101 Treatment with gonadotrophin-releasing hormone on Day 7 or 21 does not reduce pregnancy loss in dairy heifers receiving invitro-produced embryos

2020 ◽  
Vol 32 (2) ◽  
pp. 176
Author(s):  
B. J. Duran ◽  
R. V. Sala ◽  
P. L. J. Monteiro ◽  
C. Gamarra ◽  
M. Fosado ◽  
...  

Previous research has shown that induction of an accessory corpus luteum (CL) by administration of gonadotrophin-releasing hormone (GnRH) on Day 5 increases circulating progesterone and reduces pregnancy loss between Days 33 and 60 in heifers receiving invitro-produced (IVP) embryos. Therefore, the objective of the present study was to determine whether timing of induction of an accessory CL influenced pregnancy loss in IVP recipients. Holstein heifers (n=1,658) were synchronized using a modified 5-day CIDR CO-Synch protocol. Briefly, heifers received an intravaginal progesterone device (CIDR) on Day −8, Day −3: CIDR removed and first prostaglandin F2α (PGF2α) treatment, Day −2: second PGF2α, and Day 0: GnRH (G1, 100µg of gonadorelin acetate). Oestrus expression was evaluated on Day 0 with the use of an oestrus detection device. Transfer of fresh IVP embryos was performed on Day 7±1 after G1. At the time of transfer, heifers were randomly assigned, in a 2×2 factorial design, to receive GnRH (200µg of gonadorelin acetate) or remain as Control (untreated) at two different times: Day 7 (time of embryo transfer (ET)) or Day 21. Thus, the following groups were formed: Control-Control (n=410); Control-GnRH (n=409); GnRH-Control (n=419); and GnRH-GnRH (n=420). All heifers were evaluated by transrectal ultrasonography to determine number, size, and location of CL on Days 5, 14, 21, 28, 32, and 60 and pregnancy status on Days 28, 32, and 60. Data collected from each heifer included embryo stage and quality, oestrus expression, body condition score, number of transfers, and technician. Pregnancies per embryo transfer (P/ET) and pregnancy loss data were analysed by logistic regression (SAS 9.4). Ovulation to Day 7 treatment was greater (P<0.01) in GnRH treated heifers (70.0%; 585 out of 836) than in untreated Controls (2.7%; 22 of 819). Ovulation to Day 21 treatment was determined only in animals pregnant at Day 28 and was greater (P<0.01) in heifers treated with GnRH (37.6%; 153 of 407) than in untreated controls (1.0%; 4 of 390). There was no effect of Day 7 treatment (P>0.68) or Day 21 treatment (P>0.18), nor a Day 7×Day 21 treatment interaction (P>0.48) on P/ET at Day 32 or 60 (Table 1). Treatment with GnRH on Day 7 or 21 did not alter pregnancy loss between Days 32 and 60 (P>0.10). Heifers with an accessory CL present at Day 32 (11.7%; 41 of 350) had similar (P=0.55) pregnancy loss compared with heifers with no accessory CL (14.4%; 54 of 375). The number of CL present on Day 32 did not affect (P=0.23) pregnancy loss; however, heifers with 3 CL (4.9%; 3 of 61) tended (P<0.10) to have reduced pregnancy loss compared with heifers with 1 CL (14.4%; 54 of 375) or 2 CL (13.2%; 38 of 289). In conclusion, treatment with GnRH on Day 7 or 21 induced an accessory CL; however, it did not affect P/ET on Days 32 or 60 and pregnancy loss. Table 1.Pregnancies per embryo transfer (P/ET) and pregnancy loss in embryo recipients based on timing of treatment with gonadotrophin-releasing hormone (GnRH) Treatment P/ET Day 32% (n) P/ET Day 60% (n) Pregnancy loss% (n) Day 7 Day 21 Control Control 42.4 (410) 36.1 (410) 15.0 (174) GnRH 46.0 (409) 40.7 (409) 11.7 (188) GnRH Control 43.7 (419) 36.8 (419) 15.9 (183) P-value GnRH 43.1 (420) 38.8 (420) 10.0 (181) Day 7 treatment 0.68 0.72 0.96 Day 21 treatment 0.51 0.18 0.11 Interaction 0.48 0.66 0.71

2017 ◽  
Vol 29 (1) ◽  
pp. 160
Author(s):  
A. Tribulo ◽  
A. Cedeño ◽  
B. Bernal ◽  
S. Andrada ◽  
J. L. Barajas ◽  
...  

A retrospective analysis evaluated pregnancy rates and embryo losses with in vitro-produced embryos in a commercial embryo transfer program on 15 different beef farms. Recipients were beef cows and heifers (n = 1841) that were synchronized with 5 different protocols and transferred at a fixed-time (FTET). Recipients were examined by ultrasonography on Day 0, and those with a corpus luteum (CL) or a follicle ≥8 mm in diameter and with body condition score 2 to 4 (1 to 5 scale) were synchronized. The synchronization treatments were as follows. (T1) Recipients received an intravaginal device with 0.5 g of progesterone plus 2 mg of oestradiol benzoate on Day 0; device removal, plus 500 μg of cloprostenol (prostaglandin F2α), 400 IU of eCG, and 0.5 mg of oestradiol cypionate on Day 8; and FTET on Day 17. (T2) This treatment was similar to T1 but 1 mg of oestradiol cypionate was injected at device removal instead of 0.5 mg of oestradiol cypionate. (T3) This treatment was similar to T1 except that animals were tail-painted on Day 8 and observed on Day 10. Those with the tail-paint intact on Day 10 received 100 μg of gonadorelin (gonadotropin-releasing hormone) and all recipients were FTET on Day 17. (T4) Recipients received a progesterone device on Day 0; device removal, prostaglandin F2α, and eCG on Day 5; gonadotropin-releasing hormone on Day 8; and FTET on Day 15. (T5) Recipients received a progesterone device and 2 mg of oestradiol benzoate on Day 0; device removal, prostaglandin F2α, and eCG on Day 6; gonadotropin-releasing hormone on Day 9; and FTET on Day 16. On the day of FTET all recipients with CL ≥18 mm in diameter (G1), ≥16 and <18 mm in diameter (G2), and ≥14 mm and <16 mm in diameter (G3) received in vitro-produced fresh embryos. Pregnancy was diagnosed by ultrasonography at 30 and 60 days of gestation, and data were analysed by logistic regression. The overall proportion of recipients synchronized that were FTET was 80.8% (1487/1841), with a 30-day pregnancy rate to FTET (P/FTET) of 45.6% (678/1487) and the rate of 30- to 60-day embryo/fetal loses on the 528 recipients that were re-checked at 60 days was 12.8% (68/528). There were no significant differences in P/FTET among operators, animal category, time of the year, embryo stage, or body condition score; however, there was a significant effect of farm (P < 0.001) and CL diameter (P < 0.05), but no interaction between CL diameter and farm or treatment (P > 0.1). Recipients with G1 (443/953, 46%) and G2 (221/462, 47%) CL had higher pregnancy rates than those with G3 CL (23/71, 32%). There was a significant effect of synchronization treatment on the proportion of recipients transferred and on P/FTET (P < 0.01) that was highly influenced by farm (farm by treatment interaction P < 0.01). The proportions of recipients selected for embryo transfer were as follows: T1: 386/486, 79.4%; T2: 233/331, 70.3%; T3: 342/377, 90.7%; T4: 126/160, 78.7%; and T5: 400/487, 82.1%. The P/FTET were as follows: T1: 190/386, 49.2%; T2: 96/233, 41.2%; T3: 175/342, 51.1%; T4: 49/126, 38.8%; and T5: 168/400, 42.0%. Although 30- to 60-day embryo/fetal losses were not influenced by synchronization treatments, they were highly influenced by farm (P < 0.001) and ranged from 0 to 34.5%. In conclusion, P/FTET in a commercial program with beef in vitro-produced embryos was influenced by factors related to the recipient (CL diameter) and the environment (farm), whereas embryo/fetal losses were influenced by farm but not treatment or recipient factors.


2016 ◽  
Vol 28 (2) ◽  
pp. 185 ◽  
Author(s):  
A. Garcia-Guerra ◽  
R. V. Sala ◽  
G. M. Baez ◽  
M. Fosado ◽  
L. F. Melo ◽  
...  

The hypothesis was that GnRH on Day 5 of a synchronized cycle in embryo transfer recipients would increase progesterone (P4) concentrations, embryo size, and fertility. Holstein and cross-bred Holstein heifers (n = 1562) were synchronized using a modified 5-day CIDR Co-Synch as follows: Day –8 CIDR inserted; Day –3 CIDR removed; prostaglandin F2α treatment; Day –2 second prostaglandin F2α; Day 0 gonadotropin-releasing hormone (G1, 100 μg of gonadorelin acetate) to induce ovulation. On Day 5.5, heifers were assigned in a completely randomised design to 1 of 2 treatments: Control (untreated) or GnRH (200 μg of gonadorelin acetate). Transfer of fresh in vitro-produced embryos was performed between d 6 and 8 after G1. Data collected from each heifer included embryo stage and quality, body condition score, technician, interval from G1 to transfer, and number of previous transfers. All heifers were evaluated by transrectal ultrasonography on Day 5, 33, and 62 and a subset of heifers was scanned on Day 12 (n = 718; to determine ovulation to treatment) and another subset on Day 33 (n = 296; 16-s video to determine embryo and amniotic vesicle size). Serum P4 was determined from a subset of heifers on Day 12 (n = 467). Fertility data were analysed by logistic regression (LOGISTIC procedure, SAS 9.4), whereas continuous outcomes were analysed by ANOVA (MIXED procedure). Ovulation to Day 5.5 gonadotropin-releasing hormone was 83.9% (302/360) in GnRH-treated heifers v. 3.3% (12/358) in Control (P < 0.001). Progesterone on Day 12 was greater in GnRH-treated heifers 7.2 ± 0.1 ng mL–1 v. Controls 6.0 ± 0.1 ng mL–1 (P < 0.001). There was an effect of embryo stage at Day 33 and 60 of pregnancy, with Stage 7 having greater P/ET than Stage 6 embryos. Treatment with GnRH did not alter pregnancy per embryo transfer with either embryo stage but decreased pregnancy loss in Stage 7 embryos, as shown in Table 1. Embryo size measured as crown-rump length (CRL) did not differ, as shown in Table 1. Similarly, amniotic vesicle volume (AVV) was not different between GnRH (549.1 ± 16 mm3) and Control (543.5 ± 14 mm3; P = 0.86), nor was there an interaction between treatment and embryo stage (P = 0.71). In addition, neither AVV (P = 0.22) nor CRL (P = 0.41) were associated with pregnancy loss between Day 33 and 60. In conclusion, treatment with GnRH on Day 5 resulted in increased P4 and a reduction in pregnancy loss in heifers receiving a Stage 7 embryo without changing conceptus size. Table 1.Pregnancies per embryo transfer (P/ET), crown-rump length (CRL), and pregnancy loss in embryo recipients receiving gonadotropin-releasing hormone (GnRH) on Day 5.5 v. control


2020 ◽  
Vol 32 (2) ◽  
pp. 208
Author(s):  
A. Butler ◽  
H. Butler ◽  
G. Cesaroni ◽  
R. Alberio ◽  
S. Perez Wallace ◽  
...  

Oestrus expression between progesterone (P4) withdrawal and fixed-time AI (FTAI) has been shown to improve pregnancies per AI (P/AI) by 27% in beef cattle. As a result, cattle that do not express oestrus after P4 withdrawal present a challenge to the efficiency of FTAI. The present study was designed to test the hypothesis that administration of gonadotrophin-releasing hormone (GnRH) at the time of AI in heifers that do not express oestrus can improve fertility. Two-year-old Angus heifers (n=1032) with a condition score of 2.75 to 3.5 at two locations in Argentina were used. On Day 0, heifers received an intravaginal device containing 0.5g of P4 (Cronipres, Biogenesis) and 2mg of oestradiol benzoate intramuscularly (Bioestrogen, Biogenesis). On Day 8, devices were removed and heifers received 150µg of d-cloprostenol intramuscularly (Enzaprost, Biogenesis), 0.5mg of oestradiol cipionate intramuscularly (Croni-Cip, Biogenesis), and an oestrus detection patch (Estrotect, Rockway Inc.). On Day 10, AI was performed in all heifers 50 to 54h after device removal. Heifers were categorised based on oestrus expression, and those that did not express oestrus were randomised to receive 100µg of gonadorelin acetate intramuscularly (Gonasyn, Syntex; n=158) concurrent with AI or remain as untreated controls (n=151). Pregnancy was determined using ultrasonography at Days 38 and 111 after AI. Differences in fertility were evaluated using generalised linear mixed models, and the results are shown in Table 1. Oestrus expression between device removal and FTAI was 70.1% (723/1032). Pregnancies per AI at Days 38 and 111 were different between groups (P&lt;0.01). Heifers that expressed oestrus had greater P/AI than those that did not regardless of treatment (P&lt;0.01). However, in heifers that did not express oestrus, treatment with GnRH at the time of AI resulted in greater P/AI than in control heifers (P=0.004). Similarly, P/AI at Day 111 were greater in heifers that expressed oestrus than in those that did not regardless of treatment (P&lt;0.01). Heifers that did not express oestrus and were treated with GnRH had greater P/AI than those that did not express oestrus and remained as controls (P=0.02). Pregnancy loss between Days 38 and 111 was not different between groups; however, there was a tendency (P=0.06) for greater pregnancy loss in heifers that did not express oestrus and were treated with GnRH compared with heifers that expressed oestrus. In summary, treatment with GnRH at the time of AI is a suitable strategy to improve P/AI in heifers that fail to exhibit oestrus after an oestradiol-based synchronisation protocol. However, this strategy appears to increase pregnancy loss, and thus further research is needed to confirm this observation and the mechanisms underlying it. Table 1.Pregnancies per AI (P/AI) and pregnancy loss (%; no. in parentheses) in Angus heifers based on oestrus expression and treatment with GnRH Group P/AI Day 38 P/AI Day 111 Pregnancy loss Oestrus 68.9 (498/723)A 65.2 (471/723)A 5.4 (27/498) No oestrus control 29.8 (45/151)B 28.5 (43/151)B 4.4 (2/45) No oestrus + GnRH 45.6 (72/158)C 40.5 (64/158)C 11.1 (8/72) P-value &lt;0.001 &lt;0.001 0.15 A-CValues within a column with different superscripts differ significantly (P&lt;0.05).


2016 ◽  
Vol 28 (2) ◽  
pp. 183 ◽  
Author(s):  
L. C. Carrenho-Sala ◽  
R. V. Sala ◽  
M. Fosado ◽  
D. C. Pereira ◽  
S. Garcia ◽  
...  

A retrospective study was performed to evaluate factors that influence pregnancy per embryo transfer (P/ET) in an IVF-embryo transfer program. A total of 5026 fresh in vitro-produced embryos were transferred during 2014 and evaluated for effects of embryo quality, embryo stage, size of corpus luteum (CL; 18–19.9 mm or ≥20 mm), interval from GnRH to embryo transfer, number of previous embryo transfer (0, 1, 2, 3, ≥4); and interaction of embryo stage and interval from GnRH to embryo transfer. One group (n = 850) had detection of oestrus after prostaglandin F2α application but most heifers (n = 4176) received fixed timed embryo transfer after a 5-day CIDR-Synch protocol: Day –8 CIDR inserted; Day –3 CIDR removed and prostaglandin F2α; Day –2 prostaglandin F2α; Day 0 GnRH. Ultrasound was performed on Day 6 after GnRH or oestrus to measure CL size and on Day 32 and 60 to determine pregnancy. Data for P/ET were analysed by logistic regression (LOGISTIC procedure, SAS 9.4). Embryo quality influenced P/ET at Day 32 [Grade 1 48.4% (1273/2631) v. Grade 2 37.6% (900/2395); P < 0.01] and at Day 60 [Grade 1 38.9% (1023/2631) v. Grade 2 29.0% (694/2395); P < 0.01], and altered pregnancy loss [Grade 1 19.6% (250/1273) v. Grade 2 22.9% (206/900); P = 0.03]. Stage of the embryo also had an effect on P/ET at Day 32 [Stage 6 35.5%a (582/1641), Stage 7 46.3%b (1431/3092), and Stage 8 54.6%c (160/293); P < 0.01] and at Day 60 [Stage 6 28.2%a (462/1641), Stage 7 36.6%b (1131/3092), and Stage 8 41.6%b (122/293); P < 0.01], but did not affect pregnancy loss (P = 0.22). Interestingly, interval from GnRH (or oestrus) until embryo transfer did not affect P/ET at Day 32 (P = 0.10), 60 (P = 0.23), or pregnancy loss (P = 0.3), nor was there an interaction between interval and embryo stage at Day 32 (P = 0.77), 60 (P = 0.96) or pregnancy loss (P = 0.55). As shown in Table 1, embryo stage 6 was always the lowest and stage 8 always the greatest P/ET regardless of interval from GnRH to embryo transfer. Size of CL also did not affect P/ET at Day 32 (P = 0.09), 60 (P = 0.21), or pregnancy loss (P = 0.90). Number of previous embryo transfer also did not alter P/ET at Day 32 [0 = 43.3% (886/2046), 1 = 44.1% (639/1450), 2 = 43.4% (444/1024), 3 = 42.6% (146/343), and ≥4 = 35.6% (58/163); P = 0.33] or 60 (P = 0.51) or pregnancy loss (P = 0.12). In conclusion, embryo stage and quality are the major factors that impacted P/ET in this study, with surprisingly little effect of interval from GnRH to embryo transfer, size of the CL, and number of previous embryo transfer. Thus, recipient programs for IVF-embryo transfer can be designed with substantial flexibility. Table 1.Effect of embryo stage and recipient synchrony on pregnancies per embryo transfer on Day 32 in recipient dairy heifers


2010 ◽  
Vol 22 (1) ◽  
pp. 247
Author(s):  
J. R. V. Pimentel ◽  
P. H. P. Miguez ◽  
A. Kehrle ◽  
E. H. Madureira ◽  
F. A. Braga ◽  
...  

Progestagen-releasing devices are commonly used for fixed-time artificial insemination and fixed-time embryo transfer (FTET). The aim of this study was to evaluate the efficacy of a new progesterone-releasing intravaginal device made of poly(3-hydroxy)butyrate-valerate copolymer (PHB-V) and poly-ϵ-caprolactone (PCL; Progestar®, Innovare, São Paulo, Brazil) to synchronize crossbredcows and heifers for FTET by determining ovulation, conception, and pregnancy rates. A total of 205 recipients ranging in age from 2 to 8 years (heifers, n = 77; non-suckled cows, n = 69; suckled cows, n = 59) were used on 3 different farms during February, March, and April 2008. The experimental design was a generalized block design (cyclicity, body condition score, postpartum interval, and animal category, i.e. heifers, suckled, or non-suckled cows), with 2 different synchronization treatments for FTET (Progestar® v. Crestar®). Cows were assigned to either Crestar® (norgestomet ear implants; Intervet Schering-Plough, Kenilworth, NJ, USA; n = 103) or Progestar® (PHB-V; n = 102) treatments. All groups were treated with 2 mg of estradiol benzoate i.m. (Innovare) and received the intravaginal or auricular device according to their group (Day 0). Together with P4 device withdrawal (Day 8), all animals received 400 IU of eCG i.m. (Novormon®, Schering-Plough) and 500 μg i.m. of PGF2α, (Sincrocio®, Ouro Fino, Brasil). On Day 9, all cows were treated with 1 mg of estradiol benzoate (Innovare) and the innovulation was performed on Day 17. Embryos were maintained at 37°C during the innovulation period. The percentage of usable recipients (AR) based on the presence of a functional CL on the day of embryo transfer, conception rate (numbers of pregnant recipients of the total number transferred; CR), and pregnancy rate (numbers of pregnant recipients of the total number synchronized; PR) were determined by ultrasonography 23 days after embryo transfer. Data were submitted to ANOVA (PROC GLM, SAS Institute Inc., Cary, NC, USA) and significant differences were considered to have a P-value of <0.05. None of the interactions were significant and there was no effect of treatment on AR (72.8% v. 83.3%), CR (47.9% v. 42.7%), and PR (34.7% v. 35.0%) for Progestar® v. Crestar® groups, respectively. This experiment showed that Progestar® was as efficacious as Crestar® to synchronize crossbred cows and heifers for FTET. FAPESP-Fundação de Amparo à Pesquisa do Estado de São Paulo and CNPq-Conselho Nacional de Desenvolvimento Científico e Tecnológico.


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.


2014 ◽  
Vol 26 (1) ◽  
pp. 118 ◽  
Author(s):  
M. Re ◽  
J. J. de la Mata ◽  
G. A. Bo

Several studies, performed mainly in North America, have shown that the 5-day co-synch timed-AI (TAI) protocol results in similar or higher pregnancy rates than conventional 7-day co-synch protocols in beef cows and beef and dairy heifers. It is hypothesised that the 5-day co-synch protocol reduces the incidence of persistent follicles and provides for a longer proestrus, with increasing oestradiol concentrations due to continuous gonadotropin support for the dominant follicle (Bridges et al. 2008 Theriogenology 69, 843–851). Similar findings have been reported recently following the use of a shortened oestradiol-based protocol that also provides for a longer proestrus (named J-synch; de la Matta and Bo 2012 Taurus 55, 17–23). A study was designed to compare these 2 treatment protocols for synchronization of ovulation that allow for a prolonged proestrus with the conventional oestradiol-based protocol currently used for TAI in South America for heifers. Cycling Holstein heifers that were, on average, 17 months of age, weighting 350 to 380 kg, and with a body condition score of 2.5 to 3.5 (scale of 1 to 5), were randomly allocated to 1 of 3 treatment groups. Heifers in the 7-day estradiol benzoate (EB) group received a progesterone device (DIB 1 g of progesterone; Syntex SA, Buenos Aires, Argentina) and 2 mg EB (oestradiol benzoate, Syntex SA) on Day 0, 500 μg of cloprostenol (prostaglandin F2α; Sincronil, Proagro SA, Argentina) and DIB removal on Day 7, 1 mg of EB on Day 8, and TAI 30 h later (54 h after DIB removal). Heifers in the J-synch group received a DIB and 2 mg of EB on Day 0, PGF and DIB removal on Day 6, and 10 μg of buserelin acetate [gonadotropin-releasing hormone (GnRH); Receptal, Intervet, Millsboro, MD, USA] and TAI on Day 8 (72 h later). Heifers in the 5-day co-synch group received a DIB and GnRH on Day 0, DIB removal and prostaglandin F2α (twice, 12 h apart) on Day 5, and GnRH along with TAI on Day 8 (72 h later). Treatments were coordinated so that DIB were removed in all groups at the same time to decrease a possible time effect of follicle parameters and pregnancy rates. Two experiments were conducted; the first evaluated follicular dynamics and ovulation rates by twice daily ultrasonography (Honda 101 V, 5.0 MHz) in 20 heifers per group and the second determined pregnancy rates to TAI in 172 heifers. The mean (± standard error of the mean) interval from DIB removal to ovulation was shorter in the 7-day EB group (82.7 ± 3.1 h) than in the J-synch (103.8 ± 3.3 h) and 5-day co-synch (96.8 ± 3.3 h) groups (P = 0.01). However, the mean diameter of the dominant preovulatory follicle was smaller (P = 0.01) in the 7-day EB (12.8 ± 0.5 mm) and the J-synch (13.1 ± 0.6 mm) groups than in the 5-day co-synch group (14.9 ± 0.6 mm). Although pregnancy rates in the second experiment were numerically lower in the 7-day EB group (31/58; 53.4%) and 5-day co-synch group (30/55; 54.5%) than in the J-synch group (37/59; 62.7%), differences were not significant (P > 0.3). Preliminary results indicate that the 3 protocols evaluated result in comparable pregnancy rates to TAI in dairy heifers.


2020 ◽  
Vol 32 (2) ◽  
pp. 179
Author(s):  
R. Sala ◽  
L. Carrenho-Sala ◽  
V. Absalon-Medina ◽  
A. Lopez ◽  
M. Fosado ◽  
...  

Optimized fixed-time embryo transfer (FTET) protocols for synchronization of recipients have the potential to improve the overall efficiency and profitability of embryo transfer (ET) programs. The objective of the present study was to evaluate the effect of dose of gonadotrophin-releasing hormone (GnRH) at initiation of a 5-day synchronization protocol for FTET. Holstein heifers (n=2689) at two locations were synchronized using a 5-day CO-Synch protocol as follows: Day 0: CIDR inserted, Day 5: CIDR removed, prostaglandin (PG)F2α treatment (500μg cloprostenol), Day 6: PGF2α treatment, Day 8: GnRH (100μg of gonadorelin). On Day 0, at the time of CIDR insertion, heifers were assigned in a completely randomised design to the following groups: Single (a single dose of GnRH; 100μg of gonadorelin), Double (200μg of gonadorelin) or No GnRH (control). All heifers received an Estrotect patch placed on Day 5 and evaluated for signs of oestrus on Day 8. At location A, heifers were evaluated by ultrasonography 5 days after GnRH to determine presence and size of corpus luteum (CL), whereas at location B presence and location of CL were determined by transrectal palpation at the time of transfer. Heifers with a CL received an embryo 7±1 days after GnRH administration, and pregnancy was determined by ultrasonography 41 and 63 days after GnRH. Data were analysed by generalized linear mixed models. Oestrus expression was greater in heifers that received Single and Double GnRH than in the No GnRH group (P=0.001). Similarly, utilisation rate (number transferred per number treated) was greater for heifers in the Single and Double GnRH group than for those in the No GnRH group (P=0.02). Pregnancy data were analysed for a subset of recipients using data from Day 41 (n=2267) and Day 63 (n=2042). The analysis of fertility outcomes included as covariates the type of embryo (invitro fresh or frozen and invivo fresh or frozen), embryo stage, embryo quality, interval from GnRH to transfer, and oestrus expression. Pregnancies per embryo transfer (P/ET) at Days 41 and 63 were not different between treatment groups (P=0.86), and there was no interaction between type of embryo and treatment (P&gt;0.15). Pregnancy loss between Days 41 and 63 was not different (P=0.49) between treatments groups. In conclusion, the removal of the initial GnRH from a 5-day FTET protocol resulted in a slight but significant reduction in the utilisation rate and the percentage of heifers showing oestrus. However, there was no detrimental effect on fertility. As a result, the overall cost of the FTET program can be reduced by eliminating the need for the initial GnRH treatment without compromising fertility. Table 1.Reproductive performance in recipients receiving different doses of gonadotrophin-releasing hormone (GnRH) at initiation of the synchronization protocol Treatment Oestrus (n) Utilisation rate (n) P/ET1 D41 (n) P/ET D63 (n) Pregnancy loss (n) No GnRH 69.2%B (621/898) 85.0%B (763/898) 41.6% (308/740) 39.9% (268/672) 4.3% (12/280) Single GnRH 76.1%A (685/900) 88.8%A (799/900) 42.7% (329/770) 39.5% (272/689) 6.5% (19/291) Double GnRH 75.3%A (671/891) 88.7%A (790/891) 41.5% (314/757) 38.9% (265/681) 5.4% (15/280) A,BValues with different superscripts within a column differ (P&lt;0.05). 1P/ET=pregnancies per embryo transfer.


2020 ◽  
Vol 32 (2) ◽  
pp. 210
Author(s):  
R. C. Bonacker ◽  
K. R. Gray ◽  
C. A. Breiner ◽  
J. M. Anderson ◽  
D. J. Patterson ◽  
...  

An experiment was designed to evaluate the effectiveness of the recently developed 7 and 7 Synch protocol to synchronise oestrus and ovulation among recipients before embryo transfer (ET). Postpartum beef cows (n=1358) across 13 locations were assigned to either the 7-day CO-Synch + controlled internal drug release (CIDR) protocol or the 7 and 7 Synch protocol before oestrus detection and subsequent ET. Cows were pre-assigned to balanced treatments within location based on age and days postpartum, and body condition score was recorded at ET. Cows assigned to the 7-day CO-Synch + CIDR protocol were administered gonadotrophin-releasing hormone (GnRH; 100µg of gonadorelin acetate) on Day 0, an intravaginal CIDR insert (1.38g of progesterone) from Day 0 to 7, and prostaglandin F2α (PG; 25mg of dinoprost tromethamine) coincident with CIDR removal on Day 7. Cows assigned to the 7 and 7 Synch protocol were administered PG (25mg of dinoprost tromethamine) coincident with CIDR insertion on Day −7, GnRH (100µg of gonadorelin acetate) on Day 0, and PG (25mg of dinoprost tromethamine) coincident with CIDR removal on Day 7. The 7 and 7 Synch protocol was hypothesised to enhance response to GnRH administration on Day 0 among mixed groups of oestrous cycling and anestrous cows, ultimately resulting in improved oestrous response and synchrony of oestrus before ET. Cows were observed for visible signs of oestrus following oestrus synchronisation, with GnRH (100µg of gonadorelin acetate) administered to cows failing to express oestrus during the detection period. Length of the detection period varied among locations but was consistent across treatments within location. Embryo transfer was performed approximately 7 days after oestrus or GnRH administration. Presence of corpora lutea (CL) was determined via transrectal palpation by a single veterinarian blinded to treatment, and embryos were transferred only to cows with palpable CL. Embryo transfer was performed using fresh or frozen embryos staged and graded according to IETS recommended guidelines, with embryo information recorded for each recipient. Data were analysed using the PROC GLIMMIX procedure of SAS (SAS Institute Inc.). Proportion of cows expressing oestrus was improved (P&lt;0.0001) among cows assigned to the 7 and 7 Synch protocol (86% (529/615) vs. 76% (488/640)). The proportion of recipient females with palpable CL tended to be greater (P=0.07) among cows assigned to the 7 and 7 Synch protocol (82% (478/586) vs. 80% (451/565)). Among cows that expressed oestrus, a greater proportion (P&lt;0.01) had palpable CL following the 7 and 7 Synch protocol (88% (466/529) vs. 86% (418/488)). Consequently, irrespective of embryo type received, the proportion of recipients pregnant to ET was greater (P&lt;0.01) following the 7 and 7 Synch protocol (39%; 263/668) compared with the 7-day CO-Synch + CIDR protocol (33%; 228/690). In summary, the 7 and 7 Synch protocol improved the likelihood of oestrus expression in recipient cows, increased the proportion eligible to receive an embryo, and resulted in greater pregnancy rate to ET.


2020 ◽  
Vol 32 (2) ◽  
pp. 178
Author(s):  
D. Pereira ◽  
D. Moreno ◽  
R. Sala ◽  
L. Carrenho-Sala ◽  
M. Fosado ◽  
...  

Time elapsed between removal from culture and embryo transfer (ET) can have a profound effect on the success of an invitro-produced (IVP) ET program. The embryo culture medium provides the necessary nutrients for embryo development and the use of media with a different nutrient composition to load embryos into straws could negatively affect embryo viability. The objective of the present study was to evaluate the effect of type of media used for embryo loading on pregnancy establishment and maintenance. Holstein heifers (n=800) were synchronized using a modified 5-day CO-Synch + controlled internal drug release (CIDR) as follows: Day −8: CIDR inserted, Day −3: CIDR removed, prostaglandin F2α treatment (500μg cloprostenol sodium), Day 0: gonadotrophin-releasing hormone (GnRH; 100μg of gonadorelin acetate). Five days after GnRH, heifers were evaluated by ultrasonography to determine presence of a corpus luteum (CL). Embryos were removed from culture on Day 7 (Day 0=fertilization), placed into tubes containing SOFaaci, and transported in an incubator (LabMix, WTA) to the transfer facility within 1.5h. Upon arrival embryos were removed from transport tubes and randomly assigned to be loaded into 0.25-mL straws containing either holding media (Vigro Holding Plus) or SOFaaci-HEPES. After loading into straws, embryos were placed in an ET gun and AI gun warmers set at 35°C until transfer by 1 of 5 technicians. Heifers with a CL were randomised for transfer of a fresh IVP embryo loaded into a straw containing either holding media or SOFaaci-Hepes on Day 7±1. Interval from embryo loading to transfer ranged from 1 to 3h. Pregnancy was determined by ultrasonography on Days 32 and 60. Data were analysed by logistic regression and included the fixed effects of loading media, embryo stage, embryo quality, interval between GnRH and ET, and biologically relevant interactions. Pregnancies per ET (P/ET) on Day 32 were not different between the groups in which embryos were loaded using holding media and those which used SOFaaci-Hepes, nor there were interactions between loading medium and embryo stage, embryo quality, or interval from GnRH to ET (P&gt;0.10; Table 1). Pregnancies per ET (P/ET) on Day 60 were not different between the loading media groups, nor were there interactions between loading medium, embryo stage, and embryo quality, or interval from GnRH to ET (P&gt;0.10). Pregnancy loss between Days 32 and 60 was not different between groups, nor there were interactions between loading media groups and any other factor (P&gt;0.10). In conclusion, the use of either holding medium or SOFaaci-HEPES for fresh IVP embryo loading does not affect fertility; thus, both are suitable alternatives for loading of embryos into transfer straws. Table 1.Pregnancies per embryo transfer (P/ET) and pregnancy loss in recipient heifers transferred with fresh invitro-produced embryos, using either holding medium or SOFaaci-HEPES medium for loading Item P/ET Day 32 (n) P/ET Day 60 (n) Pregnancy loss (n) Loading medium Holding 47.0% (186/396) 41.3% (163/395) 11.9% (22/185) SOFaaci-HEPES 48.8% (197/404) 43.1% (174/404) 11.7% (23/197) P-value 0.77 0.22 0.84


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