97 BOER GOAT OFFSPRING BORN FOLLOWING TRANSFER OF CRYOPRESERVED EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 207
Author(s):  
K. C. Lehloenya ◽  
J. P. C. Greyling

Cryopreservation of embryos is an important technique in the whole MOET program, which could help improve the transportation of genetic material across South Africa and globally. This trial evaluated the survival rate of goat embryos following transfer with cryopreserved Boer goat embryos. Twenty-seven multiparous Boer goat recipients were synchronized with CIDR for 16 days and injected with 300 IU of eCG at CIDR withdrawal. The recipients were allocated into 3 groups (n = 9). Group 1 received fresh embryos; Group 2 received slow frozen embryos; and Group 3 received vitrified embryos. Expanded blastocysts used were surgically collected from donors superovulated with pFSH on 7 following AI. Two blastocysts were transferred laparoscopically to the uterine horn ipsilateral to functional CL. A pregnancy rate of 85.7% (6) was obtained following the transfer of fresh embryos and tended to be better than in the does receiving slow frozen and vitrified embryos, (n = 4; 50.0% and n = 3; 37.5% does pregnant, respectively) with no significant differences. The kidding rate of the recipient does declined to 57.0% (4) and 25.0% (2) for fresh and slow frozen groups, respectively. The embryo survival rate of 35.7% (n = 5) for fresh, 25.0% (n = 4) for conventional slow freezing and 31.3% (n = 5) for vitrification was obtained and was not affected by the number of CL present on the respective ovaries at the time of embryo transfer. Although the pregnancy rate following the transfer of fresh embryos was satisfactory, the embryo survival rate following the transfer of fresh or cryopreserved embryos tended to be lower. The authors acknowledge the University of the Free State for financial and facility support and National Research Foundation (Thuthuka) for financial support for conducting this trial.

2004 ◽  
Vol 16 (2) ◽  
pp. 208
Author(s):  
C. Cuello ◽  
F. Berthelot ◽  
F. Martinat-Botté ◽  
P. Guillouet ◽  
V. Furstoss ◽  
...  

The present study was designed to determine the effect of pooling embryos from two donors on the reproductive success of transfer of vitrified/warmed porcine blastocysts. Superovulated Large White hyperprolific gilts (n=24) were used as embryo donors. Gilts were artificially inseminated 12 and 24h after initial detection of estrus using fresh semen, and slaughtered on Days 5.5 to 6 of the estrous cycle (Day 0=Onset of estrus). Embryos were recovered by flushing the uterine horns, and unhatched blastocysts were selected. Vitrification and warming were performed as reported previously (Berthelot et al., 2000 Cryobiology 41, 116–124). Embryo transfers were conducted in asynchronous (−24h) Meishan gilts (n=20). Twenty vitrified/warmed blastocysts were surgically transferred into one uterine horn. Ten recipients received embryos from one donor (group 1) and the other ten transfers were performed with mixed embryos from two donors (group 2). Pregnancy was assessed ultrasonographically at Day 25 after estrus and recipients were slaughtered five days later. The pregnancy rate from the different groups was compared using Fisher exact test. The GLM procedure of SAS was used to determine the effect of the origin of embryos (one or two donors) on the number of developed fetuses and viable fetuses at Day 30 of pregnancy. The ovulation rate was 32.5±11.8 (mean±SD). The total number of embryos collected was 634, of which 57 (9.0%), 36 (5.7%), 513 (80.9%) and 28 (4.4%), were unfertilized oocytes and degenerated embryos, morulae, unhatched blastocysts and hatched blastocysts, respectively. The ratio of collected embryos to the number of corpora lutea was 81.3%. The pregnancy rate for group 1 (70%) was not different (P>0.05) than that for group 2 (90%). No significant differences were detected between group 1 and group 2 for in vivo embryo development (number fetuses/transferred embryos in pregnant recipients; 33.3% v. 40%) or in vivo embryo survival (number viable fetuses/transferred embryos in pregnant recipients; 27.9% v. 33.9%). However, the in vivo efficiency (number viable fetuses/total transferred embryos) was higher (P<0.05) when transfers were performed with embryos from two donors (19.5% v. 30.5%). These results indicate that pooling embryos from two donors increases the in vivo efficiency after transfer of vitrified/warmed porcine blastocysts. This study was supported by grant from SENECA (FPI/99, Spain).


2004 ◽  
Vol 16 (2) ◽  
pp. 213 ◽  
Author(s):  
J. Small ◽  
M. Colazo ◽  
D. Ambrose ◽  
R. Mapletoft ◽  
J. Reeb ◽  
...  

The objective was to evaluate the effect of pLH treatment on pregnancy rates in recipients receiving in vivo- or in vitro-produced bovine embryos. Heifers (n=37) and lactating (n=28) and non-lactating (n=150) beef cows were treated at random stages of the cycle with 100μg GnRH i.m. (Cystorelin, Merial Canada Inc., Victoriaville, Quebec, Canada) on Day −9, 500μg cloprostenol i.m. (PGF; Estrumate, Schering Plough Animal Health, Pointe-Claire, Quebec, Canada) on Day —2 and GnRH on Day 0 (66h post-PGF; without estrus detection). Cattle were placed at random, by class, into three groups: no further treatment (Control; n=71), or 12.5mg pLH (Lutropin-V, Bioniche Animal Health, Belleville, Ontario, Canada) on Day 5 (n=72) or on Day 7 (n=72) after the second GnRH. On Day 7, cattle with a CL >10mm in diameter (determined ultrasonically) received in vivo-produced, fresh (Simmental) or frozen (Holstein), or in vitro-produced frozen (Holstein) embryos (embryo type balanced among groups). Embryos were cryopreserved in 10% ethylene glycol; in vivo-produced frozen embryos were thawed 5 to 10s in air, 15s in a water-bath at 30°C and then “direct-transferred” nonsurgically. In vitro-produced frozen embryos (donated by IND Lifetech Inc., Delta, British Columbia, Canada) were thawed in a water-bath at 27°C for 10s and placed in ViGro Holding Plus medium (AB Technology, Pullman, WA, USA) at room temperature, evaluated and then transferred nonsurgically. Pregnancy was determined by ultrasonography on Day 35. Data were analyzed with CATMOD, chi-square and GLM procedures (SAS Institute, Cary, NC, USA.). Twenty cattle (9.3%) did not receive embryos; five heifers had cervical problems, and five heifers and 10 cows did not have a CL >10mm. Overall, 7.1% of the recipients had two CL on the day of embryo transfer. There was no effect (P>0.05) of treatment, embryo type (or interaction) or class of recipient on pregnancy rate (overall, 44.1%, 86/195; Table 1). Similarly, mean (±SD) CL diameter and luteal area did not differ (P>0.05) among groups or between pregnant and open recipients (overall, 22.0±3.4mm and 352.0±108.7mm, respectively). However, recipients with a CL diameter ≥18mm tended (P<0.1) to have a higher pregnancy rate (45.8 vs 25.0%). In a subset of 40 recipients examined ultrasonically on Day 12, 50% of those treated on Day 5 and 70% of those treated with pLH on Day 7 had two CL. In summary, overall pregnancy rate in GnRH-synchronized recipients receiving in vitro- or in vivo-produced embryos by nonsurgical transfer was 44.1%. Embryo survival to Day 35 was not affected by type of embryo or treatment with pLH 5 or 7 days after ovulation. Table 1 Pregnancy rate in recipients on Day 35 based on pLH treatment and embryo-type


2005 ◽  
Vol 17 (2) ◽  
pp. 199 ◽  
Author(s):  
B. Peachey ◽  
K. Hartwich ◽  
K. Cockrem ◽  
A. Marsh ◽  
A. Pugh ◽  
...  

Vitrification has become the method of choice for the preservation of in vitro derived embryos of a number of species, and several methods of vitrification have been developed. One such method, the cryoLogic vitrification method (CVM) yields high survival rates of warmed embryos (Lindemans W et al. 2004 Reprod. Fertil. Dev. 16, 174 abst). In this study, the post-warm viability of bovine IVP embryos following either vitrification using CVM or slow freezing using ethylene glycol (EG) was compared. In addition, the survival of embryos following triple transfer to synchronized recipients was measured and the embryo (“e”) and recipient (“r”) contributions to embryo survival was determined using the “er” model for embryo survival (McMillan WH et al. 1998 Theriogenology 50, 1053–1070). Bovine IVP methods were those of van Wagtendonk et al. 2004 Reprod. Fertil. Dev. 16, 214 (abst). On day 7 of culture (Day 0 = IVF), Grade 1 and 2 embryos that had reached at least the late morula stage were selected for vitrification (20% DMSO, 20% ethylene glycol) or freezing in 1.5 M ethylene glycol + 0.1 M sucrose (0.5°C/min to −35°C). Following storage in LN2 for at least 24 h the embryos were thawed, the cryoprotectant removed, and the embryos cultured for 72 h in mSOF medium under 5% CO2, 7% O2, 88% N2. The number of hatching embryos was recorded at 24-h intervals. In addition, blastocyst and expanded blastocyst embryos were thawed and immediately transferred nonsurgically to recipients (three embryos of the same grade to each recipient) on Day 7 of a synchronized cycle (Day 0 = heat). The recipients were ultrasound-scanned for the presence of, and number of, fetuses on Days 35 and 62, respectively. The invitro assessment of 148 CVM and 230 EG frozen embryos indicated that more vitrified than EG embryos hatched by 72 h (73% vs. 62%; CVM vs. EG, χ2 = 4.5, P < 0.05). Overall, more Grade 1 embryos hatched than Grade 2 (74% vs. 60%, χ2 = 7.2, P < 0.01). CVM embryos (105) were triple-transferred to 35 recipients, and EG embryos (30) were triple-transferred to 10 recipients. Recipient pregnancy rates at Day 62 were 80% and 70%, respectively. Overall embryo survival was 38.5% (41% for CVM and 30% for EG). The overall calculated “e” and “r” values were 0.39 and 1.0 (“e”: 0.42 and 1.0, and “r”: 0.31 and 1.0, respectively, CVM and EG groups). Survival rates of CVM embryos to Day 62 (41%) were slightly lower than that previously obtained for fresh embryos produced using an identical IVP procedure (44% – van Wagtendonk AM 2004).


2007 ◽  
Vol 19 (1) ◽  
pp. 297
Author(s):  
S. Li ◽  
W. Yu ◽  
J. Fu ◽  
Y. Bai ◽  
F. Jin ◽  
...  

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.


1993 ◽  
Vol 57 (03) ◽  
pp. 369-378
Author(s):  
M. M. Lohuis ◽  
C. Smith ◽  
J. C. M. Dekkers

AbstractA dispersed hybrid MOET nucleus project was implemented in the Canadian dairy industry. Embryo yield, pregnancy rate, and effects of management and selection practices are reported. Repeatability of viable embryo yield was 0.31. Average viable embryo yield was 6.85 (s.e. 5.35) embryos per collection, with average pregnancy rates of 0.58, 0.46 and 0.51 for grade 1 fresh, frozen, and split embryos, respectively. Of the donors selected, 80% produced enough embryos to form a family of at least 15 offspring. Based on observed results, expectations of family size and structure and generation intervals were calculated. Only 10% of completed families are expected to result in less than three milking daughters. Generation interval for sib-tested sires was 58 months of age under practical field conditions, which was shorter by a factor of 0.19 than a traditional progeny test but longer than original theoretical estimates by a factor of 0.32. Realized selection intensities for a total merit index were 2.22 for sires and 3.77 for dams of embryos. Accuracies of selection for sires and dams were 0.93 and 0.73 respectively. To improve the scheme, use of more fresh embryos, embryo splitting and less delay in transferring frozen embryos are recommended.


Author(s):  
D. Pullar ◽  
K F D Brown ◽  
A E Wrathall

Early embryo mortality results in significant inefficiency in cattle breeding. A pregnancy rate of 65-70% to a single service, AI or transfer of fresh embryos is considered to be very good indeed. A pregnancy rate of 55-60% using frozen embryos would also be considered very good. Of the 30%, or so, of cattle which do not become pregnant to a single service, approximately one quarter are accounted for by fertilisation failure (Diskin and Sreenan, 1980; Roche et al, 1981) and another quarter by cytogenetic abnormalities in the embryos, which usually occur within the first few days after fertilisation (Hare et al. 1980; Gayerie de Abreu et al, 1984). Failure in the communication between embryo and uterus probably accounts for the remaining 50% of pregnancy loss. Fertilisation failure and early deaths due to cytogenetic abnormalities are by-passed by embryo transfer (ET) so that other causes of loss become rather more significant.


2018 ◽  
Vol 53 (3) ◽  
pp. 237
Author(s):  
K. STAMATARIS (Κ. ΣΤΑΜΑΤΑΡΗΣ) ◽  
K. DELIGIANNIS (Κ. ΔΕΛΗΓΙΑΝΝΗΣ) ◽  
T. LAINAS (Θ. ΛΑΪΝΑΣ) ◽  
G. ARSENOS (Γ. ΑΡΣΕΝΟΣ)

The objective of the study was to evaluate the use multiple ovulation and embryo transfer techniques in an indigenous Greek dairy breed of sheep. We stimulated selected donor ewes of the Karagouniko breed to produce large numbers of embryos after the induction of multiple ovulations by gonadotropin treatment (superovulatory response). A total of 50 Greek Karagouniko ewes were synchronised into oestrus using progestagen pessaries and superovulated for embryo transfer using ovine FSH. Six days following laparoscopic insemination with fresh semen ewes were flushed surgically and embryos collected. Subsequently, the embryo recovery, along with embryo cryopreservation, embryo survival and quality were assessed. The Karagouniko donor ewes achieved a mean ovulation rate of 11.9 (SE. 0.89). The ova Recovery rate was 80,9% and 87,6% of the ova recovered being fertilised. A total of 327 (77,5%) of the viable embryos were assessed as being of sufficient quality for cryopreservation. The embryos ranged from late morulae to expanded blastocyst and were frozen via a 3-step process in 1.5 M ethylene glycol following repeated washing and trypsination. Cryopreserved embryos were frozen and then transported to Scotland, UK. There, embryos were thawed rapidly and re-hydrated via a 2 step sucrose/ethylene glycol gradient. A total of 92.4% of embryos frozen remained suitable for transfer semi-surgically into synchronized Scottish Blackface ewes. 183 embryos were transferred in total with a 66.1% survival rate. The survival rate of frozen thawed blastocysts(75%) was significantly greater than (P<0.01) that for morulae (48%). It was concluded that MOET could be successfully applied in Greek dairy breeds of sheep as a means for genetic improvement. Frozenembryos could be a successful medium for the transportation of ovine genetic material from and to Greece, however, most likely the choice of embryonic stage for cryopreservation is crucial.


2022 ◽  
Author(s):  
Yuling Mao ◽  
Ping Yin ◽  
Yanfen Luo ◽  
Jingda Qiao ◽  
Lei Li

Abstract Objective: To evaluate the impact of cryopreservation storage duration on embryo viability, implantation competence, pregnancy outcome and neonatal outcomes.Design: Retrospective study.Setting: Center for Reproductive Medicine,The Third Affiliated Hospital of Guangzhou Medical University.Patient(s): In vitro fertilization patients who had vitrified cryopreserved embryos and following the first frozen embryo transfer cycles from January 2004 to August 2019. A total of 31143 patients met the inclusion criteria and were grouped according to the storage time (20926 patients in Group 1 with storage time <3 months, 6472 patients in Group 2 with storage time between 3 and 6 months, 2237 patients in Group 3 with storage time between 6 and 12 months and 746 in Group 4 with storage time between 12 and 24 months, 762 patients in Group 5 with storage time >24 months).Intervention(s): None.Main Outcome Measure(s): In the total FET cycles, the embryo survival rate was decreased significantly with the increase of cryopreservation time, and the highest rate was 98. 63 % in the 1-3 months group, and the lowest was 71.13% in the >=731 days group (P <0. 01). The HCG positive rate (57.85%) and clinical pregnancy rate (55. 26%) in the 1-3 months group were the highest (P<0. 01). The >=731 group had the lowest sex ratio of 0.96. There were no significant differences in neonatal birth weight, neonatal height and congenital anomalies among the groups (P>0. 05).Result(s): Length of storage time had a significant effect on post-thaw survival and outcomes for IVF cycles. Conclusion(s): With the prolongation of cryopreservation time, the embryonic survival rate and pregnancy rate were decreased significantly. Short-term cryopreservation (<=3 months) can obtain higher clinical pregnancy rate. Therefore, although long-term hryopreservation of the embryo has no effect on the health of the new baby, but hryopreserved embryos should be recovery as soon as possible if condition allows.


2015 ◽  
Vol 27 (1) ◽  
pp. 168
Author(s):  
K. Karakas ◽  
H. Alkan ◽  
G. Onur ◽  
D. Ozen ◽  
M. Kaymaz ◽  
...  

The aims of this study were to compare the embryo recovery rate in Angora goats based on application timing; at the beginning (September – October; Group 1) and end (December; Group 2) of the breeding season and to evaluate the viability and survivability of fresh or vitrified-thawed embryos when transferred. For this purpose, nine Angora goats were used as donors and thirthy Angora goats were used as recipients. Donor goats were synchronized and superovulated with traditional protocol and were mated with fertile bucks. At the 156th hour of the mating, embryos were collected surgically and evaluated under a stereo microscope. In group 1, 103 embryos and in group 2, 63 embryos were collected from nine goats. Fresh or vitrified-thawed embryos were transferred surgically to synchronized recipients. In Group 1 fresh/thawed embryos were transferred to 15/15 goats and in group 2, fresh/thawed embryos were transferred to 8/8 goats, respectively. Each recipient received 1 or 2 embryos ipsilateral to the ovary containing one or more corpora lutea. On day 30 of the transfer, goats were examined by transrectal ultrasonography, pregnancy rates of fresh/thawed embryos were 66.6%/26.6% for group 1 and 62.5%/62.5% for group 2. On day 100 of the transfer, goats were examined again by ultrasonography, and pregnancy rates were 46.6%/0% for group 1 and 37.5%/0% for group 2, respectively. After about 50 days, goats were kidded. In group 1, 3 twins and 4 single kids were born; in group 2, 2 twins were born. The total number of collected embryos and pregnancy rates among the groups were analysed using SPSS® (version 14.01, Chicago, IL, USA) and for all comparisons, differences were considered with a minimum of 5% significance level. After statistical analyses, the numbers of collected embryos at the beginning and at the end of the breeding season were compared. There was no difference in freezable/transferable embryo quality. As a result, embryos could be collected after superovulation protocols in Angora goats both at beginning and end of the breeding season, however there might be a decrease in numbers of collected embryos and the reasons for this might not be only the seasonal factors but also the environment, care, nutrition and previous superovulation protocols. The pregnancy rate following transfer of fresh embryos was satisfactory but not all does confirmed pregnant kidded; hence, reducing the number of recipients kidding. The pregnancy rate following transfer of vitrified-thawed embryos was generally low and unsatisfactory. Further research is warrented in improving the cryopreservation techniques and thus the embryo survival rate of Angora goat embryos. This study was financed with the University of Ankara Grant.


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