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.

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

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

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

scholarly journals “Which Factors Affect Pregnancy Until Calving and Pregnancy Loss in Buffalo Recipients of in vitro Produced Embryos?”

2020 ◽  
Vol 7 ◽  
Author(s):  
Wilson Pardini Saliba ◽  
Lindsay Unno Gimenes ◽  
Roberti Martins Drumond ◽  
Henrique Xavier Salgado Bayão ◽  
Rossella Di Palo ◽  
...  
Keyword(s):  
Breeding Season ◽  
Pregnancy Loss ◽  
Fixed Time ◽  
Factors Associated ◽  
Nonbreeding Season ◽  
Two Factors ◽  
Embryo Stage ◽  
Calving Rate ◽  
Synchronization Protocol

In vitro embryo production and embryo transfer (ET) in buffaloes has been developed for decades. However, most studies are focused on the donor or laboratory improvements, and there is a lack of reports regarding the recipients. Therefore, our aim was to investigate factors associated to pregnancy (P/ET), pregnancy loss (PL), and calving rates in buffalo recipients. The studied factors were season, recipient parity, the synchronization protocol, the CL diameter, asynchrony between the embryo and the recipient, the day of the recipient estrous cycle, the embryo (fresh vs. vitrified), the day of embryo development, and the embryo stage. These retrospective data, from a program of in vitro produced embryos, were analyzed by logistic regression, and the odds ratio was also estimated. Two factors were related to P/ET and the calving rate: (1) progesterone associated to estradiol plus eCG protocol for fixed time ET tended to affect positively P/ET on day 30 (41.9 vs. 36.1%, respectively; P = 0.07; AOR = 1.28) and P/ET on day 60 (37.8 vs. 36.1%, respectively; P = 0.09; AOR = 1.08) compared to the Ovsynch protocol; and (2) the CL diameter (≥14.5 mm) at transfer increased P/ET on day 30 (47.4 vs. 32.5%; P &lt; 0.01; AOR = 1.87) and on day 60 (45.3 vs. 27.7%; P &lt; 0.01; AOR = 2.16), and also the calving rate (37.9 vs. 21.7%; P &lt; 0.01; AOR = 2.20). PL was greater when ET was done in the nonbreeding season compared to the breeding season (PL 30–60: 12.8 vs. 0.0%, P = 0.01; AOR &gt; 999.99; PL 60-calving: 26.8 vs. 3.6%, P = 0.03; AOR = 9.90; and PL 30-calving: 36.2 vs. 3.6%, P = 0.01; AOR = 15.30). In conclusion, the data of our study indicated that the synchronization protocol, the CL diameter, and ET during the breeding season impacted the reproductive efficiency of buffalo recipients.

109 COMPARISON OF METHODS FOR SYNCHRONIZING RECIPIENTS OF IN VITRO PRODUCED EMBRYOS

2016 ◽  
Vol 28 (2) ◽  
pp. 185 ◽  
Author(s):  
R. V. Sala ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
L. C. C. Tosta ◽  
R. D. Tosta ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Prostaglandin F2α ◽  
Gonadotropin Releasing Hormone ◽  
Comparison Of Methods ◽  
Releasing Hormone ◽  
Embryo Stage ◽  
Utilisation Efficiency ◽  
Potential Recipient ◽  
Second Use

The present study compared fertility, as pregnancy per embryo transfer (P/ET), and efficiency of recipient utilisation, as pregnancy per treated potential recipient (P/TX), in heifers receiving in vitro-produced embryos using synchronized oestrus after prostaglandin F2α (OESTRUS) or synchronized ovulation and fixed timed embryo transfer (FTET) with new or reused CIDR. In Expt. 1, heifers (n = 520) were randomly assigned to 1 of 3 groups: OESTRUS, FTET with new CIDR, or FTET with second-use CIDR (previously used for 5 days). Heifers in OESTRUS group (n = 166) were synchronized with two prostaglandin F2α 14 days apart and detection of oestrus performed using tail chalk during 5 days after the second prostaglandin F2α. Heifers in FTET were synchronized with a new CIDR (n = 178) or second-use CIDR (n = 176) using a modified 5-day CIDR-Synch; Day –8: CIDR inserted; Day –3: CIDR removed, prostaglandin F2α; Day –2: second prostaglandin F2α; Day 0: gonadotropin-releasing hormone to induce ovulation. In Expt. 2, heifers (n = 422) were randomly assigned to 1 of 2 groups: FTET with new CIDR or FTET with third-use CIDR (previously used twice for 5 days each time) using the FTET protocol described for Expt. 1. Fresh in vitro-produced embryos were transferred between 6 and 8 days after OESTRUS or gonadotropin-releasing hormone (FTET). All heifers were evaluated by transrectal ultrasonography on Day 32 and 60 for pregnancy detection. Measurements of P/ET and P/TX for both experiments were analysed by logistic regression (LOGISTIC procedure, SAS 9.4) using biologically meaningful covariates such as embryo stage and quality, interval from oestrus or GnRH to transfer, and technician in the statistical analyses. In Expt. 1, two preplanned contrasts were performed to compare differences between OESTRUS v. FTET, and between FTET with new v. second-use CIDR. The P/ET at Day 32 was similar (P = 0.50) with 41.3% (45/109) for OESTRUS and 43.4% (134/309) for FTET groups. Similarly, P/ET on Day 60 was 30.3% (33/109) for OESTRUS and 32.4% (100/309) for FTET groups (P = 0.37). However, P/TX heifer on Day 60 was greater (P = 0.04) in the FTET (28.2%; 100/354) compared to OESTRUS (19.9%; 33/166). This difference is attributed to a greater (P < 0.001) utilisation efficiency (transferred/treated) of heifers in FTET (87.3%) v. OESTRUS (65.6%). In the second contrast, P/ET on Day 32 were similar (P = 0.87) for FTET heifers synchronized with a new CIDR (43.9%, 69/157) v. second-use CIDR (42.8%, 65/152). In addition, P/TX heifer on Day 60 was also similar (P = 0.52) for heifers receiving a new or second-use CIDR (29.8%, 53/178 v. 26.7% 47/176). In Expt. 2, P/ET on Day 32 was similar (P = 0.73) for FTET with a new CIDR (41.0%, 77/188) or third-use CIDR (42.3%, 83/196). The P/TX heifer on Day 60 was also not different (P = 0.58) for new CIDR (28.6%, 61/213) v. third-use CIDR (31.1%, 65/209). Thus, use of FTET with new or used CIDR can produce similar P/ET and greater efficiency of recipient utilisation compared to OESTRUS.

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.

94 EFFECTS OF 6- OR 12-HOUR CULTURE IN A Micro Q STRAW INCUBATOR ON DEVELOPMENT OF IN VITRO-PRODUCED BOVINE EMBRYOS

2016 ◽  
Vol 28 (2) ◽  
pp. 177
Author(s):  
C. R. Looney ◽  
J. H. Pryor ◽  
M. Snyder ◽  
A. Ilercil ◽  
C. R. Long
Keyword(s):  
Embryo Transfer ◽  
Uv Light ◽  
Tween 20 ◽  
Treatment Groups ◽  
Cell Counts ◽  
Stage 4 ◽  
Embryo Stage ◽  
Transfer Services ◽  
In Transit

Transporting in vitro-produced (IVP) embryos can be challenging when an embryo transfer destination is more than 6 h away or electricity is not available on site to unload embryos for transfer. The objective of this study was to determine if development rates would be compromised for Day 6.5 IVP embryos when placed in warmed Vigro holding medium (Vetoquinol, Pullman, WA, USA) loaded and plugged into 1/4 cc straws (Professional Embryo Transfer Services, Canton, TX, USA) for a period of either 6 or 12 h in a 38.5°C Micro Q straw block incubator (Micro Q iQ1T 64). Bovine oocytes were shipped and matured in transit from a commercial abattoir (DeSoto Biosciences, Seymour, TN, USA), fertilized (IVF = Day 0) with frozen-thawed semen, and cultured in Bovine Evolve (Zenith Biotech, Guilford, CT, USA) supplemented with 4 mg mL–1 BSA (Probumin, EMD Millipore, Norcross, GA, USA) under oil in a 5% CO2, 5%O2, 90% N2 humidified incubator (Pryor et al. 2011 Theriogenology 75, 24–33). Cleavage rates of 87.7% (664/757) from three replicates produced 273 (36.0%) viable embryos on Day 6.5 post-IVF, which were evenly distributed by IETS stage (4–7) and grade (1 and 2) into three treatment groups (0 = control, 6 or 12 h straw incubation) before in vitro culture for an additional 24 h. For each replicate, the average embryo stage was calculated by multiplying the number of embryos in each treatment by their IETS stage and dividing by total embryos per group. The change in stage for each treatment was calculated by subtracting the initial average stage from the final average stage on Day 8. Grade 1 and 2 embryos at stage 6–8 were counted and used to calculate total viable rates. Day 8 (post-IVF) embryos were fixed in cold methanol, washed in PBS/0.1%Tween 20, mounted in 10 μg mL–1 Hoechst 33342/glycerol and viewed under UV light to count nuclei. Percentage data were transformed using arcsine square root function before analysis, and means were compared using a one-way ANOVA and Tukey’s HSD. Although viability decreased with increasing time in straw incubation, there were no statistical differences between control, 6 and 12 h treatments for total viable rates (90.8, 80.3, and 70.8%, respectively). Average embryo stage on Day 8 for control, 6 and 12 h (7.0 ± 0.66, 6.6 ± 0.24, and 6.2 ± 0.30 s.e.m., respectively) was not different, but tended to be higher in control (P = 0.08). The change in stage, however, was different between control and 12 h (1.46 ± 0.33 and 0.66 ± 0.24, respectively; P < 0.05). Likewise, cell numbers were greater in control and 6 h embryos compared with 12 h straw incubation (149.8 ± 9.14, 138.7 ± 7.94, and 101.8 ± 5.29; n = 47, 50, and 46, respectively P < 0.01). In conclusion, 6.5 day IVP embryos held in warm Vigro holding medium for 12 h in 1/4 cc straws fail to develop at the same rate and incurred lower cell counts than either control or 6 h treatments. Further research to evaluate pregnancy rates following transfer and utilising different incubation or media and/or temperature is warranted to further evaluate the utility of in straw incubation for extended periods of time.

scholarly journals Role of Low Dose Intravenous Immunoglobulin in IVF Failure - A Retrospective Analysis

2018 ◽  
Vol 3 (1) ◽  
Keyword(s):  
Pregnancy Rate ◽  
Retrospective Analysis ◽  
Intravenous Immunoglobulin ◽  
Embryo Transfer ◽  
Low Dose ◽  
Transvaginal Ultrasound ◽  
Clinical Pregnancy ◽  
Ivf Failure

Background: In developing countries the number of in vitro fertilization (IVF) attempts is often limited by the high costs of the procedure and relatively low success rates. In such a setup we have tried to evaluate the effect of low dose intravenous immunoglobulins (IVIg) administered to patients who have had previous failed IVF outcomes. Bearing in mind that the inherent fecundity of Indian population is higher and thus a lower dose of IVIg may suffice to give positive outcomes at an affordable price, in this manner providing them with the possibility of affording more attempts if required. Objective: To evaluate the role of low dose intravenous immunoglobulin in IVF failure. Design: Retrospective analysis Materials and methods: This is a retrospective study beginning from 1st January 2014 till 31st December 2014. During this period, 124 patients with two or more failed IVF cycles were included. The controlled ovarian stimulation was started on cycle day 2 using gonadotropins (225 - 450 iu daily) and GNRH antagonist was added on the day when follicle reached 13 -14 mm. When follicles reached 18mm, transvaginal ultrasound guided oocyte aspiration was performed within 36 hours of the hcg trigger. On the ovum pick up day, 5 grams IVIg was administered to the patient as a slow infusion. Embryo transfer was done on day 2 or 3. Serum beta hcg was done 14 days after the embryo transfer and pregnancy rate and clinical pregnancy were evaluated. Results: The total pregnancy rate was 46% (57/124) and clinical pregnancy rate was 42.7% (53/124). Conclusion: Our study concluded that low dose IVIg may play a significant role in improving pregnancy rates in women with previous failed IVF attempts.

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.

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