95 Successful Ovum Pick-Up-In Vitro Embryo Production in Lidia Cattle in France

2018 ◽  
Vol 30 (1) ◽  
pp. 187
Author(s):  
G. G. Lazo ◽  
S. Lacaze ◽  
D. Di Scala
Keyword(s):  
Transvaginal Ultrasound ◽  
Genetic Material ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Vitro Production ◽  
Maximum Humidity

Lidia cattle are a breed of Bos taurus that has been selected specially to produce bulls with the temperament and aggressiveness necessary to face a bullfighter in a ring. The genetic wealth of this fighting breed is divided into small lineages, traditionally called encastes, which has resulted in the risk of a loss of genetic variability (Ministerio de Medio Ambiente y Medio Rural y Marino, 2011; http://www.toroslidia.com/wp-content/uploads/2012/01/Programa-de-mejora-de-la-Raza-Bovina-de-Lidia.pdf). The technique to produce embryos in vitro may be a useful tool in the conservation of genetic material from this breed in a selection program. The aims of the study were to demonstrate the effectiveness of in vitro production of Lidia cattle embryos, and to evaluate variation in embryo production among males of the breed. Lidia cows, 7 to 13 years of age (n = 12), were used in an ovum pick-up (OPU)-in vitro production (IVP) program in the south of France. Ovarian superstimulation was induced with decreasing doses of pFSH (Stimufol; Reprobiol, Liège, Belgium) twice daily over 3 days (total dose: 350 µg). Transvaginal ultrasound-guided collection of cumulus–oocyte complexes (COC) was done 12 to 24 h after the last FSH injection. The COC were evaluated immediately after OPU and placed into 2.0-mL tubes (Corning Inc., Corning, NY, USA) containing 500 µL of maturation medium. A gas mix (5% CO2 in air) was injected into each tube and the tube was sealed tightly and placed in a portable incubator (Minitub, Tiefenbach, Germany) at 38.0°C for 12 h. On arrival in the Auriva IVP laboratory, tubes were opened and placed into an incubator with 5% CO2 at 38.5°C at maximum humidity to complete a 24-h maturation period. Semen was collected by electro-ejaculation previously from 5 different Lidia bulls (A, B, C, D, and E) and had been frozen by the same technique. The COC were fertilized with the frozen–thawed semen in TALP medium. Presumed zygotes were cultured in SOF medium (Minitub) to Day 7 (Day 0 = fertilization day) at 38.5°C in a 5% CO2, 5% O2, and 90% N2 atmosphere with maximum humidity. A total of 19 OPU/IVP sessions were performed, 5 cows were collected once, and 7 cows collected twice, and 143 COC were processed for in vitro embryo production. Blastocyst and expanded blastocyst numbers were recorded on Day 7. Oocyte recovery and embryo production by bull were analysed by ANOVA and blastocyst yield by Chi-square. The number (mean ± SEM) of oocytes allocated to each bull per IVP session was (P > 0.05): bull A (4.5 ± 1.9), bull B (5.8 ± 2.1), bull C (9.3 ± 2.5), bull D (6.5 ± 2.1), and bull E (7.0 ± 4.4). The cleavage rate differed among bulls (P < 0.05): bull A (4%), B (80%), C (89%), D (81%), and E (76%). The number (mean ± SEM) of blastocysts was lowest (P < 0.05) for bull A and highest (P < 0.05) for bull C (0, 3.7 ± 1.8, 7.0 ± 1.0, 4.3 ± 1.3, 4.7 ± 2.3 for bulls A to E, respectively). The blastocyst development rate (number of blastocysts/number of oocytes entering the IVF process) was also different among bulls (0, 63, 75, 65, and 67%, respectively; P < 0.05). Although there was a male effect on blastocyst production, our data demonstrate that successful in vitro embryo production in Lidia cattle is possible and suggests that this tool would be useful in a genetic program for the multiplication and the conservation of this breed.

scholarly journals Synchronization with estradiol benzoate in the presence of the corpus luteum in Wagyu cows increases the number of medium follicles but does not interfere with in vitro production of embryos

2021 ◽  
Vol 42 (3) ◽  
pp. 1147-1158
Author(s):  
Maria Fernanda Zamai ◽  
◽  
Fábio Luiz Bim Cavalieri ◽  
Marcia Aparecida Andreazzi ◽  
Fabio Morotti ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Estradiol Benzoate ◽  
Breeding Systems ◽  
In Vitro Production ◽  
Embryo Production ◽  
Follicular Wave ◽  
In Vitro Embryo Production ◽  
Oocyte Donors ◽  
Vitro Production

Reproductive biotechnologies are emerging as an important element for livestock; however, some strategies must be modified to adapt to different breeding systems, such as the use of follicular synchronization protocols. This study aimed to evaluate follicular synchronization using estradiol benzoate (EB), in the presence of the corpus luteum (CL) from Wagyu oocyte donors in in vitro embryo production (IVEP). Rounds of IVEP were performed in heifers and cows (n=19) that were classified into three groups: G1/CL - animals with CL, G2/WCL - animals without CL, and G3/CL + EB - animals with CL that were subjected to follicular synchronization with EB at D0. The groups G1/CL and G2/WCL were considered the control and undertook the natural process of follicular dynamics. The results showed that the synchronization of the follicular wave with the application of EB in the presence of CL, presented a smaller number of small (6.05 ± 0.55) and large follicles (0.45 ± 0.15), but increased (P < 0.05) the number of medium-sized follicles (16.20 ± 0.90). However, the results of ovum pick up showed that regardless of whether or not EB was applied, and regardless of the presence or absence of CL in the Wagyu donor, there was no difference among the groups (P > 0.05) concerning the number of viable oocytes and the viability rate. It was concluded that follicular synchronization using EB in Wagyu oocyte donors that presented a CL, increased the number of medium-sized follicles. However, there was no improvement in the efficiency of ovum pick up, in vitro embryo production, and pregnancy rate.

135 Use and dose of porcine follicle-stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro embryo production in pregnant Holstein heifers

2019 ◽  
Vol 31 (1) ◽  
pp. 192
Author(s):  
R. V. Sala ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
E. Peralta ◽  
D. C. Pereira ◽  
...  
Keyword(s):  
Limited Information ◽  
Dominant Follicle ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Oocyte Competence ◽  
Treatment Groups ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P&gt;0.05). Follicle numbers at the time of oocyte recovery were different (P&lt;0.01) between groups. Heifers in the FSH300 group had a greater (P&lt;0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P&lt;0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P&gt;0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P&lt;0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH

scholarly journals Effect of quality of sperm chromatin structure on in -vitro production of cattle embryos

2005 ◽  
Vol 48 (1) ◽  
pp. 32-39
Author(s):  
L. Kątska-Książkiewicz ◽  
M. Bochenek ◽  
B. Ryńska
Keyword(s):  
Chromatin Structure ◽  
Blastocyst Stage ◽  
Sperm Chromatin ◽  
In Vitro Production ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Bull Ejaculate ◽  
Vitro Production

Abstract. Bull effect on results of in vitro embryo production has been well documented. The aim of the present study was to find the relationship between quality of bull sperm chromatin and its effect on in vitro embryo production. Bovine in vitro matured oocytes were fertilized in vitro using capacitated spermatozoa (freshly ejaculated or frozen-thawed) of 12 bulls. Semen was simultaneously processed according to the sperm chromatin structure assay (SCSA) method and was analysed by flow cytometry. At least 3 replications of IVP with the same semen sample were done. The percentage of spermatozoa with abnormal chromatin ranged from 0.4% to 23.8%. All bulls used for the experiment were divided into three groups showing minimal (0.82% ± 6.82%), low (1.70% ± 15.82%) and high (18.16% ± 53.59%) percentages of spermatozoa with abnormal chromatin structure. Both cleavage rates and embryo development to the blastocyst stage were correlated significantly with sperm chromatin abnormalities and resulted in 23.1, 17.7 and 12.2% of blastocysts respectively for sperm with minimal, low and high percentages of chromatin abnormalities. The SCSA method may be used as a practical indicator of suitability of bull ejaculate for IVP purposes.

244 IN VITRO EMBRYO PRODUCTION WITH CONVENTIONAL OR Y-SEXED SEMEN IN BEEF CATTLE

2015 ◽  
Vol 27 (1) ◽  
pp. 211 ◽  
Author(s):  
H. E. Tribulo ◽  
J. Carcedo ◽  
R. J. Tribulo ◽  
B. Bernal ◽  
J. Garzon ◽  
...  
Keyword(s):  
Mixed Models ◽  
Random Variable ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Sexed Semen ◽  
The Mean ◽  
In Vitro Embryo Production ◽  
Follicle Aspiration

An experiment was designed to evaluate in vitro embryo production following the use of frozen-thawed conventional or Y-sexed semen from a single Brangus and a single Braford bull of proven fertility. Semen was obtained by splitting the same ejaculate to be frozen directly or sex-sorted and then frozen. Oocytes were obtained from 69 ultrasound-guided follicle aspiration (ovum pickup) sessions performed at random stages of the oestrous cycle without superstimulation in 24 Brangus and 10 Braford cows and heifers. Viable oocytes (n = 1120) were matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Frozen-thawed sperm from the Brangus and Braford bulls were selected with Percoll for IVF, capacitated in Fert Medium, and used at a final concentration of sperm per milliliter for conventional (non-sexed) semen and 2 × 106 sperm mL–1 for Y-sexed semen. After 16 h (sexed) or 18 h (conventional) of co-incubation with oocytes in Fert Medium, presumptive 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 from the Brangus donors was 538 and 318 for conventional and sexed semen, respectively. The total numbers of oocytes matured and fertilized from the Braford donors were 139 and 125 for conventional and sexed semen, respectively. Data were compared by ANOVA for mixed models, using breed and type of semen as fixed variables and cow (i.d.) as a random variable. Cleavage and blastocyst rates were first transformed by square root and then analysed by ANOVA for mixed models. Mean (± s.e.m.) number of total viable oocytes collected, cleaved zygotes, and blastocysts produced per ovum pickup session did not differ (P = 0.18) between breeds (Brangus: 17.1 ± 1.6, 10.0 ± 0.9, and 6.2 ± 0.7 v. Braford: 13.9 ± 2.8, 7.6 ± 1.5, and 4.0 ± 0.8), and there was no breed × semen interaction on the mean number of cleaved zygotes and blastocysts produced. However, the mean (± s.e.m.) number of cleaved zygotes and blastocysts produced was significantly higher (P < 0.05) when the oocytes were fertilized with conventional semen (10.7 ± 1.2 and 6.5 ± 0.8) than with sexed semen (7.7 ± 0.7 and 4.3 ± 0.6). The mean cleavage rate was also significantly higher (P < 0.05) when the oocytes were fertilized with conventional semen (76.8 ± 3.9) than with sexed semen (54.1 ± 4.2). Blastocyst rate tended to be higher (P = 0.1) with conventional semen (40.5 ± 3.3) than with sexed semen (33.6 ± 4.2). Although in vitro production may be the preferred alternative for the production of embryos of a known sex, the number of blastocysts produced might be reduced as compared with the use of non-sexed semen from the same bull.

132 Effect of treatment with follicle-stimulating hormone on in vitro embryo production of Gyr (Bos indicus) calves, pubertal heifers and adult cows

2019 ◽  
Vol 31 (1) ◽  
pp. 191 ◽  
Author(s):  
F. M. Elliff ◽  
E. C. Guimarães ◽  
L. F. Féres ◽  
B. M. Bayeux ◽  
M. H. A. Colli ◽  
...  
Keyword(s):  
Transvaginal Ultrasound ◽  
Bos Indicus ◽  
Additional Treatment ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Commercial Laboratory ◽  
Ultrasound Guide ◽  
Effect Of Treatment ◽  
In Vitro Embryo Production

The aim of this study was to evaluate the effect of FSH treatment on the in vitro embryo production of Gyr (Bos indicus) calves (3-10 months), pubertal heifers (16-21 months) and adult cows. Thirty females were used: 10 calves, 10 pubertal heifers (puberty was determined by the presence of a corpus luteum) and 10 cows. The animals were distributed as follows: calves: control (C-C, n=5) and with FSH (FSH-C, n=5); heifers: control (C-H, n=5) and with FSH (FSH-H, n=5); and adult animals: control (C-A; n=5) and with FSH (FSH-A; n=5). All animals received an intravaginal progesterone device (calves-Primer PR®, Agener União, Brazil; heifers and cows-Prociclar®, Ceva Saúde Animal, Brazil) and oestradiol benzoate (calves and heifers: 1mg, cows: 2 mg; Fertilcare Sincronização®, MSD Saúde Animal, Brazil) on Day 0. The control animals of each category received no additional treatment. The FSH-C received 80mg IM of FSH (Folltropin®, Vetoquinol, Mairipora, Brazil), performed in 4 injections twice a day in decreasing doses [25mg (Day 4, p.m.), 25mg (Day 5, a.m.), 15mg (Day 5, p.m.), and 15mg (Day 6, a.m.); coasting period: 24 h]. The FSH-H received 100mg IM of FSH, performed in 4 injections twice a day in decreasing doses [30mg (Day 4, p.m.), 30mg (Day 5, a.m.), 20mg (Day 5, p.m.), and 20mg (Day 6, a.m.); coasting period: 24 h]. The FSH-A received 140mg IM of FSH, performed in 4 injections twice a day in decreasing doses [40mg (Day 4, a.m.), 40mg (Day 4, p.m.), 30mg (Day 5, a.m.), and 30mg (Day 5, p.m.); coasting period: 48 h]. On Day 7 the intravaginal devices were removed and all animals were submitted to epidural anaesthesia (2% lidocaine) followed by ovum pickup guided by transvaginal ultrasound (guide EC9-5 Heifer, WTA, Sao Paulo, Brazil; ultrasound S8®, SonoScape, Shenzhen, China). The recovered oocytes were sent to a commercial laboratory for the in vitro embryo production. The obtained data were analysed by the GLIMIX procedure of SAS® (SAS Institute Inc., Cary, NC, USA; means are presented as standard error of the mean. The oocyte recovery rate was lower (P=0.01) for calves and heifers treated with FSH (FSH-C: 42.8%; FSH-H: 55.2%) when compared with control calves and heifers (C-C: 65.1%; C-H: 81.1%); however, no difference was observed for cows (C-A: 63.7%; FSH-A: 63.1%). The number of viable oocytes differed according to the category (P=0.001); calves and cows had higher numbers of viable oocytes (calves: 11.6±0.7; cows: 11.7±0.9) when compared with heifers (23.4±0.6). The number of cleaved oocytes increased (P=0.03) when calves (FSH-C: 8.0; C-C: 5.8) and cows (FSH-A: 8.8, C-A: 6.0) were treated with FSH. Cleavage rate was higher (P=0.05) when animals of all categories were treated with FSH (calves=52.7%; heifers=68.0%; cows=67.9%), when compared with nontreated animals (45.8%; 60.1%; 51.8%, respectively). The number of blastocysts per ovum pickup increased (P=0.04) when calves (FSH-C: 3.2; C-C: 1.5) and cows (FSH-A: 5.8; C-A: 2.8) were treated with FSH. The number of vitrified embryos (percentage of blastocysts surviving vitrification) increased (P=0.02) when calves (FSH-C: 2.6; C-C: 1.0) and cows (FSH-A: 5.0; C-A: 2.4) were treated with FSH. These results show that treatment with FSH increases the efficacy of in vitro embryo production in Gyr calves and cows.

246 INFLUENCE OF BREED AND SEASON ON IN VITRO EMBRYO PRODUCTION

2016 ◽  
Vol 28 (2) ◽  
pp. 255
Author(s):  
B. Bernal ◽  
J. Revol ◽  
J. M. Oviedo ◽  
A. Tribulo ◽  
H. Tribulo ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Significant Difference ◽  
In Vitro Embryo Production ◽  
Follicle Aspiration ◽  
Vitro Production ◽  
Difference Test

A retrospective analysis of in vitro production (IVP) data was done to determine the influence of breed and season on the production of viable oocytes and embryos. Cumulus‐oocyte complexes (COC) were obtained from 1946 ultrasound-guided follicle aspiration (ovum pickup) sessions performed at random stages of the oestrous cycle without superstimulation in Bos taurus and Bos indicus donors in commercial IVP in Argentina. Frozen-thawed conventional semen was used in beef cattle and conventional (n = 139) and sexed-selected (n = 481) semen in dairy cattle. The COC were classified, matured in B-199 medium, fertilized in IVF-SOF medium (Day 0), and cultured in SOF medium supplemented with 0.4% BSA under oil at 38.8°C, 5.5% CO2, and saturated humidity for 7 days. The number of viable COC and transferable embryos in each breed and season were compared by ANOVA and means were compared by Fisher’s Least Significant Difference test. Proportions were first transformed by arcsin and then analysed by ANOVA. To simplify the interpretation of the results, breeds were grouped as follows: dairy Bos taurus (Holstein, n = 620), beef Bos taurus (Angus and Bonsmara, n = 229), Bos taurus × Bos indicus (Brangus and Braford, n = 1045), and Bos indicus (Brahman, n = 52). There was no interaction between breed and season for any of the end points analysed (P > 0.1). Mean (± standard error of the mean) numbers of viable COC and transferable embryos were higher (P < 0.01) in Bos indicus × Bos taurus (19.3 ± 0.4 and 5.3 ± 0.2, respectively) and Bos indicus (15.8 ± 1.4 and 6.8 ± 0.9, respectively) than in beef (11.6 ± 0.5 and 3.0 ± 0.2, respectively) and dairy (8.0 ± 0.2 and 1.6 ± 0.1, respectively) Bos taurus donors. Cleavage rates were higher (P < 0.01) in Bos indicus (72%) than in the other breeds (57% for Bos indicus × Bos taurus and dairy Bos taurus and 54% for beef). Transferable embryo rates were higher (P < 0.01) in Bos indicus (41%) and Bos indicus × Bos taurus (30%) than in beef Bos taurus (26%). Dairy Bos taurus had the lowest (P < 0.01) embryo rates of all breeds (21%). In dairy Bos taurus, cleavage rates, the number of embryos produced, and transferable embryo production rates were higher (P < 0.01) when conventional semen was used (62%, 2.8 ± 0.15, and 27%, respectively) compared to sexed-selected semen (55%, 1.3 ± 0.1, and 19%, respectively). With regards to season, the number of viable COC was highest (P < 0.01) in the spring (14.3 ± 0.5), lowest in the summer (11.3 ± 1.0), and intermediate in the fall (12.2 ± 1.2) and winter (13.7 ± 1.2), which did not differ. Although not affected significantly by season, the number of embryos produced was numerically lower in the summer (2.8 ± 0.4) than in the spring (4.2 ± 0.2), winter (4.5 ± 0.5), or fall (4.6 ± 0.5). In conclusion, in vitro embryo production was directly influenced by breed and season. Bos indicus influenced cattle and the spring season were preferable for commercial IVP programs that did not include superstimulation.

135 RECOMBINANT BOVINE SOMATOTROPIN ON THE IN VITRO PRODUCTION OF BOVINE EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 181
Author(s):  
L. Berté ◽  
L. Vasconcelos ◽  
L. Hatamoto-Zervoudakis ◽  
W. Yamazaki ◽  
L. Yamazaki ◽  
...  
Keyword(s):  
Bovine Embryos ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Recombinant Bovine Somatotropin ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Vitro Production

Bovine growth hormone (bGH) has been used to improve the results for in vitro production of bovine embryos. Inclusion of bGH in the maturation medium increases both rate of cleavage and frequency of blastocyst development. Thus, the purpose of the present study was to evaluate the effect of recombinant bovine somatotropin (rBST) on cleavage and blastocyst development of bovine embryos when included during in vitro maturation (IVM) only (Group 1), during both IVM and in vitro culture (IVC; Group 2), during IVC only (Group 3), or not included during either IVM or IVC (Group 4). Specifically, in Group 1, oocytes were matured in TCM 199 (Earle's salts) supplemented with 10% FCS, LH, FSH, oestradiol, and amikacin (IVM medium), plus 100 ng mL–1 of rBST and cultured in SOFaaci supplemented with essential amino acids, tri-sodium citrate, myo-inositol, and 5% FBS. In group 2, oocytes were matured in IVM medium containing 100 ng mL–1 of rBST and cultured in SOFaaci supplemented with essential amino acid, tri-sodium citrate, myo-inositol, 5% FBS; on Day 5, rBST (50 ng mL–1) was added. In Group 3, oocytes were matured in IVM medium without rBST; on Day 5, rBST (50 ng mL–1) was added. Group 4 was the control, without rBST supplementation. The treatment groups were analysed using the SAS® (SAS Institute Inc., Cary, NC, USA) in a completely randomised design (P < 0.05). Somatotropin has receptors in cumulus cells and in the zona pellucida acting directly in the oocyte; however, the increase in cleavage rate seen in previous studies after rBST treatment was not observed in the present study. Supplementation of culture medium with rBST during Day 2 to 6 of IVC has been shown to increase the number of trophoblast and subsequent pregnancy rate following transfer. However, in the present study, addition of 50 and 100 ng mL–1 of rBST to the maturation or culture medium did not affect the cleavage rate of embryos and blastocyst production. Table 1.Analysis of the meaning and percentages related to cleavage and production of embryos

scholarly journals MATURAÇÃO E FECUNDAÇÃO IN VITRO DE OÓCITOS BOVINOS EM TUBOS PREVIAMENTE GASEIFICADOS E MANTIDOS EM BANHO-MARIA

2002 ◽  
Vol 7 (2) ◽  
Author(s):  
M. KURTZ FILHO ◽  
L. M. SILVA ◽  
B. MOREIRA ◽  
D. S. BRUM ◽  
F. G. LEIVAS ◽  
...  
Keyword(s):  
Water Bath ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Sodium Pyruvate ◽  
Embryo Production ◽  
Vitro Production

A produção in vitro (PIV) de embriões bovinos alcançada com vacas de matadouros ou de aspiração folicular in vivo (OPU) é uma prática cada vez mais difundida e a sua simplificação poderia baixar os custos de produção. O objetivo desta pesquisa foi comparar a produção in vitro de embriões em estufa com temperatura, umidade relativa e atmosfera controlada (controle), com tubos de poliestireno gaseificados e mantidos em banho-maria (tratamento). Oócitos obtidos de ovários de vacas abatidas foram maturados in vitro em TCM- 199 modificado com 25mM de N-2-hidroxietilpiperazina-N -2-ácido etanosulfônico (HEPES); 0,025mg/ml de piruvato de sódio, 0,01UI de rFSHh/ml, 0,5µg/ml de LHs e 10% de soro de vaca em estro. Na fecundação in vitro utilizou-se Talp-Fert com 0,06mg/ml de albumina sérica bovina, 0,022mg/ml de piruvato de sódio e 10µg/ml de heparina. O cultivo foi conduzido em placas de 4 poços em SOF com 5% de soro de vaca em estro, 20µl/ml de aminoácidos essenciais e 10µl/ml de aminoácidos não essenciais, sob óleo mineral, em estufa com atmosfera de 5% de CO2, umidade saturada e 39°C, por 9 dias. Na maturação não houve diferença (P>0,05) entre o tratamento e o controle. Porém, a maturação e a fecundação ou somente a fecundação in vitro em tubos mantidos em banho-maria não demonstrou ser uma alternativa recomendada para a produção de embriões bovinos. In vitro maturation and fertilization of bovine oocytes in tubes previously gasified kept in water bath Abstract In vitro bovine embryo production either obtained from oocytes of slaughtered cows or in vivo follicular aspiration (OPU) is a well-known technique and it’s simplification might reduce the cost of embryo production. The aim of this study was to compare the cleavage rate and embryo development of the in vitro production of bovine embryos using standard culture system (temperature, gas phase and controlled humidity) versus gasified polystyrene tubes kept in water bath. Oocytes obtained from ovaries of slaughtered cows were in vitro maturated in TCM- 199’modified with 25 mM of N-2-hidroxyethylpiperazine-N’-2-ethanosulfonic acid (HEPES); containing 0.01UI rFSHh/ml and 0.5µg/ml LHs, 0.025mg/ml sodium pyruvate and 10% estrous cow serum. The in vitro fertilization was carried out in Talp-Fert containing 0.06mg/ml BSA, 0.022mg/ml sodium pyruvate and 10µg/ml heparin. The culture was performed in SOF medium with 20µl/ml essential aminoacids, 10µl/ml, non-essential aminoacids and 5% estrous cow serum, with oil overlay, in 4 well dishes and incubated with 5% CO2, maximum humidity at 39°C, for 9 days. The results of this study showed no difference (P>0.05) between the treatment and control groups during the maturation process. However, the maturation and fertilization or only the fertilization in tubes do not represent a viable alternative for the in vitro production of bovine embryos.

scholarly journals In Vitro Production of Embryos from Prepubertal Holstein Cattle and Mediterranean Water Buffalo: Problems, Progress and Potential

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2275
Author(s):  
Luke Currin ◽  
Hernan Baldassarre ◽  
Vilceu Bordignon
Keyword(s):  
Water Buffalo ◽  
Holstein Cattle ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Follicular Aspiration ◽  
Current State ◽  
Widespread Implementation ◽  
Vitro Production ◽  
Mediterranean Water

Laparoscopic ovum pick-up (LOPU) coupled with in vitro embryo production (IVEP) in prepubertal cattle and buffalo accelerates genetic gain. This article reviews LOPU-IVEP technology in prepubertal Holstein Cattle and Mediterranean Water Buffalo. The recent expansion of genomic-assisted selection has renewed interest and demand for prepubertal LOPU-IVEP schemes; however, low blastocyst development rates has constrained its widespread implementation. Here, we present an overview of the current state of the technology, limitations that persist and suggest possible solutions to improve its efficiency, with a focus on gonadotropin stimulations strategies to prime oocytes prior to follicular aspiration, and IVEP procedures promoting growth factor metabolism and limiting oxidative and endoplasmic reticulum stress.

Cleavage Rate of in vitro Matured Oocytes Fertilized with Conventional Semen in Buffaloes

2021 ◽  
Author(s):  
M.H. Pitroda ◽  
K.P. Khillare ◽  
M.B. Amle ◽  
M.D. Meshram ◽  
A.B. Mali ◽  
...  
Keyword(s):  
Fertilization Rate ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Quick Recovery ◽  
Slaughter House ◽  
Maturation Rate ◽  
Aspiration Technique ◽  
Current Scenario ◽  
In Vitro Embryo Production

Background: In vitro embryo production in buffaloes has gained much importance in this current scenario due to ever increasing population and high demand of milk and meat. Slaughter house derived bubaline ovaries are a cheap and abundant source of cumulus oocyte complexes.Methods: Oocytes from the buffalo ovarian follicles were recovered by aspiration technique as it facilitates quick recovery. Total 155 ovaries were used in the present study. Surface follicles were measured using vernier calliper and categorized into three groups viz. less than 3 mm, 3-5 mm and greater than 5 mm based on follicular diameter and oocytes were processed for IVM, IVF and IVC using conventional non sorted semen.Result: Overall percentage of small, medium and large follicles in the ovaries were recorded as 16.29 ± 0.94%, 8.14±0.60%, 5.35 ± 0.76%, respectively. Overall recovery rate of COCs was 38%. The percentage of these oocytes were 16.74% (A), 15.25% (B), 25.26% (C), 18.33% (D) and 29.87% (E) respectively. Maturation rate of oocytes were 81.96 ± 2.70%. Fertilization rate was 74.98 ± 3.87%, Cleavage rate % was 40.84±2.51% and Blastocyst percentage was 21.57±1.75% respectively. Application of in vitro embryo production technique using slaughter house ovaries can salvage the genetic potential of bubaline species.

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