101 DOSE AND TIMING OF ADMINISTRATION OF PROSTAGLANDIN F2α DURING FIXED-TIME EMBRYO TRANSFER IN AN IN VITRO-PRODUCTION PROGRAM

2017 ◽  
Vol 29 (1) ◽  
pp. 158
Author(s):  
R. V. Sala ◽  
A. Garcia-Guerra ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
E. Peralta ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Prostaglandin F2α ◽  
Fixed Time ◽  
High Fertility ◽  
Full Dose ◽  
Half Dose ◽  
Utilisation Rate

Synchronization protocols for fixed-time embryo transfer (ET) contribute significantly to the overall cost of an in vitro-produced-ET program, primarily through the cost of drugs and the labour required. Optimization of synchronization protocols to reduce cost, while providing high fertility, have the potential to improve overall efficiency and profitability. The objective of the present study was to evaluate the effect of dose and schedule of administration of prostaglandin F2α (PGF) during a synchronization protocol for fixed-time ET. Holstein and cross-bred Holstein heifers (n = 3766) were synchronized using a modified 5-day CIDR Synch as follows: Day 0: CIDR inserted; Day 5: CIDR removed, PGF2α treatment; Day 8: gonadotropin-releasing hormone (GnRH; 100 μg of gonadorelin). On Day 5, at the time of CIDR removal, heifers were randomly assigned to a 2 × 2 factorial design to receive either a full or half dose of PGF (Cloprostenol; 500 v. 250 μg) and 1 (Day 5) or 2 (Day 6) administrations resulting in the following treatments: full dose Day 5 + Day 6 (n = 938); full dose Day 5 (n = 938); half dose Day 5 + Day 6 (n = 946); and half dose Day 5 (n = 944). Heifers were evaluated by ultrasonography 5 days after GnRH to determine presence and size of the corpus luteum. Heifers with a corpus luteum received a fresh in vitro-produced embryo 7 ± 1 days after GnRH administration, and pregnancy was determined by ultrasonography 32 and 60 days after GnRH. Fertility data were analysed by logistic regression and included the fixed effects of dose, time, and their interaction. Fertility results are shown in Table 1. Utilisation rate (transferred/treated) was not affected by dose (P = 0.66), time (P = 0.19), or their interaction (P = 0.17). The percentage of heifers detected in oestrus was not affected by dose (P = 0.13), time (P = 0.72), or their interaction (P = 0.89). There were no significant differences between doses of PGF (P = 0.32), time (P = 0.71), or their interaction (P = 0.80) on pregnancies per ET on Day 32. Similarly, no differences were found on pregnancies per ET on Day 60 between doses (P = 0.35), time (P = 0.96), or their interaction (P = 0.89). In addition, pregnancy loss between Day 32 and 60 was not affected by dose (P = 0.76), time (P = 0.66), or their interaction (P = 0.54). In conclusion, the use of a half dose of PGF once on Day 5 resulted in comparable utilisation rate and fertility as the observed with 2 full dose applications 24 h apart. As a result, the overall cost of the fixed-time ET program can be reduced by eliminating the need for a second PGF treatment and by decreasing the dose without compromising fertility. Table 1. Utilisation rate, oestrus expression, pregnancies per ET (P/ET), and pregnancy loss in recipients receiving either a full or half dose of prostaglandin F2α on Days (D) 5 and 6 or once on D 5

108 FACTORS AFFECTING PREGNANCY RATES IN RECIPIENTS RECEIVING IN VITRO-PRODUCED EMBRYOS BY FIXED TIME EMBRYO TRANSFER

2016 ◽  
Vol 28 (2) ◽  
pp. 184
Author(s):  
M. Pelizzari ◽  
A. Tribulo ◽  
J. Garzon ◽  
B. Bernal ◽  
R. Tribulo ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Embryo Transfer ◽  
Body Condition Score ◽  
Prostaglandin F2α ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Uterine Horn ◽  
Device Removal ◽  
Embryo Transfer Technique

A retrospective analysis of factors that affect pregnancy rates from 4214 fresh in vitro-produced (IVP) embryos that were transferred at a fixed-time (FTET) in 20 different farms. Recipients were all cycling cows or heifers that were synchronized with 1 of 3 treatments: 1) treatments with progesterone (P4) devices and 2 mg of oestradiol benzoate (EB) on Day 0 (day of insertion) and 24 h after device removal (Day 8); 2) treatments with P4 devices and EB on Day 0, but with 0.5 mg of oestradiol cypionate (ECP) at device removal (Day 8); or 3) treatments with P4 devices and GnRH on Day 0 and a second GnRH 60 h after device removal (Day 5). Cows in all treatment groups also received 500 µg of cloprostenol (prostaglandin F2α) at the time of P4 device removal and 400 IU of eCG either at device removal or 3 days before device removal. All embryos were transferred 7 or 8 days after the expected time of oestrus (24 h after EB, 48 h after ECP or at the time of the second GNRH for each synchronization treatment, respectively). On the day of embryo transfer, recipients were examined by ultrasonography and those with corpus luteum >14 mm in diameter received a fresh, IVP embryo in the uterine horn ipsilateral to the corpus luteum. Pregnancy rates were determined by ultrasonography 35 days after FTET. Data were analysed by logistic regression. Independent variables were classified into the following three categories. 1) Factors related to the recipient and the environment; there were no significant differences in pregnancy rates for corpus luteum diameter (≥14 and <16 mm, ≥16 and <18 mm, or ≥18 mm; P = 0.46), number of corpus luteum (1 or ≥2; P = 0.26), and category of recipient (cow or heifer; P = 0.21). However, there were significant effects of farm (P = 0.01) and body condition score (BCS; P = 0.01). Cows with BCS ≥4.5 (1 to 5 scale) resulted in lower pregnancy rates (4/20, 20.0%) than those with BCS 2 (74/225, 32.9%), 2.5 (502/1434, 35.0%), 3 (570/1467, 38.9%), 3.5 (193/532, 36.3%), and 4 (44/118, 37.3%). 2) Factors related to the synchronization treatment; there were no significant differences between recipients receiving eCG at device removal (84/209, 40.2%) or 3 days before device removal (874/2291, 38.1%; P = 0.35). However, recipients synchronized with P4 devices and ECP had higher (P = 0.01) pregnancy rates (232/483, 48.0%) than those treated with EB (679/1888, 36.0%) or gonadotropin-releasing hormone (47/129, 36.4%). 3) Factors related to the embryo transfer technique; day of the recipient’s oestrous cycle (P = 0.36), stage of embryo transferred (IETS stages 6 or 7; P = 0.62), and operator (P = 0.57) did not affect pregnancy rates. However transfers made in the anterior third of the uterine horn resulted in higher (649/1545, 42.0%) pregnancy rates than those in the mid-third (845/2511, 33.6%) or in the distal third (6/35, 17.1%; P = 0.01). It was concluded that factors related to the recipient and the environment (farm and BCS), the synchronization treatment (ECP), and the embryo transfer technique (site of deposition) affect pregnancy rates in recipients of embryos produced in vitro and transferred at a fixed time.

107 FACTORS THAT INFLUENCE FERTILITY IN AN IVF EMBRYO TRANSFER PROGRAM IN DAIRY HEIFERS

2016 ◽  
Vol 28 (2) ◽  
pp. 183 ◽  
Author(s):  
L. C. Carrenho-Sala ◽  
R. V. Sala ◽  
M. Fosado ◽  
D. C. Pereira ◽  
S. Garcia ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Embryo Quality ◽  
Prostaglandin F2α ◽  
Transfer Program ◽  
Dairy Heifers ◽  
Embryo Transfer Program ◽  
Embryo Stage ◽  
Major Factors

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

105 Optimization of a five-day fixed-time embryo transfer program in dairy heifers: Use of gonadotrophin-releasing hormone at initiation of the protocol

2020 ◽  
Vol 32 (2) ◽  
pp. 179
Author(s):  
R. Sala ◽  
L. Carrenho-Sala ◽  
V. Absalon-Medina ◽  
A. Lopez ◽  
M. Fosado ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Fixed Time ◽  
Transfer Program ◽  
Releasing Hormone ◽  
Utilisation Rate ◽  
Stage Embryo ◽  
Gonadotrophin Releasing Hormone ◽  
Using Data ◽  
Synchronization Protocol

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.

104 FACTORS AFFECTING PREGNANCY RATES AND EMBRYO/FETAL LOSSES IN RECIPIENTS RECEIVING IN VITRO-PRODUCED EMBRYOS BY FIXED-TIME EMBRYO TRANSFER

2017 ◽  
Vol 29 (1) ◽  
pp. 160
Author(s):  
A. Tribulo ◽  
A. Cedeño ◽  
B. Bernal ◽  
S. Andrada ◽  
J. L. Barajas ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Body Condition Score ◽  
Prostaglandin F2α ◽  
Fixed Time ◽  
Gonadotropin Releasing Hormone ◽  
Pregnancy Rates ◽  
Device Removal ◽  
Releasing Hormone ◽  
Condition Score

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.

102 UNILATERAL AND BILATERAL TRANSFER OF 2IN VITRO-PRODUCED EMBRYOS INCREASES PREGNANCY LOSS BETWEEN 30 AND 60 DAYS

2017 ◽  
Vol 29 (1) ◽  
pp. 159
Author(s):  
L. C. Carrenho-Sala ◽  
A. Garcia-Guerra ◽  
R. V. Sala ◽  
M. Fosado ◽  
D. C. Pereira ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Pregnancy Loss ◽  
Prostaglandin F2α ◽  
Poor Quality ◽  
Single Group ◽  
Treatment Groups ◽  
Bilateral Group ◽  
Conception Rates ◽  
Twin Pregnancies

Fertility of in vitro-produced embryos is affected by embryo stage and quality. Embryos quality 1 and stage 7 result in higher fertility than embryos of earlier stages and/or lower quality. The objective was to evaluate the effect of unilateral and bilateral transfer of 2 in vitro-produced embryos of earlier stages and/or poor quality on fertility. Heifers were synchronized using a 5-day CIDR Synch or 2 prostaglandin F2α injections 14 days apart followed by oestrus detection. Embryo transfer was performed 7 ± 1 day after gonadotropin-releasing hormone/oestrus and heifers were assigned randomly to 1 of 3 groups: single embryo ipsilateral to the corpus luteum (single; n = 188); 2 embryos in the uterine horn ipsilateral to the corpus luteum (unilateral; n = 138); 2 embryos bilaterally (bilateral; n = 128). Embryos stage 4 to 8 and quality 1 or 2 were randomly assigned to treatment groups. All embryos were 7-day fresh in vitro-produced embryos, and pregnancy diagnosis was performed by ultrasonography on Days 32 and 60. Data were analysed by logistic regression. Conception rates on Days 32 and 60 were not different (P > 0.10) between heifers receiving a single embryo [Day 32 = 30.9% (58/188) and Day 60 = 25% (47/188)] or those receiving 2 embryos [Day 32 = 36.5% (97/266) and Day 60 = 22.2% (59/266)]. However, pregnancy loss between Days 32 and 60 was greater (P < 0.01) in heifers with 2 embryos (39.2%; 38/97) than in those with a single embryo (18.9%; 11/58). Conception rate on Day 32 was not different between groups (P = 0.4) and was 30.9% (58/188) for single, 36.9% (51/138) for unilateral, and 35.9% (46/128) for bilateral. Similarly, there was no difference (P = 0.8) in conception rates on Day 60, single = 25% (47/188), unilateral = 23.9% (33/138), and bilateral= 20.3% (26/128). However, there was an effect of group on pregnancy loss between Days 32 and 60 (P = 0.04). Losses were higher (P = 0.01) in the bilateral group [43.5% (20/46)] compared with the single group [18.9% (11/58)], and the unilateral group was intermediate [35.3% (18/51)] and tended to be different from the single group (P = 0.1). Interestingly, when pregnancy loss was compared between heifers with twin or single pregnancies, as determined by ultrasonography, and regardless of the number of embryos transferred or their location, twin pregnancies had a greater pregnancy loss [62.1% (18/29)] compared with single pregnancies [24.6% (31/126); P < 0.01]. For twin bearing heifers, as determined by ultrasonography on Day 32, pregnancy loss did not differ between unilateral (62.5%; 10/16) and bilateral (61.5%; 8/13) transfers (P = 0.9). Similarly, there was no difference (P = 0.2) for heifers with single embryo pregnancies: single (18.9%; 11/58), unilateral (22.9%; 8/35), bilateral (36.4%; 12/33), although bilateral transfer of 2 embryos tended to be higher than single (P = 0.07). Transfer of 2 low quality in vitro-produced embryos results in similar conception rates, although pregnancy losses are greater. Interestingly, only 30% (29/97) of the pregnancies from heifers that received 2 embryos contained twins, indicating the loss of one of the embryos before Day 32. Furthermore, the increased losses observed with the transfer of 2 embryos were attributed to those heifers in which twin pregnancies were diagnosed on Day 32 regardless of distribution.

110 TREATMENT WITH GnRH ON DAY 5 REDUCES PREGNANCY LOSS IN HEIFERS RECEIVING IN VITRO-PRODUCED EXPANDED BLASTOCYSTS

2016 ◽  
Vol 28 (2) ◽  
pp. 185 ◽  
Author(s):  
A. Garcia-Guerra ◽  
R. V. Sala ◽  
G. M. Baez ◽  
M. Fosado ◽  
L. F. Melo ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Body Condition Score ◽  
Prostaglandin F2α ◽  
Gonadotropin Releasing Hormone ◽  
Crown Rump Length ◽  
Releasing Hormone ◽  
Embryo Size ◽  
Embryo Stage

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

scholarly journals Location relative to the corpus luteum affects bovine endometrial response to a conceptus

Reproduction ◽  
2020 ◽  
Vol 159 (5) ◽  
pp. 643-657
Author(s):  
Sandra Bagés-Arnal ◽  
José M Sánchez ◽  
Beatriz Fernandez-Fuertes ◽  
Michael McDonald ◽  
Susanta K Behura ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Uterine Horn ◽  
Biological Processes ◽  
Gene Ontology Analysis ◽  
Transcriptomic Response ◽  
False Discovery ◽  
Uterine Flushing

In cattle, embryo transfer into the uterine horn contralateral to the corpus luteum results in a higher incidence of pregnancy loss compared to transfer into the ipsilateral horn. We have previously reported temporal changes in the endometrial transcriptome during the estrous cycle which differ between uterine horns. The objective of this study was to compare the transcriptomic response of endometrium from the ipsilateral and contralateral horns to an elongating conceptus. Cross-bred beef heifers (n = 16) were synchronized and either used to generate day 14 conceptuses following the transfer of in vitro-produced blastocysts or to obtain day 14 endometrial explants. Conceptuses were recovered on day 14 by post-mortem uterine flushing, placed individually on top of explants collected from the ipsilateral (IPSI-D14) or the contralateral (CONTRA-D14) uterine horn of cyclic heifers, and co-cultured for 6 h. The response to a conceptus was markedly different between uterine horns, with 61 and 239 differentially expressed genes (DEGs; false discovery rate <0.05) in the ipsilateral and contralateral horns, respectively, compared to their controls. Direct comparison between IPSI-D1 and CONTRA-D14 revealed 32 DEGs, including CXCL11, CXCL10, IFIT2, RSAD2 and SAMD9. Gene Ontology analysis of these 32 genes revealed ten enriched biological processes, mainly related to immune response and response to an external stimulus. These data indicate that the endometrial response to the presence of a conceptus varies between uterine horns in the same uterus and may contribute to the higher incidence of pregnancy loss following embryo transfer to the contralateral horn.

scholarly journals 200 Improving efficiency of embryo transfer (ET) programs by optimizing fertility and management of recipients

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 116-117
Author(s):  
Milo Wiltbank ◽  
Alvaro Garcia-Guerra ◽  
Rodrigo V Sala ◽  
Meliton Fosada ◽  
Luciana Carrenho-Sala ◽  
...  
Keyword(s):  
Retrospective Analysis ◽  
Embryo Transfer ◽  
Pregnancy Loss ◽  
Fixed Time ◽  
Embryo Stage ◽  
Potential Recipient ◽  
Better Than

Abstract Multiple experiments were performed to optimize efficiency and fertility in recipients of fresh in vitro produced (IVP) embryos. In experiment 1, heifers (n = 520) were synchronized and received an embryo on d 6–8 after estrus or 6–8 d after GnRH in a fixed time ET (FTET) program using modified 5-d CIDR-Synch protocol (d-8: CIDR inserted; d-3: CIDR removed and PGF2α treatment; d-2: second PGF2α; d0: GnRH to induce ovulation). Pregnancy per ET (P/ET) at d 32 and d 60 were similar but pregnancy per treated potential recipient (efficiency of recipient utilization) was greater for FTET than estrus (+49.7%). Subsequent experiments sought to optimize FTET protocol by analyzing whether CIDRs could be used multiple times (up to 4 uses similar P/ET) and whether multiple PGF2α treatments were needed at end of program (no difference when no GnRH given at start of protocol). Thus, a simple, inexpensive FTET program has similar fertility as ET after estrus but is more efficient at recipient utilization. A large retrospective analysis (n = 12,569 ET) was performed using FTET program. Embryo stage and quality were major embryo factors impacting P/ET. Transfer of d 7 fresh embryo to d 7 or d 8 recipient was better than d 6 (+24.4%). Two experiments (GnRH or CIDR treatment) evaluated increasing circulating P4. In GnRH experiment, heifers (n = 1,562) on d5 received GnRH (200 μg) or Control (untreated). On D12, P4 was greater (P < 0.001) in GnRH-treated (7.2 ± 0.1ng/ml) vs Controls (6.0 ± 0.1ng/ml). There was greater P/ET at D33 and D60 of pregnancy for Stage 7 than 6 embryos. Treatment with GnRH did not alter P/ET but decreased pregnancy loss between D33 and D60 in heifers receiving Stage 7 embryos (11.6 vs 27.6% in recipients with accessory CL on d 33). In CIDR experiment, treatment with 2 CIDRs (from d 13; one new CIDR each 7 d) elevated circulating P4 and tended to decrease pregnancy loss (d 27 to 62; 25.9 vs 11.9%) and increase P/ET (40.4 vs 53.6% on d 62). Thus, elevating P4 decreased pregnancy loss during FTET program using fresh IVP embryos.

scholarly journals Prostaglandin F2α Represses IGF-I-Stimulated IRS1/Phosphatidylinositol-3-Kinase/AKT Signaling in the Corpus Luteum: Role of ERK and P70 Ribosomal S6 Kinase

2010 ◽  
Vol 24 (3) ◽  
pp. 632-643 ◽  
Author(s):  
Edward Arvisais ◽  
Xiaoying Hou ◽  
Todd A. Wyatt ◽  
Koumei Shirasuna ◽  
Heinrich Bollwein ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Prostaglandin F2α ◽  
Serine Phosphorylation ◽  
Phosphatidylinositol 3 Kinase ◽  
Akt Activation ◽  
Diminished Capacity ◽  
Igf I ◽  
In Vitro Treatment

Abstract Little is known about the early intracellular events that contribute to corpus luteum regression. Experiments were designed to determine the effects of prostaglandin F2α (PGF2α) on phosphatidylinositol-3-kinase (PI3K)/Akt signaling in the corpus luteum in vivo and in vitro. Treatment of midluteal-phase cows with a luteolytic dose of PGF2α resulted in a rapid increase in ERK and mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (p70S6K1) signaling and a rapid suppression of Akt phosphorylation in luteal tissue. In vitro treatment of primary cultures of luteal cells with PGF2α also resulted in an increase in ERK and mTOR/p70S6K1 signaling and a diminished capacity of IGF-I to stimulate PI3K, Akt, and protein kinase C ζ activation. Accounting for the reductions in PI3K and Akt activation observed in response to PGF2α treatment, we found that PGF2α promoted the phosphorylation of serine residues (307, 612, 636) in the insulin receptor substrate 1 (IRS1) peptide sequence in vivo and in vitro. Serine phosphorylation of IRS1 was associated with reduced formation of IGF-I-stimulated IRS1/PI3Kp85 complexes. Furthermore, treatment with inhibitors of the MAPK kinase 1/ERK or mTOR/p70S6K1 signaling pathways prevented PGF2α-induced serine phosphorylation of IRS1 and abrogated the inhibitory actions of PGF2α on Akt activation. Taken together, these experiments provide compelling evidence that PGF2α treatment stimulates IRS1 serine phosphorylation, which may contribute to a diminished capacity to respond to IGF-I. It seems likely that the rapid changes in phosphorylation events are among the early events that mediate PGF2α-induced corpus luteum regression.

Sign in / Sign up

Export Citation Format

Share Document