scholarly journals 2 CORRELATION BETWEEN OXYGEN RESPIRATION RATES AND MORPHOLOGY, SEX, DIAMETER AND DEVELOPMENTAL STAGE OF SINGLE BOVINE IVP-EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 151 ◽  
Author(s):  
A.S. Lopes ◽  
N. Ramsing ◽  
L.H. Larsen ◽  
M. Räty ◽  
J. Peippo ◽  
...  
Keyword(s):  
Developmental Stage ◽  
Respiration Rate ◽  
Developmental Stages ◽  
Glass Tube ◽  
Outer Diameter ◽  
Bovine Embryos ◽  
Embryo Viability ◽  
Concentration Gradients ◽  
Cell Stage ◽  
Respiration Rates

A simple, non-invasive, rapid and sensitive oxygen microsensor system was developed to investigate correlations between oxygen respiration rates of individual bovine embryos and their morphology, sex, diameter and developmental stage. Bovine IVP-embryos (n = 78; Holm et al. Theriogenology 52, 683–700) were analysed around the 8-cell stage (Day 3; n = 18) and at various blastocyst stages (Day 7; n = 60). Each embryo was morphologically evaluated, its outer diameter measured and was then loaded into a glass tube (i.d. 0.68 mm, length 3 mm). After 1 h, oxygen concentration gradients generated by the embryo’s respiration were measured over app. 8 min with an oxygen microelectrode (www.unisense.com). Five embryos were measured in one round together with an empty tube as control. The procedure was repeated twice for each embryo with app. 1 h interval. Individual respiration rates in nL O2/embryo/h (nL/h) were calculated from these gradients. The measurements were performed at 38.5°C under constant flow of humidified 5% CO2 in air (app. 19% O2). After this, 64 embryos (14 Day 3; 50 Day 7) were lysed for sex diagnosis by PCR. Values are given as mean ± SD. The sensitivity of the oxygen measurement system was high (controls: 0.034 ± 0.035 nL/h, n = 15) and its repeatability from 1st to 2nd measurement was 99.7 ± 9.8% (n = 71). The average embryo respiration rate was 0.39 ± 0.05 nLl/h on Day 3 (n = 18) and 1.31 ± 0.52 nLl/h on Day 7 (n = 60). For Day 7 embryos, the respiration rates varied according to their morphological quality, being 1.87 ± 0.46a (n = 18), 1.17 ± 0.32b (n = 23), 0.95 ± 0.27b,c (n = 14) and 0.72 ± 0.24c (n = 4) nL/h for quality 1, 2, 3, and 4 embryos, respectively (Proc Mixed,a,b,c: P < 0.05; values with different superscripts differ significantly). The sex ratio (male:female) was 9:5 (Day 3) and 32:18 (Day 7), and on Day 7 this ratio varied between qualities: 11:2, 12:8, 8:4, and 1:3 for quality 1, 2, 3, and 4, respectively. The average respiration rate on day 3 was the same for males and females, as it was on day 7 (1.22 ± 0.43 nL/h (females) and 1.31 ± 0.58 nL/h (males), P > 0.05). There was a correlation between embryo diameter and respiration rate (r2 = 0.65, n = 74), which was even stronger for Day 7 male embryos (r2 = 0.72, n = 32). In conclusion, a highly reliable, repeatable and sensitive system was established for measuring respiration rates in single bovine embryos, even at early developmental stages. The respiration rate was lower on day 3 compared to Day 7 embryos, and it was correlated with the morphological embryo quality on Day 7. Oxygen consumption could be a valuable supplementary indicator of embryo viability, especially in difficult evaluations (e.g. quality 2 and 3 after IVP). It remains to be demonstrated if such measurements can also reveal quality differences already at Day 3, which would be of interest in, e.g. the human field. ASL is supported by FCT, Portugal.

scholarly journals Respiration rates of individual bovine in vitro-produced embryos measured with a novel, non-invasive and highly sensitive microsensor system

Reproduction ◽  
2005 ◽  
Vol 130 (5) ◽  
pp. 669-679 ◽  
Author(s):  
A S Lopes ◽  
L H Larsen ◽  
N Ramsing ◽  
P Løvendahl ◽  
M Räty ◽  
...  
Keyword(s):  
Respiration Rate ◽  
Embryo Quality ◽  
Developmental Stages ◽  
Embryo Morphology ◽  
The Novel ◽  
Non Invasive ◽  
Respiration Rates ◽  
Morphological Evaluation ◽  
Highly Sensitive

Oxygen consumption is a useful parameter for evaluating embryo quality, since it provides a valuable indication of overall metabolic activity. Over the years, several approaches have been used to measure the respiration rates of individual embryos, but a convincing method has not yet been reported. In this study, we introduce and have validated a novel high resolution microsensor technology to determine the respiration rates of individual embryos at different developmental stages. We have employed this technology to investigate the correlation between respiration rate and embryo morphology, diameter and sex. Following morphological evaluation, individual respiration rates of day 3 (n = 18) and day 7 (n = 60) bovine in vitro-produced embryos were determined. Of the measured embryos, 64 were lysed for sex diagnosis by PCR. Average respiration rates of day 7 embryos (1.30 ± 0.064 nl/h) were 3.4-fold higher than day 3 embryos (0.38 ± 0.011 nl/h). On day 7, the average respiration rate of quality 1 blastocysts was significantly higher than the respiration rates of the lower qualities. For both day 3 and day 7 embryos, respiration rates were directly influenced by embryo diameter but did not differ between sexes. These results have demonstrated that the novel microsensor technology can be used to accurately and rapidly (8 min) measure the respiration rates of individual embryos at different developmental stages. Respiration rates were only in partial agreement with embryo morphology, suggesting a slight discrepancy between these two methods in assessing embryo quality. It is likely that a combined assessment of embryo respiration and morphology would improve embryo classification and subsequent selection.

132 DIFFERENCES IN RESPIRATION RATES BETWEEN IN VIVO- AND IN VITRO-PRODUCED BOVINE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 174
Author(s):  
A. S. Lopes ◽  
S. E. Madsen ◽  
N. B. Ramsing ◽  
L. H. Larsen ◽  
T. Greve ◽  
...  
Keyword(s):  
Metabolic Stress ◽  
Average Diameter ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Outer Diameter ◽  
Bovine Embryos ◽  
Physiological Level ◽  
Respiration Rates

In vitro-produced (IVP) bovine embryos differ (e.g. morphology and physiology) from their in vivo counterparts. Oxygen consumption is an indicator of the overall metabolic activity of a single embryo. Therefore, the aim of this study was to determine and compare respiration rates of in vivo- and in vitro-produced bovine day 7 embryos. Diameters of these two embryo types were also compared. In vivo embryos (n = 28) were recovered from 8 superovulated Holstein Frisian cows on day 7 following AI, while IVP embryos (n = 160; Holm et al. 1999 Theriogenology 52, 683-700) were used on day 7 after fertilization. Embryos were measured (outer diameter) and morphologically evaluated (Quality 1 to 4, IETS Manual, 1998). Only transferable in vivo embryos were used (i.e. excluding Quality 4). Respiration rates were measured on each embryo by Nanorespirometer technology (Lopes et al. 2005 Reprod. Fertil. Develop. 17, 151). Data were analyzed using Proc Mixed, and values are presented as mean � SEM. Values with different superscripts differ significantly (P < 0.05). The average respiration rates were 0.82 � 0.06a nL/h for in vivo vs. 1.37 � 0.06b nL/h for IVP embryos. The average respiration rates for the different morphological qualities were as follows (nL/h, numbers in brackets): IVP: 2.1 � 0.08a (38), 1.37 � 0.07b (55), 1.08 � 0.07c (48) and 0.62 � 0.11d (19) for Quality 1, 2, 3, and 4, respectively. In vivo: 1.17 � 0.21b,c,e (6), 0.80 � 0.15c,d,e (12), and 0.64 � 0.16d,f (10) for Quality 1, 2, and 3, respectively. The average diameter (mm) of in vivo and IVP embryos was 0.157 � 0.002a and 0.176 � 0.002b, respectively. Respiration rates were directly related to embryo diameter; larger embryos were associated with higher respiration rates (y = 17.55 � 1.32 nL/h � mm, n = 188). Respiration rates of in vivo embryos were significantly lower than those of IVP embryos, regardless of quality. This difference could reflect an effect of the culture conditions on IVP embryos because media components affect embryo metabolism. Moreover, the different ages (day 7 for IVP vs. approximately Day 6.5 for in vivo embryos, because in vivo embryos are less than 7 days after fertilization at recovery) and stages (IVP: up to expanded blastocyst stage; in vivo: morula or early blastocyst stage) could have influenced the results and also partly explain the smaller diameter of the in vivo embryos. Finally, respiration rates decreased proportionately to the morphological quality within embryo type, indicating that morphological differences are reflected at the physiological level. In conclusion, this study further outlines metabolic differences between in vivo and IVP bovine embryos. Whether such differences are a manifestation of metabolic stress associated to the separation from the natural environment or reflect suboptimal culture conditions is yet to be determined. ASL is supported by FCT, Portugal.

scholarly journals The effect of vitrification in open pulled straws on pregnancy rates after transfer of in vivoproduced bovine embryos

2012 ◽  
Vol 51 (No. 9) ◽  
pp. 454-460 ◽  
Author(s):  
M. Lopatarova ◽  
S. Cech ◽  
L. Holy ◽  
R. Dolezel
Keyword(s):  
Developmental Stage ◽  
Conventional Method ◽  
Developmental Stages ◽  
Pregnancy Rates ◽  
Control Group ◽  
Bovine Embryos ◽  
Group V ◽  
Freezing Method ◽  
Conventional Freezing

The aim of this study was to compare pregnancy rates after transfer of in vivo produced embryos cryopreserved using open pulled straw (OPS) vitrification (Group V) or conventional freezing method as a control (Group C). Bovine embryos (Day<sub>6.5&ndash;7.5</sub>) collected from superovulated cows were classified according to developmental stages and morphological qualities (Grade 1 and 2) before cryopreservation and they were transferred to synchronized heifers after thawing. Pregnancy rates after transfer of morulae, early blastocysts and expanded blastocysts in Group V compared to Group C (54.5%, 12/22 vs. 56.0%, 14/25; 53.3%, 16/30 vs. 58.1%, 18/31 and 57.7%, 15/26 vs. 48.3%, 14/29) were not different (P &gt; 0.05). Likewise, pregnancy rates after transfer of embryos of Grade 1 and 2 in Group V compared to Group C (55.1%, 43/78 vs. 54.1%, 46/85 and 36.4%, 12/33 vs. 32.9%, 23/70, respectively) were not different (P &gt; 0.05). The study demonstrated similar viability of embryos which were frozen by vitrification or conventional method irrespective of their quality and developmental stage after transfer into recipients.

137 EFFECT OF FLUNIXIN MEGLUMINE IN CO-CULTURE MEDIUM ON THE DEVELOPMENT OF IN VITRO MATURED AND FERTILIZED BOVINE EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 219 ◽  
Author(s):  
S. Goda ◽  
S. Hamano ◽  
M. Miyamura ◽  
O. Dochi ◽  
H. Koyama
Keyword(s):  
Embryo Transfer ◽  
Culture Medium ◽  
Cell Monolayer ◽  
Cumulus Cell ◽  
The Other ◽  
Bovine Embryos ◽  
Embryo Viability ◽  
Cell Stage ◽  
Flunixin Meglumine

PG concentration is often increased during uterine manipulation with embryo transfer. Embryo viability is affected by the increase in the PGF2α concentration accompanying manipulation of the uterus during embryo transfer. Schrick et al. (2001 Theriogenology 55, 370 abst) observed that treatment with flunixin meglumine, an inhibitor of prostaglandin, increased pregnancy rates depending on the stage and quality of embryos transferred. On the other hand, prostaglandin was secreted by a cumulus cell monolayer in an in vitro culture of bovine oocytes. The present study aimed to assess the effects of flunixin meglumine in culture medium on the development of in vitro-matured and fertilized bovine embryos. COCs were collected from ovaries of slaughtered cows by aspiration. The COCs were matured for 20 h in TCM-199 supplemented with 5% fetal bovine serum (FBS) and antibiotics at 38.5°C under an atmosphere of 2% CO2 in air. Matured COCs were inseminated with 1.0 × 107 sperm mL−1 in BO medium (Brackett and Oliphant 1975 Biol. Reprod. 12, 260–274) containing 5 mM theophillin and 5 μg mL−1 heparin for 5 h. All of the inseminated oocytes were introduced into the maturation medium that had been kept with the cumulus cells in the CO2 incubator. At 48 h after insemination, all embryos over the 4-cell stage were cultured in TCM-199 plus 5% FBS supplemented with each of five concentrations of flunixin meglumine (0, 0.0025, 0.005, 0.01, and 0.025%) with a cumulus cell monolayer. Development to the blastocyst stage and quality were examined at Days 7 to 8 (Day 0 = day of insemination) using a microscope. The experiment was replicated four times. Data were analyzed by the chi-square test. The total blastocyst rates from the over-4-cell embryos were 61.2 (52/89), 53.7 (44/89), 65.6 (59/90), 57.3 (51/89), and 33.7% (31/92) for 0, 0.0025, 0.005, 0.01, and 0.025%, flunixin meglumine, respectively. The total blastocyst rate with the flunixin meglumine concentration of 0.025% was significantly lower than those with the other concentrations (P < 0.05). The proportion of grade 1 blastocysts with the flunixin meglumine concentration of 0.005% was significantly higher than that with the 0, 0.0025, and 0.025% concentrations (27.8 vs 11.2, 14.6, and 5.4%; P < 0.05). Our present results show that the addition of 0.005% flunixin meglumine to the co-culture medium is positively associated with blastocyst quality in bovine embryos.

scholarly journals Identification and regulation of glycogen synthase kinase-3 during bovine embryo development

Reproduction ◽  
2010 ◽  
Vol 140 (1) ◽  
pp. 83-92 ◽  
Author(s):  
I M Aparicio ◽  
M Garcia-Herreros ◽  
T Fair ◽  
P Lonergan
Keyword(s):  
Embryo Development ◽  
Developmental Stages ◽  
Glycogen Synthase ◽  
Specific Inhibitor ◽  
Blastocyst Stage ◽  
Serine Phosphorylation ◽  
Glycogen Synthase Kinase ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cell Stage

The aim of this study was to examine the presence and regulation of glycogen synthase kinase-3α (GSK3A) and GSK-3β (GSK3B) in bovine embryos and their possible roles in embryo development. Our results show that GSK3A and GSK3B are present in bovine embryos at the two-cell stage to the hatched blastocyst stage. Bovine embryo development was associated with an increase in the phosphorylation of both isoforms, being statistically significant at blastocyst and hatched blastocyst stages, compared with earlier stages. Inhibition of GSK3 with CT99021 (3 μM) resulted in a significant increase in the percentage and quality of blastocysts, while inhibition of GSK3 with lithium chloride (LiCl; 20 mM) significantly reduced at the proportion of eight-cell embryos on day 3 and inhibited blastocyst formation. The use of LY294002 (10 μM), a specific inhibitor of phosphatidylinositol-3 kinase, also produced a significant decrease in embryo development. In addition, treatment with LiCl and LY294002 produced a significant decrease in the serine phosphorylation of both isoforms of GSK3. Finally, CT99021 and LiCl reduced the phosphorylation of β-catenin on Ser45 in two-cell embryos, while LY294002 increased it. Despite the fact that LiCl inhibited GSK3 activity, as demonstrated by β-catenin phosphorylation, its effects on the bovine embryo could be mediated through other signaling pathways leading finally to a decrease in the phosphorylation of GSK3 and a reduction in embryo development. Therefore, in conclusion, GSK3A/B serine phosphorylation was positively correlated with embryo development, indicating the importance of an accurate regulation of GSK3 activity during developmental stages to achieve normal bovine embryo development.

Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos

Zygote ◽  
2014 ◽  
Vol 23 (4) ◽  
pp. 485-493 ◽  
Author(s):  
A.F. Pereira ◽  
L.M. Melo ◽  
V.J.F. Freitas ◽  
D.F. Salamone
Keyword(s):  
Dna Damage ◽  
Developmental Stages ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Bovine Embryos ◽  
Dna Breaks ◽  
Embryo Production ◽  
Cell Stage ◽  
Γh2ax Foci

SummaryIn vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of γH2AX foci or area were detected among the treatments. γH2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development.

scholarly journals 252A COMPARATIVE EXPRESSION ANALYSIS OF GENES IN PREIMPLANTATION DEVELOPMENTAL STAGES OF BOVINE EMBRYOS PRODUCED IN VITRO OR IN VIVO

2004 ◽  
Vol 16 (2) ◽  
pp. 246 ◽  
Author(s):  
D. Tesfaye ◽  
K. Wimmers ◽  
M. Gilles ◽  
S. Ponsuksili ◽  
K. Schellander
Keyword(s):  
Developmental Stage ◽  
Developmental Stages ◽  
Blastocyst Stage ◽  
The Novel ◽  
Cell Stage ◽  
Novel Transcript ◽  
Stage Of Development ◽  
Significant Difference

A comparative analysis of mRNA expression patterns between embryos produced under different in vitro and in vivo culture systems allows the isolation of genes associated with embryo quality and investigation of the effect of culture environment on the embryonic gene expression. In this study, expression analysis of four known (PSCD2, TCF7L2, NADH-subunit and PAIP1) genes and one novel transcript, derived from differential display PCR, was performed in in vitro (Ponsuksili et al., 2002, Theriogenology 57, 1611–1624) or in vivo- (Moesslacher et al., 2001 Reprod. Dom. Anim. 32, 37) produced bovine 2-, 4-, 8-, 16-cell, morula and blastocyst stage embryos using real time PCR technology. Poly(A) RNA was isolated from four separate individual embryos from each developmental stage and embryo group (in vitro or in vivo) using Dynabeads mRNA kit (Dynal, Oslo, Norway). After reverse transcription, quantitative PCR was performed with sequence specific primers in an ABI PRISM® 7000 Sequence Detection System instrument (Applied Biosystems, Foster City, CA, USA) using SYBR® Green as a double-strand DNA-specific fluorescent dye. Standard curves were generated for target and endogenous genes using serial dilutions of plasmid DNA. Final quantification was done using the relative standard curve method, and results were reported as relative expression or n-fold difference to the calibrator cDNA (i.e., the blastocyst stage) after normalization with the endogenous control (Histone2a). Data were analyzed using SAS version 8.0 (SAS Institute Inc., NC, USA) software package. Analysis of variance was performed with the main effects being the developmental stage and embryo source (in vitro or in vivo) and their interactions followed by multiple pairwise comparisons using Tukey’s test. No significant difference was observed in the relative abundance of the PSCD2 gene between the two embryo groups. However, its expression was higher (20-fold) (P&lt;0.05) at the 8-cell stage than the other developmental stages among in vitro embryos. Higher expression (P&lt;0.05) of NADH-subunit mRNA was detected in vivo than in vitro at the 2-cell stage of development. The TCF7L2 mRNA was expressed in the in vitro embryos but not in the in vivo ones. PAIP1 mRNA was higher (P&lt;0.05) in in vitro (1500-fold) than in the in vivo embryos (500-fold) at the 2-cell developmental stage compared to the calibrator. The novel transcript was also detected at higher level (P&lt;0.05) in the in vitro than in the in vivo embryos at the 2-cell stage of development. However, the PAIP1 and the novel transcript showed no significant difference in their expression between the two embryo groups beyond the 2-cell developmental stage. Both PAIP1 and the novel transcript were detected only up to 8-cell stage in both embryo groups, suggesting their maternal origin. In conclusion, the variations in the expression of studied genes between in vitro and in vivo may reflect the effect of the two culture systems on the transcriptional activity of early embryos.

EFFECT OF VARIOUS HORMONES ON THE KIDNEY RESPIRATION RATES IN THE CUTTHROAT TROUT (SALMO CLARKI CLARKI)

1960 ◽  
Vol XXXIII (III) ◽  
pp. 428-436 ◽  
Author(s):  
W. N. Holmes
Keyword(s):  
Single Dose ◽  
Respiration Rate ◽  
Cutthroat Trout ◽  
Adaptive Mechanism ◽  
Respiration Rates ◽  
Salmo Clarki

ABSTRACT Relatively large doses of vasopressin administered intraperitoneally to the trout significantly enhanced the kidney respiration rate. In contrast to vasopressin a single dose of oxytocin depressed the kidney Qo2 value. This depression continued throughout the observed 24 hour period after injection. Cortisol enhanced the kidney Qo2 values significantly and to a greater extent than vasopressin. These results are discussed in relation to possible adaptive mechanism in euryhaline species of teleosts.

Validation of On-line Respiration Meter and its Applications by Computer Simulation

1992 ◽  
Vol 26 (5-6) ◽  
pp. 1355-1363 ◽  
Author(s):  
C-W. Kim ◽  
H. Spanjers ◽  
A. Klapwijk
Keyword(s):  
Steady State ◽  
Activated Sludge ◽  
Respiration Rate ◽  
Oxygen Demand ◽  
Operating Conditions ◽  
Mass Balances ◽  
Short Term ◽  
Respiration Rates ◽  
On Line ◽  
Made In

An on-line respiration meter is presented to monitor three types of respiration rates of activated sludge and to calculate effluent and influent short term biochemical oxygen demand (BODst) in the continuous activated sludge process. This work is to verify if the calculated BODst is reliable and the assumptions made in the course of developing the proposed procedure were acceptable. A mathematical model and a dynamic simulation program are written for an activated sludge model plant along with the respiration meter based on mass balances of BODst and DO. The simulation results show that the three types of respiration rate reach steady state within 15 minutes under reasonable operating conditions. As long as the respiration rate reaches steady state the proposed procedure calculates the respiration rate that is equal to the simulated. Under constant and dynamic BODst loading, the proposed procedure is capable of calculating the effluent and influent BODst with reasonable accuracy.

scholarly journals Evaluation of Bovine Embryo Biopsy Techniques according to Their Ability to Preserve Embryo Viability

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
M. Cenariu ◽  
E. Pall ◽  
C. Cernea ◽  
I. Groza
Keyword(s):  
Pregnancy Rate ◽  
Genetic Diagnosis ◽  
Needle Aspiration ◽  
Embryo Biopsy ◽  
Embryo Cryopreservation ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Embryo Viability ◽  
Biopsy Methods ◽  
Biopsy Method

The purpose of this research was to evaluate three embryo biopsy techniques used for preimplantation genetic diagnosis (PGD) in cattle and to recommend the least invasive one for current use, especially when PGD is followed by embryo cryopreservation. Three hundred bovine embryos were biopsied by either one of the needle, aspiration or microblade method, and then checked for viability by freezing/thawing and transplantation to recipient cows. The number of pregnancies obtained after the transfer of biopsied frozen/thawed embryos was assessed 30 days later using ultrasounds. The results were significantly different between the three biopsy methods: the pregnancy rate was of 57% in cows that received embryos biopsied by needle, 43% in cows that received embryos biopsied by aspiration, and 31% in cows that received embryos biopsied by microblade. Choosing an adequate biopsy method is therefore of great importance in embryos that will undergo subsequent cryopreservation, as it significantly influences their viability after thawing.

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