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<0.05]. There was no difference in pregnancy rate between grade 1B [37% (10/27)] and EB [44.1% (15/34)] embryos (P>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.

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'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 > 0.05). The proportion of heifers treated for 9 days that exhibited standing estrus (93%, 40/43) was significantly higher than the proportion of heifers treated for 7 days that exhibited standing estrus (66%, 23/35, P < 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%, 37/40) than for heifers that were treated for 7 days (65%, 15/23; P < 0.01). Pregnancy rates of heifers treated for 9 days (84%, 32/38) and 7 days (81%, 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 PGF2α 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 < 0.01). These results suggest that a combination of PRID treatment for 9 days and injection of PGF2α 2 days before PRID removal successfully synchronized estrus in recipient heifers and led to high pregnancy rates following embryo transfer.

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

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 < 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 (>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 < 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 < 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.

scholarly journals PREGNANCY RATE OF RECIPIENT COWS AFTER TRANSFER OF IN VITRO-PRODUCED NELLORE EMBRYOS

2019 ◽  
Vol 32 (4) ◽  
pp. 1087-1091 ◽  
Author(s):  
MÔNICA ZUCHELLI JAGUSZESKI ◽  
ADALGIZA PINTO NETO ◽  
WILLIAM DE OLIVEIRA ◽  
JONATAS CATTELAM ◽  
HELTON APARECIDO GARCIA GREGIANINI
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Embryo Development ◽  
Development Stage ◽  
Blastocyst Stage ◽  
Private Company ◽  
Chi Square ◽  
Development Stages ◽  
Advanced Development

ABSTRACT The objective of this study was to evaluate the pregnancy rate (PR) of cows of six farms in Bolivia and in the states of Acre and Rondônia in Brazil after the transfer of 4,321 in vitro Nellore embryos produced by a private company, in 2015 to 2016. The effects of the farm location, year of embryo transfer (ET), season of ET, number of previous ET per recipient, and embryo development stage on the PR of the recipient cow (chi square p<0.05) were evaluated. The PR of the six evaluated farms were similar, with an average of 45.15%. The farm location, year of ET (2015 or 2016), and season of ET (rainy or dry season) did not affect the PR (p>0.05). The PR found after embryo transfer for recipient cows that had already been used once (45.58%) or twice (43.40%) for this purpose were higher than that of recipient cows used three to eight times (29.01%) (p<0.05). ET at expanded or hatched blastocyst stage resulted in higher PR (47.35%) when compared to the ET at morula or initial blastocyst (41.06%) and at blastocyst stage (43%) (p<0.05), which did not differ from each other (p>0.05). Under the conditions of this study, the transfer of in vitro-produced Nellore embryos at advanced development stages (expanded or hatched blastocyst) results in higher PR in cows that had been used once or twice for this purpose.

191 INFLUENCE OF THE DIFFERENT TIME COMPONENTS BETWEEN FLUSHING AND TRANSFER ON PREGNANCY RATES OF FROZEN CATTLE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 203
Author(s):  
C. Ponsart ◽  
H. Quinton ◽  
A. Rohou ◽  
J. Kelhembo ◽  
G. Bourgoin ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Chi Square ◽  
Frozen Embryos ◽  
Multivariate Logistic Model ◽  
Parity Number ◽  
Embryo Stage ◽  
Chi Square Analysis ◽  
Fresh Embryos ◽  
Donor Cows

Previous studies have shown that the time between flushing and freezing of bovine embryos can influence pregnancy rates (PRs) following embryo transfer (ET). The aim of this study was to determine which time components can influence ET results. Time components between flushing of a superovulated donor and freezing of the collected embryos were investigated under field conditions. Embryos were frozen in 1.5 M ethylene glycol (EG) for direct transfer. During January 2003, ET technicians (EmbryoTop, Rennes cedex, France) recorded systematically times corresponding to each step comprising the time spent in vitro (TIV) from 153 recovery sessions (RS) with freezing: end of flushing, beginning and end of search of embryos, start of equilibration in EG, beginning and end of straw loading, introduction to −7°C in the freezer, and seeding. Numbers of donor cows and ET technicians doing the freezing (n = 5) were noted for each RS. Embryo (stage, quality) and recipient (breed, parity) characteristics were also noted. A total of 548 frozen embryos were transferred and PRs were assessed. Variability of time components was investigated (Bourgoin et al. 2004 Reprod. Fertil. Dev. 16, 207). The influence of time components and other variation factors was tested on PRs (t-tests and chi-square analysis). The TIV averaged 210 ± 80 min and did not influence PR (≤4 h = 51.9% (n = 393) vs. >4 h = 55.5% (n = 155); P > 0.05), as well as duration of flushing (32 ± 8 min), interval between end of flushing and search (31 ± 27 min), duration of search (45 ± 25 min) and interval between end of search and beginning of freezing (101 ± 63 min). Only significant factors were kept for further analysis. The effects of recipient parity, number of donor cows per RS, and interval between introduction of straw to −7°C, and seeding were tested in a multivariate logistic model. PR varied strongly with parity of recipient (+25% in heifers vs. cows; P = 0.001). PRs were higher when the interval between straw introduction in the freezer and seeding lasted at least 5 min (2–4 min = 48.0% (n = 254) vs. 5–8 min = 57.1% (n = 294); P = 0.009). Time and operator effects were confounded. Overall PR results for the two technicians who used mostly 2–4 min intervals averaged 47% (operator values = 35.6, 48.9, and 54.5) whereas PRs were 54.9 and 60.5% for those waiting 5 min or more before inducing seeding (n = 2). PRs were higher when at least two donor cows were collected per RS (1 donor cow = 49% (n = 259) vs. ≥2 donor cows = 56.4% (n = 289); P = 0.003). This was not in agreement with previous observations in fresh embryos (Bourgoin et al. 2004). However, the number of donor cows strongly influenced the number of viable embryos per RS (1 donor cow = 11 ± 5 vs. ≥2 donor cows = 18 ± 8.5; P < 0.05) and could permit the choice of more embryos to be frozen. These results show that good PR may be achieved with a delay of several hours between flushing and freezing, when heifers are used as recipients. Moreover, confirmed from higher numbers of operators, these data show that it is better to wait at least 5 min to achieve equilibration of the embryo before seeding.

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.

scholarly journals 181COMPARISON OF THE PREGNANCY RATES AFTER SYNCHRONIZATION OF OVULATION USING GnRH AND PGF2± IN RECIPIENT DAIRY CATTLE

2004 ◽  
Vol 16 (2) ◽  
pp. 212 ◽  
Author(s):  
T. Nishisouzu ◽  
M. Sugawara ◽  
S. Aoki ◽  
K. Kishida ◽  
M. Moriyoshi ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Control Group ◽  
Corpora Lutea ◽  
Transfer Program ◽  
Chi Square ◽  
Follicle Diameter

Treatments with GnRH and PGF2α for synchronization of ovulation has resulted in acceptable pregnancy rates after fixed-time artificial insemination in dairy cows without estrus detection. The objective of the present study was to evaluate the practicability of ovulation synchronization (Ovsynch, Pursley JR et al. 1995 Theriogenology 44, 915–923) in dairy cattle using GnRH and PGF2α for the embryo transfer recipients. Dairy cattle (cows; n=100, heifers; n=88) were randomly allocated to one of two groups. The control group (cows; n=45, heifers; n=37) was composed of cows in natural estrus. The ovulation synchronization group (cows; n=55, heifers; n=51) was treated with an intramuscular injection of 100μg of GnRH at a random stage of the estrous cycle. Seven days later, the cattle received PGF2α (Cows; 25–30mg) or PGF2α analog (Heifers; 0.5mg) in order to regress the corpora lutea (CL). Forty-eight hours later, cows and heifers received a second injection of 100μg GnRH. Embryo transfer was carried out 7 days after the second injection of GnRH in the ovsynch group and 7 days after estrus in the control group. The cattle judged to have CL 17mm were classified as acceptable recipients. The size of the follicles and the CL were determined to be of estrus stage and embryo transfer by means of ultrasonography. The mean numbers of follicles and CL were analyzed by ANOVA, while pregnancy rates were analyzed by chi-square test. The results are presented in the Table. The proportion of cows and heifers determined to be acceptable embryo transfers was not different between the control group and the ovsynch group. There were no differences in the proportion of acceptable embryo transfers between the control group and the ovsynch group. Follicle diameter at the time of estrus in the control group (cows; 20.7±0.7mm, heifers; 16.8±0.5mm) were significantly larger than that of the ovsynch group (cows; 18.0±1.0mm, heifers; 14.7±0.2mm) (P&lt;0.05). Although CL diameter at the time of embryo transfer in heifers showed no differences between the control group and the ovsynch group (25.0±1.0mm v. 22.8±1.5mm), The CL diameter of the control cow group was larger than that of the ovsynch group (29.8±0.7mm v. 26.1±1.0mm, P&lt;0.05). However, no differences in pregnancy rate were seen between the control group and the ovsynch group. These results suggest that ovsynch can be effectively applied in an embryo transfer program for cattle. Table 1 Proportion of acceptable embryo transfer recipients and pregnancy rate in dairy cattle in the control ovsynch groups

204 RELATIONSHIP BETWEEN RESPIRATORY ACTIVITY AND THE PREGNANCY RATE OF BISECTED BOVINE EMBRYOS IN VIVO

2007 ◽  
Vol 19 (1) ◽  
pp. 219 ◽  
Author(s):  
S. Moriyasu ◽  
H. Hirayama ◽  
K. Sawai ◽  
S. Kageyama ◽  
S. Aoyagi ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Respiratory Activity ◽  
Pregnancy Rates ◽  
Bovine Embryos ◽  
Chi Square ◽  
Important Indicator ◽  
Consumption Rates

Oxygen consumption is an important indicator of the metabolic activity of living cells, which may provide valuable information for evaluating embryo quality. We have found that the bovine embryos with high oxygen consumption possess stronger potential for further development. However, the relationship between respiratory activity and the pregnancy rate of embryos is still unclear. In this study, we investigated the respiration rates of bisected bovine embryos and the pregnancy rates of demi-embryos after embryo transfer. Compact morula-stage embryos were bisected evenly by micro glass needle. One hundred bisected embryos were incubated for 24 h in embryo culture medium (IVD101; Research Institute for the Functional Peptides, Yamagata, Japan) at 39�C under 5% CO2, 5% O2, 90% N2. After the incubation, demi-embryos were classified into 2 groups: blastocoel-formed (BC) and blastocoel-not-formed (CM) embryos. Oxygen consumption rates of demi-embryos were measured by scanning electrochemical microscopy (SECM; Hokuto Denko Corporation, Tokyo, Japan). Within 3 h after the measurement, 80 demi-embryos were transferred into recipient cows (one demi-embryo/one recipient) at 7–8 days after estrus. Recipient cows were diagnosed for pregnancy by ultrasonography approximately 40 days after estrus. Statistical difference was analyzed by Tukey's post-hoc test and chi-square test. A total of 27 recipient cows became pregnant; the pregnancy rates for cows with CM and BC demi-embryos were 40.6% (13/32) and 29.2% (14/48), respectively. Mean oxygen consumption rates (� 10-14 mol s-1) in pregnant and non-pregnant cows were 0.47 and 0.39 for CM demi-embryos and 0.63 and 0.52 for BC demi-embryos, respectively. Retrospective analysis showed that the respiratory activity of demi-embryos in the pregnant group was higher than those in the non-pregnant group. In particular, the pregnancy rates for demi-embryos with respiratory activity higher than 0.35 in CM and 0.40 in BC groups were 52.0% (13/25) and 35.9% (14/39), respectively. On the other hand, cows with demi-embryos having an oxygen consumption rate under 0.35 in CM (n = 7) and 0.40 in BC (n = 9) groups did not become pregnant. These results demonstrated that bovine demi-embryos with higher respiratory activity showed a high pregnancy rate after embryo transfer. It is generally known that the pregnancy rate after the transfer of bisected embryos is lower than that of whole embryos. The measurement of oxygen consumption by SECM procedures is a useful tool to assess the quality of pre-implantation embryos and may contribute to the improvement of the success rate for bisected embryo transfer.

241 EFFECT OF PRODUCTION EFFICIENCY IN THE LIKELIHOOD OF PREGNANCY OF IN VITRO-DERIVED BOVINE EMBRYOS

2015 ◽  
Vol 27 (1) ◽  
pp. 210
Author(s):  
L. F. Feres ◽  
L. S. A. Camargo ◽  
M. P. Palhao ◽  
F. Z. Brandao ◽  
J. H. M. Viana
Keyword(s):  
Pregnancy Rate ◽  
Production Efficiency ◽  
Production Systems ◽  
Bos Indicus ◽  
Pregnancy Rates ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Chi Square

Improving in vitro culture systems to optimize embryo yield has been a major research goal. The relationship between the efficiency of embryo production systems and the pregnancy outcomes, however, remain controversial. The aim of the present study was to evaluate the likelihood of pregnancy of in vitro-produced embryos derived from batches with different relative efficiency indexes. Data of 702 ovum pick-up (OPU) and in vitro embryo production (IVEP) sessions, and of 2456 embryo transfers, recorded from 2008 to 2012, were evaluated. All donors were from the same herd, and were of the same breed (Gir, Bos indicus), as well as the semen used for IVF. The cumulus-oocycte complex (COC) recovery and IVEP were performed by the same team, in a single IVF laboratory, and using standard medium and procedures. Only data from embryos transferred as fresh were used, and records from 97 OPU/IVEP sessions in which no embryo was produced, or embryos were frozen or discharged due to lack of recipients, were discharged. The remaining 605 sessions were stratified in quartiles (I to IV, each one corresponding to 25% of total data) according to COC production of the donors, or stratified in ranges (0–25%, 26–50%, 51–75%, and 76–100%) according to COC quality (percentage of viable COC or of grade I COC) and to embryo production efficiency endpoints (cleavage rate, blastocyst rate). Pregnancy rates were compared among quartiles or ranges by the chi-square method. On average, the Gir donors produced 24.8 ± 0.6 COC per OPU, from which 14.4 ± 0.4 were classified as viable (57.8%), and 3.2 ± 0.1 as grade I (12.9%). On average 6.1 ± 0.2 embryos (morulas and blastocysts) were produced per OPU per donor, and mean pregnancy rate was 30.9%. As expected, donors with greater total COC yield (quartile I) also produced more viable oocytes (25.5 ± 0.7 v. 15.7 ± 0.3, 10.5 ± 0.2 and 5.8 ± 0.2), more COC grade I (4.8 ± 0.4 v. 3.9 ± 0.3, 2.6 ± 0.2 and 1.6 ± 0.1), and more embryos (9.0 ± 0.4 v. 6.9 ± 0.3, 5.0 ± 0.2 and 3.3 ± 0.1) than donors from quartiles II, III, or IV, respectively (P < 0.0001). Nevertheless, there was no difference (P > 0.05) in pregnancy rates for embryos produced from donors ranked in the different quartiles (30.9 v. 29.3, 31.5, and 30.5% for quartiles I to IV, respectively). Similarly, there was no difference (P > 0.05) in the pregnancy rate of embryos derived from OPU sessions in which there was a high or low percentage of viable or grade I COC. In vitro production efficiency (cleavage and blastocyst rates) also had no effect (P > 0.05) on further pregnancy rates. In conclusion, these results suggest that there is no relationship among the average number or quality of the COC recovered by OPU, the efficiency of IVEP, and the likelihood of pregnancy of in vitro-derived embryos.Research was supported by Fazendas do Basa, CNPq, and Fapemig.

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.

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