scholarly journals 168FACTORS AFFECTING ON EMBRYO TRANSFER PREGNANCY RATES OF IN VITRO-PRODUCED BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 206 ◽  
Author(s):  
S. Aoki ◽  
S. Murano ◽  
M. Miyamura ◽  
S. Hamano ◽  
Y. Terawaki ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Estrous Cycle ◽  
Embryo Transfer ◽  
Calf Serum ◽  
Pregnancy Rates ◽  
Factors Affecting ◽  
Frozen Embryos ◽  
Rectal Palpation ◽  
Day Of Embryo Transfer

The objective of this study was to analyze factors affecting the pregnancy rates after transfer of IVF-derived Japanese Black embryos. Holstein cows and heifers (n=7250) were selected as recipients, and embryo transfers were performed for 3yr (between 1998 and 2000). The IVM-IVF procedure was performed according to a method previously described (Hamano S and Kuwayama M 1993 Theriogenology 39, 703–712). IVF-derived embryos that developed into expanded blastocysts (grade 1, manual of IETS) after 7 to 8 days (insemination=Day 0) were used for this study. Some of these embryos were frozen in TCM-199 supplemented with 1.4M glycerol, 20% calf serum, and 0.25M sucrose. The embryos were seeded at −6°C, held at −6°C for 10min, and then cooled to −25°C at a rate of 0.33°Cmin−1. Frozen embryos were thawed in a 30 to 35°C water bath after 10s of air thawing. Fresh (n=3952) or frozen-thawed (n=3298) embryos were nonsurgically transferred to recipients on Days 6 to 9 of the estrous cycle. Data collected at the time of embryo transfer included recipient parity (cow or heifer), whether recipient estrus was natural or synchronized with PGF2α, cloprostenol or CIDR, methods of estrous confirmation (showing standing heat, rectal palpation of ovary without standing heat, or showing only mucous vulvular discharge), number of examinations of the CL by palpation per rectum (twice on the day before embryo transfer and the day of embryo transfer, or once on the day of embryo transfer), type of embryos (fresh or frozen), and day of the estrous cycle at the time of embryo transfer. CATMOD procedures of SAS were used to determine the factors affecting the pregnancy rate. Overall pregnancy rates were 37.3% (n=2704). Whether recipient estrus was natural or synchronized and the type of embryos did not influence the pregnancy rates. Heifers had significantly higher pregnancy rates than cows (44.0% v. 33.0%, respectively, P<0.05). Pregnancy rates among the subset of heifers and cows showing standing heat were significantly higher than those showing only mucous vulvular discharge (39.5% v. 33.5%, respectively, P<0.05). Examining the CL twive had a significantly higher pregnancy rate than did a single examination of the CL (41.1% v. 35.6%, respectively, P<0.05). Pregnancy rate on Day 8 (38.4%, 1358/3533) of the estrous cycle at the time of embryo transfer was significantly higher than on Days 6 (27.7%, 23/83) and 7 (36.2%, 1235/3408) (P<0.05), and the pregnancy rate on Day 6 of the estrous cycle at the time of embryo transfer tended to be lower than on Day 9 (38.9%, 88/226) (P<0.08). These results demonstrate that confirming standing heat, performing CL examination twice before embryo transfer, freezing high quality embryos, and performing embryo transfers on Day 8 resulted in an improved pregnancy rate for the transfer of IVF-derived embryos.

188 EFFECT OF TRANSFER OF FROZEN-THAWED IVF EMBRYOS ON PREGNANCY IN REPEAT-BREEDER INSEMINATED OR NON-INSEMINATED HOLSTEIN CATTLE

2006 ◽  
Vol 18 (2) ◽  
pp. 202 ◽  
Author(s):  
O. Dochi ◽  
M. Tanisawa ◽  
S. Goda ◽  
H. Koyama
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Calf Serum ◽  
Pregnancy Rates ◽  
Holstein Cattle ◽  
Chi Square ◽  
Vitro Fertilization ◽  
Chi Square Test ◽  
Repeat Breeder

Repeat-breeding is one of the important factors that affect dairy management. The objective of this study was to investigate the effect of transfer of frozen–thawed IVF embryos on pregnancy in repeat-breeder Holstein cattle. Cumulus–oocyte complexes (COCs) were collected by aspiration of 2–1-mm follicles from ovaries obtained at a local abattoir. COCs were matured for 20 h in TCM-199 supplemented with 5% calf serum (CS) and 0.02 mg/mL of FSH at 38.5°C under a 5% CO2 atmosphere in air. Matured oocytes were inseminated with spermatozoa of 5 × 106/mL in BO solution (Brackett and Oliphant 1975 Biol. Reprod. 12, 260–274) containing 10 mM hypotaurine and 4 units/mL heparin. After 18 h of gamete co-culture, presumptive zygotes were cultured in CR1aa (Rosenkrans et al. 1991 Theriogenology 35, 266) supplemented with 5% CS for 8 days at 38.5°C under 5% CO2, 5% O2, 90% N2 atmosphere in air. After in vitro fertilization, Day 7 and Day 8 blastocysts were frozen in 1.5 M ethylene glycol (EG) in Dulbecco's PBS (DPBS) supplemented with 0.1 M sucrose and 20% CS. Embryos were transferred into a freezing medium, loaded into 0.25-mL straws, and allowed to stand for 15–20 min for equilibration. The straws were then plunged into a −7°C methanol bath of a programmable freezer for 1 min, seeded at −7°C, maintained at −7°C for 15 min, cooled to −30°C at the rate of −0.3°C/min, and then plunged into liquid nitrogen. Recipient animals (43 heifers, 131 cows) included those that did not conceive after being artificially inseminated (AI) 3 to 15 times. The frozen–thawed IVF embryos were directly transferred to the recipient animals 7 days after estrus or AI. Pregnancy rates were analyzed by chi-square test. The results are presented in Table 1. There were no significant differences in the pregnancy rates between treatments. However, a slightly higher pregnancy rate was achieved by embryo transfer after AI. These results suggest that embryo transfer may increase the pregnancy rate in repeat-breeder Holstein cattle. Table 1. Pregnancy rates after transfer of IVF frozen–thawed embryos in repeat-breeder Holstein cattle

100 FACTORS AFFECTING PREGNANCY RATES IN THE TRANSFER OF IN VITRO-PRODUCED JAPANESE BLACK CATTLE EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 157
Author(s):  
T. Nishisouzu ◽  
A. Abe ◽  
S. Matoba ◽  
O. Dochi ◽  
K. Okamura
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Pregnancy Rates ◽  
Oestrous Cycle ◽  
Rapid Expansion ◽  
Japanese Black Cattle ◽  
Factors Affecting ◽  
Increased Activity

Despite the rapid expansion of in vitro embryo production (IVP) technology for genetic improvement in the cattle industry in the last decades, pregnancy rates by the transfer of IVP embryos are still lower than those derived from in vivo-produced embryos. The objective of this study was to analyse factors affecting pregnancy rates after the transfer of IVP Japanese Black cattle embryos under farm conditions. Holstein heifers (n = 4,475) and cows (n = 8,541) were selected as recipients. Most cows (80%) were managed in tie-stall barns and most heifers (80%) were managed in pens. Embryo transfers were performed for 9 years, from 2004 to 2012. The embryos were produced from oocytes derived from a local abattoir and semen from 14 proven bulls by the Livestock Improvement Association of Japan (Hamano and Kuwayama 1993 Theriogelogy 39, 703–712). The fresh IVP embryos (quality; IETS code 1) that reached the blastocyst stage after 7 to 8 days (insemination = Day 0) were transported by an airplane (2 h) and subsequently by a car (1.5 h). Embryos were non-surgically transferred to each recipient on Day 7 to 9 of their natural oestrous cycle on farms. Pregnancy was diagnosed on Day 40 to 60 after oestrus. Pregnancy results were statistically analysed using the GLM procedures of SAS. The following variables were included in the model: recipient parity (0, 1, 2, or 3), day (7, 8, or 9) of the oestrous cycle at the time of embryo transfer, oestrus behaviour (increased activity observed by farmers), presence of mucus at oestrus, presence of blood after oestrus, and year (1, 2, 3, 4, 5, 6, 7, 8, or 9) and season (April–June as spring, July–September as summer, October–December as fall, or January–March as winter) of embryo transfer. The Bonferroni correction was used to counteract the problem of multiple comparisons. Heifers had significantly higher pregnancy rates than cows (51.0% v. 37.9%, respectively; P < 0.01), and first parity cows had higher pregnancy rates than third parity cows (42.9% v. 35.7%, respectively; P < 0.01). Pregnancy rates were significantly higher in recipients that received an embryo transfer on Day 8 of their oestrous cycle, than on Day 7 (46.6% v. 42.4%, respectively; P < 0.01). Recipients without oestrus behaviour had higher pregnancy rates than those with oestrus behaviour (46.3% v. 43.4%, respectively; P < 0.01). The presence of mucus and/or blood after oestrus and the season of transfer were not found to significantly affect pregnancy rates. The results of this study indicated that performing IVP embryo transfers on Day 8 of a recipient’s oestrous cycle will improve the pregnancy rate, season does not have an effect on pregnancy rate, and the detection of oestrus by monitoring increased activity is not always reliable and instead should be determined by multiple symptoms on farm conditions.

scholarly journals Is Endometrial Thickness on the Day of ET Really Predictive of IVF Outcome?

2012 ◽  
Vol 3 (2) ◽  
pp. 40-47 ◽  
Author(s):  
MS Srinivas ◽  
Kamini Rao ◽  
Richa Sharma ◽  
Theodre Jones
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Endometrial Thickness ◽  
Pregnancy Rates ◽  
Ivf Outcome ◽  
Male Factor ◽  
Cleavage Rate ◽  
Multiple Variables ◽  
Day Of Embryo Transfer

ABSTRACT Background The effect of endometrial thickness on pregnancy rates in assisted reproductive technology (ART) patients has been evaluated by many authors, with controversial results. Endometrial thickness has been utilized as an indirect indicator for endometrial receptivity. Objective To evaluate relationship between endometrial thickness on day of embryo transfer and pregnancy outcome in in vitro fertilization and embryo transfer (IVF-ET) cycles. Should we cancel cycles based on endometrial thickness only? Material and methods A prospective analysis was conducted at Dr Kamini Rao Hospital, Bangaluru, of 239 patients. Various parameters were compared between pregnant and nonpregnant patients to see whether there is any cut-off for endometrial thickness on day of embryo transfer by which we can predict good prognosis in form of pregnancy and what effect other variables on endometrial thickness and pregnancy respectively and should we cancel embryo transfer, if endometrial thickness is not within certain range? Results In the study population, 174 (73%) had primary and 65 (27%) had secondary infertility, Ovarian stimulation was performed with long protocol in 37% cases, antagonist protocol in 47% and other protocols like microflare, short, ultralong, ultrashort in 15%. Mean age of patients was 31.04 ± 3.79 years. Among causes of infertility male factor was present in 39%, tubal factor was seen in 18%, unexplained were 13%, polycystic ovarian syndrome in 11%, poor ovarian reserve in 4.1% and mixed causes in 13%. Majority of our patients were in normal and overweight as per body mass index (BMI). ET were easy in 90% of cases and 14 (5.8%) ETs were cancelled. The reason for cancellation was ovarian hyperstimulation syndrome (OHSS) in 9 cases, fluid in cavity in 2 cases, one patient had hyperpyrexia on day of ET and 2 cases of failed fertilization. Endometrial thickness was >10 mm in 35% cases. Overall clinical pregnancy rate was 39% with implantation rate of 21%, fertilization rate of 92% and cleavage rate of 95% and live birth rate of 26%. There were more follicles, oocytes and embryos, the endometrium was >10 mm and embryo quality was higher among women who became pregnant when compared with nonpregnant women after assisted reproduction though not statistically significant (p > 0.05). The pregnancy rate improved as endometrial thickness increased showing a linear association. Conclusion Increased endometrial thickness is associated with higher pregnancy rates, but as such a cut off cannot be decided. In our study we have seen pregnancies at both thin and thick endometrium so we should not cancel ET merely on the basis of endometrial thickness as pregnancy is affected by multiple variables and not by endometrial thickness alone. How to cite this article Sharma R, Rao K, Srinivas MS, Jones T. Is Endometrial Thickness on the Day of ET Really Predictive of IVF Outcome? Int J Infertility Fetal Med 2012;3(2): 40-47.

206 EFFECTS OF A COMBINATION OF A PRID, PGF2α AND eCG, ON ESTRUS SYNCHRONIZATION AND PREGNANCY RATE FOLLOWING EMBRYO TRANSFER IN HOLSTEIN HEIFERS

2007 ◽  
Vol 19 (1) ◽  
pp. 220 ◽  
Author(s):  
Y. Aoyagi ◽  
A. Ideta ◽  
M. Matsui ◽  
K. Hayama ◽  
M. Urakawa ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Plasma Concentrations ◽  
Pregnancy Rates ◽  
Bovine Embryo ◽  
Estrus Synchronization ◽  
Chi Square ◽  
End Of Treatment ◽  
Chi Square Analysis ◽  
Day Of Embryo Transfer

Successful bovine embryo transfer requires synchronization of luteolysis, estrus and ovulation. The objective of the present study was to evaluate the effect of a combination of a PRID, PGF2� and eCG, on estrus synchronization and pregnancy rate in recipient heifers. A PRID� (ASKA Pharmaceutical Co., Ltd., Tokyo, Japan) was inserted into the vagina at random days of the estrous cycle for 7 (n = 35) or 9 (n = 43) days. Two days before removal of the PRID, the heifers were injected with PGF2� IM (2 mL Resipron�-C containing 0.25 mg mL-1 cloprostenol; ASKA). About half of the heifers in each group received 250 IU eCG IM (Serotropin�; ASKA) at the time of PRID removal. Blood was collected several times from the start of treatment for 7 (n = 9) or 9 (n = 9) days and on the day of embryo transfer by jugular venipuncture; plasma was immediately separated and stored at -20�C until assayed for plasma concentrations of estradiol-17α (E2) and progesterone (P4). The E2 and P4 determinations were performed by enzyme immunoassay after extraction by diethyl ether. Pregnancy was determined by ultrasonography on Day 30 (Day 0 = estrus). The rates of successful standing estrus (no. in estrus/PRID inserted), embryo transfer (no. transferred/estrus), and pregnancy (no. pregnancy/transferred) were compared between groups. Data were analyzed by chi-square analysis or Fisher&apos;s PLSD test following ANOVA. Injection of eCG at the time of PRID removal had no significant effect on the rates of successful standing estrus, embryo transfer, or pregnancy (P &gt; 0.05). The proportion of heifers treated for 9 days that exhibited standing estrus (93&percnt;, 40/43) was significantly higher than the proportion of heifers treated for 7 days that exhibited standing estrus (66&percnt;, 23/35, P &lt; 0.01). Of the heifers that were treated for 9 days, the proportion of heifers exhibiting standing estrus within 2 days after the end of treatment was significantly higher (93&percnt;, 37/40) than for heifers that were treated for 7 days (65&percnt;, 15/23; P &lt; 0.01). Pregnancy rates of heifers treated for 9 days (84&percnt;, 32/38) and 7 days (81&percnt;, 17/21) were not significantly different. The E2 : P4 ratio normally increases during follicle growth and CL regression. The plasma E2 : P4 ratio between the time of injection of PGF2&alpha; and the time of PRID removal was significantly higher for heifers that were treated for 9 days than it was for heifers that were treated for 7 days (P &lt; 0.01). These results suggest that a combination of PRID treatment for 9 days and injection of PGF2&alpha; 2 days before PRID removal successfully synchronized estrus in recipient heifers and led to high pregnancy rates following embryo transfer.

289 EFFECTS OF YEAR PERIOD, TECHNICAL TEAM, AND BREED ON THE PREGNANCY RATES AFTER TRANSFER OF IVF BOVINE EMBRYOS IN THE SOUTHERN REGION OF RIO DE JANEIRO

2010 ◽  
Vol 22 (1) ◽  
pp. 301
Author(s):  
B. G. Moura ◽  
J. Almeida ◽  
F. L. Lima ◽  
G. Balbi ◽  
R. Calmerani ◽  
...  
Keyword(s):  
Beef Cattle ◽  
Dairy Cattle ◽  
Embryo Transfer ◽  
Pregnancy Rates ◽  
Chi Square ◽  
Chi Square Test ◽  
Few Data ◽  
Rectal Palpation ◽  
Fresh Embryos

The aim of the work was to study the effects of year period, technical team, breed, beef cattle and dairy cattle on the pregnancy rates in fresh embryos used in bovine transfer of IVF programs. The study was carried out at the fertilization laboratory In Vitro Nyltta Britto de Carvalho, in partnership with In Vitro Brazil, located at the Boa Vista farm, Barra do Pirai, during August 2007 to September 2008, seeking subsidies to improve the use of the technique in the field. During that period, aspirations and inovulations in 3 different periods I (August to December), II (January to April), and III (May to September) were carried out. The jobs were accomplished by 9 technical teams (A, B, C, D, E, F, G, H, and I) rendering services to the laboratory, by working with 2 beef breeds (Brahman and Nelore) and 3 dairy breeds (Gir, Girolando, and Holstein). The different breed receivers were synchronized, and in general, from 6 to 8 days after heat, they received embryo transfer, the cervical way, under low epidural anesthesia, where each female received 1 fresh embryo of IVF. All cows were submitted to gestation diagnosis by rectal palpation and ultrasonography, in general, 42 days after embryo transfer. The numbers of embryo transferred and pregnancy rates were submitted to the chi-square test, which presented significant differences (P < 0.05). There were pregnancy rates of 36.25%a (n = 960), 39.83%a (n = 1180), and 32.59%b (n = 919) in the I, II, and III periods, respectively. Among the 9 technical teams, there were verified pregnancy rates (%) of 33.51d (n = 1313), 30.30d (n = 330), 35.00cd (n = 405), 39.24cd (n = 1060), 59.25a (n = 7), 33.33d (n = 24), 53.57bc (n = 28), 43.31c (n = 157), and 58.33ab (n = 12) for A, B, C, D, E, F, G, H, and I teams, respectively. Among breeds there were rates (%) of 36.89ab (n = 412), 34.68b (n = 1286), 35.13ab (n = 74), 38.94a (n = 1140), and 37.80ab (n = 82) for Brahman, Nelore, Gir, Girolando, and Holstein, respectively. In the study, pregnancy rates (%) of 35.21b (n = 1698) in beef cattle and 38.65a (n = 1296) in dairy cattle were observed. The differences in pregnancy rates with respect to the evaluated factors, may be explained by individual, breed, and nutritional variations of the animals. There are few data in the literature with results on the embryo transfer use of IVF bovine under field conditions.

109 RELATIONSHIP BETWEEN ULTRASONIC MORPHOLOGY OF CORPUS LUTEUM IN HOLSTEIN HEIFERS AND PREGNANCY RATE AFTER EMBRYO TRANSFER

2012 ◽  
Vol 24 (1) ◽  
pp. 167
Author(s):  
A. Shirasawa ◽  
Y. Nakamura ◽  
A. Ideta ◽  
Y. Oono ◽  
M. Urakawa ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Blood Clot ◽  
Pregnancy Rates ◽  
Bovine Embryo ◽  
Tissue Area ◽  
The Mean ◽  
Rectal Palpation ◽  
Experimental Group

Recipient animals for bovine embryo transfer (ET) are routinely selected according to the morphology of the corpus luteum (CL) estimated by rectal palpation. However, rectal palpation is not a precise method of diagnosing the functional status of a CL. Ovarian ultrasonography (US) may be used to improve such diagnoses. The aim of this study was to evaluate the relationship between ultrasonographic images of CL and pregnancy rates after ET in Holstein heifers to determine whether US can be used to select recipients for ET. Recipient heifers (n = 285) were selected by detection of natural oestrus or following oestrus synchronization using a progesterone-releasing intravaginal device (PRID; ASKA Pharmaceutical, Tokyo, Japan). Transrectal US was performed immediately before ET, on Days 6 to 8 of the oestrous cycle (oestrus = Day 0), using a B-mode scanner (HS1500V; Honda Electronics Co. LTD, Aichi, Japan) equipped with a 7.5-MHz linear-array transducer designed for intrarectal placement. A cross-sectional image of the maximal area of the CL and luteal cavity was obtained. The areas of the CL and luteal cavity were each calculated using the formula for the area of an ellipse (height/2 × width/2 × π). (1) Ultrasonic morphology of CL was classified into 3 types: without cavity (n = 128), with cavity (n = 145) and with blood clot (n = 12). (2) The luteal cavity was categorized into 3 groups: small (<100 mm2, n = 93), medium (100 ≤ x < 200 mm2, n = 32) and large (≥200 mm2, n = 20). (3) Luteinized tissue area (total area of CL minus the area of the luteal cavity) was categorized into 3 groups: small (<250 mm2, n = 61), medium (250 ≤ x < 350 mm2, n = 128) and large (≥350 mm2, n = 84). In vivo–produced embryos were transferred nonsurgically into the uterine horn ipsilateral to the CL. Pregnancy was determined by transrectal US on Days 30 to 40 of gestation. The pregnancy rates of each experimental group were analysed by logistic regression. In this study, the pregnancy rate did not differ significantly in each experimental group: (1) without cavity: 77.3% (99/128), with cavity: 75.2% (109/145) and blood clot: 75.0% (9/12); (2) small cavity: 73.1% (68/93), medium: 75.0% (24/32) and large: 85.0% (17/20). The mean area of the cavity was 100.8 ± 110.3 mm2 (mean ± standard deviation) and recipients with 0 to 539.7 mm2 sized cavities had successful pregnancies (observational range was 0 to 539.7 mm2). (3) The pregnancy rates of recipients that had small, medium and large luteinized tissue were 77.0% (47/61), 75.0% (96/128) and 77.4% (65/84), respectively. The mean area of luteinized tissue was 318.9 ± 90.3 mm2 and 155.0 to 620.0 mm2 sized luteinized tissue had pregnancy success (observational range was 132.8 to 620.0 mm2). In conclusion, the results from this study indicate that the presence of a luteal cavity or blood clot has no detrimental effect on pregnancy success after ET in Holstein heifers. Furthermore, no relationship was found between luteinized tissue area at the time of ET and pregnancy rate.

216 THE RELATIONSHIP BETWEEN OXYGEN CONSUMPTION RATE AND PREGNANCY RATE OF BOVINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 225 ◽  
Author(s):  
N. Sakagami ◽  
K. Akiyama ◽  
Y. Nakazawa
Keyword(s):  
Oxygen Consumption ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Embryo Quality ◽  
Calf Serum ◽  
Pregnancy Rates ◽  
Bovine Embryos ◽  
Significant Difference ◽  
Consumption Rates ◽  
The Relationship

A precise evaluation of embryo quality is important to estimate the suitability of embryo transfer to recipient animal. Recently, an objective evaluation method was reported for bovine embryos, in which the oxygen consumption of embryos can be noninvasively determined by scanning electrochemical microscopy (SECM) (Shiku et al. 2001 Anal. Chem. 73, 3751–3758). Trimarchi et al. (2000 Biol. Reprod. 62, 1866–1874) suggested that the oxygen consumption reflects the cell number and mitochondrial activity of embryos. The objectives of this study were (1) to examine the oxygen consumption of in vivo-derived embryos by SECM, (2) to investigate the relationship between oxygen consumption and morphological estimation of embryos, and (3) to assess the correlation among the oxygen consumption, embryo viability, and pregnancy rates. Fifty-six embryos were collected from Japanese Black cattle, which were superovulated with a total dose of 20 mg porcine FSH (FSH-R; Kawasaki Pharmaceutical Co., Ltd., Tokyo, Japan) followed by AI. The qualities of collected embryos at the stage of compacted morulae (CM), early blastocysts (EB), and blastocysts (BL) on Day 7 after AI were categorized as grade 1 and grade 2, according to the IETS manual (2002). The oxygen consumption rates of embryos were evaluated by SECM, as previously described by Abe et al. (2004 J. Mamm. Ova Res. 21). Embryos were frozen by programmable freezer in Dulbecco&apos;s PBS containing 1.5 M ethylene glycol, 0.1 M trehalose, and 20&percnt; calf serum. They were thawed by holding the straws in air for 8 s and then immersing them in a 30&deg;C water bath for 15 s. After thawing, the embryos were examined for oxygen consumption. Twenty-eight embryos were then cultured in TCM-199 supplemented with 20&percnt; fetal bovine serum and 0.1 mM &beta;-mercaptoethanol for 24 h to assess the viability of embryos by re-expansion of blastocole. The remaining 28 embryos were transferred to recipients. The pregnancy rates were determined by rectal palpation on Day 70. Data were analyzed by ANOVA. The consumption rates of BL embryos on Day 7 were significantly higher (P &lt; 0.05) than those of CM collected on the same day (0.84 vs. 1.29 &times; 10&minus;14 mol s&minus;1, respectively). A significant difference was also observed in consumption rates between grade 1 and 2 embryos at the BL stage (P &lt; 0.05). After freezing&ndash;thawing, the average oxygen consumption rates of embryos were 0.52 &times; 10&minus;14 mol s&minus;1 for CM (n &equals; 9), 0.67 &times; 10&minus;14 mol s&minus;1 for EB (n &equals; 8), and 0.96 &times; 10&minus;14 mol s&minus;1 for BL (n &equals; 11). The CM embryos with rates of &lt; 0.5 &times; 10&minus;14 mol s&minus;1 and the EB and BL embryos with those &lt; 0.6 &times; 10&minus;14 mol s&minus;1 did not show good morphological appearance after 24 h in culture. Pregnant animals were not obtained from embryos with rates &lt;0.5 &times; 10&minus;14 mol s&minus;1 for CM (n &equals; 5) and &lt;0.7 &times; 10&minus;14 mol s&minus;1 for EB (n &equals; 9). A high pregnancy rate (67&percnt;) was obtained from embryos with rates &gt;1.0 &times; 10&minus;14 mol s&minus;1 for BL (n &equals; 14). These results suggest that the measurement of oxygen consumption of embryos after embryo freezing and prior to embryo transfer may be useful for estimating embryo quality and suitability of embryo transfer.

144 PREGNANCY RATES FOLLOWING THE TRANSFER OF IN VITRO FERTILIZED EMBRYOS TO RECIPIENTS ON DAY 6, 7, OR 8 AFTER OESTRUS

2015 ◽  
Vol 27 (1) ◽  
pp. 164
Author(s):  
R. C. Fry ◽  
K. L. Fry ◽  
H. A. McCartney ◽  
W. R. Geddes ◽  
K. Geddes
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Pregnancy Rates ◽  
Transfer Program ◽  
In Vitro Production ◽  
Chi Squared ◽  
Grade 3 ◽  
Vitro Production ◽  
Embryo Grade

The aim of this experiment was to investigate the effect of day of synchrony on the pregnancy rate of recipients following the transfer of Day 7 IVF embryos. In addition, the effect of IVF embryo grade and corpus luteum (CL) grade of recipients was determined. A total of 317 cumulus-oocyte complexes collected from 24 dry Brahman cows by TVR were matured, fertilized, and cultured under standard in vitro production procedures (Fry et al. 2003 Theriogenology 59, 446). A total of 89 (44 Grade 1, 43 Grade 2, and 2 Grade 3, IETS classification) in vitro-produced embryos were transferred to parous 4- to 9-year-old dry Brahman cross recipient cattle 7 days after IVF. Two groups of recipient cows were synchronised one day apart with an 8-day CIDR/pg protocol so that oestrous would be concentrated over 3 days with the middle day aligning with the day of IVF (Day 0). Donors that produced a large number of IVF embryos had these divided and transferred into recipients either on Day –1 or Day +1 of synchrony, and those producing less than 4 IVF embryos were transferred into recipients on Day 0. At embryo transfer the ovaries of the recipient were palpated and then scanned by rectal ultrasound and the grade of CL noted (Grade 1 = large distinct CL by palpation, Grade 2 = small distinct CL by palpation, Grade 3 = CL not distinguishable by palpation). Pregnancy was diagnosed by ultrasound scanning on Day 92. Although recipient numbers were low, differences in pregnancy rate between groups were analysed by Chi-squared. Data from the 2 Grade 3 embryos transferred were not included in the analysis (0/2 pregnant). Similar (P > 0.05) pregnancy rates were found when Day 7 IVF embryos were transferred to either Day 6 (17/32 = 53%), Day 7 (9/24 = 38%), or Day 8 (14/31 = 45%) recipients. Furthermore, neither the grade of the embryo (Grade 1: 20/44 = 45%, Grade 2: 20/43 = 47%) nor the grade of recipient CL (Grade 1: 17/45 = 38%, Grade 2: 17/29 = 59%, Grade 3: 6/13 = 46%) effected pregnancy rate (P > 0.05). This experiment demonstrates the flexibility of the IVF embryo to achieve an acceptable pregnancy rate over a range of recipient stages thereby allowing a high usage rate of good-quality recipients in an IVF embryo transfer program.

363 FACTORS INFLUENCING PREGNANCY RATES FOLLOWING TRANSFER OF BOVINE IN VIVO EMBRYOS BIOPSIED FOR SEX DETERMINATION

2007 ◽  
Vol 19 (1) ◽  
pp. 297
Author(s):  
S. Li ◽  
W. Yu ◽  
J. Fu ◽  
Y. Bai ◽  
F. Jin ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Pregnancy Rates ◽  
Chi Square ◽  
Embryo Survival ◽  
Chi Square Test ◽  
First Time ◽  
Fresh Embryos

Data collected from commercial embryo transfer programs in 63 farms in China during June 2002 to December 2005 was analyzed to examine the effects of various factors (biopsy, freezing, sample size, embryo development and quality, in vitro culture, and recipient quality) on pregnancy rates of in vivo-biopsied embryos. Embryos were flushed from superovulated dairy cattle and subjected to a biopsy for sexing determination using protocols and sexing kits supplied by AB Technology Ltd. Fresh embryos were implanted on the same day or frozen with AG freeze medium (AB Technology Ltd., Pullman, WA, USA) for later transfer. Recipients were synchronized with CIDA + PG protocols. Embryos were cultured in 6-well dishes containing 1.3 mL of holding medium (AB Technology Ltd.) in each well at room temperature (20–25�C) for examination of embryo survival in vitro. The chi-square test was used in statistic analysis. The implantation of fresh embryos after biopsy did not affect pregnancy rates (49.6%, 257/518) compared to that of non-biopsied fresh and frozen–thawed embryo groups (52.9%, 47/140 and 46.6%, 177/380, respectively). However, for biopsied embryos subjected to frozen and thawed procedures before implantation, particularly for those subjected to the removal of a larger biopsy, a reduced pregnancy rate was observed (41.8%, 297/710; P &lt; 0.01). Pregnancy rates among biopsied embryos at 3 different development stages (morula-early blastocyst, blastocyst, and expanded blastocyst) were not different. Similar results were found between embryo groups of grade 1 and 2. A significant decrease in pregnancy rate (0/10) was observed with embryos held in vitro for a longer period of time (&gt;5 h), suggesting detrimental effects of in vitro conditions on embryo survival. The highest pregnancy rate (68.0%) was observed in recipients synchronized for the first time before being implanted with biopsied embryos. Significant decreases in such rates were found in recipients synchronized for the second or third times or those with an abortion history at the first or second synchronization-implantation treatment (P &lt; 0.01). Better pregnancy rates (45.6%, 41/90; 46.1%, 76/165; and 45.5%, 5/11) were obtained for recipients implanted with biopsied embryos at Days 7.5, 8.0, and 8.5 post-heat detection, respectively, compared to 16% at Day 7 (3/18, P &lt; 0.05). It is concluded that mechanical treatment (cutting) does not reduce the survival of biopsied embryos; however, cryopreservation reduces their ability to survive in vivo. The analyses also suggest that holding embryos in vitro should not be longer than 5 h unless more favorable in vitro conditions can be provided. To achieve better results of implantation of biopsied embryos, embryo transfer should be performed during 7.5–8.5 days post-estrus, and the healthy recipients synchronized for the first time should be used.

110 First ovum pickup-in vitro-produced lidia breed calves using Lidia breed recipients: influence of age and state of the recipients and in vitro-produced embryos on pregnancy rates

2019 ◽  
Vol 31 (1) ◽  
pp. 181
Author(s):  
G. Gamarra Lazo ◽  
D. Di Scala ◽  
S. Maunas ◽  
R. Chaubet ◽  
S. Lacaze
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Development Stage ◽  
Pregnancy Rates ◽  
Chi Square ◽  
Related Factors ◽  
Single Operator ◽  
Pregnancy Status ◽  
Chi Square Analysis

We previously demonstrated the success of in vitro embryo production (IVP) in Lidia breed cattle (Gamarra Lazo et al. 2017 Reprod. Fertil. Dev. 30, 187). As in other species, the success of IVP is linked to the birth of calves from this technique. In the Lidia breed, an important factor to consider is the use of Lidia recipients in order to keep the temperament characteristic of this breed to next generations. The aim of the study was to produce ovum pickup (OPU)-IVP calves in the Lidia breed and to assess the effects of recipient and embryo related factors (status of the recipients; development stage of IVF embryos) on pregnancy rate following embryo transfer. Ovum pickup-IVP embryos from Lidia breeds were produced by a standard protocol (Gamarra Lazo et al. 2017 Reprod. Fertil. Dev. 30, 187). Numbers of blastocysts and expanded blastocysts were recorded on Day 7. A total of 27 blastocysts (B) and 34 expanded blastocysts (EB) of excellent quality (grade 1 according to IETS classification) were selected for fresh transfer. All embryos were transferred to Lidia breed recipients (heifers or cows) by a single operator under similar environmental and field conditions. Recipients were synchronized by subcutaneous insertion of an ear implant of 3.3mg of Norgestomet (Crestar®, MSD, Courbevoie, France) for 9 days. Two days before implant withdrawal, 0.5mg of Cloprostenol (Estrumate®, MSD) was injected. No oestrous detection was performed and synchronized females were selected as recipients when they presented a well developed corpus luteum at Day 9 after implant withdrawal (Day 6 to 7 after the expected oestrus). Blood samples were collected from recipients to determine pregnancy status using the bovine pregnancy associated glycoprotein (Idexx, Westbrook, ME, USA) 50-60 days after transfer. Pregnancy rates were analysed by chi-square analysis to compare results between heifers and cows and between B and EB embryo stages. The overall pregnancy rate after transfer of IVP fresh embryos from Lidia breed averaged 41.0% (n=25). A higher pregnancy rate was achieved in cows compared to heifers [51.2% (21/41) v. 20.0% (4/20) respectively, P&lt;0.05]. There was no difference in pregnancy rate between grade 1B [37% (10/27)] and EB [44.1% (15/34)] embryos (P&gt;0.05). Surprisingly, these results suggest that Lidia breed cows are the best recipients for OPU-IVP embryos. This may be related to the limited feasibility of manipulating the uterine horn during the embryo transfer in Lidia breed heifers, which have a low weight (less than 280kg) and present a narrow rectum diameter. It has been also observed that the cervix is very thin and difficult to cross, thus increasing the stress and potentially inflammatory and immune products secretion. Development stage of embryos did not affect pregnancy rate. To our knowledge, no OPU-IVP Lidia breed calves have been reported previously following transfer into Lidia breed recipients. In the current work, 13 OPU-IVP Lidia breed calves were born. Therefore, we confirmed the possibility of applying OPU-IVP and embryo transfer techniques in this breed within a genetic program.

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