Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle

2013 ◽  
Vol 25 (1) ◽  
pp. 17 ◽  
Author(s):  
Ciro M. Barros ◽  
Rafael A. Satrapa ◽  
Anthony C. S. Castilho ◽  
Patrícia K. Fontes ◽  
Eduardo M. Razza ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Embryo Quality ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Treatment Protocol ◽  
Cumulus Cells ◽  
Fixed Time ◽  
Treatment Protocols ◽  
Oocyte Competence

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the ‘P-36 protocol’ for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT2 receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus–oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.

P–178 Mural granulosa cells of the human follicles indirectly show death molecular signals not depending on different ovarian stimulation protocols

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Ruvolo ◽  
F Geraci ◽  
M C Roccheri ◽  
R Alessandro ◽  
L Bosco
Keyword(s):  
In Vitro Culture ◽  
Granulosa Cells ◽  
Pregnancy Outcomes ◽  
Protein Level ◽  
Ovarian Stimulation ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Oocyte Competence

Abstract Study question Could the expression of the anti-apoptotic molecules AKT, p-Akt and ERK1/2 in Mural Granulosa Cells (MGC) be considered as marker of oocyte quality? Summary answer MGCs activate cell death pathways in analyzed follicles and it is not influenced by different stimulation protocols and it is not correlated to oocyte competence. What is known already It has been previously demonstrated that apoptosis rate of mural granulosa or cumulus cells (CC) were correlated with follicular oocyte number, age, embryo numbers in IVF/ICSI and also clinical pregnancy. Moreover, our previous data demonstrated that in selected patients, who received recombinant LH associated with recombinant FSH (rFSH), the DNA fragmentation in cumulus cells was significantly lower and pregnancy rate was higher, compared to patients treated with rFSH alone. However, to date little is known about the differences between MGC and CC regarding death/survival pathways and whether the two cell types respond in the same way. Study design, size, duration Molecular study on MGCs to investigate the role of the surviving/apoptotic molecules AKT, p-Akt and ERK1/2 and their relationship with the administration of exogenous r-LH combined with r-FSH administration in ovarian stimulation comparing with r-FSH alone. We analyzed also the oocyte competence, for each follicle, according to the embryo development during in vitro culture and the pregnancy outcome. We included fifty-three normo-responder women undergoing ICSI in two years. Participants/materials, setting, methods Patients were divided into two groups: 1) 34 women were stimulated with r-FSH and used as control group, 2) 19 women were stimulated with r-FSH combined with r-LH. Mural granulosa cells isolated singularly from 255 MII oocytes of the 53 patients were used for the study. The study was conducted in public university laboratory. MGCs obtained from each single follicle were suspended in medium, without serum. For immunoreaction anti-AKT, p-AKT, ERK1/2 antibodies were used. Main results and the role of chance Out of 255 MII oocytes collected, 197 were fertilized and the derived embryos had the following evolution: 117 transferred, 57 vitrified and 23 arrested during in vitro culture. 58 oocytes were not analyzed because of failed fertilization or because of their immature condition (GV or MI). In the MGCs isolated from the follicle of each oocyte generating an embryo, the expression AKT, pAKT and ERK1/2 was analyzed and associated with embryo quality and pregnancy outcomes. Immunoblot analysis on granulose cells showed no statistically significant differences in protein level in MGCs isolated from oocytes that have generated transferred embryos (blastocyst at day 5 or 6) comparing with embryos who arrested during in vitro culture. No differences were found also in MGCs collected from the follicles derived from r-FSH ovarian stimulation compared to r-FSH+r-LH. Moreover, no difference was highlighted even between protein level and pregnancy outcomes. The results seem to demonstrate that the MGCs primarily have an endocrine function and support the growth of the follicle, and finally follow a specific death pathway. This condition is not influenced by different ovarian stimulation protocol, in contrast with CC, and is not correlated to oocyte competence, embryo quality and clinical outcomes. Limitations, reasons for caution Only a limited number of patients have been observed. Wider implications of the findings: Our current and past results suggest that the evaluation of cumulus cells and mural granulosa cells in the same follicle show different expression of molecules involved in the apoptotic pathway and therefore they cannot be used, as molecular markers, in the same way, to assess the competence of oocytes. Trial registration number Not applicable

scholarly journals Investigating the Intrafollicular Factors Affecting Oocyte Developmental Competency

2021 ◽  
Author(s):  
◽  
Zaramasina Clark
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Significant Finding ◽  
High Quality ◽  
Final Maturation

<p>The number of cycles of assisted reproductive technologies (ART) performed increased by ~9.5 % globally between 2008 and 2010. In spite of this, the success rate in terms of delivery was only ~19.0 % (Dyer et al., 2016). This discrepancy between the demand for, and success of, these technologies necessitates the development of tools to improve ART efficiency. To facilitate this, a better understanding of how the microenvironment changes within the developing follicle to culminate in a mature, developmentally-competent oocyte is required. This study employed an in vivo and in vitro ovine model to investigate the relationship between the surrounding microenvironment and oocyte maturation, and in particular, the attainment of oocyte developmental competency and high-quality embryos.  The first objective of this PhD study was to comprehensively investigate the changing microenvironment of in vivo matured, presumptive preovulatory (PPOV) follicles from wild-type (++) and high ovulation rate (OR; I+B+) ewes. The high OR ewes were heterozygous carriers of mutations in BMP15 (I+) and BMPRIB (B+). Functional differences in follicular somatic (granulosa and cumulus) cells between these genotypes, including differential gonadotropin responsiveness of granulosa cells, composition of follicular fluid and gene expression profiles in cumulus cells were evident. These differences emerged as part of a compensatory mechanism by which oocytes from smaller follicles, containing fewer granulosa cells, achieved developmental competency in I+B+ ewes.  The second objective of this PhD study was to develop new approaches for improving current in vitro maturation (IVM) strategies. The first approach utilised in this study focused on developing biomarkers that could be used to improve prediction of developmental competency in oocytes and in vitro produced embryos. This involved interrogating the hypothesis that a combination of molecular and morphokinetic biomarkers would better predict the developmental competency of oocytes and embryos compared to using these biomarkers alone. The second approach utilised in this PhD study tested the effects of modulating IVM conditions to better mimic the follicular microenvironment of a high, compared to a low, OR species on oocyte developmental competency and embryo quality. This involved supplementing IVM media with different ratios of two oocyte-secreted growth factors, i.e. GDF9:BMP15, that were representative of low or high OR species. These approaches demonstrated significant potential and warrant further investigation.  The most significant finding of this study was that despite variances in the surrounding microenvironment during in vivo and in vitro oocyte maturation that culminated in differential gene expression patterns in cumulus cells, and divergent gonadotropin-responsiveness of granulosa cells, the gene expression signatures of developmentally-competent oocytes and the morphokinetics of high-quality embryos were unaltered. This confirms the value of developing such biomarkers for oocyte development competency and embryo quality that remain unaltered despite a changing surrounding environment. Interestingly, simulating the ratio of GDF9:BMP15 that oocytes from high OR species are exposed to during maturation improved developmental competency in oocytes as demonstrated by increased blastocyst rates. Furthermore, this study has demonstrated that combinations of molecular (cumulus cell gene expression) and morphokinetic biomarkers improved the ability to predict developmental competency in oocytes and embryos. Overall, this study revealed novel information regarding the follicular microenvironment during final maturation and identified several novel approaches to improving the efficiency of ART.</p>

207 INFLUENCE OF THE INTERVAL BETWEEN THAWING TO TRANSFER ON PREGNANCY RATES OF FROZEN - THAWED DIRECT-TRANSFER EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 220
Author(s):  
G. A. Bo ◽  
L. C. Peres ◽  
D. Pincinato ◽  
M. de la Rey ◽  
R. Tribulo
Keyword(s):  
Bos Taurus ◽  
Body Condition Score ◽  
Bos Indicus ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Direct Transfer ◽  
Condition Score ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

An experiment was designed to evaluate the effect of the interval between thawing to deposition of the embryo into the uterine horn on pregnancy rates of in vivo-produced frozen–thawed embryos in 1.5 M ethylene glycol (direct transfer). Data were collected from 1122 embryo transfers performed in the same farm (Estancia El Mangrullo, Lavalle, Santiago del Estero, Argentina) during the spring and summer of 2004/05 and 2005/06 (6 replicates, ambient temperature between 20 and 40�C). Recipients used in all replicates were non-lactating, cycling, multiparous Bos taurus � Bos indicus crossbred cows with body condition score between 3 and 4 (1 to 5 scale) that were synchronized using fixed-time embryo transfer protocols. Briefly, the synchronization treatments consisted of the insertion of a Crestar ear implant (Intervet, Sao Paulo, Brazil) or a progesterone-releasing device (DIB; Syntex SA, Buenos Aires, Argentina), plus 2 mg of estradiol benzoate (EB; Syntex) intramuscularly (IM) on Day 0, and 400 IU of eCG (Folligon 5000; Intervet, or Novormon 5000; Syntex) IM plus 150 �g d-cloprostenol IM (Preloban; Intervet, or Ciclase; Syntex) on Day 5. Progestin devices were removed on Day 8 and all cows received 1 mg of EB IM on Day 9. All cows were examined by ultrasonography on Day 16 and those with a luteal area &gt;76 mm2 (by calculating the area of the CL minus the area of the cavity) received, on Day 17, frozen–thawed embryos by nonsurgical transfer. All embryos were Grade 1, and all were frozen in 1.5 M ethylene glycol at the Embryo Plus Laboratory (Brits, South Africa). After being stored in liquid nitrogen, the embryos were plunged directly (no air thawing) in a 30�C water bath for 30 s, and then transferred to the recipient cows by either one of two technicians. Based on the interval between thawing and transfer, the transfers were classified as being in one of 3 groups: Group 1: &lt;3 min; Group 2: 3 to 6 min; and Group 3: 6 to 16 min. The main reason for delayed transfers beyond 6 min was the replacement of one recipient for another because of difficulty in threading the cervix (1% of the total transfers) or a recipient falling down into the chute or with very bad disposition and behavior. Pregnancy was determined by ultrasonography 28 to 35 days after fixed-time embryo transfer, and data were analyzed by logistic regression. There were no effects of replicate, technician, CL area, recipient body condition score, embryo stage, and time from thawing to transfer on pregnancy rates. Pregnancy rates in the 3 thawing to transfer intervals were: Group 1: 215/385, 55.8%; Group 2: 372/655, 56.8%; Group 3: 42/82, 51.2%; P &gt; 0.6. These results may be interpreted to suggest that there is no significant effect of time from thawing to transfer (up to 16 min) in direct transfer embryos using Bos taurus � Bos indicus recipients transferred at a fixed time.

155 CHARACTERIZATION OF LIPID DROPLETS IN BOS INDICUS AND BOS TAURUS EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 226 ◽  
Author(s):  
E. P. López-Damián ◽  
T. Fiordelisio ◽  
M. A. Lammoglia ◽  
M. Alarcón ◽  
M. Asprón ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Embryo Quality ◽  
Developmental Stages ◽  
Bos Taurus ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Bos Indicus ◽  
Nile Red ◽  
Blastocyst Stage ◽  
Transfer Program

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.

121 DIFFERENTIAL mRNA EXPRESSION BETWEEN IN VIVO AND IN VITRO-DERIVED BOS INDICUS AND BOS TAURUS EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 160
Author(s):  
L. Nasser ◽  
P. Stranieri ◽  
A. Gutiérrez-Adán ◽  
M. Clemente ◽  
L. Jorge de Souza ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Repeated Measures ◽  
Bos Taurus ◽  
Receptor Gene ◽  
Expression Patterns ◽  
Bos Indicus ◽  
Growth Factor Receptor ◽  
P53 Gene

Brazil is a leading country in the world of commercial use of in vitro-produced bovine embryos with 200 000 transfers per year. The majority of in vitro-produced embryos are pure breed Nelore and are transferred fresh with 40% pregnancy rate. However, pregnancies are drastically reduced with frozen in vitro embryos. This experiment is part of our effort to learn more about molecular composition and morphology of in vitro-derived embryos that may be responsible for such discrepancy. We examined molecular expression of mRNA transcripts of 6 selected genes; apoptosis Bax,TP53(p53), SHC1SHC(p66), insulin growth factor receptor (IGF2R), stabilization of the plasma membrane PLAC8 and glucose conversion H6PD in in-vivo (control) and in-vitro Nelore and Bos taurus embryos. In vivo embryos were collected from superovulated cows at Day 7. In vitro embryo was produced from oocytes aspirated from live cows. A total of 284 oocytes (4 replicates) were matured and fertilized by standard IVF procedures. Presumptive zygotes were cultured in CR2 medium with 5% BSA in 50 μL drops (25 zygotes per drop) at 39°C under paraffin oil and 5% CO2 in humidified air. Embryos that developed on Days 7 to blastocyst were transferred to recipients, and 10 blastocysts from each replicate were frozen for evaluation of gene expression patterns. Poly(A) mRNA was prepared from 3 groups of pools of 10 in vitro embryos and 10 of control in vivo-derived embryos. The quantification of all gene transcripts was carried out by real-time quantitative RT-PCR using the comparative CT method. Data on mRNA expression were normalized to the endogenous H2a.z and was analyzed by one-way repeated-measures ANOVA. The cleavage rates at Day 2 and number of blastocysts developed at Day 7 were 80.3 ± 3.2 and 42.2 ± 6.4, respectively. The level of expression of IGF2R was significantly (P < 0.05) higher in in vivo-derived embryos than in both groups of in vitro embryos. The expression of all 3 apoptosis genes were lower (P < 0.05) in in vivo than in vitro embryos with exception of p53 gene that was not different between Nelore in vitro and in vivo embryos but was significantly higher (P < 0.05) in Bos taurus in vitro embryos. There was no difference in expression of PLAC8 gene among any tested group of embryos and in expression of H6PD gene between Nelore in vitro and in vivo embryos. We concluded that significant differences in molecular makeup between in vitro and in vivo-derived Nelore embryos exist. Of particular importance seems to be pattern of expression of IGF2R receptor gene known as a good indicator of embryo quality, which promotes proliferation and differentiation. Similarly, higher expression of 2 BAX and p66 genes of apoptosis in in vitro embryos seems to be a further indication of inferior quality of Nelore in vitro-derived embryos that showed to be more profound in Bos taurus in vitro-derived embryos.

39 EFFECT OF TIME OF FIXED-TIME AI ON PREGNANCY RATES IN ZEBU CROSS-BRED BEEF HEIFERS TREATED WITH PROGESTERONE RELEASING DEVICES AND ESTRADIOL CYPIONATE

2010 ◽  
Vol 22 (1) ◽  
pp. 177 ◽  
Author(s):  
M. Ramos ◽  
L. Cutaia ◽  
P. Chesta ◽  
G. A. Bó
Keyword(s):  
Bos Taurus ◽  
Body Condition Score ◽  
Bos Indicus ◽  
Fixed Time ◽  
Estradiol Benzoate ◽  
Pregnancy Rates ◽  
Beef Heifers ◽  
Treatment Groups ◽  
Condition Score ◽  
To Receive

Two experiments were designed to evaluate the effect of the timing of fixed-time AI (FTAI) in relation to the removal of an intravaginal progesterone-releasing device (1 g of progesterone, DIB, Syntex SA, Buenos Aires, Argentina) on pregnancy rates in Bos indicus × Bos taurus cross-bred heifers. In experiment 1, 285 Bonsmara × zebu cross-bred heifers, between 18 and 24 months of age and with a body condition score (BCS) between 3.0 and 3.5 (1-5 scale) were used. On the day of initiation of treatment (Day 0), the heifers’ ovaries were palpated (92% of them had a CL) and they received a new DIB plus 2 mg of estradiol benzoate (EB; Syntex SA) and 250 μg of cloprostenol (Ciclase DL, Syntex SA). On Day 8, DIB devices were removed and all heifers received 250 μg of Ciclase plus 0.5 mg of estradiol cypionate (ECP; Cipiosyn, Syntex SA). At that time the heifers were randomly divided to receive FTAI between 48 to 49 h, 53 to 54 h, or 58 to 59 h after DIB removal. The heifers underwent FTAI with semen from 4 bulls by 2 inseminators. In experiment 2, 260 heifers from the same group as those used in experiment 1 (87% with a CL) were treated exactly as those in experiment 1, except that previously used DIB was inserted on Day 0. Pregnancy diagnosis was performed 30 days post-fixed-time AI by ultrasonography. The data were analyzed by logistic regression, taking into account the effect of time of FTAI, semen, and inseminator on pregnancy rates. In experiment 1, pregnancy rates were lower (P = 0.04) in the heifers undergoing FTAI between 48 and 49 h after DIB removal (46/95, 48.4%) than those undergoing FTAI 53 to 54 h (61/99, 61.6%) or 58 to 60 h (57/91, 62.6%) after DIB removal. However, no differences in pregnancy rates were found (P = 0.72) in experiment 2 between the 3 treatment groups, with 39/91 (42.9%) for the 48 to 49 h group, 45/89 (50.6%) for the 53 to 54 h group, and 35/89 (43.8%) for the 58 to 59 h group. There was no effect of the semen or inseminator (P > 0.2) in either experiment. We conclude that when Bos indicus × Bos taurus beef heifers are synchronized with new DIB devices and ECP, higher pregnancy rates are obtained in heifers undergoing FTAI late (between 53 to 60 h after DIB removal) than in those undergoing FTAI early (48 to 49 h after DIB removal). However, time of insemination does not apparently affect pregnancy rates when Bos indicus × Bos taurus beef heifers are synchronized with previously used DIB devices and ECP.

179 COMPARISON OF OOCYTE AND EMBRYO PRODUCTION AMONG BOS TAURUS, BOS INDICUS, AND INDICUS-TAURUS DONOR COWS

2010 ◽  
Vol 22 (1) ◽  
pp. 248 ◽  
Author(s):  
J. H. F. Pontes ◽  
K. C. F. Silva ◽  
A. C. Basso ◽  
C. R. Ferreira ◽  
G. M. G. Santos ◽  
...  
Keyword(s):  
High Efficiency ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Cumulus Cells ◽  
Holstein Cows ◽  
Embryo Production ◽  
Total N ◽  
The Mean ◽  
Donor Cows

In recent years, Brazil has become the leading country in the world for the number of embryos produced in vitro (Thibier M 2009 IETS Embryo Transfer Newsletter 22, 12-19). This is partly due to the large numbers of Bos indicus animals in Brazil, making up about 80% of the total cattle. The mean oocyte production per ultrasound-guided follicular aspiration from Bos indicus is higher than those for European breeds (Pontes JHF et al. 2009 Theriogenology 71, 690-697). In the present study, we analyzed 5407 ovum pick ups (OPU) and compared the average production of total (n = 90,086) and viable (n = 64,826) oocytes and the number of embryos produced in vitro from Gir (Bos taurus indicus), Holstein (Bos taurus taurus), 1/4 Holstein × 3/4 Gir, and 1/2 Holstein-Gir crossbreed cows. To obtain oocytes, OPU was repeated from 4 to 7 times (mean = 5.7 ± 2.4) in each donor cow aged from 3 to 7 years (mean = 5.0 ± 2.3) during a 12-mo period. COCs (n = 90,086) obtained were classified according to the presence of cumulus cells and the oocyte cytoplasm aspect (homogeneous or heterogeneous/fragmented). The viable oocytes (n = 64,826) were in vitro matured for 24 h at 38.8°C in an atmosphere of 5% CO2 in air. Since this was a commercial programm, frozen sexed semen (2 × 106 mL-1) from Gir (n = 8) or Holstein (n = 7) sires previously tested for high efficiency was used for IVF. Fertilization was carried out (18-20 h) and the presumed embryos were cultured for 7 days in the same conditions as were used for IVM. Data were analyzed by ANOVA. On average, 16.7 ± 6.2 oocytes were obtained per OPU/IVF procedure and 71.96% were considered viable. The mean numbers of total oocytes per OPU/IVF procedure were 17.1 ± 4.4 for Gir cows (n = 617), 11.4 ± 3.9 for Holstein cows (n = 180), 20.4 ± 5.8 for 1/4 Holstein × 3/4 Gir (n = 44), and 31.4 ± 5.6 for 1/2 Holstein-Gir crossbreed females (n = 37, P < 0.01). The mean numbers of viable oocytes per OPU/IVF procedure were 12.1 ± 3.8 for Gir cows, 8.0 ± 2.6 for Holstein cows, 16.8, ± 5.0 for 1/4 Holstein × 3/4 Gir, and 24.3 ± 4.7 for 1/2 Holstein-Gir crossbreed females (P < 0.01). The average number of embryos produced by OPU/IVF were 3.2 (n = 12,243/3378) for Gir cows, 2.2 (n = 2426/1138) for Holstein cows, 3.9 (n = 1033/267) for 1/4 Holstein × 3/4 Gir, and 5.5 (n = 1222/224) for 1/2 Holstein-Gir. The average number of embryos produced per IVF session from 1/2 taurus × indicus donor cows was greater (P < 0.01) than from Bos indicus cows. The number of recoverable and viable oocytes and the number of embryos produced in vitro from Bos indicus donors were higher than from Bos taurus females. Therefore, the highest oocyte yield and the greatest embryo production were obtained from 1/2 taurus × indicus females. This work was supported by In Vitro Brasil.

117 Subclinical Mastitis Reduces Ovulation and Oocyte Quality in Milk-Producing Cows

2018 ◽  
Vol 30 (1) ◽  
pp. 198
Author(s):  
G. Santos ◽  
M. P. Bottino ◽  
M. B. D. Ferreira ◽  
J. C. Silveira ◽  
A. C. F. C. M. Avila ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Target Genes ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Subclinical Mastitis ◽  
Oocyte Quality ◽  
Ovulation Rate ◽  
Follicular Dynamics

The aim was to evaluate the effect of subclinical mastitis by somatic cell count (SCC) on follicular dynamics, ovulation, oocyte and cumulus cell quality, exosome size and concentration in milk-producing cows. Crossbred cows (Bos taurus × Bos indicus; that is, Holstein × Gyr) were randomly allocated to control (SCC <200,000 cells mL−1] and mastitis (SCC >400,000 cells mL−1) groups. In experiment 1 (follicular dynamics), cows (n = 57) were submitted to ultrasonographic evaluations every 24 h, after removal of an intravaginal progesterone device (Day 8) up to Day 10. From Day 10, ultrasound evaluations were performed every 12 h, until ovulation or until 96 h after progesterone device withdrawal, in order to follow final dominant follicle growth and ovulation. In experiment 2 (oocyte, cumulus cells, and follicular fluid evaluation), cows (n = 23) were submitted to follicular aspirations, preceded by synchronization of the emergence of the follicular wave. The levels of target genes in cumulus cells (BCL2, BAX, PI3K, PTEN, FOXO3) were evaluated by RT-qPCR. In the follicular fluid, the exosomes were isolated for evaluation of particle size. Data were analysed by the Glimmix procedure of SAS (SAS Institute Inc., Cary, NC, USA). Ovulation rate (P = 0.09) was higher in control cows [control 77.42% (24/31) and mastitis 57.69% (15/26)]. Viable oocyte rate (P = 0.01) was also higher in control cows [control 59.1% (130/220) and mastitis 41.9% (125/298)]. The dynamics of follicular growth did not differ between groups. The number of degenerate oocytes (P = 0.001) was higher in cows of the mastitis group. In the evaluation of cumulus cell gene expression, there was a higher abundance of BAX transcripts (P = 0.003) in cells of mastitis cows. Additionally, the mean and mode of exosome diameter in mastitis cows were smaller (P = 0.03 and P = 0.02, respectively). In conclusion, ovulation rate, oocyte quality, and follicular fluid exosome diameter were lower in cows with subclinical mastitis, demonstrating a link between mammary gland sanitary status and reproduction.

scholarly journals Progesterone production in superovulated holstein heifers and in crossbred recipient of embryo supplemented with betacarotene and tocopherol

2011 ◽  
Vol 35 (4) ◽  
pp. 817-825
Author(s):  
José Nélio de Sousa Sales ◽  
Lilian Mara Kirsch Dias ◽  
Celso Rodrigues Franci ◽  
Alexandro Aluísio Rocha ◽  
Guilherme Gastão Cardoso ◽  
...  
Keyword(s):  
Intramuscular Injection ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Fixed Time ◽  
Corpora Lutea ◽  
Estrus Synchronization ◽  
Plasma Progesterone ◽  
Blood Samples ◽  
Implant Placement ◽  
Intravaginal Device

Two experiments were conducted to evaluate the effect of the intramuscular injection of betacarotene associated to tocopherol on the plasma concentration progesterone of superovulated Holstein heifers (experiment 1) and in crossbred (Bos taurus x Bos indicus) heifers submitted to fixed-time embryo transfer (FTET, experiment 2). In experiment 1, after estrus synchronization and superovulation animals were inseminated 12 and 24 hours after estrus onset and embryos flushed 7 days later. Heifers were allocated randomly to one of three treatments: Control; T800 (800 mg of betacarotene plus 500 mg of tocopherol) and T1200 (1,200 mg of betacarotene plus 750 mg of tocopherol). The treatments were given on the day of ear implant placement and repeated on the first day of superovulation. Blood samples were collected on D0, D5, D9, D12 and D16. In experiment 2, treatments were imposed at intravaginal device insertion (D0). The same experimental design, as in experiment 1, was used. Blood samples were collected on D17 (embryos implanted) for progesterone determination by radioimmunoassay. In experiment 1, average plasma progesterone concentrations after corpora lutea formation (D12 plus D16 means) were 13.7±1.8 ng/ml, 14.5±2.3 ng/ml and 10.8±2.3 ng/ml for control, T800 and T1200, respectively, and did not differ (P=0.44). In experiment 2, progesterone concentrations on D17 in Control (8.88±0.57 ng/ml), T800 (7.48±0.64 ng/ml) and T1200 (5.90±1.33 ng/ml) groups were similar (P=0.11). Results indicate that the supplemental betacarotene and tocopherol injections did not influence peripheral progesterone concentrations in superovulated Holstein donors and crossbreed recipients heifers.

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