229 OOCYTE RECOVERY, IN VITRO FERTILIZATION AND EMBRYO TRANSFER IN THE SERVAL (LEPTAILURUS SERVAL)

2006 ◽  
Vol 18 (2) ◽  
pp. 223 ◽  
Author(s):  
C. E. Pope ◽  
M. C. Gómez ◽  
A. Cole ◽  
C. Dumas ◽  
B. L. Dresser
Keyword(s):  
Embryo Transfer ◽  
Oocyte Retrieval ◽  
Sub Saharan Africa ◽  
Medium Size ◽  
Embryo Survival ◽  
Frozen Semen ◽  
Old Donor ◽  
Fresh Embryos

Servals are medium size (9 to 18 kg) spotted cats found in sub-Saharan Africa that are protected by CITES under Appendix II regulations. There are at least six sub-species, one of which is listed as Endangered by the U.S. Endangered Species Act. In vitro-derived embryos have been produced in at least one-half of the 36 species of nondomestic cats, and kittens have been born after embryo transfer in six species. In the present study we evaluated (1) ovarian response of servals to repeated exogenous gonadotropin stimulation, and (2) in vitro and in vivo developmental ability of in vitro-derived embryos. One two-year-old and one five-year-old female were treated six and three times, respectively, over a 3.5-year period, with a total of 20 or 25 IU of porcine FSH (i.m.; Sioux Biochem, Sioux City, IA, USA) administered daily over four days during interestrus. On Day 5, 15 IU of porcine LH (i.m.; Sioux Biochem) was given, and laparoscopic oocyte retrieval was performed 24 h later. A total of 234 preovulatory oocytes were recovered: 182 (mean = 30.3) from the two-year-old and 52 (mean = 17.3) from the five-year-old female. A total of 91 and 91 oocytes were recovered at retrievals 1 through 3 and 4 through 6, respectively, from the two-year-old donor. Eighty oocytes from the two-year-old donor were inseminated with cooled (24 h, 4°C) semen. Frozen semen from the same male was used to inseminate 102 oocytes from the two-year-old female and 52 oocytes from the five-year-old female. Overall, 136 embryos (58% cleavage frequency) were produced: 119 (65% cleavage frequency) from the two-year-old and 17 (33% cleavage frequency) from the five-year-old female. Cleavage frequency of oocytes from the two-year-old female inseminated with cooled or frozen semen was similar, 68% (54/80) and 64% (65/102), respectively. Embryos were cultured for 5 or 6 days before controlled rate cryopreservation or uterine transfer (Gómez et al. 2003 Theriogenology 60, 239–251). On Day 5, 66 early to mid-stage morulae were cryopreserved at a slow controlled rate. Sixty Day 5 and 18 Day 6 embryos were auto-transferred to a recipient (8 to 26/transfer) in a total of six surgical procedures, of which five were with fresh embryos (n = 70) and one was with cryopreserved embryos (n = 8). The sixth embryo transfer procedure (26 fresh embryos) resulted in the unassisted birth of a live male kitten on Day 77 of gestation. We have shown that in vitro-derived embryos can be generated in the serval and that oocyte retrieval rates and cleavage frequencies are comparable to those reported for other species of mid-sized nondomestic cats. The nominal incidence of pregnancy and frequency of embryo survival may be improved by transferring early cleavage staged embryos into the oviduct, as demonstrated in the African wildcat (Felis silvestris lybica; Gómez et al. 2004 Cloning and Stem Cells 6, 247–258). This work was partially funded by the Dan Heard Conservation Challenge Grant.

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.

66 DEVELOPMENT OF IN VITRO-PRODUCED CAT EMBRYOS AFTER VITRIFICATION AND NONSURGICAL EMBRYO TRANSFER: PRELIMINARY RESULTS

2009 ◽  
Vol 21 (1) ◽  
pp. 133
Author(s):  
E. Iacono ◽  
B. Merlo ◽  
M. Regazzini ◽  
D. Zambelli
Keyword(s):  
Ethylene Glycol ◽  
Embryo Transfer ◽  
Domestic Cat ◽  
Embryo Survival ◽  
Abdominal Ultrasonography ◽  
Preliminary Results ◽  
Pregnancy Status ◽  
Viable Embryo

There are no refereed reports on vitrification of domestic cat embryos derived from in vitro-matured oocytes and transferred using a nonsurgical embryo transfer technique. The aim of this study was to verify the effects of vitrification on the in vitro and in vivo developmental ability of in vitro-produced (IVP) cat blastocysts. Oocytes recovered from minced ovaries were matured, fertilized, and cultured in vitro as previously reported (Merlo B et al. 2005 Theriogenology 63, 2032–2039). On Day 7 of in vitro culture (IVC), blastocysts were selected and vitrified in straws (Cristal ET 0.25 mL, 133 mm, IMV-Technologies, Paillette Crista, France). For vitrification (modified from Campos-Chillòn LF et al. 2006 Theriogenology 65, 1200–1214), the embryos were transferred in 1 mL of V1 [ethylene glycol 3.5 m in HEPES synthetic oviductal fluid (HSOF)] for 3 min, and then in 10 μL of V2 (ethylene glycol 7 m, galactose 0.5 m, Ficoll 70 18% in HSOF) for 20 s. Finally, the embryos were loaded in straws preloaded with 190 μL of dilution solution (galactose 0.5 m in HSOF). Straws were heat sealed and immediately plunged into liquid nitrogen. Vitrified embryos were warmed in air for 10 s, and then in a waterbath at 37°C for 30 s. For developmental ability and in vitro evaluation, 27 embryos were warmed and immediately examined: 25 re-expanded, 2 did not re-expand, and 1 had damaged zona pellucida. Re-expanded embryos were cultured in SOF plus amino acids, 16 mg mL–1 BSA, and 5% fetal bovine serum at 38.5°C in 5% O2, 5% CO2, 90% N2. After 24 h of IVC, only 4 blastocysts were expanded, and after 48 h, embryos were clearly degenerated or shrunk. in vivo developmental ability was tested by nonsurgical embryo transfer of 8 vitrified-warmed embryos and 6 IVP fresh embryos into 2 natural estrus queens, injected with 200 IU of hCG i.m. (Day 0) for induction of ovulation. Ovulation was confirmed by plasmatic progesterone assay on Day 5. Nonsurgical embryo transfer was made on Day 8 using the catheter proposed by Zambelli et al. 2001 for transcervical insemination in the cat. The catheter was connected to a 1-mL syringe and loaded with the embryos. Then, it was inserted in the vagina and transrectally guided into the uterus, where the embryos were deposited. To assess pregnancy status, abdominal ultrasonography was done on recipients on Day 13, 25, and 40. On Day 13, an embryonic vesicle was observed in both queens, although a smaller diameter than expected was detected in the recipient of the vitrified embryos. On Day 25, a viable embryo was detected only in the recipient of fresh IVP embryos. On Day 40, the gestational chamber was still present but no sign of a viable embryo was detected. Further studies are in progress to improve the nominal incidence of pregnancy and frequency of embryo survival after vitrification. Nevertheless, the preliminary results obtained using an AI catheter for nonsurgical embryo transfer are encouraging, and the improvement of the technique could make it reliable in the cat. Supported by Animal Stem Cells Laboratory, Regione Emilia Romagna, PRRIITT Project Number M-404AIWTSV.

Establishing twin pregnancies in cattle by embryo transfer

1995 ◽  
Vol 1995 ◽  
pp. 140-140
Author(s):  
K D Sinclair ◽  
P J Broadbent ◽  
D F Dolman ◽  
R G Watt ◽  
J S Mullan
Keyword(s):  
Embryo Transfer ◽  
Survival Rates ◽  
Embryo Survival ◽  
Transfer Method ◽  
Twin Pregnancies

Various methods of creating twin pregnancies in cattle have been investigated by other authors (see review by Sreenan and Diskin, 1987). However, virtually all of these methods have involved in vivoproduced embryos which, in separate studies, have employed either surgical or non-surgical transfer techniques, where embryos were transplanted either unilaterally or bilaterally in recipients which may or may not have been previously artificially inseminated. There have been no studies where all of these factors were examined collectively, and included with the transplantation of either frozen-thawed in vivoor in vitroproduced embryos. The objectives of the current study were, therefore, to compare pregnancy, twinning and embryo survival rates of recipients in which twin pregnancies were induced by various combinations of embryo source and transfer method to animals inseminated or not prior to embryo transfer, and the distribution of the embryos in the uterus.

Establishing twin pregnancies in cattle by embryo transfer

1995 ◽  
Vol 61 (1) ◽  
pp. 25-33 ◽  
Author(s):  
K. D. Sinclair ◽  
P. J. Broadbent ◽  
D. F. Dolman ◽  
R. G. Watt ◽  
J. S. Mullan
Keyword(s):  
Embryo Transfer ◽  
Twin Pregnancy ◽  
Beef Cows ◽  
Pregnancy Rates ◽  
Uterine Horn ◽  
Embryo Survival ◽  
Pregnancy Status ◽  
Twin Pregnancies

AbstractAn experiment zoas conducted to assess differing methods of twin pregnancy establishment in Hereford × British Friesian beef cows and heifers. The experiment was 2 × 2 × 2 × 2 factorial design in which the factors were (i) source of embryos (in vivo or in vitro produced); (ii) pregnancy status of recipient (inseminated or non-inseminated); (Hi) method of embryo transfer (surgical or cervical); and (iv) uterine location of a native and transferred embryo, or two transferred embryos (both located in the ipsilateral, or one in each of the ipsi and contralateral uterine horns). Pregnancy and twinning rates for 285 animals used for embryo transfer were initially diagnosed at day 56 after induced oestrus by transrectal ultrasonography. Subsequently, calving rate and birth weiglit at calving were recorded.Pregnancy rates at day 56 after induced oestrus were similar for both surgical and cervical transfers (58·6% v. 55·2%), as was the case for twinning rate (36·2% v. 30·0%). Similarly, there were no differences between these two methods of transfer (50·0% v. 46·9%) and (26·1% v. 17·7%) for calving and twin calving rates respectively. Recipients which had two embryos located in the ipsilateral uterine horn had higher (P < 0·001) pregnancy rates (66·6% v. 47·3%) but similar twinning rates (32·6% v. 33·4%) at day 56 after induced oestrus to recipients which had one embryo located in each horn. A greater (P < 0·05) percentage of recipients with two embryos originally located in the ipsilateral horn calved (56·0% v. 41·0%) but fewer (P > 0·05) produced twins (17·8% v. 25·7%) than was the case for recipients which originally had one embryo located in each horn. In vivo produced embryos resulted in higher (P < 0·001) pregnancy rates (74·4% v. 39·7%) and twinning rates (48·3% v. 18·0%) at day 56, and higher (P < 0·001) calving rates (64·5% v. 32·7%) and twin calving rates (36·3% v. 7·6%) than did in vitro produced embryos. Inseminated (Al + ET) recipients had slightly greater (P>0·05) pregnancy rates (61·6% v. 51·6%) and twinning rates (36·9% v. 28·7%) than non-pregnant recipients which received two embryos. A greater (P<0·05) percentage of inseminated recipients (Al + ET) calved (54·3% v. 42·0%) than was the case for non-pregnant recipients which received two embryos. The percentage producing twins at calving were similar for these two methods of twin pregnancy establishment.Embryo survival to day 56 after induced oestrus averaged 45·0% and was found to be non-independent of its co-twin. From day 56 to parturition foetal loss averaged 21·0% and foetal survival was found to be independent of the fate of its co-foetus. Twin foetuses located in the same uterine horn were lighter at birth than twin foetuses located in separate uterine horns (33·0 v. 35·2 kg; P < 0·05).

scholarly journals 332IN VIVO DEVELOPMENTAL CAPACITY OF IN VITRO-PRODUCED EMBRYOS DERIVED FROM SEX-SORTED AND RE-CRYOPRESERVED FROZEN-THAWED RAM SPERM

2004 ◽  
Vol 16 (2) ◽  
pp. 286 ◽  
Author(s):  
J.K. O'Brien ◽  
F.K. Hollinshead ◽  
G. Evans ◽  
W.M.C. Maxwell
Keyword(s):  
Potential Application ◽  
High Speed ◽  
Fisher Exact Test ◽  
Embryo Survival ◽  
Food Dye ◽  
Frozen Semen ◽  
Developmental Capacity ◽  
Ram Sperm

The ability to sort and re-freeze frozen-thawed sperm would significantly increase the potential application of sperm sexing technology to species management. Frozen-thawed, sorted, re-frozen and then thawed ram sperm appear fully functional in vitro with blastocyst production greater than that for frozen-thawed, non-sorted sperm (Hollinshead FK et al. 2003 Theriogenology 59, 209 abst). The aim of this study was to evaluate the in vivo capacity of in vitro-produced embryos derived from frozen-thawed sperm after sorting and a second cryopreservation/thawing step. Frozen semen from 2 rams (n=3 ejaculates per ram) was used throughout. Post-thaw sperm treatments comprised (i) non-sorted (Control); (ii) sorted (Froz-Sort) and (iii) sorted, then re-frozen (Froz-Sort-Froz). X and Y sperm were separated using a high-speed sorter (SX MoFlo®, DakoCytomation, Fort Collins, CO, USA) after incubation with Hoechst 33342 and food dye to eliminate nonviable sperm. Reanalysis revealed high levels (mean±SEM) of purity for X- and Y-enriched samples for all treatments (89±1.2%). At Day 6 post-insemination, 2 embryos (blastocyst stage or greater) were transferred per recipient. Data were analyzed by chi-square and Fisher Exact Test. In vivo embryo survival was similar across sperm treatments (28/64, 43.8% overall) and 20 of 23 (87.0%) sexed lambs were of the predicted sex (Table 1). These results demonstrate high in vivo developmental capacity of in vitro-produced sexed embryos derived from frozen-thawed ram sperm after sorting and a second cryopreservation/thawing step, and increase the potential application of sperm sexing technology. Research support by XY, Inc., Australian Research Council and Zoological Parks Board of NSW. Table 1 In vivo survival of transferred in vitro produced embryos derived from frozen-thawed non-sorted (Control), frozen-thawed and sorted (Froz-Sort) and frozen-thawed, sorted, then frozen-thawed (Froz-Sort-Froz) ram sperm.

scholarly journals 88 PREGNANCY RATES FOR IN VITRO AND IN VIVO PRODUCED OVINE EMBRYOS VITRIFIED USING THE MINIMUM VOLUME COOLING CRYOTOP METHOD

2005 ◽  
Vol 17 (2) ◽  
pp. 194
Author(s):  
J. Kelly ◽  
D. Kleemann ◽  
M. Kuwayama ◽  
S. Walker
Keyword(s):  
Pregnancy Rate ◽  
Sucrose Solution ◽  
Bovine Embryo ◽  
Minimum Volume ◽  
Embryo Viability ◽  
Embryo Survival ◽  
First Order ◽  
Fresh Embryos

Previously we reported that, using the minimum volume cooling (MVC) cryotop vitrification method, in vitro-produced ovine and bovine embryo survival after thawing was similiar to that of fresh embryos (Kelly et al. 2004 Reprod. Fert. Dev. 16, 172). While survival of vitrified embryos after thawing can be indicative of embryo viability, this assessment does not always correlate with embryo survival after transfer. This study assesses the effect of vitrification using the MVC cryotop method on the survival after transfer of in vitro- and in vivo-produced ovine embryos. Fresh or vitrified Day 6 ovine embryos (expanded blastocysts, blastocysts, compact morulae) were used in this study. Ovine cumulus–oocyte complexes were obtained and matured, fertilized (Day 0), and cultured in vitro (Walker et al. 1996 Biol. Reprod. 55, 703–708). In vitro embryos for vitrification were produced and vitrified (Kelly et al. 2004 Reprod. Fert. Dev. 16, 172) 10 days prior to the day of transfer. In vivo embryos were recovered from donor Merino ewes and vitrified 7 days prior to the day of transfer while fresh in vivo embryos were collected and transferred on the same day. Semen used for both in vivo and in vitro embryo production was from the same sire. On the day of transfer, vitrified embryos were thawed directly into 1.25 M sucrose solution, followed by stepwise dilution of the cryoprotectants. Embryos were transferred as singles into synchronized recipient ewes on a randomized basis. Fetal number was detected at Day 50. Variables were assessed using the CATMOD procedure in SAS. Pregnancy rate for in vivo-derived embryos was higher (P < 0.01) than for in vitro-derived embryos. Embryo treatment (fresh vs. vitrified) did not significantly affect pregnancy rate. Pregnancy rate for ewes detected (by vasectomized rams) in estrus within 48 h of progesterone pessary removal was higher (P < 0.05) than for both the 48–68 h and unmarked groups. The latter two groups did not differ significantly. None of the first-order interactions were significant (P > 0.05). This study demonstrates that ovine embryos (in vitro and in vivo) can be vitrified, thawed, and transferred without compromising embryo viability. However, the differences in pregnancy rate between the recipient groups warrant further investigation. The MVC cryotop method is a vitrification technique that can be adapted to routine field use. Table 1. Pregnancy rate of fresh and vitrified in vivo and in vitro ovine embryos after embryo transfer

ECONOMIC PERSPECTIVES OF BIOTECHNOLOGIES USING IN ANIMAL FARMING

1970 ◽  
pp. 57-60
Author(s):  
I. F. Gorlov ◽  
A. A. Mosolov ◽  
G. V. Komlatskiy ◽  
M. A. Nesterenko ◽  
K. D. Nimbona ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Economic Effect ◽  
Dairy Herd ◽  
State Level ◽  
Modern Level ◽  
Economic Perspectives ◽  
The Cost ◽  
Short Time

The article presents materials on the study of the possibility of reproduction and increase in the herd of highly productive cows through the use of embryo transplantation technology. The classical (in vivo) and more modern, developing (in vitro) methods of embryotransfer, their positive and negative sides are considered in detail. The possibility of accelerating the breeding process by using the method of transplantation, in which from one cow can be obtained from 10 to 100 calves, which will allow for 4-5 years, almost any herd (of any size and breed) with the help of biotechnology to turn into a cattle-breeding enterprise of the most modern level. At the same time, heifers obtained from unproductive cows can be used as "surrogate" mothers who are transplanted with the best donor embryos, which allows to obtain a full-fledged offspring adapted to local environmental conditions. A detailed scheme of obtaining, evaluation, storage, as well as the cost and economic effect of embryo transplantation was calculated, the market was evaluated, the required annual volume of transplants and the number of donor cows for large livestock farms were determined. As a positive example of "Scientific-production enterprise "Centre of biotechnology and embryo transfer" in 2014, implemented a project for accelerated replacement and genetic improvement of the dairy herd, engraftment averaged 57-69%, and the economic effect of the enterprise from getting a single animal by the method of embryo transfer, compared with imports of similar close in quality, ranged from 60 to 100 thousand rubles on his head. It is shown that it is necessary to organize at the state level a developed service for embryo transplantation to reduce the cost of embryo transfer and the possibility of creating in a short time in the country's own highly productive breeding nucleus of dairy and beef cattle, which will reduce, and in the future completely eliminate, import dependence on cattle products.

Effects of rate of development of in vitro-produced ovine embryos on sex ratio and in vivo survival after embryo transfer

1997 ◽  
Vol 48 (8) ◽  
pp. 1369-1378 ◽  
Author(s):  
S.L. Catt ◽  
J.K. O'Brien ◽  
W.M.C. Maxwell ◽  
G. Evans
Keyword(s):  
Sex Ratio ◽  
Embryo Transfer ◽  
Rate Of Development

scholarly journals Schistosoma mansoni eggs induce Wnt/β-catenin signaling and activate the protooncogene c-Jun in human and hamster colon

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jakob Weglage ◽  
Friederike Wolters ◽  
Laura Hehr ◽  
Jakob Lichtenberger ◽  
Celina Wulz ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Colorectal Carcinogenesis ◽  
Sub Saharan Africa ◽  
Interleukin 4 ◽  
Health Burden ◽  
Sub Saharan ◽  
Considerable Morbidity ◽  
First Time

AbstractSchistosomiasis (bilharzia) is a neglected tropical disease caused by parasitic flatworms of the genus Schistosoma, with considerable morbidity in parts of the Middle East, South America, Southeast Asia, in sub-Saharan Africa, and particularly also in Europe. The WHO describes an increasing global health burden with more than 290 million people threatened by the disease and a potential to spread into regions with temperate climates like Corsica, France. The aim of our study was to investigate the influence of S. mansoni infection on colorectal carcinogenic signaling pathways in vivo and in vitro. S. mansoni infection, soluble egg antigens (SEA) and the Interleukin-4-inducing principle from S. mansoni eggs induce Wnt/β-catenin signaling and the protooncogene c-Jun as well as downstream factor Cyclin D1 and markers for DNA-damage, such as Parp1 and γH2a.x in enterocytes. The presence of these characteristic hallmarks of colorectal carcinogenesis was confirmed in colon biopsies from S. mansoni-infected patients demonstrating the clinical relevance of our findings. For the first time it was shown that S. mansoni SEA may be involved in the induction of colorectal carcinoma-associated signaling pathways.

scholarly journals 183PREGNANCY RATE FOLLOWING TRANSFER OF IN VITRO- AND IN VIVO-PRODUCED BOVINE EMBRYOS TO LH-TREATED RECIPIENTS

2004 ◽  
Vol 16 (2) ◽  
pp. 213 ◽  
Author(s):  
J. Small ◽  
M. Colazo ◽  
D. Ambrose ◽  
R. Mapletoft ◽  
J. Reeb ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Animal Health ◽  
Beef Cows ◽  
Water Bath ◽  
Bovine Embryos ◽  
Chi Square ◽  
Embryo Survival ◽  
Frozen Embryos

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 &gt;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 &gt;10mm. Overall, 7.1% of the recipients had two CL on the day of embryo transfer. There was no effect (P&gt;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&gt;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&lt;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

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