Fixed-time Embryo Transfer Research Articles

10 Awardee Talk: Pregnancy losses in Bos indicus-influenced cowherds

Abstract Pregnancy loss is recognized as a major reproductive challenge in beef cattle operations. Most pregnancy losses are associated with early embryonic mortality, but limited research evaluated pregnancy losses throughout the gestation period. Research is warranted to address this gap in knowledge, particularly fetal losses in Bos indicus-influenced females. This subspecies predominates in tropical regions of the planet, which contain more than 80% of the world’s cattle population and supply a major portion of the global demand for beef products. Our group recently evaluated pregnancy losses from d 30 of gestation to calving in 4,926 Nelore females assigned to an ovulation synchronization + fixed-time artificial insemination (FTAI) protocol. Pregnancy loss was greater (P < 0.01) from d 60 to 150 of gestation (10.2%) compared with d 30 to 60 of gestation (6.0%) and with d 150 of gestation to calving (7.4%) and differed (P = 0.04) between these latter two periods. Pregnancy loss from d 30 to 60 of gestation did not differ (P = 0.26) among cow parities (nulliparous, primiparous, and multiparous), whereas total pregnancy losses (d 30 to calving) were greater (P < 0.01) in precocious (28.4%) and conventional (27.1%) heifers compared with primiparous (16.4%) and multiparous (13.0%) cows. In a second experiment, pregnancy losses from fixed-time embryo transfer (FTET) to calving were evaluated in 4,366 FTET events using Nelore × Angus recipient heifers. Pregnancy losses were greater (P < 0.01) from FTET to d 32 of gestation compared with d 32 to 100 and with d 100 to calving (58.7, 39.5, and 36.7%, respectively), and did not differ (P = 0.56) between these latter periods. Recipients that lost the pregnancy from FTET to d 32 gained less (P < 0.01) body condition score after FTET compared with recipients that maintained the pregnancy. Pregnancy losses from d 32 to calving were greater (P < 0.01) in recipients reared in drylots and moved to pastures on d 32 compared with recipients reared on pasture. Collectively, these results provide novel information about pregnancy losses in Bos indicus females, with a substantial proportion of losses occurring as fetal mortality.

Pregnancy losses in Bos indicus-influenced beef and dairy recipients assigned to a fixed-time embryo transfer protocol

Pregnancy losses from fixed-time embryo transfer (FTET) to calving were evaluated in Bos indicus-influenced beef and dairy recipients. Data from 4366 FTET events were collected from Nelore × Angus recipient heifers, and from 38538 FTET events in Gir × Holstein recipient heifers and cows. In beef recipients, pregnancy losses were greater (P < 0.01) from FTET (day 7 of the experiment) to day 32 compared with day 32–100 and with day 100 to calving (58.7, 39.5, and 36.7%, respectively), and did not differ (P = 0.56) between these latter periods. Recipients that lost the pregnancy from FTET to day 32 gained less (P < 0.01) body condition score after FTET compared with recipients that maintained the pregnancy. Pregnancy losses from day 32 to calving were greater (P < 0.01) in recipients reared in drylots and moved to pastures on day 32 compared with recipients reared on pasture. In dairy recipients, pregnancy losses from FTET (day 7) to day 32 were greater (P < 0.01) than losses from day 32 to calving (50.4 and 29.4%). Pregnancy losses throughout gestation were greater (P < 0.01) when the FTET event was performed during the warm season, and greater (P < 0.01) in recipients with < 3/8 Gir influence. Recipients with ≥ 3/8 Gir influence experienced a lesser (P ≤ 0.05) increase in pregnancy losses during the warm season compared with recipients with < 3/8 Gir influence. Collectively, this experiment provides novel information about pregnancy losses in B. indicus-influenced herds receiving FTET.

Efeito do tratamento com gonadotrofina na transferência de embriões nos resultados de prenhez em bovinos

Abstract The objectives were to evaluate the effects of recipient category, season and administration of a gonadorelin analogue at the time of fixed-time embryo transfer (FTET) on pregnancy per FTET at 30 and 60 days after oestrus and on pregnancy losses (PL). Recipients were randomly assigned to: treated group (n = 624), in which recipients received an intramuscular injection of 0.2 mg of gonadorelin (Fertagyl®) at FTET; or a control group (n = 687) that remained untreated. Recipients were previously treated with a synchronisation protocol based on progesterone and oestradiol. All embryos were produced in vitro. The data with binomial distribution were analysed by multivariate logistic regression, using the GLIMIX procedure of SAS. Higher P/FTET was observed at 30 days (45.8 vs. 40.0%; P = 0.03) and 60 days (43.0 vs. 37.0%; P = 0.01) in the treated group. There was a tendency toward reduced PL in the treated group (4.0 vs. 7.0%; P = 0.09). Dry cows (2.70%) and lactating cows (2.47%) had less PL (P = 0.001) compared with heifers (10.42%). In the spring/summer season the P/ FTET at 60 days was smaller (P =0.024). Greater PL tended to occur in the warmer season. Treatment with gonadorelin at the time of bovine ET increased the pregnancy per ET at days 30 and 60 and reduced PL. Additionally, dry and lactating recipient cows showed a lower PL rate compared to heifers. Furthermore, ET performed in the warmer seasons of the year resulted in a lower pregnancy rate at day 60 and greater PL.

114 Effect of length of progesterone device administration during a modified CO-Synch for fixed-time embryo transfer in heifers

114 Effect of length of progesterone device administration during a modified CO-Synch for fixed-time embryo transfer in heifers

Effects of long-acting injectable progesterone supplementation at early dioestrus on pregnancy maintenance in beef and dairy recipient cattle.

We tested in the present study the hypothesis that supplementation with long-acting P4 (iP4) at different times of the initial dioestrus improves pregnancy rates in dairy and beef recipients submitted to fixed-time embryo transfer (FTET). Recipients from commercial farms had their oestrous cycle synchronized with an E2/P4-based protocol in three experiments (Exp. 1 to 3). In Exp. 1, dairy heifers (n = 76) and cows (n = 104) were randomly assigned to two experimental groups: the control group (n = 89) and the iP4D4 group (n = 91). For Exps. 2 and 3, suckled beef recipients were used. In Exp. 2, recipients were assigned to two experimental groups: Control group (n = 147) and iP4D7 group (n = 144); whereas in Exp. 3, recipients were randomly assigned to three experimental groups: Control group (n = 85), iP4-D4 group (n = 86) and iP4D7 group (n = 81). Recipients in the iP4D4 and iP4-D7 groups received an i.m. administration of 150 mg iP4, on D4 or D7 (D0 was the day of expected oestrus). On D7, all recipients were evaluated by transrectal ultrasonography and those that had a CL received a fresh or vitrified invitro-produced embryo. In Exp. 2 and 3, the CL area was also determined by ultrasonography at the time of FTET. The pregnancy diagnosis was performed at 30 days in Exp. 1, 57 days in Exp. 2, and between 40 and 72 days of pregnancy in Exp. 3. In Exp. 1, the pregnancy rate did not differ (p > .1) between the Control group (38.2% [34/89]) and iP4D4 group (49.5% [45/91]); yet, a parity effect indicated a greater (p < .05) pregnancy rate in heifers (57.9% [44/76]) than cows (30.8% [32/104]). In Exp. 2, the pregnancy rate was greater (p < .05) in the iP4D7 group (45.0% [65/144]) than in the Control group (34.0% [50/147]). Also, a greater (p = .08) pregnancy rate was observed for recipients with a small CL (≤2.75 cm2 ) that were treated with iP4 on the day of FTET than the control recipients (46.4% [32/69] vs. 32.6% [28/86]). In Exp. 3, no significant effects (p > .1) of the treatment group or CL size were detected on pregnancy rates at days 30 and 60. In conclusion, the beneficial effects of iP4 supplementation at early dioestrus on pregnancy maintenance may vary according to the experimental conditions, but its use at the time of FTET can be used as an alternative to enhance the fertility of beef recipients in challenging conditions in commercial herds.

Conception rate and pregnancy loss in fixed-time cattle embryo transfer programs are related to the luteal blood perfusion but not to the corpus luteum size

This study aimed to evaluate the effect of luteal blood perfusion and corpus luteum (CL) area on the conception rate and occurrence of pregnancy loss of recipients in a large-scale fixed-time embryo transfer (FTET) program. Multiparous Brangus cows (n = 1700) at 45 days postpartum and body condition scores (BCS) between 2.5 and 4.0 (3.0 ± 0.3) were used in this study. On a random day of the estrous cycle (day −10), the females received progesterone and estradiol based on the FTET protocol. On day 7, 1465 recipients had at least one CL and were evaluated using B-mode ultrasound for the CL area (cm2) and color Doppler for the luteal blood perfusion score (I/low-vascularization area <40% of the CL; II/medium-vascularization >45% to < 50%; and III/high-vascularization >50%). Immediately after CL evaluation, each recipient received a single fresh embryo (blastocyst stage) ipsilateral to the CL, in vitro produced from a commercial laboratory. Pregnancy diagnosis was performed at 30 days and repeated 60 days later to evaluate pregnancy loss (30–90 days). Ultrasound evaluation and embryo transfer were performed by a single technician. For data analysis, in addition to luteal blood perfusion groups, recipients were retrospectively ranked according to CL area into small (<3 cm2; 2.63 ± 0.01), medium (>3 to < 4 cm2; 3.44 ± 0.01), and large (>4 cm2; 4.77 ± 0.03). Data were analyzed using a logistic regression model (P < 0.05). The overall conception rate was 44.2% (648/1465), influenced by the luteal blood perfusion score [P = 0.03; high 48.4%a (134/277), medium 44.6%a (427/958), and low 37.8%b (87/230)] but not by CL area ranking [P = 0.37; large 41.8% (225/538), medium 45.2% (276/610), and small 46.4% (147/317)]. There was no interaction between the luteal blood perfusion score and CL area ranking (P = 0.81), and the BCS did not affect the results of this study (P = 0.51). In terms of pregnancy loss up to 90 days, there was no effect on the CL area ranking (P = 0.77), but the flow score showed an effect [P = 0.03; high 3.6%b (5/139), medium 9.3%a (44/471), and low 10.3%a (10/97)]. The conception rate and occurrence of pregnancy loss in the FTET program in beef cattle are related to luteal blood perfusion but not CL size.

Review: Current status of corpus luteum assessment by Doppler ultrasonography to diagnose non-pregnancy and select embryo recipients in cattle.

A number of potentials uses of Doppler ultrasonography have been explored in the last decades, both as research tools in reproductive physiology investigations and for the reproductive management of farm animals. The objective of this review was to address some of the recent strategies developed in fixed-time reproductive programs and resynchronization of ovulation in cattle, based on the evaluation of corpus luteum function by color-Doppler ultrasound imaging. Recent studies in dairy and beef cattle pointed out to a high accuracy when Doppler ultrasonography is used to assess the functionality of the corpus luteum and identify non-pregnant females at 20-24days after breeding. Therefore, super-early resynchronization programs starting in the second week after timed-artificial insemination or embryo transfer have been developed and are being implemented in commercial assisted reproduction programs; thus, anticipating conception with proven semen or genetically superior embryos. In addition, assessment of corpus luteum blood perfusion can be used for identifying high fertility embryo recipients in fixed-time embryo transfer programs.

Bioclimatic interference in fertility in cows in the Amazon biome

The objective was to evaluate the correlation between environmental temperature-humidity index (THI), rectal temperature (RT), plasma cortisol concentration and pregnancy rate in embryo recipient cows in the Amazon biome. Embryo recipients (n=235) underwent estrus synchronization via simplification of the P36 protocol for Fixed-Time Embryo Transfer (TETF). On days zero (D0), eight (D8) and day 16 (D16), the rectal temperature of the recipient cows was measured, as well as the ambient temperature and relative humidity. On the 16th day after the start of the synchronization protocol (D16), each recipient received an embryo transferred after being diagnosed with a corpus luteum (CL) in one of the ovaries by ultrasonography (US) and, from each of them, a blood sample was collected by venipuncture of the coccygeal vein, in tubes with ethylenediaminetetraacetic acid (EDTA) as an anticoagulant to obtain blood plasma, where cortisol was measured by radioimmunoassay (RIA) in solid phase, using commercial kits. It was found that in the group of pregnant cows, the mean values of the parameters RT, plasma cortisol and THI (P &lt; 0.05) were lower than those presented in the group of non-pregnant cows. Moreover, between the two groups there were positive and significant correlations between RT and THI (P &lt; 0.05), as well as in relation to plasma cortisol parameters and THI (P &lt; 0.05). It is concluded that it is possible to validate the use of RT and THI as an auxiliary tool for estimating plasma cortisol concentrations.

PROGRAMAS DE INSEMINACIÓN Y TRANSFERENCIA DE EMBRIONES: EFECTO DEL CELO SOBRE LA PREÑEZ Y PÉRDIDAS EMBRIONARIAS Y FETALES EN BOVINOS

The primary objective of implementing embryo transfer in meat 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 the nutrition, the management and the synchronization of the estrus. As a result of research conducted in the last 20 years, the use of the receptor has increased through the application of protocols that synchronize ovulation and allow the transfer of embryos without the need for oestrus detection, generally known as fixed-time embryo transfer. (FTET). Although these protocols have functioned well for several years, attention has recently been directed to the effect of estrus expression and estradiol concentrations during pre-ovulatory follicle growth on embryonic growth and pregnancy. The experiments reviewed in this document demonstrate that oestrus expression is positively associated with high pregnancy rates and reduced pregnancy losses in recipients receiving frozen / thawed bovine embryos produced in vitro and in vivo.

Evolución del conocimiento sobre la fisiología ovárica y su contribución a la aplicación de las biotecnologías reproductivas en Sudamérica

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 elective induction of luteal regression. More recently, follicular wave dynamics have been studied and protocols have been designed to induce follicular wave emergence and ovulation, and the need to detect oestrus has even been eliminated. The addition of progesterone, estradiol, GnRH and equine chorionic gonadotropin (eCG) releasing devices has provided opportunities for fixed-time AI (IATF) and possibilities for achieving higher pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need to detect oestrus, allowing fixed-time embryo transfer (TETF) and the initiation of superstimulatory treatments regardless of the day of the estrous cycle. Taken together, the new protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the past 20 years, the use of AI in South America has increased, due in large part to the use of the IATF. The use of the IATF has been multiplied by more than 10 times in Brazil, with more than 11 million treatments in 2016, which represents 85% of all AI. Similar trends are occurring in Argentina and Uruguay. In vivo embryo production produced by superovulation (DIV) has remained relatively stable over the years, but in vitro embryo production (PIV) has increased dramatically in the last 10 to 15 years, especially in Brazil, where More than 300,000 PIV embryos were produced in 2010. Worldwide, over 990,000 bovine PIV embryos were produced in 2017, of which more than% were produced in South America. The use of assisted reproductive technologies has facilitated the spread of genetic improvements and increased reproductive performance; These practices are spreading to many South American countries.

GnRH Administration in the Synchronization of Receptors Cows for Fixed-Time Embryo Transfer

The bovine production chain in Brazil is highlighted worldwide; however, researchers are seeking to improve production even further by evaluating the use of reproductive biotechnologies, such as the transfer of embryos at a fixed time. The aim of this study was to analyze the expression of estrus associated or not with the use of gonadotropin releasing hormone (GnRH), on the diameter of the dominant follicle, diameter of the corpus luteum, utilization rate and conception rate of receptors cows. The experiment was carried out in southern Brazil and 400 receptors cows were used, distributed in a 2x2 factorial scheme, considering estrus expression and GnRH administration. All receptors were synchronized and, on Day 10, their ovaries were evaluated and, based on estrus expression, they were distributed into 2 subgroups: with and without GnRH application. On Day 18, they had their ovaries evaluated and they were inovulated, and on Day 41, the pregnancy diagnosis was made. The expression of estrus increased the diameter of the dominant follicle and corpus luteum and the utilization rate of the recipient cows; however, it did not improve the conception rate. The administration of GnRH, with or without the expression of estrus, did not affect any of the studied variables.

Doppler evaluation of recipient cows treated with progesterone and estradiol

Context Due to the advances in the field of biotechnology of reproduction, success rates in embryo transfer (ET) programs have been growing continuously. Such rates are influenced by several factors, among which fertility and success in synchronising the estrus of the recipients stand out. Doppler ultrasound is emerging as a useful tool fot the assessment of fertility in recipients and for the early diagnosis of pregnancy. Aims This study aimed to verify whether luteal vascularisation, uterine haemodynamics and corpus luteum (CL) volume, evaluated only on the day of fixed-time ET (FTET), are useful parameters for the selection of Girolando recipients synchronised with progesterone and estrogen. Methods Girolando recipients received a hormonal protocol for FTET and were evaluated on the day of embryo transfer by B-mode and Doppler ultrasonography. The volume of the CL, the side of the CL, the estimation of luteal vascularisation and the Doppler indexes of the right and left uterine arteries were evaluated in real time. The ultrasound images were saved and used for later pixel counting and determination of the vascularised luteal area using specific software. Blastocysts (n = 92) produced in vitro (IVF) with Grade 1 morphology on Day 7 of development were transferred to all animals with corpus luteum. The pregnancy diagnosis was made 24 days after embryo transfer and the animals were separated into two groups, namely, pregnant and non-pregnant. Key results Retrospective analyses of luteal vascularisation; haemodynamics of uterine arteries and corpus luteum volume did not differ (P &gt; 0.05) between the groups. The means of the Doppler indexes of the uterine arteries were not affected (P &gt; 0.05) by the presence of CL ipsilateral to the assessed vessel. Conclusions It is concluded that the assessments of uterine haemodynamics, size and vascularisation of the CL, through Doppler ultrasonography on the day of ET, under the proposed conditions, were not useful to predict recipients with greater chances of establishing pregnancy. Implications Although Doppler is a useful tool for real-time fertility assessment, its accuracy if affected by protocols that do not guarantee synchrony between ovulation times. This suggests that other fertility assessment methods be used under these field conditions.

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Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Gestational losses after innovating with two embryos in a fixed-time embryo transfer program

The aim of this study was to quantify the pregnancy rate after implantation of two embryos after FTET protocols, as well as to monitor pregnancy losses until parturition, evaluating, mainly, if this strategy results in more number of animals born. Therefore, 423 multiparous recipients were selected, standardized in terms of body score, who had high-quality corpora lutea. Animals were randomly divided into two groups according to one or two embryos transferred (1 embryo = Control, n = 237; 2 embryos = Group 1, n = 186). All recipients received the same hormonal treatment, which consisted of administering, on Day 0, 2 mL of estradiol benzoate (Gonadiol, ZOETIS) + 1.9 g multidose 1st use progesterone implant (CIDR, ZOETIS); on Day 8 the implants were removed + injected 0.4 mL of estradiol cypionate (E.C.P, ZOETIS) + 1.5 mL of eCG (Novormon, ZOETIS) + 1 mL of dinoprost tromethamine (Lutalyse, ZOETIS). The animals were evaluated by ultrasonography at 30 and 60 days after embryo transfer, to diagnose the success rate and embryo losses during this period. Furthermore, information was collected on births, length of gestation, number of twin births, number of childbirth assistance and the weight of the calves. The results showed that Group 1 had better success than the Control, with higher conception rates at 30 days (68.3% vs. 53.2%, P&lt;0.001) and at 60 days (62.9% vs. 52.3%; P&lt;0 .05). The number of animals born was also higher for Group 1 (53.3% vs. 43.3%, P&lt;0.01). The percentage of twins born in Group 1 was 17.9%, and the animals had lower weight compared to the Control (34.29 + 7.36 vs 37.63 + 5.73, P&lt;0.05). The length of pregnancy and the number of assistances were similar between groups. In conclusion, the strategy adopted in this experiment suggests a considerable increase in the calf birth rate, but losses during pregnancy and their mechanisms need to be elucidated.

Reproductive seasonality influences oocyte retrieval and embryonic competence but not uterine receptivity in buffaloes

Since buffaloes are a seasonal, polyestrous species, optimizing reproduction during the non-breeding season is a key factor in increasing the reproductive and productive efficiency of herds. Ovum pick-up associated with in vitro embryo production and embryo cryopreservation is an alternative to reduce seasonal impacts. We studied the effects of seasonality in buffalo oocyte donors and embryo recipients during the favorable and non-favorable breeding seasons. Donors were evaluated for oocyte recovery and blastocyst production rate as dFBS (donors in favorable breeding season) or dNBS (donors in non-favorable breeding season). Embryos produced from dFBS or dNBS were cryopreserved by vitrification or the slow-freeze method for direct transfer and transferred to recipients in the favorable (rFBS) or non-favorable breeding season (rNBS). The heifers or cows were subjected to a fixed-time embryo transfer protocol and conception rates were determined on day 30 and on day 60. The oocyte recovery was lower in dFBS than in dNBS (7.6 vs. 10.0 oocyte/OPU, p = 0.0262); while no difference was found comparing blastocyst production rate (23.7% vs. 30.9% of blastocysts, respectively). Embryos from dFBS resulted in greater (p = 0.0013) conception rates on day 30 compared to dNBS (46.5% vs. 22.4%, respectively), despite the breeding season. The rFBS and rNBS treatments had similar (p = 0.6714) conception rates on day 30 (38.0% vs. 33.0%, respectively), indicating similar uterine receptivity. However, heifers on FBS had higher (p = 0.0003) conception rates on day 30 than cows (73.9% vs. 13.3%, respectively) when receiving embryos from dFBS. Vitrification and direct transfer had similar (p = 0.1698) conception rates on day 30 (30.4% vs. 41.4%, respectively). In conclusion, in vitro-produced embryos derived from dFBS were more competent in establishing pregnancy than dNBS counterparts, independent of recipients' reproductive seasonality. Heifers achieved better conception rates than cows during the favorable breeding season when the embryo came from dFBS. Cryopreserved in vitro produced embryos represent a reliable alternative to reduce seasonal variations in buffalo reproduction. The data elucidate the seasonal effects on embryo competence and on recipients’ uterine receptivity, affording new strategies to implement ovum pick-up associated with in vitro embryo production programs in buffalo herds.

Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and B. taurus suckled beef cows

Recent research from our group demonstrated that Bos indicus-influenced suckled beef cows had greater resilience to withstand nutrient restriction and establish pregnancy compared with B. taurus cows exposed to the same conditions. To further understand these findings, differences in metabolic profile between these same B. indicus-influenced and B. taurus females were explored. Suckled beef cows (n = 134) were enrolled in a completely randomized design with a 2 × 2 factorial arrangement of treatments. On day −21, Angus (AN; Bos taurus) and Brangus (BN; B. indicus-influenced) cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the daily energy maintenance requirements (RESTR). Cows were exposed to an estrus synchronization protocol and received an embryo 7 d after ovulation was pharmacologically induced on day 0. Blood samples were collected on days −21 and 19 to determine circulating concentrations of non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), insulin, glucose, and IGF-1. Pregnancy status after embryo transfer was determined on day 28. As a consequence of the proposed diets, cows in the RESTR diet had less body condition score (BCS) on day 19 (P = 0.008) across breed types. Moreover, BCS change from day −21 to 19 was included as independent covariate into subsequent analyses, allowing for the comparison of breed types under an equivalent level of body reserve mobilization. A breed × diet interaction was observed for plasma insulin (P = 0.03) and IGF-1 (P = 0.04) on day 19, where AN–RESTR cows had less plasma concentrations on day 19 compared with AN–MAINT cows. Diets did not impact (P > 0.10) plasma insulin and IGF-1 concentrations in BN cows. No diet or breed effects were observed in circulating concentrations of NEFA, BHB, and glucose (P > 0.10). Across breed types and nutritional treatment, there was positive linear effect (P ≤ 0.04) of plasma concentrations of insulin and IGF-1 on the probability of pregnancy to fixed-time embryo transfer. In summary, the negative impacts of nutrient restriction on the somatotropic axis, independently of body tissue mobilization, were heightened in Bos taurus females compared with B. indicus-influenced cohorts, which corroborate with the differences observed in fertility between these subspecies.

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.

Optimization of a 5-day fixed-time embryo transfer (FTET) protocol in heifers I. Manipulation of circulating progesterone through reutilization of intravaginal progesterone devices during FTET

The objectives of the present study were to: 1) compare the reproductive efficiency of embryo transfer (ET) recipients after synchronization of estrus or a 5-day synchronization of ovulation protocol for fixed time ET (FTET), and 2) determine the effect of reutilization of intravaginal P4 devices (CIDRs), up to four times, in a 5-day FTET protocol. In Experiment 1, 817 dairy heifers were assigned to one of three groups: PGF + estrus detection, 5-d FTET protocol with new (1.38 g P4) or 2nd use CIDR (previously used once for 5 d). Fresh in vitro produced embryos were transferred 7 ± 1 day after estrus (PGF + estrus) or GnRH (5-day FTET). Utilization rate (transferred/treated) was greater (P < 0.001) in heifers submitted to FTET compared to ET after estrus, however pregnancies per ET (P/ET) were not different (P > 0.10). As a result, pregnancy per treated (P/treated) recipient was greater (P < 0.05) in heifers in the 5-day FTET protocol. In Experiment 2, 40 dairy heifers without a corpus luteum (CL) were randomly allocated into one of four groups using new, 2nd use, 3rd use (previously used twice for 5 d/each), or 4th use (previously used thrice for 5 d/each) CIDRs. Circulating P4 was reduced (P < 0.01) with each reutilization. In Experiment 3, ovarian follicular dynamics were evaluated in 238 dairy heifers submitted to a 5-day protocol with either new, 2nd use, 3rd use or 4th use CIDRs at random stages of the estrous cycle. Prostaglandin F2α (PGF) was administered at CIDR removal and again 24 h later. Ovulation was induced by GnRH treatment 72 h after CIDR removal. Preovulatory follicle diameter increased (P < 0.001) progressively with increasing CIDR reutilization. Ovulation rate did not differ between treatments, however, interval from CIDR removal to ovulation decreased (P < 0.001) in heifers receiving 3rd and 4th use CIDRs compared to new or 2nd use. Finally, in Experiments 4 and 5, 1203 heifers submitted to a 5-day FTET protocol were randomly assigned to receive either a new CIDR, a 3rd use CIDR (Experiment 4) or a 4th use CIDR (Experiment 5). Despite the increase in CL volume on D5 in heifers treated with 3rd use (P = 0.03) or 4th use CIDRs (P < 0.01), there were no differences (P > 0.05) in utilization rate, P/ET, or P/treated. Thus, use of a 5-day FTET synchronization protocol improves reproductive efficiency by increasing recipient utilization, and reutilization of CIDRs up to four times in recipient dairy heifers does not compromise reproductive performance.

Influence of Injectable Progesterone on the Pregnancy Rate of Heifers Receiving Bovine Embryos

Background: In vitro embryo production (IVEP) allows the spread of superior animal genetics, but pregnancy rates show a high variability with this biotechnique. In the initial stage of pregnancy, progesterone plays a fundamental role in uterine preparation, acting on embryonic growth, implantation, and development. However, on the day of the IVEP transfer to the recipients, progesterone levels may be lower than that expected, influencing the uterine environment and, consequently, the pregnancy rate. Therefore, the objective of this study was to evaluate the pregnancy rate in heifers after the administration of injectable progesterone (P4) in the fixed-time embryo transfer (FTET) protocol.Materials, Methods &amp; Results: The experiment was conducted inside a rural property near the city of Rio Branco, Acre, Brazil. The experimental group consisted of 232 animals, including 78 zebuine (Bos indicus) and 154 mixed (½ blood B. indicus and ½ blood B. taurus) animals, aged between 16 and 24 months, with a mean weight of 300 and 330 kg for zebuine and mixed animals, respectively. The selected animals were previously synchronized using the progesterone-estrogen-prostaglandin-estrogen protocol. Embryo transfer was performed on day 18 of the protocol, which was 9 days after the removal of intravaginal progesterone implant. On day 15 of the protocol, that is, 144 h (6 days) after the device removal, the animals were randomly distributed into two experimental groups: Control Group (CG; 0.5 mL of 0.9% saline solution, intramuscular) and Treated Group (P4G; 0.5 mL of injectable P4, 150 mg, intramuscular). Chi-square test was used for the statistical analysis of the pregnancy rate at a 5% probability. After 23 days of embryo transfer, pregnancy was diagnosed by ultrasonography. The general pregnancy rate, considering all groups (CG and P4G) and breeds included, was 55.17% (128/232). The pregnancy rates of the P4G and CG groups, regardless of breeds, were 55.08% (65/118) and 55.26% (63/114), respectively, with no statistical difference (P = 0.8344). Angus animals presented a similar pregnancy rate in the P4G and CG groups of 54.93% (39/71) and 57.83% (48/83), respectively. Furthermore, similar results were found for the Nellore breed, with no difference in pregnancy rate between the CG (55.81%, 24/43) and P4G (48.57%, 17/35) groups.Discussion: The strategy of using P4 to increase the pregnancy rate is very widespread in fixed-time artificial insemination (FTAI) protocols, although differences still exist. The literature presents zero or negative effects, as in this study, when exogenous P4 was used to increase endogenous P4 concentrations and, consequently, the pregnancy rate. However, some studies demonstrated the beneficial effects of increasing blood P4 concentrations, and that the increased fertility depends on the method and time of P4 supplementation and the animal’s physiological state. In this context, the fact that the injectable progesterone supplementation did not increase the pregnancy rate in this study is justified mainly by factors such as the animal category used (heifers), time of P4 supplementation (day 4 after ovulation), biotechnology used (FTET), and the heterogeneous characteristics related to phases of the estrous cycle of heifers. Supplementation with 150 mg of injectable long-acting progesterone intramuscularly did not interfere in the pregnancy rate of Angus and Nellore heifers receiving bovine embryos.

Factors associated with pregnancy rate in fixed-time embryo transfer in cattle under humid-tropical conditions of México.

The objective was to determine the effect of some factors on pregnancy rate of fixed-time embryo transfer (FTET), in cows and heifers kept under Mexican tropical conditions. Recipients females (n=405) grazing in pastures were selected according to breed group (Zebu and crosses), parity (nulliparous and multiparous), body condition score (BCS) and the presence of a corpus luteum (CL). The females were synchronized on day 0 using a progesterone vaginal device and 2 mg estradiol benzoate (EB), two groups were established. Group 1 (conventional protocol) were animals in which the progesterone device was removed on day 7. At this time, also received an injection of 50 mg cloprostenol sodium and 1 mg estradiol cypionate. Animals also received 300 IU (heifers) or 360 IU (cows) of eCG. Group 2 (J-Synch protocol) were animals in which the progesterone device was removed on day 6. Cloprotenol and eCG injections were applied as in Group 1. Additionally, on day 9, animals of group 2 received 0.01 mg buserelin acetate. Embryo transfer of in vivo or in vitro was done on day 16 and pregnancy diagnosis was realized by ultrasonography on days 23 and 53 after FTET. Statistical analyses were carried out using Chi-square tests and logistic regression. Pregnancy rate varied between farms (P<0.05). The highest pregnancy rate was for multiparous cows (66%). The recipient utilization rate was better in the J-Synch protocol (85%), and in vivo embryos (75%) had higher pregnancy rate. The diameter of the follicle and the CL had no effect on pregnancy rate (P>0.05). However, the logistic regression determined that the only significant factor on pregnancy rate was the type of embryo. In conclusion, pregnancy rate in FTET females was higher for in vivo embryos than for in vitro embryos in cows evaluated under humid tropical conditions in Mexico.