Treatment alternatives to induce follicular wave emergence for timed-AI in lactating dairy Cows.

Two experiments evaluated the effect of different hormonal treatments to synchronize follicle wave emergence on follicle dynamics and pregnancies per AI (P/AI) in estradiol (E2)/progesterone (P4) timed-AI (TAI) protocols in lactating dairy cows. In Experiment 1, lactating, primiparous Holstein cows (n = 36) received a P4 releasing device (Day 0) and were allocated at random to one of the following three treatment groups: Group EB received 2 mg E2 benzoate (EB) intramuscularly (i.m.), Group EB + GnRH received 2 mg EB+20 µg buserelin (GnRH) i.m., or Group EB + P4 received 2 mg EB + 100 mg of injectable P4 (iP4) in oil i.m. All cows received 0.150 mg D-Cloprostenol on Days 7 and 8 followed by P4 device removal, 400 IU eCG and 1 mg ECP on Day 8. Daily ultrasound examinations revealed that although the interval from P4 device removal to ovulation was not affected by treatment, cows that received EB + GnRH had an earlier (P < 0.05) emergence of the new follicular wave (Day 2.6 ± 0.2) than the other two treatment groups (Days 3.5 ± 0.3 and 6.1 ± 0.3, for EB and EB + P4, respectively). In Experiment 2, 808 lactating cows were assigned randomly to the three treatments evaluated in Experiment 1, and all the cows were TAI to determine P/AI. Cows in the EB + GnRH group had greater P/AI (57.4 %, P < 0.01) than those in the EB (44.6 %) or EB + P4 (45.7 %) groups. In conclusion, the administration of GnRH, but not iP4, on the day of insertion of a P4 device improves P/AI in lactating dairy cows synchronized for TAI with an estradiol/P4-based protocol.

The Economic Evaluation of Mastitis Control Strategies in Holstein-Friesian Dairy Herds.

The economic evaluation of mastitis control is challenging. The objective of this study was to perform the economic evaluation of mastitis control, under different intervention scenarios, quantifying the total cost of mastitis caused by S. aureus in Holstein cows in Argentina. A model was set for a dairy herd of Holstein cows endemically infected with S. aureus. A basic mastitis control plan including proper milking procedures, milking machine test, dry cow therapy, and treatment for clinical mastitis, was compared against other more complex and costly interventions, such as segregation and culling of chronically infected cows. Sensitivity analysis was performed by modifying the intramammary infection transition probabilities, economic parameters, and efficacy of treatment strategies. The basic mastitis control plan showed a median total cost of USD88.6/cow per year, which was close to the infected cows culling scenarios outputs. However, the segregation scenario was the most efficient, in which the total cost was reduced by about 50%. Such cost was more sensitive to probabilities and efficacy than the economic parameters. The model is flexible and can be customized by producers and veterinarians according to different control and herd settings.

Effect of the addition of GnRH and a second prostaglandin F2α treatment on pregnancy per artificial insemination in lactating dairy cows submitted to an estradiol/progesterone-based timed-AI protocol.

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 with >50% 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.

Success of artificial insemination based on expression of estrus and the addition of GnRH to an estradiol/progesterone-based protocol on pregnancy rates in lactating dairy cows.

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 F2α, estradiol cypionate and eCG at device removal. In Experiment 1, P/AI was less (P < 0.01) when all cows were inseminated at 48 h after device removal (31%) than when cows showing estrus at 48 h were inseminated at that time and those not in estrus received GnRH and were inseminated 12 h later (53%). In Experiment 2, cows in estrus by 48 h ovulated earlier than cows that were not in estrus but received GnRH and those that did not receive GnRH. Cows that were in estrus and inseminated at 48 h (47.8%) and those not in estrus at 48 h but inseminated at 60 h (53.4%) had greater P/AI (P < 0.05) than cows that were in estrus at 48 h but were inseminated at 60 h (29.7%) or inseminated at 48 h in the absence of estrus (19.4%). In Experiment 3, GnRH treatment of cows not showing estrus at 48 h and inseminated at 60 h had greater P/AI (58.5%; P < 0.05) than those not treated with GnRH but inseminated at 60 h (39.5%). Artificial insemination based on estrus expression and the inclusion of GnRH in those not in estrus increased P/AI in lactating dairy cows.

Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells.

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.

Effect of expression of estrus and treatment with GnRH on pregnancies per AI in beef cattle synchronized with an estradiol/progesterone-based protocol.

Three experiments evaluated the effects of expression of estrus and gonadotropin releasing hormone (GnRH) treatment on pregnancies per AI (P/AI) in beef cattle that were treated with an estradiol/progesterone (P4)-based protocol for fixed-time artificial insemination (FTAI). In Experiment 1, 20 non-lactating beef cows were treated with 2 mg estradiol benzoate (EB) and an intravaginal device containing 0.5 g of P4. Seven days later, P4 devices were removed and all cows received prostaglandin F2 alpha (PGF2α) and 0.5 mg estradiol cypionate (ECP). Estrus was detected using tail paint and cows that did not show estrus by 48 h after P4 device removal were randomized to receive GnRH or no treatment. Ovulation, as determined by ultrasonography, occurred earlier in cows that showed estrus (68.0 ± 2.5 h) than in cows that did not (82.0 ± 2.1 h, P < 0.05). Furthermore, cows that received GnRH ovulated earlier (78.0 ± 2.6 h) than those that did not (86.0 ± 2.0, P < 0.05). Experiment 2 determined whether expression of estrus and the administration of GnRH to animals that did not show estrus increased P/AI. Non-lactating beef cows and heifers (n = 1356) were treated as in Experiment 1 (P4 device removal, PGF2α and ECP administration on Day 7) or extended until Day 8. All animals in estrus by 48 h after P4 device removal were inseminated and those not showing estrus received GnRH or no treatment and were FTAI 8 h later (i.e., at 56 h). P/AI were greater (P < 0.01) in animals that were observed in estrus by 48 h (56.4%) than in those that did not show estrus (46.5%). Likewise, animals that did not show estrus but were treated with GnRH had greater P/AI (53.8%, P < 0.04) than those that did not receive GnRH (37.9%). Experiment 3 was designed to determine the effect of delaying GnRH treatment to the time of FTAI (at 56 h) in cows not showing estrus by 48 h after P4 device removal. Suckled beef cows (n = 969) were treated as in Experiment 1, except that all cows also received 400 IU of eCG at the time of P4 device removal on Day 7. Cows that showed estrus by 48 h or 56 h had greater P/AI (62.3%, P < 0.05) than those did not show estrus (51.5%). Furthermore, when cows that did not show estrus by 48 h were analyzed separately, P/AI were greater (P < 0.05) in those that received GnRH at 48 h and were FTAI by 56 h (64.9%) than in those that received GnRH concurrent with FTAI by 56 h after device removal (54.6%). In summary, expression of estrus was associated with earlier ovulations and resulted in greater P/AI in cows and heifers treated with an estradiol/P4-based protocol for FTAI. Furthermore, GnRH treatment in animals that did not show estrus hastened the time of ovulation and increased P/AI.

Effect of estrus expression or treatment with GnRH on pregnancies per embryo transfer and pregnancy losses in beef recipients synchronized with estradiol/progesterone-based protocols.

Two experiments were designed to determine the effect of expression of estrus or GnRH treatment on pregnancies per embryo transfer (P/ET) and pregnancy losses in beef recipients that were synchronized with estradiol/progesterone based protocols for fixed-time embryo transfer (FTET). Experiment 1 evaluated the effect of expression of estrus and GnRH treatment in the absence of estrus on P/ET. Beef cows (n = 729) were treated with 2 mg estradiol benzoate (EB) and an intravaginal device containing 0.5 g of progesterone. Devices were removed 8 d later and all cows received prostaglandin F2α (PGF2α), 400 IU eCG, and 0.5 mg estradiol cypionate (ECP) at that time. Expression of estrus was determined at 48 and 56 h after device removal using tail-paint and cows that did not show positive signs of estrus by 48 h received GnRH or no treatment at random. The overall estrus rate was 76.0% (554/729); 68.0% had positive signs of estrus by 48 h after progesterone device removal and 28.0% of those not in estrus by 48 h showed estrus by 56 h. The proportion of recipients receiving in vivo-derived (IVD) or in vitro-produced (IVP) embryos and P/ET were greater in recipients that showed estrus by 48 and 56 h (94.0% and 48.4%, respectively) than in those that did not show estrus 41.0% and 29.0%, respectively; P < 0.01). However, GnRH treatment of recipients not showing estrus by 48 h did not improve P/ET. Experiment 2 evaluated the effect of expression of estrus on P/ET and pregnancy loses up to parturition in recipients synchronized with two estradiol-based protocols. Beef cows (n = 403) were divided at random to receive the same synchronization protocol as in Experiment 1 (ECP) or a J-Synch protocol (device removal on day 6 and without using estradiol cypionate to induce ovulation). In this experiment, pregnancy was determined at 30 and 60 d by ultrasonography, and all pregnant recipients were followed until parturition to determine pregnancy losses during gestation. Although the number of recipients receiving IVP embryos was greater in the ECP group (90.5% vs. 83.5%; P = 0.03), P/ET did not differ (ECP: 37.0% and J-Synch: 39.0%; P = 0.43). Overall, 88.0% (357/407) of the recipients synchronized showed estrus and a greater P/ET (P = 0.05) was found in the recipients that showed estrus (39.0%) vs. those that did not show estrus (26.0%), regardless of treatment group. Pregnancy losses were lower (P = 0.004) and the calving rate was higher (P = 0.01) in recipients that showed estrus (25.0% and 29.3%, respectively) than in those that did not (88.8% and 2.9%, respectively). In summary, expression of estrus was associated with a greater P/ET in recipients treated with two different estradiol/P4-based synchronization protocols. The expression of estrus was associated with a greater proportion of recipients receiving embryos, P/ET and calving rate. Treatment with GnRH did not improve P/ET in the recipients that did not show estrus, questioning the its use in recipients synchronized with estradiol/progesterone based FTET protocols.

Use of embryo transfer to alleviate infertility caused by heat stress.

Heat stress (HS) has a pronounced deleterious effect on fertility in dairy herds throughout the world, especially in hot and humid summer months in tropical and subtropical areas. Summer HS reduces feed intake and increases negative energy balance, induces changes in ovarian follicular dynamics, reduces estrus detection rates and alters oviductal function leading to fertilization failure and early embryonic death. Furthermore, oocytes harvested from lactating cows during summer HS have a decreased ability to develop to the blastocyst stage after in vitro fertilization when compared with oocytes harvested during winter. The present manuscript describes the detrimental effect of HS on reproduction, with emphasis on preovulatory oocytes and carry over effects of HS on embryo development and P/AI. Embryo transfer (ET) has been an effective tool to reestablish fertility during HS because it bypasses the damage to the oocyte and early embryo caused by hyperthermia. Therefore, a management strategy to maintain increased fertility throughout the year would be to produce embryos during the cooler months, when oocyte quality is greater, and use them to produce pregnancies during the periods of HS, when oocyte quality is compromised. However, this strategy only can be implemented using cryopreserved embryos, what is still limiting. During the warmer months, the use of heifers or non-lactating cows as oocyte or embryo donors may facilitate embryo production, mainly because of the lesser deleterious effects of HS comparing to lactating cows. Also, genetic selection of donors for thermoregulation ability is one potential strategy to mitigate effects of HS and increase embryo production during the warmer months. These alternatives enable the transference of fresh embryos with more efficiency during HS periods. Additionally, the application of timed ET protocols, which avoid the need for estrus detection in recipients, has facilitated management and improved the efficiency of ET programs during HS.

Pursuit of a means of manipulating ovarian function in the cow: An adventure of serendipity, collaboration and friendship.

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.

Effect of FSH treatment on cumulus oocyte complex recovery by ovum pick up and in vitro embryo production in beef donor cows.

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.

Superstimulation of ovarian follicles in cattle: Gonadotropin treatment protocols and FSH profiles.

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.

Strategies to increment in vivo and in vitro embryo production and transfer in cattle.

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.

Expression of estrus as a relevant factor in fixed-time embryo transfer programs using estradiol/progesterone-based protocols in cattle.

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.

Pursuit of a method for single administration of pFSH for superstimulation in cattle: What we have learned.

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.

Programs for fixed-time artificial insemination in South American beef cattle.

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.

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle.

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 in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.

Fitting milk production curves through nonlinear mixed models.

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.

Alternative programs for synchronizing and resynchronizing ovulation in beef cattle.

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.

Historical perspectives and recent research on superovulation in 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.

Superovulation of beef cattle with a split-single intramuscular administration of Folltropin-V in two concentrations of hyaluronan.

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.