple FSH administrations induced similar follicular stimulation for OPU. Furthermore, with both FSH treatments there was induction of development of a larger number of follicles to be aspirated and COCs recovered by OPU compared with these values for donor beef cows with no FSH treatment for follicular stimulation.

Three experiments were designed to evaluate the effect of the length of insertion of a intravaginal progesterone (P4) device on follicular and corpus luteum (CL) diameter, time of ovulation, and pregnancy rates following fixed-time AI (P/FTAI) in Bos indicus cows treated with an oestradiol/P4-based treatment with a lengthened proestrus, named J-Synch. Multiparous suckled Bos indicus beef cows that were 35 to 60 days postpartum were used in Experiments 1 and 2, whereas non-lactating cows were used in Experiment 3. All cows had a CL or a follicle of at least 8 mm in diameter and a body condition score between 2.5 and 4 (1 to 5 scale) at the time of treatment. On Day 0, all cows received 2 mg of oestradiol benzoate (EB; Sincrodiol, Ourofino, Brazil) and a previously used intravaginal device (1 g of P4; Sincrogest, Ourofino). The P4 device was removed on Day 6 in the J-Synch 6-day group and on Day 7 in the J-Synch 7-day and Control groups. All cows received 500 µg of cloprostenol (Sincrocio, Ourofino) and 400 IU of equine chorionic gonadotrophin (SincroeCG, Ourofino) at the time of P4 device removal, and cows in the Control group also received 0.5 mg of oestradiol cypionate (ECP, SincroCP, Ourofino) at the same time. In Experiment 1, cows (n = 20 per group) were examined every 12 h, from P4 device removal to determine the time of ovulation and 7 days after ovulation to measure the CL. In Experiment 2, all cows (n = 200 per group) were tail-painted for oestrous detection. Cows with = 50% of the tail-paint rubbed-off by 72 h (J-Synch groups) or 48 h (Control group) after P4 device removal, were FTAI at that time. Cows not designated as showing oestrus at that time received 10 µg Buserelin (GnRH, Sincroforte, Ourofino) and were FTAI 8 h later. In Experiment 3, P/FTAI was compared in cows that received the J-Synch 7-day treatment (n = 265) or the 7-day Control treatment (n = 250). Data were analysed using the GLM mixed procedure (InfoStat). In Experiment 1, the diameter of the ovulatory follicle was larger (P

The objectives of this study were to evaluate the reproductive and productive performance of dairy cows with and without puerperal metritis and to evaluate the effectiveness of using a long-acting ceftiofur preparation. Dairy cows in one dairy farm, calving from July 2009 to January 2010, were examined between 3 and 14 days postpartum and classified on the basis of vaginal discharge into three groups: cows with normal discharge (control; C); cows with a bloody mucus purulent or pathologic nonfetid discharge (PnFD), and cows with bloody mucopurulent or purulent fetid discharge (PFD). Cows in C and PnFD groups were not treated, whereas those in the PFD group were randomly allocated to receive 2.2 mg/kg of ceftiofur subcutaneously behind the ear (PFD-T) or remain untreated (PFD-No T). From the 640 cows examined, 58.2% formed the C group, 13.4% formed the PnFD group, and 28.4% formed the PFD group. Survival curves differed between cows in the C group and PFD-No T group (P = 0.0013) and between PFD-No T versus PFD-T group (P = 0.0006). Survival curves of PnFD were intermediate and did not differ from those in the C group (P = 0.2) and PFD-T group (P = 0.1) but tended to be different from the PFD-No T group (P = 0.056). The postpartum interval to achieve a 25% pregnancy rate was 72 days for cows in the C group, 73 days for the PFD-T group, 83 days for PnFD group, and 95 days for the PFD-No T group. The chance of pregnancy in a cow in the C group was 1.98 times higher (95% confidence interval = 1.33, 3.08) and in cows in the PFD-T group was 2.16 times higher (95% confidence interval = 1.37, 3.50) than that in the PFD-No T group. Finally, the chance of pregnancy in cows in the PnFD group tended to be higher (P = 0.08) than that in the PFD-No T group but did not differ from the other two groups. Cumulative 305-day milk production was higher (P < 0.0001) in C group than those with vaginal discharge, regardless of fetidness and regardless of treatment. It is concluded that puerperal metritis affects the reproductive and productive performance of dairy cows and the treatment with ceftiofur was effective in reducing the adverse effects on reproductive performance but not on milk production.

The aim of the present study was to evaluate the effect of brilliant cresyl blue (BCB) selection, the type of oocyte (immature or matured) and the use of hyaluronan in the vitrification solution on further embryo developmental competence. Oocytes (n = 1308) obtained from abattoir ovaries were classified by BCB stain. Control oocytes were maintained in holding media for 90 min and then subdivided to be placed into maturation media without any treatment or were vitrified. Immature or matured oocytes were vitrified by the solid-surface technique using two different vitrification solutions. VS1: composed of 10% ethylene glycol (EG) for 10 min followed by 20% EG + 0.2M trehalose for 30 sec and finally into 30% EG + 0.5M trehalose for 30 sec, or VS2 composed by 10% EG for 10 min, followed by 20% EG + 0.2M trehalose for 30 sec, and finally into 30% EG+ 0.5M trehalose + 0.1 g/ml hyaluronan for 30 sec. Oocytes were then loaded into Fyberplugs™ and vitrified. After one week, Fyberplugs™ were open and placed directly into (37°C) 0.5M sucrose solution for 5 min, then into 0.25M of sucrose for another 5 min and finally placed into maturation medium for in vitro production. Cleavage and development rates were examined on days 2 and 7 after fertilization, respectively. The blastocyst rate of vitrified oocytes selected as BCB + (5.5 ± 0.6%) were higher than those selected as BCB - (1.0 ± 0.4%) and those that were not selected by BCB (2.0 ± 1.1%; P < 0.001). Furthermore, immature vitrified oocytes had greater (P < 0.05) cleavage and blastocyst rates (44.8 ± 1.9% and 4.0 ± 0.6%) than matured vitrified oocytes (38.3 ± 2.8% and 2.5 ± 0.6%). Finally, the addition of hyaluronan to the vitrification solution had no significant effect on development rates. In conclusion, the selection of oocytes by BCB and the use of immature oocytes increase the development rates of vitrified-warmed oocytes.

An experiment was designed to evaluate the effect of the length of insertion of a progesterone (P4)-releasing device on follicular and luteal characteristics and pregnancy rates to fixed-time AI (P/FTAI) in Bos indicus cows treated with oestradiol/P4-based treatments with a prolonged pro-oestrus. Multiparous Bos indicus suckled beef cows (n = 659), 60-90 days postpartum, with a corpus luteum (CL) or at least one follicle ≥8 mm in diameter and with a body condition score between 2 and 3.5 (1-to-5 scale) were synchronised using three treatments. On Day 0, all cows received 2 mg of oestradiol benzoate (Sincrodiol, Ourofino) and an intravaginal device with 1 g of P4 (Sincrogest, Ourofino). The P4 device was removed on Day 6 in cows in the J-Synch 6 group (n = 195) and on Day 7 in cows in the J-Synch 7 group (n = 262) and conventional group (n = 202). All cows received 500 μg of cloprostenol (Sincrocio, Ourofino) and 400 IU of equine chorionic gonadotrophin (SincroeCG 6000UI, Ourofino) at the time of P4 device removal. Cows in the conventional treatment group also received 0.5 mg of oestradiol cypionate (SincroCP, Ourofino) at the same time. In addition, all cows were tail-painted for oestrous detection (CeloTest, Biotay). Cows that had lost ≥50% of the tail paint 62-66 h (J-Synch groups) or 48-52 h (conventional group) after device removal were FTAI at that time. Cows not showing oestrus by 62-66 h (J-Synch groups) or 48-52 h (conventional group) received 10 μg of GnRH (Sincroforte, Ourofino) at the same time and were FTAI 8 h later. A subset of 20 cows per group was also examined using ultrasonography (Mindray DP50 Vet) every 12 h from the time of device removal to determine the time of ovulation, 6 days after ovulation to determine CL diameter, and 30 days after FTAI to determine P/AI. Data were analysed using the GLM Mixed procedure (InfoStat). The results are shown in Table 1. The diameter of the dominant preovulatory follicle was larger and the interval from device removal to ovulation was longer in cows in the J-Synch groups compared with those in the conventional group (P

An experiment was designed to evaluate pregnancy rates per AI (P/AI) in suckled beef cows synchronised with a progesterone (P4)/oestradiol-based protocol and AI with non-sexed (conventional) semen and with a sexed-sorted semen with 65% X-bearing sperm and 35% Y-bearing sperm that was named SuperConventionalTM. Angus and Hereford suckled cows (n=558), 60-90 days postpartum, with a body condition score of 2 to 3.5 (1-to-5 scale) and 48% (269/558) with a corpus luteum, were randomly allocated to be AI with non-sexed (conventional) or SuperConventional semen. The experiment was performed on 3 farms using straws with sexed-sorted SuperConventional semen containing 6 or 8 million sperm and non-sorted conventional semen with 25 million sperm per straw. Semen was from split ejaculates from two Angus bulls. All cows received a P4 device with 0.6g of P4 (Pluselar, Calier) and 2mg of oestradiol benzoate (Calier) on Day 0. All P4 devices were removed on Day 8, and all cows received 400IU of equine chorionic gonadotrophin (Vetegon, Calier), 150µg of d+cloprostenol (Veteglan, Calier), and 1mg of oestradiol cipionate (Calier) at the same time. All cows were also tail-painted and observed for signs of oestrus. Cows with >30% of the tail paint rubbed off by 48-50h after P4 device removal were AI at that time with either SuperConventional or conventional semen. Those not showing oestrus by 48-50h received 10µg of buserelin (Pluserelina, Calier) at that time and were AI with either SuperConventional or conventional semen 50-52h after P4 device removal. Pregnancy was diagnosed using ultrasonography 60 days after AI, and all pregnancies were sexed to determine the proportion of female pregnancies. Data were analysed using GLM for binary data. The P/AI were different between cows showing or not showing oestrus (P

An experiment was designed to evaluate the effect of the length of insertion of a progesterone (P4)-releasing device and the length of pro-oestrus on follicular and luteal characteristics and pregnancy rates to AI (P/AI) in Bos indicus heifers treated with oestradiol/P4-based treatments. Bos indicus beef heifers (n=374), 22-26 months of age, with a corpus luteum (CL) or at least one follicle ≥8mm in diameter and with a body condition score between 2.5 and 3.5 (1-to-5 scale) were synchronised using three treatments for fixed-time AI (FTAI). On Day 0, all heifers received 2mg of oestradiol benzoate (Sincrodiol, Ourofino) and an intravaginal device with 1g of P4 (Sincrogest, Ourofino). The P4 device was removed on Day 6 in heifers in the J-Synch 6 group (n=120) and on Day 7 in heifers in the J-Synch 7 group (n=105) and conventional group (n=165). All heifers received 500μg of cloprostenol (Sincrocio, Ourofino) and 300IU of equine chorionic gonadotrophin (SincroeCG 6000UI, Ourofino) at the time of P4 device removal. Furthermore, heifers in the conventional treatment group received 0.5mg of oestradiol cypionate (SincroCP, Ourofino) at the same time. In addition, all heifers were tail-painted for oestrus detection (CeloTest, Biotay). Heifers that had lost ≥50% of the tail paint by 70-74h (J-Synch groups) or 48-52h (conventional group) after device removal were FTAI at that time. Heifers not showing oestrus by 70-74h (J-Synch groups) or 48-52h (conventional group) received 10μg of gonadotrophin-releasing hormone (Sincroforte, Ourofino) at the same time and were FTAI 8h later. All heifers were also examined using ultrasonography (Mindray DP50 Vet) every 12h from the time of device removal to determine the time of ovulation, 6 days after ovulation to determine the diameter of the CL, and 28 days after FTAI for P/AI determination. Data were analysed using the MLGM procedure (InfoStat) for normal data families (follicular dynamics) and binary data family (P/AI). The results are shown in Table 1. The diameter of the dominant preovulatory follicle and the CL did not differ among groups (P>0.12). However, the interval from device removal to ovulation was longer in heifers in the J-Synch groups than in heifers in the conventional group (P

Fixed-time artificial insemination (FTAI) has been widely applied in South America within the last 20 years for the genetic improvement of commercial beef herds. Most FTAI treatments for beef cattle used in South America are based on the use of progesterone (P4) releasing devices and estradiol to synchronize follicle wave emergence, with pregnancies per AI (P/AI) ranging from 40 to 60%. More recent protocols focusing on extending the interval from device removal to FTAI (i.e. increasing the growing period of the ovulatory follicle) have been reported to improve P/AI in beef cattle. These new protocols and the more traditional FTAI protocols have also been adapted for use with sexed-sorted semen with acceptable P/AI in beef cattle. Finally, color-flow Doppler ultrasonography has been incorporated recently to determine the vascularity of the CL and thereby detect pregnancy as early as Day 22 after the first AI for re- synchronization of ovulation for a second FTAI in non- pregnant animals. In summary, FTAI protocols have facilitated the widespread application of AI in South American beef cattle by allowing for the insemination and re-insemination of herds during a defined breeding season, without the necessity of clean up bulls to achieve high pregnancy rates.

Although the first mammalian embryo transfers were done more than 100 years ago, commercial bovine embryo transfer came into being in the early 1970s with the importation of European breeds of cattle into North America. Since that time commercial bovine embryo transfer has grown throughout the world, and in 2016, approximately one million bovine embryos were transferred, and several thousands of embryos were transported internationally. Because in vivo-derived bovine embryos can be made specified pathogen-free by washing procedures, they provide the ideal means of moving animal genetics around the world. Embryo transfer techniques have improved over the years so that new methods of controlling ovarian function facilitate superstimulation of donors and synchronization of recipients and nonsurgical procedures facilitate on-farm embryo transfer.

The aim of this review is to compare the performance of different reproductive programs using natural service, estrus synchronization treatment before natural service (timed natural breeding (TNB)), artificial insemination (AI) following estrus detection and timed artificial insemination (TAI) in beef herds. It is well known that after parturition the beef cow undergoes a period of anestrous, when they do not exhibit estrus, eliminating the opportunity to become pregnant in the early postpartum by natural mating or by AI after detection of estrus. Hormonal stimulation is already a consistent and well-proven strategy used to overcome postpartum anestrus in beef herds. Basically, hormones that normally are produced during the estrous cycle of the cow can be administered in physiological doses to induce cyclicity and to precisely synchronize follicular growth, estrus and ovulation. Furthermore, two options of mating may be used after hormonal stimulation: natural service (i.e. utilization of bull service after synchronization, referred to as TNB) and TAI. These strategies improve the reproductive efficiency of the herds compared with natural service without estrus induction or synchronization. After the first synchronized service, the most common strategy adopted to get non-pregnant cows pregnant soon is the introduction of clean-up bulls until the end of the breeding season. However, methods to resynchronize non-pregnant cows after the first service are already well established and offer a potential tool to reduce the time for subsequent inseminations. Thus, the use of these technologies enable to eliminate the use of bulls by using resynchronization programs (i.e. two, three or four sequential TAI procedures). The dissemination of efficient reproductive procedures, such as TNB, TAI and Resynch programs, either isolated or in combination, enables the production of a greater quantity (obtaining increased pregnancy rates early in the breeding season) and quality (maximization of the use of AI with superior genetic sires) of beef calves. These technologies can contribute to improve the production efficiency, and consequently, improve livestock profitability.

The aim of this work was to fit and compare three non-linear models (Wood, Milkbot and diphasic) to model lactation curves from two approaches: with and without cow random effect. Knowing the behaviour of lactation curves is critical for decision-making in a dairy farm. Knowledge of the model of milk production progress along each lactation is necessary not only at the mean population level (dairy farm), but also at individual level (cow-lactation). The fits were made in a group of high production and reproduction dairy farms; in first and third lactations in cool seasons. A total of 2167 complete lactations were involved, of which 984 were first-lactations and the remaining ones, third lactations (19 382 milk yield tests). PROC NLMIXED in SAS was used to make the fits and estimate the model parameters. The diphasic model resulted to be computationally complex and barely practical. Regarding the classical Wood and MilkBot models, although the information criteria suggest the selection of MilkBot, the differences in the estimation of production indicators did not show a significant improvement. The Wood model was found to be a good option for fitting the expected value of lactation curves. Furthermore, the three models fitted better when the subject (cow) random effect was considered, which is related to magnitude of production. The random effect improved the predictive potential of the models, but it did not have a significant effect on the production indicators derived from the lactation curves, such as milk yield and days in milk to peak. Fil: Piccardi, Mónica Belén. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Departamento de Desarrollo Rural. Area de Estadística y Biometría; Argentina Fil: Macchiavelli, Raúl. Universidad de Puerto Rico; Puerto Rico Fil: Funes, Ariel Capitaine. DAIRYTECH; Argentina Fil: Bó, Gabriel A.. Universidad Nacional de Villa María; Argentina. Instituto de Reproducción Animal Córdoba; Argentina Fil: Balzarini, Monica Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Departamento de Desarrollo Rural. Area de Estadística y Biometría; Argentina

The aim of this work was to compare productive and reproductive performance between pure Holstein (H) and Swedish Red & White×Holstein crossbred (SRB/H) in first lactation dairy cows from the central region of Argentina. This study included 450 lactations from a commercial dairy of central-southern Argentina. The differences in mean daily milk yield, days to first breeding (DFB) and number of services per pregnancy (NS/P) between genotypes were evaluated. A Cox proportional hazard regression model was fitted to evaluate the breed effects on days open (DO) and adjusting by calving season and calving year. Days open were estimated trough survival curves for H breed and SRB/H crossbred using Kaplan–Meier method. Finally, average lactation curves were modeled for each genotype using a non-linear mixed model. Daily milk yield was not significantly different between genotypes. However, the cumulative 305-d yield and peak milk yield for pure H was higher than for SRB/H. Days to first breeding was lower for SRB/H than pure H. The NS/P was higher for pure H than for SRB/H. Regarding DO, SRB/H breed had 1.00–1.71 higher likelihood to become pregnant than pure H. Days open of 50% of pure H was 30 d higher than for SRB/H. The results showed that SRB/H crossbred were superior to pure H in terms of reproductive performance but inferior in terms of productive performance. Further studies may help to determine if the economics of dairying justifies the use of crossbreeding as a herd fertility enhancing tool. Fil: Piccardi, Mónica Belén. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Matemáticas. Cátedra de Estadística y Biometria; Argentina Fil: Pipino, D. Agroveterinaria Pipino. Cordoba; Argentina Fil: Bo, Gabriel Amilcar. Instituto de Reproduccion Animal Córdoba; Argentina. Universidad Nacional de Villa María; Argentina Fil: Balzarini, Monica Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Matemáticas. Cátedra de Estadística y Biometria; Argentina

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) administration associated to fixed-time AI (FTAI) on follicular dynamics, ovulation, corpus luteum (CL) development and serum progesterone concentrations. Multiparous suckled Hereford cows (n = 46) in anestrus with 60–75 days postpartum were used. Females received an intravaginal device containing 0.5 g of progesterone during 8 days and 2 mg of estradiol benzoate i.m. at device insertion. At device removal 500 μg of cloprostenol and 0.5 mg of estradiol cypionate were administered i.m., and FTAI was performed 52–56 h later. Cows were divided into two experimental groups to receive 400 IU of eCG i.m. at device removal (n = 23), while control group did not receive eCG (n = 23). Daily ovarian ultrasonography (7.5 MHz transducer) and progesterone concentrations determined by RIA were assayed from device removal until 30 or 14 days after FTAI, respectively. Treatment with eCG increased ovulation rate [65.2% (15/23) vs. 30.4% (7/23); P = 0.018], ovulatory follicle diameter (14.5 ± 0.4 vs. 13.1 ± 0.7 mm, mean ± SEM; P = 0.081), CL area from 6 to 14 days after FTAI (344.3 ± 25.1 vs. 274.2 ± 23.9 mm2; P = 0.045) and mean serum progesterone concentrations from FTAI to 14 days later (3.0 ± 0.2 vs. 1.8 ± 0.2 ng/ml; P = 0.001), in comparison with control cows. In conclusion, the addition of eCG to a progesterone and estradiol’ based treatment for FTAI improves ovulation rate and luteal function in anestrous cows. These findings have implications in order to increase pregnancy rates in FTAI treatments in Bos taurus beef cattle.

Superovulation protocols have evolved greatly over the past 40 to 50 years. The development of commercial pituitary extracts and prostaglandins in the 1970s, and partially purified pituitary extracts and progesterone-releasing devices in the 1980s and 1990s have provided for the development of many of the protocols that we use today. Furthermore, the knowledge of follicular wave dynamics through the use of real-time ultrasonography and the development of the means by which follicular wave emergence can be controlled have provided new practical approaches. Although some embryo transfer practitioners still initiate superstimulatory treatments during mid-cycle in donor cows, the elective control of follicular wave emergence and ovulation has had a great effect on the application of on-farm embryo transfer, especially when large groups of donors need to be superstimulated at the same time. The most common treatment for the synchronization of follicular wave emergence for many years has been estradiol and progestins. In countries where estradiol cannot be used, practitioners have turned to alternative treatments for the synchronization of follicle wave emergence, such as mechanical follicle ablation or the administration of GnRH to induce ovulation. An approach that has shown promise is to initiate FSH treatments at the time of the emergence of the new follicular wave after GnRH-induced ovulation of an induced persistent follicle. Alternatively, it has been suggested recently that it might be possible to ignore follicular wave status, and by extending the treatment protocol, induce small antral follicles to grow and superovulate. Recently, the mixing of FSH with sustained release polymers or the development of long-acting recombinant FSH products have permitted superstimulation with a single or alternatively, two gonadotropin treatments 48 hours apart, reducing the need for animal handling during superstimulation. Although the number of transferable embryos per donor cow superstimulated has not increased, the protocols that are used today have increased the numbers of transferable embryos recovered per unit time and have facilitated the application of on-farm embryo transfer programs. They are practical, easy to administer by farm personnel, and more importantly, they eliminate the need for detecting estrus.

The objective of the present study was to evaluate the superovulatory response and embryo production in beef donors using 8 twice-daily injections of FSH or an alternative protocol in which the last 4 FSH injections were replaced with a single injection of eCG. In Exp. 1, 12 mature Bonsmara donor cows, with a body condition score between 3 and 4 (1 to 5 scale) were superstimulated twice every 46 days in a crossover design (i.e. in each experiment all cows received 2 treatments and the 2 treatments were equally represented in each replicate). On Day 0 a.m., all donors received an intravaginal device with 1.2g of progesterone (Diprogest 1200®, Zoovet, Santa Fe, Argentina), along with 50mg of progesterone i.m (Progestar®, Zoetis, Buenos Aires, Argentina) and 5mg of oestradiol-17β (17βOestradiol®, Rio de Janeiro, Argentina) IM. On Day 4 a.m., the superstimulatory treatments were initiated and donors in the control group received 8 applications of FSH (Folltropin-V, Vetoquinol, Lure, France), IM (total dose: 300mg NIH-FSH-P1) in a twice-daily decreasing dosage schedule over 4 days (i.e. 60, 60, 50, 50, 30, 30, 10, and 10mg, respectively). Donors in the FSH+800 eCG group received only the first 4 applications of FSH (i.e. 60, 60, 50 and 50mg, respectively) and on Day 6 a.m. they received 800IU of eCG (Novormón®, Zoetis) IM in a single dose. All donors received 500μg of cloprostenol (Ciclase DL®, Zoetis) IM on Day 6 a.m. and p.m., and the intravaginal devices were removed on Day 7 a.m. All cows also received 100μg of gonadorelin acetate (GnRH, Gonasyn gdr, Zoetis) on Day 8 a.m. and were inseminated with frozen-thawed semen from 2 bulls 12 and 24h later. On Day 15, ova/embryos were collected and evaluated according to the IETS standards. The data were analysed by GLMM (Infostat, 2018). In Exp 2., 18 Bonsmara donors with similar conditions as those in Exp. 1 were superovulated twice in a crossover design. Cows in both groups received similar treatments to those in the FSH+eCG treatment of Exp. 1, except that the total dosage of FSH was 200mg (i.e. 60, 60, 40, and 40mg, respectively) and the eCG given on Day 6 a.m. was either 600IU (group 1) or 800IU (group 2). In Exp 1., the FSH (control) group had a higher (P

An experiment was designed to evaluate pregnancy rates (P/AI) in suckled beef cows synchronized with a progesterone (P4)/oestradiol-based protocol (J-Synch) and inseminated (AI) with sexed or nonsexed (conventional) semen. Angus or Angus crossbred cows (n=877), 60 to 90 days postpartum, with a body condition score of 2.5 to 3.5 (scale of 1 to 5) and a corpus luteum or a follicle >8mm in diameter detected by ultrasonography, were randomly allocated to be AI with sexed-sorted or nonsexed (conventional) semen. The experiment was performed in 4 farms, and the sexed-sorted (SexedUltra™ 4M, with 4 million sperm; ST Genetics, Navasota, TX, USA) and conventional (25 million sperm) semen were from split ejaculates from 5 Angus bulls. All cows received a P4 device (DIB 0.5, Zoetis, Villa Adelina, Argentina) and 2mg of oestradiol benzoate (Gonadiol, Zoetis) on Day 0. On Day 6, cows received 400IU of eCG (Novormon 5000, Zoetis) and 500µg of cloprostenol (Ciclase DL, Zoetis) and DIB was removed. All cows were also tail painted and observed for signs of oestrus (i.e. >30% of the tail paint rubbed off) at 60, 72, and 84h after DIB removal. Cows with the tail paint rubbed off by 60 or 72h after DIB removal were artificially inseminated at 72h with either sexed or conventional semen. Those not showing oestrus by 72h received 100µg of gonadorelin acetate (Gonasyn GDR, Zoetis) at that time and were artificially inseminated with either sexed-sorted or conventional semen 12h later (i.e. 84h). Pregnancy was diagnosed by ultrasonography 30 days after AI. Data were analysed by general linear model for binary data with a logit link. Overall, 76.4% (678/877) of cows showed oestrus 60 to 84h after DIB removal, and there were differences in P/AI between sexed-sorted and conventional semen (P

Contents Over the last several decades, a number of therapies have been developed that manipulate ovarian follicle growth to improve oocyte quality and conception rates in cattle. Various strategies have been proposed to improve the responses to reproductive biotechnologies following timed artificial insemination (TAI), superovulation (SOV) or ovum pickup (OPU) programmes. During TAI protocols, final follicular growth and size of the ovulatory follicle are key factors that may significantly influence oocyte quality, ovulation, the uterine environment and consequently pregnancy outcomes. Progesterone concentrations during SOV protocols influence follicular growth, oocyte quality and embryo quality; therefore, several adjustments to SOV protocols have been proposed depending on the animal category and breed. In addition, the success of in vitro embryo production is directly related to the number and quality of cumulus oocyte complexes harvested by OPU. Control of follicle development has a significant impact on the OPU outcome. This article discusses a number of key points related to the manipulation of ovarian follicular growth to maximize oocyte quality and improve conception rates following TAI and embryo transfer of in vivo- and in vitro-derived embryos in cattle.

Three experiments were designed to evaluate the superovulatory response of beef cows following two intramuscular (IM) administrations 48 h apart of Folltropin-V diluted in reduced concentrations of hyaluronan (Split-single IM administrations; Experiment 1-300 mg Folltropin-V on the first day and 100 mg 48 h later; Experiment 2-200 mg Folltropin-V on the first day and 100 mg 48 h later). In Experiments 1 and 2, superovulatory response and ova embryo/embryo production did not differ between donors receiving twice daily IM of Folltropin-V over 4 days and those given a Split-single IM administration of Folltropin-V diluted in 10 mg/mL hyaluronan solution. Experiment 3 compared Split-single IM administration of Folltropin-V diluted in two hyaluronan concentrations (5 or 10 mg/mL) with Folltropin-V diluted in saline and administered twice-daily over 4 days. Beef cows (17 Angus and 12 Simmental) were randomly assigned to one of three treatment groups to be superstimulated three times in a cross-over design, so that all cows received all treatments. A total dose of 300 mg Folltropin-V was divided into twice-daily IM over 4 days, or in two IM treatment 48 h apart (200 mg on first day and 100 mg 48 h later) in the hyaluronan groups. Mean (± SEM) numbers of transferable embryos did not differ among treatment groups (Control: 4.0 ± 0.8; 10 mg/mL hylauronan: 5.0 ± 0.9; 5 mg/mL hyaluronan: 6.1 ± 1.3). We concluded that the Split-single IM administration of Folltropin-V diluted in either concentration of hyaluronan resulted in a comparable superovulatory response to the traditional twice-daily protocol.

Fixed-time artificial insemination (FTAI) has been regarded as the most useful method to increase the number of cows inseminated in a given herd. The main treatments for FTAI in beef cattle are based on the use of progesterone-releasing devices and GnRH or estradiol to synchronize follicle wave emergence, with a mean pregnancy per AI (P/AI) around 50%. However, more recent protocols based on GnRH (named 5-day Co-Synch) or estradiol (named J-Synch) that reduce the period of progesterone device insertion and extend the period from device removal to FTAI have been reported to improve P/AI in beef cattle. Furthermore, treatments to resynchronize ovulation for a second FTAI in nonpregnant cows have provided the opportunity to do sequential inseminations and achieve high P/AI in a breeding season, reducing or even eliminating the need for clean-up bulls. In summary, FTAI protocols have facilitated the widespread application of AI in beef cattle, primarily by eliminating the necessity of estrus detection in beef herds.

El propósito de este estudio fue estimar, a través de una regresión logística múltiple, la contribución relativa de factores que afectan la probabilidad de preñez a los 100 días en lactancia en rodeos lecheros argentinos. Los factores analizados fueron: nivel productivo, estación de parto, número de lactancia,tipo de servicio utilizado inseminación artificial (IA), natural o combinación de ambas y la interacción del nivel productivo y el tipo de servicio. Los 291 tambos (162.116 lactancias) son usuarios del software Dairy Comp305, del cual se obtiene la producción estimada que se espera alcance la vaca adulta(305 equivalente maduro). Todos los factores resultaron estadísticamente significativos. Las vacas con bajo nivel productivo tuvieron mayores chances depreñarse que vacas con niveles productivos mayores. Las vacas con parición en estaciones más frías tuvieron mayores chances de preñarse a los 100 días.La relación resultó cuadrática entre la probabilidad de preñarse a los 100 días y el número de lactancia. Los bovinos que recibieron servicio natural presentaron mayor tasa de preñez acumulada; sin embargo, la IA estuvo asociada a niveles productivos mayores. Los resultados sugieren la necesidad de diseños alternativos y evaluación del manejo reproductivo según niveles productivos específicos, como herramienta para incrementar la eficiencia reproductiva global.

The present study aimed to evaluate the efficacy of different inducers of new follicular wave emergence (FWE) and ovulation in fixed-time artificial insemination (FTAI) synchronization protocols using norgestomet ear implants (NORG) in Bos indicus cattle. In Experiment 1, the synchronization of FWE was evaluated when two different estradiol esters in different doses [2mg estradiol benzoate (EB), 2.5mg EV or 5mg estradiol valerate (EV)] were administered with NORG implant insertion in B. indicus cattle (estrous cyclic heifers and cows with suckling calves; n=10 per treatment). After estradiol treatment, ovarian ultrasonic exams were performed once daily to detect the interval between treatment and FWE. There were significant treatment-by-animal category interaction (P=0.05) on the interval from the estradiol treatment to FWE. An earlier (P0.0001) and less variable (P=0.02) interval from estradiol treatment to FWE was observed in heifers treated with EB (2.5±0.2; mean±SE) than in those treated with 2.5mg EV (4.2±0.3) or 5mg EV (6.1±0.6). Cows treated with 5mg EV (4.0±0.5) had longer (P=0.05) interval than cows receiving EB (2.5±0.2), however, there was an intermediate interval in those cows treated with 2.5mg EV (3.1±0.4). In Experiment 2, the number of uses of the NORG implant (new; n=305 or previously used once; n=314) and three different ovulation induction hormones [0.5mg estradiol cypionate (EC) at implant removal (n=205), 1mg EB given 24h after implant removal (n=219), or 100μg gonadorelin (GnRH) given at FTAI (n=195)] were evaluated in Nelore heifers (2×3 factorial design). Similar pregnancy per AI (P/AI; 30 days after FTAI; P0.05) were achieved using each of the three ovulation induction hormones (EB=40.6%; EC=48.3%, or GnRH=48.7%) and with a new (47.2%) or once-used NORG implant (44.3%). In Experiment 3, the effect of different ovulation induction hormones for FTAI [1mg EC at NORG implant removal (n=228), 10μg buserelin acetate at FTAI (GnRH; n=212) or both treatments (EC+GnRH; n=215)] on P/AI was evaluated in suckled beef cows treated with a once-used NORG implant and EB to synchronize the FWE. Similar P/AI (P=0.71) were obtained using GnRH (50.9%), EC (51.8%) or both treatments (54.9%) as ovulation induction hormones. Therefore, both doses of EV (2.5 or 5.0mg) with NORG implant delayed and increased the variation of the day of new FWE compared with EB in B. indicus cattle. These effects were more pronounced in B. indicus heifers than cows. Synchronization protocols for FTAI with either a new or once-used NORG implant with EB at insertion to induce a new FWE and either the use of EB, EC or GnRH as ovulation induction hormones may be successful in B. indicus heifers. Also, when a once-used NORG implant was used, either the administration of EC, GnRH or both as ovulation inducers resulted in similar P/AI in suckled B. indicus cows, showing no additive effect of the combination of both ovulation induction hormones.

Two experiments were conducted to investigate the effects of equine chorionic gonadotropin (eCG) at progestin removal and gonadotropin-releasing hormone (GnRH) at timed artificial insemination (TAI) on ovarian follicular dynamics (Experiment 1) and pregnancy rates (Experiment 2) in suckled Nelore (Bos indicus) cows. Both experiments were 2 × 2 factorials (eCG or No eCG, and GnRH or No GnRH), with identical treatments. In Experiment 1, 50 anestrous cows, 134.5 ± 2.3 d postpartum, received a 3 mg norgestomet ear implant sc, plus 3 mg norgestomet and 5 mg estradiol valerate im on Day 0. The implant was removed on Day 9, with TAI 54 h later. Cows received 400 IU eCG or no further treatment on Day 9 and GnRH (100 μg gonadorelin) or no further treatment at TAI. Treatment with eCG increased the growth rate of the largest follicle from Days 9 to 11 (means ± SEM, 1.53 ± 0.1 vs. 0.48 ± 0.1 mm/d; P < 0.0001), its diameter on Day 11 (11.4 ± 0.6 vs. 9.3 ± 0.7 mm; P = 0.03), as well as ovulation rate (80.8% vs. 50.0%, P = 0.02), whereas GnRH improved the synchrony of ovulation (72.0 ± 1.1 vs. 71.1 ± 2.0 h). In Experiment 2 (n = 599 cows, 40 to 120 d postpartum), pregnancy rates differed (P = 0.004) among groups (27.6%, 40.1%, 47.7%, and 55.7% for Control, GnRH, eCG, and eCG + GnRH groups). Both eCG and GnRH improved pregnancy rates (51.7% vs. 33.8%, P = 0.002; and 48.0% vs 37.6%, P = 0.02, respectively), although their effects were not additive (no significant interaction). In conclusion, eCG at norgestomet implant removal increased the growth rate of the largest follicle (LF) from implant removal to TAI, the diameter of the LF at TAI, and rates of ovulation and pregnancy rates. Furthermore, GnRH at TAI improved the synchrony of ovulations and pregnancy rates in postpartum Nelore cows treated with a norgestomet-based TAI protocol.

The objective of this study was to evaluate the effect of synchronization treatment and oestrus expression on conception rates and pregnancy losses in recipients receiving in vitro-produced (IVP) embryos. Crossbred beef cows (n = 407) with a corpus luteum (CL) determined by ultrasonography and body condition between 2.5 and 4.5 (1 to 5 scale) received a progesterone-releasing device (DIB 0.5 g, Zoetis, Argentina) and 2 mg of oestradiol benzoate (EB, Gonadiol, Zoetis) on Day 0 and were randomly allocated to 1 of 2 treatments. Recipients in the Conventional group (n = 201) had their devices removed and received 500 µg of cloprostenol (PGF, Ciclase, Zoetis), 0.5 mg of oestradiol cypionate (ECP, Cipiosyn, Zoetis), and 400 IU of eCG (Novormón, Zoetis) on Day 8. Recipients in the J-Synch group (n = 206) had their devices removed and received PGF and eCG on Day 6. All recipients were tail-painted at device removal to determine the presence of oestrus on Day 10 (a.m.) in the Conventional group and on Day 9 (a.m.) in the J-Synch group. All recipients not in oestrus in both groups received 100 µg of gonadorelin (GnRH, Ovurelin, Bayer, New Zealand) at that time. In addition, the paint was observed again in the p.m. and it was recorded. Recipients were examined by ultrasonography 7 days after oestrus or GnRH treatment (Day 16 in the J-Synch group and Day 17 in the Conventional group) and those with a CL = 16 mm in diameter received an IVP fresh embryo non-surgically. Pregnancy was determined by ultrasonography 23 and 53 days after embryo transfer and calving rates were recorded. Data were analysed using generalized linear models for binary data and a logit link (InfoStat, https://www.infostat.com.ar/). The proportion of recipients transferred was higher (P 0.11), but they were lower (P

The aim of the present study was to investigate the effects of a strategy for extending pro-oestrus (the interval between luteolysis and ovulation) in an oestrus synchronisation protocol (named J-Synch) in beef heifers on follicular growth, sexual steroid concentrations, the oestrogen receptor ERα and progesterone receptors (PR) in the uterus, insulin-like growth factor (IGF) 1 and pregnancy rates. In Experiment 1, heifers treated with the new J-Synch protocol had a longer pro-oestrus period than those treated with the conventional protocol (mean (± s.e.m.) 93.7 ± 12.9 vs 65.0 ± 13.7 h respectively; P

High temperature vulcanizing silicone elastomers have been widely used in controlled delivery systems of steroid hormones with the aim of controlling estrous cycle in livestock. This paper reports experiments conducted to evaluate the possibility of using room temperature vulcanizing (RTV) silicone elastomers for the intravaginal administration of progesterone to cattle. In vitro studies showed that RTV silicones and high-temperature vulcanizing silicone release progesterone at a similar rate. Y-shaped inserts made of different polymers were designed as supports of RTV silicone sheaths to test the in vivo release of progesterone. Field evaluation showed that RTV silicone sheaths containing 0.75 g of progesterone were at least as effective at estrous synchronization as commercially available intravaginal inserts.

Protocols that controlled follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer, as they permitted the initiation of superstimulatory treatments at a self-appointed time. However, the most commonly used approach for synchronization of follicular wave emergence involved estradiol, which cannot be used in many countries. Therefore, alternative treatments are required. Mechanical removal of the dominant follicle by ultrasound-guided follicle aspiration was effective, but required the use of specialized equipment and trained technical staff, which made it difficult to utilize in the field. Exogenous GnRH or pLH have also been used to induce ovulation of a dominant follicle, synchronizing follicular wave emergence, but their efficacy was dependent on the stage of the dominant follicle at treatment; thus, the emergence of the ensuing follicular wave may be too variable for superstimulation. An alternative approach could be initiating treatments at the time of emergence of the first follicular wave, but the need to synchronize ovulation may be a disadvantage in groups of donors at random stages of the estrous cycle. The final alternative may be to use FSH or eCG to initiate a new wave, without regard to the presence of a dominant follicle, followed by superstimulatory treatment at a predetermined time. All alternatives need to be thoroughly investigated in order to confirm their utility in the superstimulation of donor cows, regardless of the stage of the estrous cycle and without compromising ova/embryo production.

The adaptation of Bos indicus cattle to tropical and subtropical environments has led to their widespread distribution around the world. Although artificial insemination (AI) is one of the best alternatives to introduce new genetics into Bos indicus herds, the peculiarity of their temperament and the tendency to show short oestrus (many of them during the night) greatly affects the effectiveness of genetic improvement programs. Therefore, the most useful alternative to increase the number of females that are inseminated is the use of protocols that allow for AI without the need for oestrus detection, usually called fixed-time AI (FTAI). Besides, the development of protocols to advance the resumption of cyclicity during the early postpartum period has a great impact on beef production and will allow for the inclusion of a significantly larger population of animals into genetic improvement programs. Fixed-time AI protocols using progestin devices, oestradiol and eCG have resulted in consistent pregnancy rates in suckled Bos indicus and Bos indicus x Bos taurus cows. Furthermore, fertility in the successive cycles and the overall pregnancy rates at the end of the breeding season, have been shown to be improved by the use of progestin devices at the beginning of the breeding season. In summary, exogenous control of luteal and follicular development has facilitated the application of assisted reproductive technologies in Bos indicus-influenced cattle, by offering the possibility of planning programs without the necessity of oestrus detection and may provide the opportunity to improve reproductive performance of beef cattle in tropical climates.

Made available in DSpace on 2018-12-11T16:40:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-03-15 Plasma FSH profiles, in vitro embryo production (IVP) after ovum pickup (OPU), and establishment of pregnancy with IVP embryos were compared in untreated Holstein oocyte donors and those superstimulated with multiple injections or a single intramuscular (IM) injection of porcine FSH (pFSH) in hyaluronan (HA). Plasma FSH profiles were determined in 23 heifers randomly allocated to one of four groups. Controls received no treatment, whereas the F200 group received 200 mg of pFSH in four doses, 12 hours apart. The F200HA and F300HA groups received 200- or 300-mg pFSH in 5 mL or 7.5 mL, respectively of a 0.5% HA solution by a single IM injection. Plasma FSH levels were determined before the first pFSH treatment and every 6 hours over 96 hours. All data were analyzed by orthogonal contrasts. Circulating FSH area under curve (AUC) in pFSH-treated animals was greater than that in the control group (P = 0.02). Although the AUC did not differ among FSH-treated groups (P = 0.56), the total period with elevated plasma FSH was greater in the F200 group than in the HA groups (P < 0.0001). However, the F300HA group had a greater AUC than the F200HA group (P = 0.006), with a similar total period with elevated plasma FSH (P = 0.17). The IVP was performed in 90 nonlactating Holstein cows randomly allocated to one of the four treatment groups as in the first experiment. A greater proportion of medium-sized (6-10 mm) follicles was observed in cows receiving pFSH, regardless of the treatment group (P < 0.0001). Also, numbers of follicles (P = 0.01), cumulus-oocyte complexes (COCs) retrieved (P = 0.01) and matured (P = 0.02), cleavage rates (P = 0.002), and blastocysts produced per OPU session (P = 0.06) were greater in cows receiving pFSH, regardless of the treatment group. Cows in the F200HA group had a greater recovery rate (P = 0.009), number of COCs cultured (P = 0.04), and blastocysts produced per OPU session (P = 0.06) than cows in the F300HA group. Similar pregnancy rates were observed 50 to 60 days after transferring IVP embryos from donors in the different treatment groups (P > 0.05). In conclusion, a single IM injection of pFSH combined in 0.5% HA resulted in similar plasma FSH profiles as twice-daily pFSH treatments. Treatment of nonlactating donors with pFSH, with or without HA, resulted in increased IVP over untreated controls. A single dose of 200 mg of pFSH in 0.5% HA resulted in greater IVP than 300-mg pFSH in HA. Finally, pregnancy rates with IVP embryos were similar, regardless donor treatment. Departamento de Reprodução Animal FMVZ/USP Clínica Veterinária Samvet São Carlos FinInVitro Reprodução Animal Departamento de Reprodução Animal UNESP NeoGen Reprodução Assistida Instituto de Reproducción Animal Córdoba (IRAC) Department of Large Animal Clinical Sciences Western College of Veterinary Medicine University of Saskatchewan Departamento de Reprodução Animal UNESP

Compared to Bos taurus breeds, Bos indicus breeds of cattle present several differences in reproductive physiology. Follicular diameter at deviation and at the time of ovulatory capability are smaller in B. indicus breeds. Furthermore, B. indicus breeds have a greater sensitivity to gonadotropins, a shorter duration of estrus, and more often express estrus during the night. These differences must be considered when setting up embryo transfer programs for B. indicus cattle. In recent studies, we evaluated follicular dynamics and superovulatory responses in B. indicus donors with the objective of implementing fixed-time AI protocols in superstimulated donors. Protocols using estradiol and progesterone/progestrogen releasing devices to control follicular wave emergence were as efficacious as in B. taurus cattle, allowing the initiation of superstimulatory treatments (with lower dosages of FSH than in B. taurus donors) at a self-appointed time. Furthermore, results presented herein indicate that delaying the removal of progesterone/progestogen-releasing devices, combined with the administration of GnRH or pLH 12 h after the last FSH injection, results in synchronous ovulations, permitting the application of fixed-time AI of donors without the necessity of estrus detection and without compromising the results.

Most of the world's bovine herd is found in tropical regions. Bos indicus predominates, due to their adaptation to the climate and management conditions. Anestrous is the main factor that negatively affects reproductive performance of animals bred in these regions of the globe. Several factors affect postpartum anestrous, including suckling and maternal-offspring bond, and pre- and postpartum nutritional status. The short duration of estrus and the tendency to show estrus during the night, greatly affect the efficiency of artificial insemination (AI) programs in B. indicus cattle managed in tropical areas. Several restricted suckling or weaning procedures (temporary or permanent), and hormonal treatments have been used to induce ovulation and cyclicity in postpartum cows. Most hormonal treatments are based on progesterone/progestogen (P4) releasing devices associated with estradiol benzoate (EB), or a combination of GnRH/PGF(2alpha)/GnRH (Ovsynch). Treatments with GnRH/PGF(2alpha)/GnRH has presented inconsistent results, probably due to the variable number of cows in anestrous. Treatments using P4 devices and EB have resulted in apparently more consistent results than Ovsynch programs in B. indicus cattle; however, pregnancy rates are low in herds presenting high anestrous rates and moderate to low body condition. The addition of an eCG treatment at the time of device removal, which increased plasma progesterone concentrations and pregnancy rates in anestrous postpartum suckled B. indicus cows, may be useful to improve reproductive performance of beef cattle in tropical climates.

We have previously shown that expression of oestrus significantly influences the time of ovulation and pregnancy rates to timed-AI (P/TAI) in beef cattle synchronized with progesterone devices and oestradiol (Bó et al. 2016 Theriogenology 86, 388–396). Furthermore, delaying TAI from 48 h to 54–58 h after device removal improved P/TAI of those animals not showing oestrus by 48 h. The objective of the present study was to determine whether the administration of GnRH to those animals not showing oestrus by 48 h after device removal had an impact on P/TAI. Non-lactating and suckled beef cows and heifers (Bonsmara, Brangus, and Braford; n = 868), with a corpus luteum (CL) or a follicle ≥8 mm in diameter detected by ultrasonography (Mindray DP 30 Vet, 7.5 MHz, China) and body condition score 2 to 4 (1 to 5 scale) were synchronized in 3 replicates. On Day 0, animals received a progesterone device (DIB 0.5 g, Zoetis, Argentina) and 2 mg of oestradiol benzoate (Von Franken, Argentina). On Day 8, DIB were removed and cows received 0.150 mg of D+cloprostenol (Río de Janeiro, Argentina) plus 0.5 mg oestradiol cypionate (Cipiosyn, Zoetis). In addition, all cows were tail-painted on the sacrococcygeal area (CeloTest, Biotay, Argentina) at DIB removal and were observed to determine the expression of oestrus, according to the percentage of paint loss (PL) 48 h later. The animals that showed oestrus (PL > 30%) were recorded and TAI at that time while those with PL ≤ 30% (not in oestrus) were randomly allocated to receive 100 μg of gonadorelin (GnRH; Gonasyn, Zoetis) or no treatment and TAI between 54 to 58 h after DIB removal. Pregnancy was diagnosed by ultrasonography at 45 days after TAI, and data were analysed by logistic regression. There were no significant differences among replicates, inseminators, semen source or animal class (dry cows, suckled cows or heifers; P > 0.11). However, animals showing oestrus by 48 h after DIB removal had higher P/TAI (371/666; 55.7; P

To describe the epidemiology of cystic ovarian disease (COD), to find possible risk factors associated with the incidence of cysts and to analyse the impact of COD on the reproductive performance of dairy cows, databases from 22 dairy herds from the main dairy region in Argentina were retrospectively evaluated throughout a 3-year period (2009-2011). A total of 248 COD cases over 9156 parturitions were recorded, resulting in a cumulative incidence rate of 2.7%. Cystic ovarian disease incidence density was lower during the first 100 days post-partum (DPP) than during later stages of lactation. Seasonality had a significant influence on the disease presentation with higher incidence rates during winter and spring. Cows with a previous diagnosis of clinical mastitis showed 2.72 times more chances of developing ovarian cysts. Cystic cows had longer calving to first service and calving to conception intervals and lower conception rate than controls.