289 IN VITRO PRODUCTION OF OVINE EMBRYOS FROM WOOL AND HAIR BREEDS

2006 ◽  
Vol 18 (2) ◽  
pp. 252
Author(s):  
E. A. Ordóñez-León ◽  
J. A. Medrano ◽  
V. O. Mejía ◽  
J. De Lucas ◽  
Y. C. Ducolomb ◽  
...  
Keyword(s):  
Physiological Saline ◽  
Relevant Information ◽  
In Vitro Production ◽  
Concentration Gradients ◽  
Independent Variables ◽  
Genetic Potential ◽  
Significant Difference ◽  
Tropical Areas ◽  
Vitro Production

In tropical areas, wool-less or haired breeds of sheep are more prolific than wool breeds. There are no reports about IVF in tropical breeds; therefore, it is not known how these respond under IVF conditions. Developing protocols for in vitro production of embryos in haired breeds could contribute to the preservation and use of their genetic potential in tropical countries where they are economically important. The aim of this study was to determine differences in IVM, IVF, and in vitro (IVD) between wool and hair breeds of sheep. A protocol for IVF (Wani 2002 Small Rum. Res. 44, 89-95) was used in wool (W) and hair (H) breeds. A total of 251 W and 251 H ewes were used. The ovaries were obtained after slaughter and transported to the laboratory in physiological saline (25�C). A total of 411 ovaries from W and 440 from H ewes were used, and 805 follicles of W and 790 of H ewes were aspirated. From these, 663 (82%) cumulus-oocyte complexes (COCs) of Wand 597 (76%) of H ewes were obtained and then used for the procedures of IVM, IVF, and IVD. The average number of COCs recovered per ovary was 1.6 forW and 1.4 for H. The average number of follicles per ovary was 1.9 for W and 1.7 for H. For IVM, COCs were incubated in TCM-199 supplemented with 20% serum from estrous ewe (SEE) for 24 h. All incubations were performed at 38.5�C in a humidified atmosphere of 5% CO2 in air. After this period, COCs were placed in fertilization medium (TALP supplemented with 200�g/mL heparin, 3�g/mL penicillamine, and 1�g/mL hypotaurine). For insemination, frozen-thawed semen from H and W rams was washed by centrifugation in two concentration gradients of a silicone solution. Oocytes and semen from the corresponding breed types were co-incubated for 18 h. For IVD, presumptive zygotes were incubated in SOF medium supplemented with 20% SEE for 7 days. Eight replicates were made. The rates of IVM, IVF, and IVD were analyzed by logistic regression, using as response variables: IVM, IVF, and IVD results, and as independent variables: breed and replicate. The percentage of recovered oocytes was 82% for W and 76% for H. For IVM and IVF, the recovered oocytes produced 535 fertilized oocytes of W and 446 of H. From these, 81% of the W oocytes and 75% of H were fertilized. Oocytes from W showed a higher percentage of IVM and IVF, with a statistically significant difference (P < 0.05). The percentage of division was 63% for W (n = 419) and 52% for H (n = 312). There were no statistically significant differences between the two groups for embryo IVD (P > 0.05). No statistical differences were found between replicates and no interactions were observed for breed � replicate (P > 0.05). It is concluded that IVM, IVF, and IVD procedures used for the development of embryos in W ewes can be used with similar results in H ewes. This is the first report of sheep IVF in Mexico that provides relevant information about the procedures of IVM, IVF, and IVD in hair and wool sheep, andsets a precedent for future investigations on in vitro embryo production in haired sheep breeds in Mexico. Funding for E.A. Ord��ez-Le�n was provided by CONACYT and UNAM.

scholarly journals Use of Melatonin in the In Vitro Production of Bovine Embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Alan da Silva LIRA ◽  
Ricardo de Macedo CHAVES ◽  
Felipe de Jesus MORAES JUNIOR ◽  
Sergio Henrique COSTA JUNIOR ◽  
Brenda Karine Lima do AMARAL ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Control Group ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Vitro Production

ABSTRACT We aimed to assess the effects of melatonin in the in vitro production of bovine embryos. Our experiment was conducted at the Laboratório de Reprodução Animal of the Universidade Estadual do Maranhão. The cumulus-oocyte complexes (COCs) were distributed among treatments at concentrations of 0, 10-1, 10-3 and 10-5 µMol/L melatonin. Our experiment was further divided into two: the first was to assess the effect of different concentrations of melatonin (treatments) on the maturation rate of COCs, and the second was to assess the effects of melatonin treatments on the in vitro production of bovine embryos. The results from the first experiment demonstrated no significant difference between the in vitro maturation rate of the cultivated COCs in treatments with melatonin. In the second experiment, however, melatonin treatments yielded statistically higher cleavage, morula and blastocyst rates in the 10-5 µM group (52.9%, 52.9%, and 35.3%, respectively), and lower rates in the 10-1 µM group (19.5%, 19.5% and 7.8%, respectively), compared to the others. The control group (no melatonin) and the 10-3 µM group showed similar results. We concluded that supplementation of melatonin in the in vitro maturation medium resulted in no improvement in the oocyte maturation rate, but in the in vitro production of embryos at different concentrations, the 10-5 µM group displayed better results, but with no improvement in the variables (P < 0.05).

180 DEVELOPMENTAL CAPACITY OF SINGLE CULTIVATED EMBRYOS IN SMALL AND BIG DROPLETS

2007 ◽  
Vol 19 (1) ◽  
pp. 206
Author(s):  
I. G. F. Goovaerts ◽  
J. B. P. De Clercq ◽  
M. Nichi ◽  
P. E. J. Bols
Keyword(s):  
Blastocyst Stage ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Significant Difference ◽  
Group Treatments ◽  
Developmental Capacity ◽  
Droplet Sizes ◽  
The Individual ◽  
Vitro Production

An in vitro production system where a single oocyte can be followed from the ovary to the blastocyst stage would be a useful tool for studies concerning developmental competence or follicular environment. Unfortunately, until now, only low blastocyst rates could be obtained after single embryo production, and there is still discussion about the ideal droplet size. The objective of the present experiment was to compare the developmental competence of single cultivated zygotes in 20- and 500-µL droplets. Cumulus–oocyte complexes were obtained from slaughterhouse ovaries and were matured and fertilized in groups of 100 for 22 h; the presumptive zygotes were divided into 4 groups. In treatment 1, 25 zygotes were transferred into 50 µL of SOF medium supplemented with 5% serum under oil, whereas in treatment 2, 25 zygotes were transferred into 500 µL of medium. Zygotes were cultivated separately in treatments 3 and 4: in treatment 3 in 20 µL of medium under oil and in treatment 4 in 500 µL of medium. Cleavage rates and division stages were assessed after 3 days of cultivation (5% CO2, 5% O2, 90% N2); blastocyst rates were determined after 7 days. Statistical analysis was performed by logistic regression using SAS (PROC LOGISTIC). There was no difference in cleavage rates between the 2 group treatments or between the 2 single treatments. Also, the division stages were not different between the 2 single treatments (16-cell: 2.0 vs. 1.3%; 8-cell: 25.8 vs. 31.6%; 4-cell: 41.2 vs. 38.0%; and 2-cell: 31.0 vs. 29.1% for the 20 µL and the 500 µL droplet sizes, respectively). Group cultivation after 7 days in 50 µL was significantly better than in 500 µL; however, both treatments resulted in significantly higher blastocyst rates compared with the individual cultures in 20 or 500 µL, between which no significant difference could be found. Noteworthy, only 4-cell and 8-cell stages on Day 3 resulted in blastocysts on Day 7 of cultivation. In conclusion, these results indicate that cultivation in groups gives higher blastocyst rates, although the same embryo density is used as in individual cultivation (1 embryo 20 µL in treatments 2 and 3). Moreover, no significant difference could be found between single cultivation in small and big droplets. This is confirmed by the cleavage stages on Day 3, which indicate no difference in timing of cleaving between small and big droplets; time of cleaving is indicative of further developmental capacity. Table 1.Cleavage and blastocyst rates after single and group cultivation

The In Vitro Production of Thromboxane B2 by Platelets of Diabetic Patients Is Normal at Physiological Concentrations of lonized Calcium

1993 ◽  
Vol 70 (03) ◽  
pp. 389-392 ◽  
Author(s):  
Cristina R Falcon ◽  
Marco Cattaneo ◽  
Alberto Ghidoni ◽  
Pier Mannuccio Mannucci
Keyword(s):  
In Vitro Studies ◽  
Thrombin Inhibitor ◽  
Diabetic Patients ◽  
In Vitro Production ◽  
Significant Difference ◽  
Patients With Diabetes ◽  
Vitro Production ◽  
Induced Aggregation

SummaryPlatelets of patients with diabetes and no evidence of macroangiopathy produce normal amounts of thromboxane (Tx) B2 in vivo, whereas they usually show increased production in vitro. Since in vitro studies have been usually performed in citrated PRP, we tested the hypothesis that the discrepancy between in vivo and in vitro studies is due to the low concentration of plasma ionized calcium ([Ca 2+ ]o) that is present in citrated PRP. In fact, low [Ca 2+ ]o artifactually potentiates the platelet TxB2 production in vitro. Forty patients with diabetes mellitus and 37 matched Controls were studied. Blood was anticoagulated with citrate, the thrombin inhibitor D-phenylalanyl-l-prolyl-l-chloromethylketone (PPACK) or both anticoagulants. Platelet aggregation, release of 14C-serotonin and TxB2 production were induced in platelet rieh plasma (PRP) by several agonists. The following results were obtained: i) Citrated PRP: Arachidonic acid induced aggregation (p <0.01) and TxB2 production (p <0.02) were significantly greater in patients than in Controls. No statistically significant differences were found with other agonists. ii) PPACK PRP: No statistically significant difference was found between diabetic platelets and Controls, iii) PPACK plus citrate PRP: The results were not different from those obtained with citrate alone. Therefore, our results show that diabetic platelets produce normal amounts of TxB2 in vitro when the [Ca2+]o is physiological.

scholarly journals In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Anita Soares Barbosa GUIMARÃES ◽  
Laiara Fernandes ROCHA ◽  
Ronival Dias Lima de JESUS ◽  
Gisvani Lopes VASCONCELOS ◽  
Gabriela ANGHINONI ◽  
...  
Keyword(s):  
Culture Medium ◽  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Vitro Production ◽  
In Vitro Performance

ABSTRACT In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P < 0.0001; n = 681 vs. 509 for grade 2 COC, P = 0.010, for zebu and taurine donors, respectively). There were differences in embryo production percentages obtained from OM (0.44% from zebu and 6.42% from taurine, P = 0.017), BL (14.18% from zebu and 3.74% from taurine, P < 0.0001), and BX (81.43% from zebu and 75.13% from taurine, P < 0.0001). No significant difference was observed for embryo production from BI and pregnancy rate (P > 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.

158 EFFECTS OF APOTRANSFERRIN ON IN VITRO MATURATION OF CUMULUS - OOCYTES COMPLEXES (COCs) IN HANWOO, KOREAN NATIVE COWS

2006 ◽  
Vol 18 (2) ◽  
pp. 187
Author(s):  
S.-H. Choi ◽  
S.-R. Cho ◽  
M.-H. Han ◽  
H.-J. Kim ◽  
C.-Y. Choe ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Basic Fuchsin ◽  
In Vitro Production ◽  
Control Groups ◽  
Significant Difference ◽  
Native Cattle ◽  
Vitro Development ◽  
Vitro Production

For in vitro production of embryos, animal sera have been used as energy sources, maturation promoters, vitamins, growth factors, and antioxidative compounds. However, the sera had risk of virus and mycoplasma infections which could result in too big offspring and cause dystocia in ovine and bovine. Apotransferrin (apo-Tf) is a component of mammalian sera and has played a role as an antioxidant in media. A study was conducted to investigate the effects of apo-Tf on in vitro maturation of cumulus-oocytes complexes (COCs) in Hanwoo, Korean native cows. Ovaries were collected from a slaughterhouse and COCs were taken from 2-6-mm antral follicles. The collected COCs were washed three times with 0.1M polyvinyl alcohol (PVA)-TCM199 and matured in 0, 1, 10, or 100 �g/mL apo-Tf with TCM-199 at 39�C, 5% CO2, 95% air for 6, 12, or 24 h. Mature COCs were fertilized with frozen-thawed Korean native cattle semen treated with BO medium (Brackett and Oliphants 1975 Biol. Reprod. 12, 260-274) containing 5 mM caffeine and 1 �g/mL heparin for 8 h and developed to the blastocyst stage in 5% FBS and 0.3% BSA in TCM199-IVMD (IFP, Japan). To evaluate the morphology of nuclear types, the matured COCs were fixed in 1:3 acetic acid-ethanol for 30 s and stained with 3% basic Fuchsin. IVM and IVF were replicated three times. All of the results were analyzed by ANOVA using the STATVIEW program. The maturation rates of control were 34.2%, 37.3%, and 45.8% for 6, 12, and 24 h, respectively. There were no differences among the concentrations of apo-Tf, and nuclear types at 78.3-87.0% GVBD for 6 h, 82.8-91.3% MI for 12 h, and 88.9-100.0% MII for 24 h, with 1, 10, and 100 �g/mL apo-Tf, respectively. Conversely, there was significant difference between 1 �g/mL and 10 �g/mL in terms of cleavage rates, although the others did not vary significantly (P < 0.05). There were significant differences among the concentrations of apo-Tf for blastocyst formation (P < 0.05). Blastocysts matured with 1, 10, and 100 �g/mL apo-TF and developed in 5% FBS and 0.3% BSA in TCM199-IVMD showed rates of 8.8-21.6%, 9.4-35.3%, and 9.1-19.1%, respectively. The control groups developed to the blastocyst stage showed rates of 8.6%, 10.8%, and 10.5% in 5% FBS and 0.3% BSA in TCM199-IVMD, respectively. These results suggest that apo-Tf is an important factor for the in vitro maturation and in vitro development of bovine COCs.

290 CYTOKINE PROFILE IN THE AMNIOTIC FLUID OF NELORE CALVES CONCEIVED BY IN VITRO PRODUCTION AND EMBRYO TRANSFER

2010 ◽  
Vol 22 (1) ◽  
pp. 301
Author(s):  
C. F. Moya-Araujo ◽  
N. C. Preste ◽  
M. Piagentini ◽  
G. H. M. Araujo ◽  
M. G. Silva
Keyword(s):  
Amniotic Fluid ◽  
Embryo Transfer ◽  
In Vitro Production ◽  
Plastic Tube ◽  
Tnf Α ◽  
Significant Difference ◽  
Ifn Γ ◽  
The Moment ◽  
Vitro Production

Cytokines have several functions in many cells and their appropriate balance in the maternal-fetal connections is necessary to maintain a successful pregnancy. The incidence of fetal problems reported at the delivery moment in cows with conceptus conceived by in vitro production (IVP) and lack of literature in the immunology gestation area in bovine prompted this study. The aim of the present study was to determine the levels of cytokines such as tumor necrosis factor a (TNF-α), interferon-y (IFN-γ), interleukin-6 (IL-6), and IL-8 in amniotic fluid of calves from IVP and conventional embryo transfer at the moment of delivery. Forty animals were used and divided into 2 groups: 1 - Twenty Nelore cross-breed cows pregnant with Nelore calves conceived by conventional embryos transfer method (ET group); 2 - Twenty Nelore cross-breed cows pregnant with Nelore calves obtained by IVP after follicular aspiration (IVP group). Around labor, cows were transferred to a maternal paddock to permit observation of delivery. During the expulsion phase the amnion was punctured and 15 mL of fluid was collected in a plastic tube and stored in a freezer for later analysis. The cytokine levels were measured by immunoenzymatic assay (ELISA). The commercial kits used were ESS0011 (Bovine TNF Alpha), ESS0026 (Bovine IFN-gamma), and ESS0029 (Bovine IL-6) from Pierce Biotechnology™ (Rockford, IL, USA) and kit D8000C (Human IL-8) from R&D Systems™ (Minneapolis, MN, USA). The protocol was performed according to the manufacturer’s recommendations. The densities were evaluated by using an automatic optical reader ELISA. The well plates were read at 450 nm. The ANOVA on ranks was used for statistical analysis because of nonparametric distribution of data, with 5% of significance. No calves in this experiment showed signs of sepsis or death during the peripartum period. There was no significant difference in the concentration of TNF-α between the animals of the ET (250.40 pg mL-1) and IVP (398.70 pg mL-1) groups (P > 0.05). There was no difference statistically for IFN-γ; the medians of this cytokine were 35.7 pg mL-1 for the ET group and 48.22 pg mL-1 for the IVP group (P > 0.05). Only samples 15 and 18 of the IVP group had detectable concentrations of IL-6 (2227.47 and 2686.30 pg mL-1, respectively) and for IL-8 only 15 of the samples in the same group (577.6 pg mL-1). The results for TNF-α and INF-γ in this experiment can be adopted as standards for normal pregnancy in cattle, because there are no reports of such data in the literature. However, additional research in the area of gestation immunology area is necessary to describe the role of cytokines in the maintenance of pregnancy and at the moment of delivery in cattle. Supported by FAPESP, Brazil.

304 SHORT-PERIOD SPERM - OOCYTE INCUBATION AND FERTILIZATION MEDIA EFFECTS ON THE IN VITRO PRODUCTION OF SWINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 267
Author(s):  
M. G. Marques ◽  
A. B. Nascimento ◽  
L. F. Martins ◽  
F. R. O. Barros ◽  
M. D. Goissis ◽  
...  
Keyword(s):  
Incubation Period ◽  
In Vitro Production ◽  
Standard Medium ◽  
Sodium Pyruvate ◽  
Short Period ◽  
Significant Difference ◽  
Incubation Periods ◽  
Chi Squared ◽  
Vitro Production

The aim of this study was to evaluate the effect of 2 sperm–oocyte incubation periods (30 min and 6 h) and 3 IVF media (TCM-199, TCM-199 with Ca-lactate, or mTBM). Cumulus–oocyte complexes (COCs) from abattoir-derived swine ovaries were matured in TCM-199 supplemented with 3.05 mM glucose, 50µg mL−1 gentamycin, 0.91 mM sodium pyruvate, 10% swine follicular fluid, 0.57 mM cysteine, 10 ng mL−1 of epidermal growth factor, 10 IU mL−1 eCG, and 10 IU mL−1 of hCG for 22 h, followed by a 22-h incubation without hormones. After in vitro maturation, oocyteswere allocated into 3 IVF media: TCM-199 (supplemented with 3.05 mM glucose, 0.91 mM sodium pyruvate, 0.57 mM cysteine, 50µg mL−1 gentamycin, 1 mg mL−1 BSA, and 3.06 mM mL−1 caffeine-standard medium);TCM-199 Ca (standard medium with 2.92 mM Ca lactate); or mTBM (supplemented with 5 mM mL−1 sodium pyruvate, 0.57 mM cysteine, 50µg mL−1 entamycin, 1 mg mL−1 BSA, and 2 mM mL−1 caffeine). The refrigerated semen at 15°C was centrifuged and capacitated for 2 h at 38.5°C in an atmosphere of 5% CO2 in air and under high humidity conditions in the respective IVF media. Oocytes were denuded, washed with the respective IVF media, and inseminated with the capacitated spermatozoa at a concentration of 6 × 105 sperm mL−1 in 90-µL microdroplets. After 30 min, a group of oocytes from each fertilization medium was gently washed to remove nonbound spermatozoa and returned to a new medium droplet for another 5.5 h (30-min group). The other group of oocytes remained in the same droplet with spermatozoa for 6 h (6-h group). After IVF, oocytes were cultured in porcine zygote medium-3 for 7 days. Data were analyzed by chi-squared test (P &lt; 0.05).With regard to cleavage rates on the third day of embryo development (Day 3), no significant difference was observed among sperm–oocyte incubation periods; however, there was a difference in the TCM-199 group, compared with the mTBM and TCM-199 Ca groups, at the 30-min incubation period. With 6 h of incubation, no differences were observed among groups. When blastocyst (Day 7) rates were evaluated, no significant differences were observed between the 2 TCM-199 media groups; however, the mTBM groups showed higher blastocyst rates. The sperm–oocyte incubation period of 30 min or 6 h of IVF did not interfere with the blastocyst rate. In conclusion, the IVF medium mTBM was more efficient than the TCM-199 media when considering embryo production. Moreover, 30 min of IVF was sufficient for the spermatozoa responsible for oocyte fertilization to bind to the zona pellucida. Table 1.Cleavage (Day 3) and blastocyst (Day 7) rates of embryos incubated in two sperm–oocyte periods (30 min or 6 h) and cultivated in three IVF media This work was supported by the FAPESP 05/01420-7.

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.

scholarly journals The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn

2014 ◽  
Vol 34 (3) ◽  
pp. 277-280 ◽  
Author(s):  
Lílian R. Martins ◽  
Claudia B. Fernandes ◽  
Ana I.S.B. Villaverde ◽  
Fernanda C. Landim-Alvarenga ◽  
Maria D. Lopes
Keyword(s):  
Breeding Season ◽  
Culture Medium ◽  
Domestic Cat ◽  
Blastocyst Formation ◽  
In Vitro Production ◽  
Cumulus Oocyte Complex ◽  
Significant Difference ◽  
Vitro Production ◽  
Fresh Semen

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. Ovaries were classified according to their status and were sliced in PBS for cumulus oocyte complex (COC) releasing. Grade I COC were washed three times in H-MEM supplemented with BSA, glutamine, sodium pyruvate, cysteine, streptomycin and penicillin. Oocytes were incubated in groups of 20-30 in 400µL of DMEM supplemented with FSH, LH, estradiol, IGF-I and basic fibroblast growth factor under mineral oil for 30 or 36 hours at 38°C in humidified environment of 5% de O2, 5% CO2 and 90% N2. COC were fertilized in Ham's F-10 medium supplemented with BSA, cysteine, pyruvate and streptomycin/penicillin (culture medium) with fresh semen selected through swim up technique. Eighteen hours later, the presumptive zygotes were denuded, the percentage of cleavage was determined and every 10 zygotes were transferred to 100mL drops of culture medium for culture during three days. After 72 hours of culture the percentage of morulae formation was evaluated and these structures were transferred to drops of the same culture medium. At the eighth day of culture blastocyst formation was analyzed. During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.

scholarly journals BIOCHEMICAL AND HORMONAL PROFILE OF THE AMNIOTIC FLUID FROM NELORE CALVES CONCEIVED BY DIFFERENT REPRODUCTION BIOTECHNOLOGIES

2012 ◽  
Vol 10 (2) ◽  
pp. 205
Author(s):  
Marcelo Piagentini ◽  
Nereu Carlos Prestes ◽  
Carla Fredrichsen Moya-Araujo ◽  
Gustavo Henrique Marques Araujo ◽  
Regina Kiomi Takahira ◽  
...  
Keyword(s):  
Amniotic Fluid ◽  
Chloride Concentration ◽  
In Vitro Production ◽  
Significance Level ◽  
Significant Difference ◽  
Group 3 ◽  
Lower Chloride ◽  
Commercial Kits ◽  
Vitro Production

This study aimed to quantify biochemical constituents and hormonal concentrations in the amniotic fluid of Nelore calves conceived by different reproduction biotechnologies. Sixty animals were used and divided into: 1 – Twenty Nelore cows pregnant of calves obtained by artificial insemination; 2 – Twenty cows pregnant with Nelore calves obtained by embryo transference; 3 – Twenty cows pregnant with Nelore calves obtained by in vitro production. During the expulsion phase, the amnion was punctured and 15 mL of fluid were collected. The biochemical evaluation was performed with commercial kits in accordance to the manufacturer’s recommendations. It was used the radioimmunoassay to assess the progesterone and testosterone levels. The statistical analysis used was the variance analysis and Tukey (5% significance level). Mean urea and creatinine when comparing the three groups showed that group 3 had higher values of these two metabolites, suggesting that the kidney maturity is suitable. For gamma-glutamyltransferase, group 3 had a lower value compared to group 1. This enzyme is related to metabolism, may be inferred that the metabolic activity is high in animal production in vitro. The glucose values did not differ between the groups, suggests that the maternal-fetal exchanges were suitable for the power supply to the fetus. Lower chloride concentration was showed in group 3. Sodium and potassium were found in greater quantities in group 3 when compared to other groups. There was no significant difference for the testosterone and progesterone. The results obtained in biochemical analysis can conclude that the renal and liver activity and maternal-fetal exchange are normal in the different groups.

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