106 COMPARISON OF IN VITRO EMBRYO PRODUCTION IN PANAMA USING HOLSTEIN OOCYTES WITH BOS INDICUS SEXED SEMEN FROM DONORS IN DIFFERENT LOCATIONS: FIELD DATA

2016 ◽  
Vol 28 (2) ◽  
pp. 183
Author(s):  
S. J. R. Rodriguez ◽  
Y. E. Ramirez ◽  
E. Gomes ◽  
L. F. Nasser ◽  
J. H. F. Pontes ◽  
...  
Keyword(s):  
Embryo Development ◽  
Bos Indicus ◽  
Embryo Production ◽  
University Of Illinois ◽  
Oocyte Collection ◽  
Sexed Semen ◽  
The Usa ◽  
In Vitro Embryo Production ◽  
The University

The objective of this work was to compare in vitro embryo production of Bos taurus × Bos indicus cross embryos using oocytes from Holstein donors under different production and environment systems. This study also examined the possibility for in vitro production using oocytes imported and transported fresh between the USA and Panama. All animals were mature Holstein cows going through a normal lactation. The first group of donors was from the University of Illinois dairy herd and went through 3 ovum pickup sessions. The second group of donors were Holstein cows already adapted to Panama and went through 10 ovum pickup sessions. The Panamanian herd of Holstein donors were born and raised in Panama in an area of mountains, on average 1300 m above sea level. This environment does not have the typical hot and humid tropical weather seen in other regions of Panama. Both groups of donors were aspirated without stimulation during the years 2013 and 2014. Oocytes recovered from donors in Illinois were imported fresh under a special sanitary research protocol between Panama and the University of Illinois. The transport of fresh oocytes from the USA to Panama was done using a portable incubator set at 39°C (Minutube of America). Oocytes were matured during transport in 5-mL tubes (~30–35 oocytes per tube) containing 400 µL of maturation media (TCM-199) that had been equilibrated with 5% CO2. Oocytes recovered from donors in Panama were matured using the same media. For both groups, oocytes were inseminated 24 h after ovum pickup using sexed semen from the same bull. All embryo production procedures followed the protocols of the In vitro Brasil™ commercial system. At 72 h postinsemination, cleavage was evaluated. On Day 7 after insemination, embryo development to the blastocyst stage (early to expanded) was recorded. Data were analysed using Chi-squared. As shown in Table 1, there was no effect of oocyte collection location on embryo development. These results indicate that it is possible to produce a viable in vitro-produced embryo using fresh oocytes collected and transported from different countries. This work opens the possibility to access superior genetics and improve herds in countries seeking to increase their production systems and potentially improve their quality of life. Table 1.Effect of oocyte collection location on embryo development This project was supported by Programa de Competitividad ProCom Senacyt, Panama.

Effects of sexed semen and interactive effects on commercial in vitro embryo production when oocytes are collected from cows of Bos indicus, and Bos taurus breeding and crossbred cows of these subspecies

2015 ◽  
Vol 156 ◽  
pp. 58-63 ◽  
Author(s):  
William Omar Contreras Lopez ◽  
Hernando Raphael Alvis-Miranda ◽  
Andres Felipe Gamarra ◽  
Bibiana Rendon ◽  
Diego Armando Vega Borda ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Interactive Effects ◽  
Embryo Production ◽  
Crossbred Cows ◽  
Sexed Semen ◽  
In Vitro Embryo Production

169 OVUM PICK-UP, IN VITRO EMBRYO PRODUCTION, AND EMBRYO TRANSFER IN CATTLE IN TROPICAL AND SUBTROPICAL MEXICO

2016 ◽  
Vol 28 (2) ◽  
pp. 215
Author(s):  
S. Castañeda ◽  
S. Romo ◽  
M. E. Kjelland
Keyword(s):  
Embryo Transfer ◽  
Production Efficiency ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Hormonal Treatment ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Sexed Semen ◽  
In Vitro Embryo Production

Biotechnology continues to evolve rapidly, allowing the development of artificial reproductive techniques (ART) to increase reproductive efficiency and contribute to the genetic improvement of domestic animals. The present study examines the results obtained after 30 months of starting a commercial practice for ovum pickup (OPU) in vitro embryo production (IVP) and embryo transfer (ET) in cattle in tropical and subtropical Mexico. This research was conducted from 2013–2015 in beef and dairy cattle kept under different environmental and management conditions in 6 states (Chiapas, Oaxaca, Tabasco, Tamaulipas, Veracruz, and Yucatan). Oocytes were collected by OPU, without hormonal treatment, from 10 donor cows: 2 Bos taurus (Bt), 5 Bos indicus (Bi), and 3 Bt × Bi. A total of 98 oocyte recovery sessions were performed on 756 cows and produced 12 524 viable oocytes (1349 GI, 3383 GII, 7792 GIII), which were sent to a central laboratory for IVP. Both conventional and sexed semen were used for IVF, from 9 breeds: 2 Bt, 5 Bi and 2 Bt × Bi. The overall cleavage rate was 69% (8587/12 524). The embryo production efficiency rate was 31% (3905/12 524). Fresh sexed and conventional embryos were transferred to recipients synchronised with the following protocol: Day 0, application of an intravaginal device (ID) with progesterone and 2 mg of oestradiol benzoate IM; Day 8, removal of the ID, 400 IU eCG IM, 0.5 mg cloprostenol sodium IM, and 0.5 mg oestradiol cipionate IM. Day 10 was considered the day of oestrus. Pregnancy rate after ET was 38% (945/3905). The average number of viable oocytes per donor cow was 16.57; the average number of transferred embryos per donor cow was 5.17, and the average number of pregnancies per donor cow was 1.25. The OPU-IVP were successful in producing pregnancies even under several adverse conditions, such as a tropical environment, many donors being prepuberal females and the majority of the adult cows having a previous non-productive history due to hormonal mishandling in superovulation programs or because of overfeeding for show purposes. The consequences of these factors can be observed in the lower overall cleavage rate obtained (69%), compared to the expected (75%). Some variables require further analysis (e.g. different OPU and ET technicians, time of year, cow age, cow breed, and use of conventional and sexed semen). A successful IVP practice has to face and overcome multiple problems that may arise in some geographic locations but, perhaps, not in others. It is interesting to note the use of sexed versus conventional embryos, of the 98 ET sessions, 25 involved sexed embryos versus 73 for conventional embryos. The use of sexed semen for OPU-IVP and ET in this region of Mexico continues to expand. We thank Genemex Internacional and the ranch owners that were involved with this research.

238 KNOCKING DOWN OF THYROID HORMONE RECEPTORS INHIBITS DEVELOPMENT OF EARLY BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 267
Author(s):  
N. Y. Rho ◽  
F. A. Ashkar ◽  
T. Revay ◽  
P. Madan ◽  
W. A. King
Keyword(s):  
Embryo Development ◽  
Reference Genes ◽  
Messenger Rna ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Embryo Production ◽  
Mrna Transcripts ◽  
In Vitro Embryo Production

Thyroid hormones (TH) play an important role in the physiology of vertebrates, ranging from the regulation of metabolic processes to cell proliferation, differentiation, and embryo development. We have previously shown a beneficial effect of supplementing TH in in vitro embryo production media. Recently, detection of TH receptors (TR) in oocytes and early stages of pre-implantation embryos indicated a possible regulatory role for TH in these stages (unpublished data). The objective of this study was to investigate the importance of TR expression in the pre-attachment bovine embryo in vitro. Bovine embryos, produced by standard in vitro embryo production procedures, were microinjected at the zygote stage with small interfering RNA (siRNA) specifically designed for knocking down either TR-α or TR-β. In addition, groups of zygotes were microinjected with scrambled siRNA (SI) or were not injected (NI), and these groups served as controls. Embryo developmental rates were assessed using light microscopy for blastocyst formation rates and expression of TR messenger RNA (mRNA) transcripts at the blastocyst stage was assessed by quantitative PCR across all groups. Expression of TR mRNA was normalized against glyceraldehyde 3-phosphate dehydrogenase, H2a, and 18S as reference genes. There was a significant decrease in blastocyst formation rates in both embryo groups injected with either TR-α (P < 0.002) and TR-β (P < 0.001) siRNA compared with the NI and SI groups. Moreover, the TR-β knockdown group exhibited a lower developmental rate than the TR-α knockdown group, which indicates a stronger inhibitory role for TR-β. Quantification of the level of TR mRNA expression in four groups normalized with three different reference genes shows a consistent significant reduction in the levels of TR-α (P < 0.05) and TR-β (P < 0.02) mRNA transcripts compared with the NI and SI groups. However, TR-β expression was inhibited more than was TR-α expression. In conclusion, the results indicate that knocking down either TR-α or TR-β restrains embryo development. This suggests that TH play a vital role in the regulation of embryo development through their receptors during bovine early embryogenesis. The specific role of each of these receptors and their mechanism of action in mediating development needs to be further elucidated. Funding was provided by CRC, NSERC, and the EmbryoGENE network.

scholarly journals Effects of a high-energy diet on oocyte quality and in vitro embryo production in Bos indicus and Bos taurus cows

2015 ◽  
Vol 98 (5) ◽  
pp. 3086-3099 ◽  
Author(s):  
J.N.S. Sales ◽  
L.T. Iguma ◽  
R.I.T.P. Batista ◽  
C.C.R. Quintão ◽  
M.A.S. Gama ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Oocyte Quality ◽  
High Energy ◽  
Embryo Production ◽  
In Vitro Embryo Production

Effects of superovulation on repeated ultrasound guided oocyte collection and in vitro embryo production in pregnant heifers

1997 ◽  
Vol 47 (1) ◽  
pp. 157 ◽  
Author(s):  
C.Guyader Joly ◽  
S. Ponchon ◽  
J.M. Thuard ◽  
M. Durand ◽  
M. Nibart ◽  
...  
Keyword(s):  
Ultrasound Guided ◽  
Embryo Production ◽  
Oocyte Collection ◽  
In Vitro Embryo Production

scholarly journals 287IN VITRO PRODUCTION OF HOLSTEIN EMBRYOS USING SEX SORTED SEMEN

2004 ◽  
Vol 16 (2) ◽  
pp. 263
Author(s):  
R.D. Wilson ◽  
K.A. Weigel ◽  
P.M. Fricke ◽  
M.L. Leibfried-Rutledge ◽  
D.L. Matthews ◽  
...  
Keyword(s):  
Low Cost ◽  
Laboratory Data ◽  
Blastocyst Stage ◽  
Embryo Production ◽  
Cull Cows ◽  
Sexed Semen ◽  
Dairy Cattle Breeding ◽  
In Vitro Embryo Production ◽  
Freeze Date

Our objective was to explore the synergy between sexed semen and in vitro embryo production and assess benefits of these technologies on commercial farms. Ovaries were collected from high genetic merit Holstein cull cows via colpotomy or at the time of slaughter. Oocytes were aspirated from the ovaries, fertilized 20–24h later, and matured to the morula or blastocyst stage. Embryos were transferred into recipient Holstein cows and heifers on the same farms. Seven Wisconsin herds participated, and 365 embryos were produced from 104 donor cows. Only 272 of these embryos were transferred due to limited availability of recipients. Sexed semen from three Holstein sires was used. On average, 3.5±0.37 transferable embryos were produced per donor, including 1.4±0.18 grade 1 embryos and 1.5±0.20 grade 2 embryos. Individual farms averaged from 1.6 to 5.8 transferable embryos per donor. Laboratory data also revealed interesting results. On average 43.7±4.0 oocytes were collected per donor, and the number of usable oocytes (33.9±3.4), and percent embryos cleaved (52.1±1.9), were significant predicators of the number of blastocysts developed. We divided the usable oocytes and embryos cleaved per donor into quartiles. The fourth quartile for embryos cleaved was significantly greater (P&lt;0.05) than the lower three quartiles, and the usable oocyte quartiles all significantly differed from each other. Semen freeze date was also a significant predicator of the number of blastocysts developed, suggesting significant variation in the quality of sorted semen per ejaculate. To preliminarily test the effect of sorting on the percentage of embryos developing to blastocyst stage, oocytes were recovered from ovaries collected at a slaughterhouse and fertilized using non-sorted semen or sex-sorted semen from the same sires. Oocytes (n=3312) fertilized using non-sorted semen tended (P=0.06) to produce more embryos developing to blastocyst stage than oocytes (n=1577) fertilized using sex-sorted semen (20.1±2.9% v. 12.2±2.3%, respectively). Preliminary pregnancy results show strong farm and sire effects. Overall conception rate was 36% for heifer recipients and 18milking cow recipients. These results suggest that low cost in vitro embryo production may have promise as an early system for utilizing sexed semen in dairy cattle breeding programs.

97 IN VIVO AND IN VITRO EMBRYO PRODUCTION WITH Y-SEXED SORTED OR CONVENTIONAL SEMEN IN BEEF CATTLE

2014 ◽  
Vol 26 (1) ◽  
pp. 162
Author(s):  
H. Tribulo ◽  
J. Carcedo ◽  
R. Tribulo ◽  
J. Menajovsky ◽  
B. Bernal ◽  
...  
Keyword(s):  
Animal Health ◽  
T Test ◽  
High Fertility ◽  
In Vitro Production ◽  
Embryo Production ◽  
Sexed Semen ◽  
Chi Squared ◽  
In Vitro Embryo Production

An experiment was designed to evaluate in vivo and in vitro embryo production following the use of frozen–thawed conventional or Y-sexed semen from a Brangus bull with known high fertility. For in vivo embryo production, Brangus heifers (n = 12) were superovulated twice in a crossover design and inseminated with sexed or conventional semen. On Day 0, all heifers received an intravaginal progesterone device (DIB 1 g, Syntex S.A., Buenos Aires, Argentina) and 2.5 mg oestradiol benzoate and 50 mg progesterone (Progestar, Syntex S.A.) by intramuscular injection (IM). On Day 4, heifers were superstimulated with 200 mg of NIH-FSH-P1 Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) in twice-daily decreasing doses over 4 days. In the a.m. and p.m. of Day 6, all heifers received PGF2a (Ciclase, Syntex) and DIBs were removed in the p.m.. In the a.m. of Day 8, heifers received 100 μg de Gonadolerin (Gonasyn, Syntex S.A.) and were randomly allocated to receive either one straw of conventional semen (24 × 106 sperm per dose) 12 and 24 h later or two straws of sexed semen (2.4 × 106 sperm per dose) 18 and 24 h after GnRH. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared by t-test. Mean ( ± s.e.m.) number of ova/embryos, fertilized ova, and transferable embryos were 14.8 ± 2.7, 9.4 ± 1.8, and 7.1 ± 1.7 v. 16.8 ± 3.1, 9.9 ± 2.5, and 8.1 ± 2.0 for donors inseminated with conventional or sexed semen, respectively (P > 0.6). For in vitro production, oocytes were obtained from 50 ultrasound-guided follicle aspiration (OPU) sessions that was performed at random stages of the oestrous cycle and without superstimulation in 22 Brangus cows and heifers. Oocytes were classified and matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Semen samples from the same bull used for in vivo embryo production were selected using Percoll and capacitated in Fert medium and used at a final concentration of sperm/mL for nonsexed semen and 2 × 106 sperm mL–1 for sexed semen. After 16 h (sexed) or 18 h (conventional) in Fert medium, zygotes were denuded and cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2 and saturated humidity for 7 days. The total number of oocytes matured and fertilized was 528 and 318 for conventional and sexed semen, respectively. Means were compared by t-test and proportions by chi-squared test. Mean (± s.e.m.) number of cleaved zygotes and blastocysts produced per OPU session did not differ between conventional (11.0 ± 1.4 and 7.1 ± 1.0) and sexed (8.7 ± 0.8 and 4.9 ± 0.7; P > 0.2) semen. However, the proportion of cleaved zygotes and blastocysts produced were significantly higher (P < 0.05) with conventional semen (61.2%; 329/538 and 39.4%; 212/538) than with sexed semen (54.4%; 173/318 and 30.8%; 98/318), respectively. In conclusion, comparable number of embryos can be obtained in vivo with sexed or conventional semen from a bull with proven high fertility. However, the proportion of blastocysts produced in vitro is likely to be reduced following the use of sexed as compared with conventional semen from the same bull.

Ovarian stimulation protocols for oocyte collection and in vitro embryo production from 8 to 9 week old lambs

1995 ◽  
Vol 43 (1) ◽  
pp. 203 ◽  
Author(s):  
C.R. Earl ◽  
B.J. Irvine ◽  
J.M. Kelly ◽  
J.P. Rowe ◽  
D.T. Armstrong
Keyword(s):  
Ovarian Stimulation ◽  
Embryo Production ◽  
Oocyte Collection ◽  
In Vitro Embryo Production

Ovum pick up, in vitro embryo production, and pregnancy rates from a large-scale commercial program using Nelore cattle (Bos indicus) donors

2011 ◽  
Vol 75 (9) ◽  
pp. 1640-1646 ◽  
Author(s):  
J.H.F. Pontes ◽  
F.A. Melo Sterza ◽  
A.C. Basso ◽  
C.R. Ferreira ◽  
B.V. Sanches ◽  
...  
Keyword(s):  
Large Scale ◽  
Bos Indicus ◽  
Pregnancy Rates ◽  
Embryo Production ◽  
Commercial Program ◽  
In Vitro Embryo Production ◽  
Nelore Cattle
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