scholarly journals Development and pregnancy rates of Camelus dromedarius-cloned embryos derived from in vivo- and in vitro-matured oocytes

2021 ◽  
Author(s):  
Young-Bum Son ◽  
Yeon Ik Jeong ◽  
Yeon Woo Jeong ◽  
Per Olof Olsson ◽  
Mohammad Shamim Hossein ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Camelus Dromedarius

scholarly journals Reproductive Outcomes and Endocrine Profile in Artificially Inseminated versus Embryo Transferred Cows

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1359
Author(s):  
Jordana S. Lopes ◽  
Estefanía Alcázar-Triviño ◽  
Cristina Soriano-Úbeda ◽  
Meriem Hamdi ◽  
Sebastian Cánovas ◽  
...  
Keyword(s):  
Neonatal Mortality ◽  
Culture Medium ◽  
Culture Conditions ◽  
Pregnancy Rates ◽  
Gestation Length ◽  
Safe Production ◽  
Endocrine Profile ◽  
Hormonal Levels

The increasing use of in vitro embryo production (IVP) followed by embryo transfer (ET), alongside with cryopreservation of embryos, has risen concerns regarding the possible altered pregnancy rates, calving or even neonatal mortality. One of the hypotheses for these alterations is the current culture conditions of the IVP. In an attempt to better mimic the physiological milieu, embryos were produced with female reproductive fluids (RF) as supplements to culture medium, and another group of embryos were supplemented with bovine serum albumin (BSA) as in vitro control. Embryos were cryopreserved and transferred while, in parallel, an in vivo control (artificial insemination, AI) with the same bull used for IVP was included. An overview on pregnancy rates, recipients’ hormonal levels, parturition, and resulting calves were recorded. Results show much similarity between groups in terms of pregnancy rates, gestation length and calves’ weight. Nonetheless, several differences on hormonal levels were noted between recipients carrying AI embryos especially when compared to BSA. Some calving issues and neonatal mortality were observed in both IVP groups. In conclusion, most of the parameters studied were similar between both types of IVP derived embryos and the in vivo-derived embryos, suggesting that the IVP technology used was efficient enough for the safe production of calves.

Fertilizing capacity of epididymal and testicular sperm using intracytoplasmic sperm injection (ICSI)

1995 ◽  
Vol 7 (2) ◽  
pp. 281 ◽  
Author(s):  
SJ Silber ◽  
P Devroey ◽  
H Tournaye ◽  
Steirteghem AC Van
Keyword(s):  
Pregnancy Rate ◽  
Intracytoplasmic Sperm Injection ◽  
Pregnancy Rates ◽  
Obstructive Azoospermia ◽  
Motile Sperm ◽  
Epididymal Sperm ◽  
Testicular Sperm ◽  
Cleavage Rate

For men with uncorrectable obstructive azoospermia, their only hope of fathering a child is microsurgical epididymal sperm aspiration (MESA) combined with in vitro fertilization (IVF). In 1988, proximal epididymal sperm were demonstrated to have better motility than senescent sperm in the distal epididymis, and it was thought that retrieval of motile sperm from the proximal epididymis would yield reliable fertilization and pregnancy rates after conventional IVF. However, the results to date have been poor, and although a minority of patients achieved good fertilization rates with IVF, the vast majority (81%) had consistently poor or no fertilization and the pregnancy rate averaged only 9%. Recently, intracytoplasmic sperm injection (ICSI) has been successfully used to achieve fertilization and pregnancies for patients with extreme oligoasthenozoospermia. ICSI has therefore been applied to cases of obstructive azoospermia and, in this report, 67 MESA-IVF cases are compared with 72 MESA-ICSI cases. The principle that motile sperm from the proximal segments of the epididymis should be used for ICSI was followed, although in the most severe cases in which there was an absence of the epididymis (or absence of sperm in the epididymis), testicular sperm were obtained from macerated testicular biopsies. These sperm only exhibited a weak, twitching motion. In 72 consecutive MESA cases, ICSI resulted in fertilization and normal embryos for transfer in 90% of the cases, with an overall fertilization rate of 46%, a cleavage rate of 68%, and ongoing or delivered pregnancy rates of 46% per transfer and 42% per cycle. The pregnancy and take-home baby rates increased from 9% and 4.5% with IVF to 53% and 42% with ICSI. There were no differences between the results for fresh epididymal, frozen epididymal or testicular sperm, and the number of eggs collected did not affect the outcome. The results were also unaffected by the aetiology of the obstruction such as congenital absence of the vas deferens or failed vasoepididymostomy. The only significant factor which affected the pregnancy rate was female age. It is concluded that although complex mechanisms involving epididymal transport may be beneficial for conventional fertilization of human oocytes (in vivo or in vitro), none of these mechanisms are required for fertilization after ICSI. Given the excellent results with epididymal and testicular sperm, ICSI is obligatory for all future MESA patients. Finally, the use of ICSI with testicular sperm from men with non-obstructive azoospermia is also discussed.

108 THE EFFECTS OF FORSKOLIN ON BRAHMAN IN VIVO-PRODUCED EMBRYOS AND SUBSEQUENT PREGNANCY RATES POST-THAW

2012 ◽  
Vol 24 (1) ◽  
pp. 166
Author(s):  
J. H. Pryor ◽  
J. A. Trant ◽  
C. R. Long ◽  
D. W. Forrest ◽  
C. R. Looney
Keyword(s):  
Animal Health ◽  
Survival Rates ◽  
Slow Cool ◽  
Subsequent Pregnancy ◽  
Pregnancy Rates ◽  
Phase Iii ◽  
Study Phase ◽  
Grade One

It has been hypothesised that lower survival rates post-thaw of Brahman (BR) embryos is due in part to higher intracellular lipids. The objective of this study was to determine whether pregnancy rates of in vivo–produced BR embryos could be enhanced by exposing them to 10 μM of forskolin (FSK; lipolytic agent) before slow cool freezing, thawing and direct transfer. A previous report (Phase II) indicated that the addition of FSK to culture 24 h before freezing 7 days in vitro–produced, BR-sired embryos can increase survival and blastocyst hatching rates (Pryor et al. 2010 Reprod. Fertil. Dev. 22, 214). For this study (Phase III), 11 BR cows were injected with 25 mg of Lutalyse® (Pfizer Animal Health, New York, NY, USA) to synchronize oestrus. On Day 0 (7 days after synchronized oestrus), all donors were inserted with a CIDR (Controlled Intravaginal Releasing Device; Pharmacia Co., Kalamazoo, MI, USA) and injected with 50 mg of progesterone plus 2.5 mg of oestradiol 17 β (Medshop, Longview, TX, USA). On Day 4, decreasing doses of Folltropin (Bioniche, Pullman, WA, USA) ranging from 160 to 264 mg were given twice daily for 3 days along with 0.625 mg of cloprostenol (Estrumate; Merck/Schering-Plough Animal Health, Whitehouse Station, NJ, USA) given twice on Day 6 (AM and PM) and CIDR removal on Day 7 AM. Donors were artificially inseminated 12 and 24 h post-oestrus (oestrus = Day 8) with frozen/thawed BR semen. On Day 14 (embryo age 6 days), donors were nonsurgically collected, producing a total of 75 grade-one morulae, which were randomly allocated and cultured for 24 h in Evolve (Zenith Biotechnology, Canada) supplemented with 4 mg mL–1 of Probumin BSA (Millipore, Norcross, GA, USA; control) or with FSK (Sigma, St. Louis, MO, USA) at 38.5°C under a 5% CO2, 5% O2 and 90% N2 humidified atmosphere. Immediately following treatment, 7 days compact morula/blastocysts were washed in Vigro Holding Plus medium (Bioniche, Belleville, Ontario, Canada) and submitted to Vigro Ethylene Glycol Freeze Plus medium (Bioniche) for 5 to 7 min before being frozen at 0.5°C min–1 from –6°C to –32°C and plunged in liquid nitrogen. Frozen embryos (n = 35 FSK; n = 35 control) were air thawed for 7 s and then immersed in 30°C H2O for 10 s before being nonsurgically transferred into synchronized recipients. Pregnancy rates were assessed by ultrasonography via rectal palpation 30 to 60 days post-transfer. Contingency analysis was performed using forskolin treatment, technician and embryo stage and location as independent variables (JMP 8.0, SAS Institute Inc., Cary, NC, USA). There were no statistical differences or interactions for any of the analysed variables. Pregnancy rates between control and FSK treatments did not vary (34.3 and 31.4%, respectively; chi-square P = 0.80). In conclusion, treating in vivo–produced BR embryos with 10 μM of forskolin for 24 h did not alter pregnancy rates. The authors acknowledge support from the American Brahman Breeders Association.

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.

29 Co-Incubation of Equine Cloned Embryos with Sialic Acid: Effect on Pregnancy Rate

2018 ◽  
Vol 30 (1) ◽  
pp. 154
Author(s):  
D. Vichera ◽  
R. Olivera ◽  
V. Arnold ◽  
J. Vergara ◽  
R. Jordan ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Pregnancy Rate ◽  
Chemical Activation ◽  
Transrectal Ultrasound ◽  
Morphological Characteristics ◽  
Pregnancy Rates ◽  
Adhesive Properties ◽  
Acid Effect

In vitro-produced equine embryos have certain morphological characteristics that differ from embryos produced in vivo. One of them is the absence or inadequate formation of the embryo capsule. This capsule is composed of mucin-like glycoproteins produced by the trophectoderm with high proportion of sialic acid, which confers anti-adhesive properties. This characteristic is necessary for the intrauterine embryo migration process to occur, which is vital and fundamental for pregnancy recognition. For this reason, inadequate formation of the glycoprotein capsule could result in a lower pregnancy rate. In this study, we aimed to evaluate the effect of cloned equine embryo co-incubation with sialic acid on blastocyst and pregnancy rates. To achieve this, equine oocytes obtained from slaughterhouse ovaries were matured in TCM-199 HEPES medium with 2% fetal bovine serum, 2% antibiotics, and 1 μg mL−1 FSH, incubated at 39°C for 24 h. Matured oocytes were denuded with pronase, enucleated, and fused to donor bone marrow mesenchymal cells according to Olivera et al. (2016 PLoS One 11, e0164049, 10.1371/journal.pone.0164049). Chemical activation was induced using 8.7 μM ionomycin for 4 min and embryos were incubated with 1 mM 6-DMAP and 5 mg mL−1 cycloheximide for 4 h. Afterwards embryos were cultured in microwells for 8 days in DMEM-F12 medium. On Day 6, cloned equine embryos were exposed to 5 μM sialic acid for 48 h (SA group). On Day 8, blastocysts were transferred to recipient mares 5 days post-ovulation and pregnancy was confirmed 15 days post-transfer by transrectal ultrasound. Embryo clones generated without sialic acid exposure were used as a control (C) group. Fisher test was used to analyse both blastocyst and pregnancy rates. Blastocyst rates were 14% (46/328) and 15% (62/413) and pregnancy rates were 30.4% (7/23) and 19.4% (6/31) for SA and C groups, respectively. No statistical differences were observed between groups for the analysed parameters, even though pregnancy rates tended to be higher in the SA group. This effect could be a consequence of higher concentrations of the glycoprotein involved in the formation of the embryo capsule.

Biopsy of bovine embryos produced in vivo and in vitro does not affect pregnancy rates

2017 ◽  
Vol 90 ◽  
pp. 25-31 ◽  
Author(s):  
Regivaldo Vieira de Sousa ◽  
Célia Regina da Silva Cardoso ◽  
Guilberth Butzke ◽  
Margot Alves Nunes Dode ◽  
Rodolfo Rumpf ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Bovine Embryos

scholarly journals 68 REPRODUCTIVE PERFORMANCE OF CLONED BULLS

2005 ◽  
Vol 17 (2) ◽  
pp. 184
Author(s):  
R.T. Tecirlioglu ◽  
M.A. Cooney ◽  
N.A. Korfiatis ◽  
R. Hodgson ◽  
M. Williamson ◽  
...  
Keyword(s):  
Reproductive Performance ◽  
Embryo Cell ◽  
Pregnancy Rates ◽  
Small Data ◽  
Data Set ◽  
Cell Numbers ◽  
Cell Counts ◽  
Original Donor

The generation of cloned animals by somatic cell nuclear transfer has been reported in a number of countries worldwide. However, progress has been impeded by the extremely low efficiency of cloning and health of some of the cloned animals. Surprisingly, little is known of the reproductive performance of viable clones when compared to the original cell donor, given that a major motivation of cloning is dissemination of superior genotypes. The aim of this study was to compare semen collected from three cloned bulls (Clones A1, B1, and B2) to that of the original donor bulls (Donors A and B). Parameters examined included ejaculate volume, sperm concentration and the motility characteristics of frozen/thawed semen using computed-assisted semen analysis (Hamilton Thorne Biosciences, Inc., Beverly, MA, USA). The fertilization ability of each semen sample was examined using in vitro matured oocytes derived from abattoir-source ovaries. Frozen/thawed semen samples from donor and cloned bulls were prepared on a Percoll gradient and diluted with fertilization medium to a concentration of 1 million sperm/mL prior to fertilization (IVF). The number of blastocysts and total cell counts were analyzed on Day 7 of culture. Finally, in vitro-fertilized blastocysts (Day 7) were transfer to synchronized recipients (n = 49) to examine in vivo development. Proportional data for the in vitro development of embryos and subsequent pregnancy rates were analyzed by chi-square test, and embryo cell numbers were analyzed using Student's t-test. Progressive motility percentage between donor and cloned bull did not differ: Donor A (62.25 ± 3.89, n = 12); Donor B (66.69 ± 4.47, n = 13); Clone A1 (71.37 ± 8.57, n = 8); Clone B1 (73.75 ± 2.42, n = 12); Clone B2 (72.41 ± 3.26, n = 12). No obvious differences in kinetic motility parameters were evident between cloned and non-cloned donor animals. However, blastocyst rates were significantly higher in cloned bulls (Clone A1: 30.9%, 81/262; Clone B1: 34.4%, 98/285; and Clone B2: 42.9%, 120/280) compared to donor bulls (Donor A: 20.7%, 54/261; Donor B: 20.9%, 76/364). Total embryo cell numbers did not differ significantly between donor bulls (Donor A: 138.3 ± 5.3, n = 39; Donor B: 133.2 ± 5.2, n = 47) and cloned bulls (Clone A1; 126.3 ± 4.4, n = 45; Clone B1: 134.4 ± 7.1, n = 26; and Cloned B2: 140.1 ± 3.9, n = 46). Initial pregnancy rates on Day 30 were also not different between Donor A (42.3%, 11/26) and Clone A1 (47.8%, 11/23). Preliminary observations from the small data set on postpubertal cloned bulls indicate that semen production, semen morphology, and reproductive performance (in vitro and in vivo) were similar in terms of semen characteristics and reproductive performances when compared to their original donor bulls.

Role of progesterone in embryo development in cattle

2016 ◽  
Vol 28 (2) ◽  
pp. 66 ◽  
Author(s):  
Pat Lonergan ◽  
Niamh Forde ◽  
Thomas Spencer
Keyword(s):  
Corpus Luteum ◽  
Embryo Development ◽  
Early Pregnancy ◽  
Pregnancy Rates ◽  
In Vitro Experiments ◽  
Conceptus Development

Progesterone (P4) from the corpus luteum is critical for the establishment and maintenance of pregnancy and plays a major role in regulating endometrial secretions essential for stimulating and mediating changes in conceptus growth and differentiation throughout early pregnancy in ruminants. Numerous studies have demonstrated an association between elevated systemic P4 and acceleration in conceptus elongation. A combination of in vivo and in vitro experiments found that the effects of P4 on conceptus elongation are indirect and mediated through P4-induced effects in the endometrium. Despite effects on elongation, data on the effects of post-insemination supplementation with P4 on pregnancy rates are conflicting. This review highlights the effects of P4 on conceptus development and examines strategies that have been undertaken to manipulate P4 concentrations to increase fertility.

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