Comparison of four synchronization protocols for fixed-time bovine embryo transfer in Bos indicus × Bos taurus recipients

2004 ◽  
Vol 62 (9) ◽  
pp. 1577-1584 ◽  
Author(s):  
L.F. Nasser ◽  
E.L. Reis ◽  
M.A. Oliveira ◽  
G.A. Bó ◽  
P.S. Baruselli
2010 ◽  
Vol 22 (1) ◽  
pp. 67 ◽  
Author(s):  
Pietro S. Baruselli ◽  
Roberta M. Ferreira ◽  
Manoel F. Sá Filho ◽  
Luiz F. T. Nasser ◽  
Carlos A. Rodrigues ◽  
...  

Numerous studies have shown that it is possible to manipulate follicular and luteal dynamics, thereby eliminating the need for oestrus detection in embryo transfer (ET) programmes. Fixed-time ET (FTET) protocols are based on the use of gonadotrophin-releasing hormone (GnRH) and prostaglandin (PG) F or progesterone/progestogen (P4)-releasing devices and oestradiol. The FTET protocols increases the proportion of recipients transferred, and therefore pregnancy rates, compared with the use of PGF followed by ET 7 days after oestrus. Furthermore, the addition of equine chorionic gonadotrophin (eCG) to the P4 and oestradiol-based FTET protocols results in an even higher proportion of recipients transferred, and thus higher pregnancy rates. The beneficial effect of eCG treatment may be related to increased growth of the dominant follicle and increased plasma P4 concentrations during the subsequent luteal phase. In Bos taurus × Bos indicus recipients, pregnancy rates were positively correlated with the diameter of the corpus luteum (CL) and the number of CL at ET. When repeat-breeder Holstein cows were used as recipients, FTET protocols increased number of recipients transferred and pregnancy rates compared with the traditional PGF-based synchronisation protocols. In conclusion, the use of FTET protocols eliminates the need for the detection of oestrus and results in a greater proportion of recipients transferred and satisfactory pregnancy rates. Thus, FTET optimises the use of recipients, reducing labour and animal handling and facilitating the use of ET.


2004 ◽  
Vol 16 (2) ◽  
pp. 212
Author(s):  
L.F. Nasser ◽  
E.L. Reis ◽  
A.M. Oliveira ◽  
G.A. Bo ◽  
P.S. Baruselli

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus×Bos indicus beef heifers (n=301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had >1 CL or a single CL with diameter ≥18mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2+P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5=2.4±0.3 v. Day 8=1.7±0.2; P=0.03) and the number of CL (Day 5=1.4±0.1 v. Day 8=1.1±0.0; P=0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P=0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.


2007 ◽  
Vol 19 (1) ◽  
pp. 220
Author(s):  
G. A. Bo ◽  
L. C. Peres ◽  
D. Pincinato ◽  
M. de la Rey ◽  
R. Tribulo

An experiment was designed to evaluate the effect of the interval between thawing to deposition of the embryo into the uterine horn on pregnancy rates of in vivo-produced frozen–thawed embryos in 1.5 M ethylene glycol (direct transfer). Data were collected from 1122 embryo transfers performed in the same farm (Estancia El Mangrullo, Lavalle, Santiago del Estero, Argentina) during the spring and summer of 2004/05 and 2005/06 (6 replicates, ambient temperature between 20 and 40�C). Recipients used in all replicates were non-lactating, cycling, multiparous Bos taurus � Bos indicus crossbred cows with body condition score between 3 and 4 (1 to 5 scale) that were synchronized using fixed-time embryo transfer protocols. Briefly, the synchronization treatments consisted of the insertion of a Crestar ear implant (Intervet, Sao Paulo, Brazil) or a progesterone-releasing device (DIB; Syntex SA, Buenos Aires, Argentina), plus 2 mg of estradiol benzoate (EB; Syntex) intramuscularly (IM) on Day 0, and 400 IU of eCG (Folligon 5000; Intervet, or Novormon 5000; Syntex) IM plus 150 �g d-cloprostenol IM (Preloban; Intervet, or Ciclase; Syntex) on Day 5. Progestin devices were removed on Day 8 and all cows received 1 mg of EB IM on Day 9. All cows were examined by ultrasonography on Day 16 and those with a luteal area >76 mm2 (by calculating the area of the CL minus the area of the cavity) received, on Day 17, frozen–thawed embryos by nonsurgical transfer. All embryos were Grade 1, and all were frozen in 1.5 M ethylene glycol at the Embryo Plus Laboratory (Brits, South Africa). After being stored in liquid nitrogen, the embryos were plunged directly (no air thawing) in a 30�C water bath for 30 s, and then transferred to the recipient cows by either one of two technicians. Based on the interval between thawing and transfer, the transfers were classified as being in one of 3 groups: Group 1: <3 min; Group 2: 3 to 6 min; and Group 3: 6 to 16 min. The main reason for delayed transfers beyond 6 min was the replacement of one recipient for another because of difficulty in threading the cervix (1% of the total transfers) or a recipient falling down into the chute or with very bad disposition and behavior. Pregnancy was determined by ultrasonography 28 to 35 days after fixed-time embryo transfer, and data were analyzed by logistic regression. There were no effects of replicate, technician, CL area, recipient body condition score, embryo stage, and time from thawing to transfer on pregnancy rates. Pregnancy rates in the 3 thawing to transfer intervals were: Group 1: 215/385, 55.8%; Group 2: 372/655, 56.8%; Group 3: 42/82, 51.2%; P > 0.6. These results may be interpreted to suggest that there is no significant effect of time from thawing to transfer (up to 16 min) in direct transfer embryos using Bos taurus � Bos indicus recipients transferred at a fixed time.


2010 ◽  
Vol 22 (1) ◽  
pp. 177 ◽  
Author(s):  
M. Ramos ◽  
L. Cutaia ◽  
P. Chesta ◽  
G. A. Bó

Two experiments were designed to evaluate the effect of the timing of fixed-time AI (FTAI) in relation to the removal of an intravaginal progesterone-releasing device (1 g of progesterone, DIB, Syntex SA, Buenos Aires, Argentina) on pregnancy rates in Bos indicus × Bos taurus cross-bred heifers. In experiment 1, 285 Bonsmara × zebu cross-bred heifers, between 18 and 24 months of age and with a body condition score (BCS) between 3.0 and 3.5 (1-5 scale) were used. On the day of initiation of treatment (Day 0), the heifers’ ovaries were palpated (92% of them had a CL) and they received a new DIB plus 2 mg of estradiol benzoate (EB; Syntex SA) and 250 μg of cloprostenol (Ciclase DL, Syntex SA). On Day 8, DIB devices were removed and all heifers received 250 μg of Ciclase plus 0.5 mg of estradiol cypionate (ECP; Cipiosyn, Syntex SA). At that time the heifers were randomly divided to receive FTAI between 48 to 49 h, 53 to 54 h, or 58 to 59 h after DIB removal. The heifers underwent FTAI with semen from 4 bulls by 2 inseminators. In experiment 2, 260 heifers from the same group as those used in experiment 1 (87% with a CL) were treated exactly as those in experiment 1, except that previously used DIB was inserted on Day 0. Pregnancy diagnosis was performed 30 days post-fixed-time AI by ultrasonography. The data were analyzed by logistic regression, taking into account the effect of time of FTAI, semen, and inseminator on pregnancy rates. In experiment 1, pregnancy rates were lower (P = 0.04) in the heifers undergoing FTAI between 48 and 49 h after DIB removal (46/95, 48.4%) than those undergoing FTAI 53 to 54 h (61/99, 61.6%) or 58 to 60 h (57/91, 62.6%) after DIB removal. However, no differences in pregnancy rates were found (P = 0.72) in experiment 2 between the 3 treatment groups, with 39/91 (42.9%) for the 48 to 49 h group, 45/89 (50.6%) for the 53 to 54 h group, and 35/89 (43.8%) for the 58 to 59 h group. There was no effect of the semen or inseminator (P > 0.2) in either experiment. We conclude that when Bos indicus × Bos taurus beef heifers are synchronized with new DIB devices and ECP, higher pregnancy rates are obtained in heifers undergoing FTAI late (between 53 to 60 h after DIB removal) than in those undergoing FTAI early (48 to 49 h after DIB removal). However, time of insemination does not apparently affect pregnancy rates when Bos indicus × Bos taurus beef heifers are synchronized with previously used DIB devices and ECP.


2018 ◽  
Vol 30 (1) ◽  
pp. 182
Author(s):  
J. Duran ◽  
D. Argudo ◽  
S. Bravo ◽  
C. Soria ◽  
G. Guevara ◽  
...  

Recipient handling during embryo transfer (ET) induces prostaglandin F2α (PGF2α) production in 2 periods: an early transient and rapid increase around the time of ET, followed by another 2 to 4 h later. This PGF2α is associated with embryonic loss during early gestation by affecting both the embryo and the corpus luteum. To control this, antiprostaglandins such as flunixin meglumine (FM) have been applied IM at the time of ET with varying results. In such studies, the interaction of IM administration of FM and difficulty of transfer has not always been evaluated, possibly confusing the interpretation of the results. Furthermore, IV FM injection at ET and its relationship with pregnancy rates (PR) has not been determined. The objectives were (1) to determine the relationship between difficulty of ET and PR; and (2) to evaluate the efficacy of IM v. IV FM on pregnancy outcomes. One hundred and ten crossbred (Bos taurus × Bos indicus) heifers (18-24 months old) from 3 farms were used as recipients. Two evaluation systems of ET difficulty were used: (1) duration of transfer (objective determination of the elapsed time measured in seconds between the introduction of the catheter and embryo release), and (2) level of difficulty experienced by the practitioner (subjective determination; 1 = minimum and 2 = medium to extreme manipulation). Quality 1 and 2 fresh embryos from superovulated cows were transferred by the same practitioner. At ET, recipients were randomly divided into 3 groups: (1) Control (no treatment, n = 31); (2) FM-IM (n = 39): injected IM with 2.2 mg kg−1 FM at ET; and (3) FM-IV (N = 40): injected with 2.2 mg kg−1 FM IV at ET. Pregnancy was diagnosed at 30 to 40 and 60 to 90 days after ET. Spearman’s test was performed to determine the correlation between duration and difficulty at ET and Chi-square test was used to compare PR. The mean duration of transfer for all heifers was 62.3 ± 57.5 s (11 to 357 s; median: 44.5 s). There was a high correlation (0.8; P < 0.001) between the ET difficulty evaluation systems. Overall, ET difficulty 1 had higher PR than ET difficulty 2 (64.2 v. 40.7; P = 0.013). The PR was significantly improved (P < 0.01) in the FM-IV group (75 and 70% at 30 and 60 days after ET) compared with control (45.2 and 32.3%) and FM-IM (33.3 and 30.7%). In conclusion, results indicate that the difficulty of transfer affects PR achieved following the transfer of in vivo-derived bovine embryos. Treatment with FM-IV following transfer resulted in significantly higher PR compared with control and FM-IM recipients. The IV injection of FM may antagonize the very early and transient increase of PGF2α caused by genital tract manipulation (even gently performed) at embryo transfer. Further research is necessary to confirm the results of the present study.


2005 ◽  
Vol 17 (2) ◽  
pp. 234 ◽  
Author(s):  
R. Tribulo ◽  
E. Balla ◽  
L. Cutaia ◽  
G.A. Bo ◽  
P.S. Baruselli ◽  
...  

Although several studies have investigated the relationship between circulating progesterone and pregnancy rates in cattle, the beneficial effect of treatments that increase progesterone concentrations, by insertion of a progesterone (P4) releasing device or induction of an accessory CL with hCG, GnRH, or LH treatment, has resulted in inconsistent effects on pregnancy rates in embryo recipients. An experiment was designed to evaluate the effect of hCG or GnRH treatment, given at the time of embryo transfer without estrus detection, on pregnancy rates in recipients treated with intrauterine P4-releasing devices, estradiol benzoate (EB), and eCG. The experiment was performed in two replicates; non-lactating Bos taurus × Bos indicus crossbred beef cows with a body condition score between 2.5 to 3.5 (1-to-5 scale) were used (replicate 1, n = 180; replicate 2, n = 140). All cows received 1 g of P4 via a P4-releasing device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex) on Day 0, and 400 IU of eCG i.m. (Novormon 5000, Syntex) plus 150 μg d(+)cloprostenol i.m. (Ciclase, Syntex) on Day 5. DIBs were removed on Day 8 and all cows received 1 mg EB i.m. on Day 9. Recipients were not observed for signs of estrus, and those >1 CL, or a single CL with an area >256 mm2, received 195 Grade 1 and 46 Grade 2 frozen/thawed “direct transfer” embryos on Day 17. At the time of embryo transfer, recipients were randomly allocated to 1 of 3 treatment groups to receive 1500 IU hCG (Ovusyn, Syntex), 50 μg Lecirelina (GnRH, Gonasyn, Syntex), or no treatment (control) at that time. Ovarian ultrasonography was performed on Day 0 to determine ovarian status (only cows with a CL or a follicle >10 mm and uterine tone were used), on Day 17 to measure CL area, and 40 days after embryo transfer to determine pregnancy status. Data were analyzed by logistic regression and the effects of replication, technician, treatment, and embryo quality were considered in the model. From the 320 recipients treated with a DIB plus EB and eCG, 241 (75.3%) were selected to receive an embryo. Nine (3.7%) and 1 (0.4%) of the selected recipients had 2 and 3 CL, respectively. Pregnancy rates did not differ between replicates (replicate 1: 80/140, 57.1%; and replicate 2: 57/101, 56.4%; P = 0.84), technicians (technician 1: 65/118, 55.1%; and technician 2: 72/123, 58.5%; P = 0.64), or treatments (hCG: 43/80, 53.8%; GnRH: 45/83, 54.2%; and control: 49/78, 62.8% P = 0.99). However, pregnancy rates were higher (P = 0.001) in recipients receiving Grade 1 embryos (121/195, 62.1%) than in those receiving Grade 2 embryos (16/46, 34.8%). GnRH or hCG treatment at the time of embryo transfer did not increase pregnancy rates in recipients synchronized with P4 releasing devices, EB, and eCG. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.


2011 ◽  
Vol 35 (4) ◽  
pp. 817-825
Author(s):  
José Nélio de Sousa Sales ◽  
Lilian Mara Kirsch Dias ◽  
Celso Rodrigues Franci ◽  
Alexandro Aluísio Rocha ◽  
Guilherme Gastão Cardoso ◽  
...  

Two experiments were conducted to evaluate the effect of the intramuscular injection of betacarotene associated to tocopherol on the plasma concentration progesterone of superovulated Holstein heifers (experiment 1) and in crossbred (Bos taurus x Bos indicus) heifers submitted to fixed-time embryo transfer (FTET, experiment 2). In experiment 1, after estrus synchronization and superovulation animals were inseminated 12 and 24 hours after estrus onset and embryos flushed 7 days later. Heifers were allocated randomly to one of three treatments: Control; T800 (800 mg of betacarotene plus 500 mg of tocopherol) and T1200 (1,200 mg of betacarotene plus 750 mg of tocopherol). The treatments were given on the day of ear implant placement and repeated on the first day of superovulation. Blood samples were collected on D0, D5, D9, D12 and D16. In experiment 2, treatments were imposed at intravaginal device insertion (D0). The same experimental design, as in experiment 1, was used. Blood samples were collected on D17 (embryos implanted) for progesterone determination by radioimmunoassay. In experiment 1, average plasma progesterone concentrations after corpora lutea formation (D12 plus D16 means) were 13.7±1.8 ng/ml, 14.5±2.3 ng/ml and 10.8±2.3 ng/ml for control, T800 and T1200, respectively, and did not differ (P=0.44). In experiment 2, progesterone concentrations on D17 in Control (8.88±0.57 ng/ml), T800 (7.48±0.64 ng/ml) and T1200 (5.90±1.33 ng/ml) groups were similar (P=0.11). Results indicate that the supplemental betacarotene and tocopherol injections did not influence peripheral progesterone concentrations in superovulated Holstein donors and crossbreed recipients heifers.


2013 ◽  
Vol 25 (1) ◽  
pp. 17 ◽  
Author(s):  
Ciro M. Barros ◽  
Rafael A. Satrapa ◽  
Anthony C. S. Castilho ◽  
Patrícia K. Fontes ◽  
Eduardo M. Razza ◽  
...  

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the ‘P-36 protocol’ for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT2 receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus–oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.


2017 ◽  
Author(s):  
◽  
James William Cameron Locke

The Bos indicus species of beef cattle presents a number of unique reproductive challenges, especially with regard to those that influence reproductive efficiency including timely attainment of puberty, response to certain pharmaceutical drugs used to control estrus and ovulation, and response to physiological stressors. Cattle of this biological type comprise a substantial percentage of the beef cattle population in southern regions of the United States because of their ability to withstand high ambient temperatures and relative humidity. Additionally, Bos indicus cattle are regarded to exhibit increased parasite and disease resistance, resulting in improved production in subtropical climates as compared to Bos taurus cattle in those same regions. Research utilizing composite breeds of Bos taurus and Bos indicus beef heifers has been limited in the United States, and adoption of reproductive technologies such as artificial insemination and estrus synchronization has been sluggish compared to Bos taurus cattle in other regions of the country. To help answer a number of questions about this biological type, an experiment was designed to evaluate reproductive performance of Bos indicus-influenced heifers (n = 1,456). Weights and reproductive tract scores (RTS; Scale 1-5) were obtained for heifers prior to assignment of one of five treatments: Non-synchronized + natural service (NS); melengestrol acetate + natural service (MGA + NS; 0.5 mg x animal-1 x d-1); 14-d controlled internal drug release + natural service (CIDR + NS; 1.38g progesterone); 14-d MGA-PG + fixed-time AI (FTAI); and 14-d CIDR-PG + FTAI. Heifers in the three NS treatments were exposed to fertile bulls for 65 d, beginning 10 d after progestin removal for CIDR + NS and MGA + NS groups. Heifers in FTAI treatments were administered PGF2[alpha] (PG; 25 mg, IM) 16 or 19 d following CIDR or MGA removal. Fixed-time AI was performed 66 and 72h after PG for CIDR-PG and MGA-PG treatments, respectively. Gonadotropin-releasing hormone (GnRH; 100[mu]g, i.m.) was administered at FTAI. Estrus detection aids were applied at PG for heifers in FTAI treatments and evaluated at AI. Heifers in FTAI treatments were exposed to fertile bulls 12 d following FTAI. Blood samples were collected and ovarian ultrasounds performed at PG administration and FTAI to compare serum concentrations of estradiol and progesterone and to evaluate follicular dynamics among a subset of heifers assigned to FTAI treatments. Pregnancy status was determined at the end of a 65-d breeding period. Data were analyzed using PROC FREQ and GLIMMIX procedures of SAS. Mean concentrations of estradiol at AI differed between MGA- versus CIDR-treated heifers (P = 0.04; 8.2 versus 6.6 pg/ml), however estrous response after PG (52% versus 53%) and pregnancy rates after FTAI (40%) did not differ between MGA- and CIDR-PG treatments, respectively. Across all treatments, pregnancy rates were compared on 21, 30 and 60 d of the breeding period based on pubertal status (prepubertal RTS = 1 and 2; peripubertal RTS = 3; pubertal RTS = 4 and 5). Pregnancy rates differed at each time point based on pretreatment pubertal status (P [less than or equal to] 0.02) and weight (P [less than or equal to] 0.05). No differences were apparent with regard to progestin type at any time point, although higher pregnancy rates (P [less than or equal to] 0.004) were observed among NS treatments than FTAI followed by NS exposure at Days 30 and 60. This experiment is the largest comprehensive field trial conducted in the United States and reported in the literature involving Bos indicus-influenced beef heifers utilizing estrus synchronization prior to natural service or FTAI. This study reinforced results from previous studies, however raises a number of questions yet to be answered for heifers of this biological type. These data should be used as a basis for improvements in reproductive management of Bos indicus-influenced beef heifers prior to their first breeding season, and creates a number of possibilities for future research trials that build upon results reported in this thesis. Selection, management, and post-weaning to pre-breeding development of Bos indicus-influenced replacement beef heifers should involve the cooperative efforts of beef producers, veterinarians, and research scientists to further investigate methods to improve reproductive efficiency and genetic merit of these herds.


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