Effect of body condition and season on yield and quality ofin vitroproduced bovine embryos

Zygote ◽  
2014 ◽  
Vol 23 (6) ◽  
pp. 893-899 ◽  
Author(s):  
Peter Chrenek ◽  
Elena Kubovičová ◽  
Lucia Olexíková ◽  
Alexander V. Makarevich ◽  
Silvia Toporcerová ◽  
...  

SummaryThe aim of our study was to examine the effects of cow's body condition score (BCS; scale 1–5) and season on the quality of bovinein vitroproduced embryos. The proportion of good quality oocytes (Q1 and Q2) was higher (P< 0.05) in the BCS 2 (57.60%) and BCS 3 (60.90%) groups compared with the BCS 1 (43.60%) group. There were no statistical differences in embryo cleavage and blastocyst rate among the BCS groups. The highest total cell number (TCN, DAPI stain) of blastocysts (P< 0.05), recorded in BCS 1 (122.27 ± 6.90) in comparison with BCS 2 (101.8 ± 3.60) or BCS 3 (105.44 ± 3.70) groups, was related to higher dead cell (DCI, TUNEL) index in this group (7.07%) when compared with BCS 2 (6.54%) or BCS 3 (6.06%), respectively. The yield of good quality oocytes during spring was lower (P< 0.05) compared with the summer season. There were significant differences (P< 0.05) in maturation and cleavage rates between autumn and summer (73.42%, 76.2% vs. 85.0%, 41.8%, respectively). The highest (P< 0.01) blastocyst rate was noted during spring and summer months. Significant difference (P< 0.05) in the TCN among spring (99.38 ± 3.90), autumn (110.1 ± 4.58) or summer (108.96 ± 3.52) was observed. The highest proportion of embryos with the best (grade I) actin cytoskeleton (phalloidin–TRITC) quality was noted during the summer months. Our results indicate that body condition affects the initial quality of oocytes, but does not affect embryo cleavage, blastocyst rate and actin quality. This finding may suggest that developmentin vitrocan mask the influence of BCS. The season affects yield and quality of blastocysts in the way that the autumn period is more favorable for embryo development.

2007 ◽  
Vol 19 (1) ◽  
pp. 147
Author(s):  
H. T. Lee ◽  
J. M. Jang ◽  
S. H. Lee ◽  
M. K. Gupta

In vitro production of cloned porcine embryos by somatic cell nuclear transfer (SCNT) has become routine in several laboratories but the efficiency and quality of the resultant blastocysts remains sub-optimal. Cloned porcine blastocysts show low cell number, high fragmentation rate, and apoptosis which results in lower pregnancy rates upon embryo transfer. Earlier we reported that supplementation of culture media with amino acids benefit pre-implantation embryo development of in vivo- as well as in vitro-fertilized porcine embryos (Koo et al. 1997 Theriogenology 48, 791–802). This study evaluated how exogenous amino acids could affect pre-implantation development and quality of cloned or parthenogenetic porcine embryos. The effects of commercially available amino acids, referred to as Eagle&apos;s non-essential amino acids (NEAA), added or not added (control) to NCSU23 medium containing fatty acid-free BSA were studied. Oocytes recovered from abattoir-derived prepubertal porcine ovaries were matured in vitro and parthenogenetically activated (PA) or nuclear-transferred with fetal fibroblasts (SCNT), as described earlier (Uhm et al. 2000 Mol. Reprod. Dev. 57, 331–337). At 168 h post-activation, blastocysts were harvested for assessment of embryo quality by TUNEL labeling, Hoechst 33342 staining, and gene expression analysis. Results showed that, in the PA group, the cleavage rate was not affected by the supplementation of NEAA. However, the blastocyst rate was significantly improved when NEAA was present in the medium compared to that of the control group (38.9 &plusmn; 0.3 vs. 27.5 &plusmn; 0.3&percnt;, respectively) throughout the culture period. The supplementation during the pre-compaction period alone gave better results than during the post-compaction period alone (59.5 &plusmn; 0.9 vs. 33.4 &plusmn; 0.3&percnt;, respectively). In the SCNT group, however, both cleavage (73.6 &plusmn; 0.2 vs. 64.2 &plusmn; 0.4&percnt;) and blastocyst rate (18.7 &plusmn; 0.2 vs. 13.8 &plusmn; 0.3&percnt;) were improved by NEAA supplementation. Furthermore, these blastocysts had higher hatching ability (30.0 &plusmn; 1.8 vs. 14.6 &plusmn; 4.9&percnt;) than those of control group (P &lt; 0.05). Supplementation of NEAA also increased the mean nuclei number of PA-derived (76.1 &plusmn; 4.9 vs. 66.5 &plusmn; 3.3) as well as SCNT-derived (43.1 &plusmn; 2.6 vs. 31.8 &plusmn; 1.9) blastocysts and reduced the time during which blastocysts formed. TUNEL assay revealed that incidence of nuclear fragmentation and apotosis was reduced by NEAA. Real-time qRT-PCR for Bax and Bcl-XL transcripts revealed that the relative abundance of Bax was reduced while that of Bcl-XL was increased. These effects were more pronounced when NEAA was present during the pre-compaction period alone. Thus, our data suggest that NEAA improves the yield and quality of cloned porcine embryos by enhancing blastocyst expansion and positively modulating the total cell number and apoptosis. These data may have implications for understanding the nutritional needs of cloned porcine embryos produced in vitro and for optimizing the composition of culture media to support their development. This work was supported by the Research Project on the Production of Bio-Organs (No. 200503030201), Ministry of Agriculture and Forestry, Republic of Korea.


Zygote ◽  
2015 ◽  
Vol 24 (3) ◽  
pp. 378-388 ◽  
Author(s):  
A.V. Makarevich ◽  
L. Stádník ◽  
E. Kubovičová ◽  
Z. Hegedüšová ◽  
R. Holásek ◽  
...  

SummaryThis study examined the impact of cow body condition on the quality of bovine preimplantation embryos. The embryos (n = 107) were flushed from dairy cows and classified according to a five-point scale body condition score (BCS2 n = 17; BCS3 n = 31; BCS4 n = 11) on the 7th day after insemination and then analyzed for development, dead cell index (DCI), cell number and actin cytoskeleton quality. The highest embryo recovery rate (P < 0.05) was recorded in the BCS3 group and the lowest in the BCS4 group. More transferable (morula, blastocyst) embryos were obtained from the BCS4 cows (79%), compared with the BCS2 (64%) or BCS3 (63%) animals. However, cell numbers were higher in the BCS2 and BCS3 groups (P < 0.05) compared with the BCS4 embryos. Conversely, the DCI was lowest in the BCS2 (3.88%; P < 0.05) and highest in the BCS4 (6.56%) embryos. The proportion of embryos with the best actin quality (grade I) was higher in the BCS2 and BCS3 cows compared with the BCS4 group. Almost 25% of all embryos showed fragmented morphology and a higher DCI (5.65%) than normal morulas (1.76%). More fragmented embryos were revealed in the BCS2 (28.6%) and BCS4 (31.25%) groups, and less (19.15%) in the BCS3 group. The cell numbers in such embryos were lower in the BCS4 (22.57) than in the BCS2 (46.25) or BCS3 (42.4) groups. In conclusion, the body condition of dairy cows affects the quality of preimplantation embryos. A BCS over 3.0 resulted in a higher incidence of poor (fragmented) embryos.


2013 ◽  
Vol 25 (1) ◽  
pp. 220
Author(s):  
R. Li ◽  
Y. Liu ◽  
H. Pedersen ◽  
P. Kragh ◽  
P. Hyttel ◽  
...  

During in-vitro handling, oocytes and embryos are always exposed to light. This has been shown to affect embryonic development and quality in different species, i.e. hamster and mouse (Takenaka et al. 2007 PNAS, 104, 14 289–14 293) and human (Tatsuo et al. 2010 J. Assist. Reprod. Genet. 27, 93–96). However, similar experiments have not been made on porcine embryos, so our aim was to test effects of different types of light on the development and quality of porcine parthenote embryos. Cumulus–oocyte complexes from slaughterhouse-derived sow ovaries were aspirated and matured (38.5°C, 5% CO2, maximum humidity, 42 h). Parthenogenetic activation was made (Day 0) first by an electric pulse (1.26 kV cm–1, 80 µs) and then by incubation with 5 µg mL–1 cytochalasin B and 10 µg mL–1 cycloheximide in porcine zygote medium-3 (PZM-3) for 4 h. During these processes, the oocytes would be exposed to ~30 min of light. After activation, the oocytes were either directly cultured in incubator (0 h), or experimentally exposed to two types of light (DAY: near window, no direct sunlight; LAB: app. 40 cm from warm white lamps (12 V, 40 W) in PZM-3 while placed on a heating plate (38.5°C) with the culture dish covered by a crystal plastic foil filled with the appropriate gas (5% O2, 5% CO2) for different periods (1 h, 4 h, 24 h), and then cultured in PZM-3 in incubator. On Day 6, total and good blastocysts were counted. Good blastocysts were defined as blastocysts having expanded to 1.5 times the oocytes’ size, having cells of uniform color and distribution, and having formed a regular blastocyst cavity. The total number of cells and the apoptotic cells were detected on Day 6 blastocysts with TUNEL assay to evaluate embryonic quality. All statistics analysis were performed by ANOVA test (R). The results are summarized in Table 1. The developmental rates were decreased with both types of light: the decrease appeared earlier for good blastocyst rates, only after 1 h exposure; however, a clear adverse effect was also found on total blastocyst rates after 24 h exposure. The total cell number decreased after 4 h light exposure for both types of light. The rate of apoptotic cells tended to increase with both types of light (from approximately 7 to 9.5%) when embryos were exposed during 24 h, but no significant difference was found between groups. We conclude that the blastocyst morphology would be altered already after 1 h extra exposure to both light types, and their quality would further decrease after 4 h exposure. However, under normal working conditions, this should not represent a real problem. Table 1.Effect of light exposure on development of porcine parthenote embryos


Zygote ◽  
2016 ◽  
Vol 24 (6) ◽  
pp. 890-899 ◽  
Author(s):  
A.L.S. Guimarães ◽  
S.A. Pereira ◽  
M. N. Diógenes ◽  
M.A.N. Dode

SummaryThe aim of this study was to evaluate the effect of adding a combination of insulin, transferrin and selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation (IVM) and in vitro culture (IVC) on in vitro embryo production. To verify the effect of the supplements, cleavage and blastocyst rates, embryo size and total cell number were performed. Embryonic development data, embryo size categorization and kinetics of maturation were analyzed by chi-squared test, while the total cell number was analyzed by a Kruskal–Wallis test (P < 0.05). When ITS was present during IVM, IVC or the entire culture, all treatments had a cleavage and blastocyst rates and embryo quality, similar to those of the control group (P < 0.05). Supplementation of IVM medium with ITS and AA for 12 h or 24 h showed that the last 12 h increased embryo production (51.6%; n = 220) on D7 compared with the control (39.5%; n = 213). However, no improvement was observed in blastocyst rate when less competent oocytes, obtained from 1–3 mm follicles, were exposed to ITS + AA for the last 12 h of IVM, with a blastocyst rate of 14.9% (n = 47) compared with 61.0% (n = 141) in the control group. The results suggest that the addition of ITS alone did not affect embryo production; however, when combined with AA in the last 12 h of maturation, there was improvement in the quantity and quality of embryos produced. Furthermore, the use of ITS and AA during IVM did not improve the competence of oocytes obtained from small follicles.


Zygote ◽  
2009 ◽  
Vol 17 (1) ◽  
pp. 57-61 ◽  
Author(s):  
M. Popelková ◽  
Z. Turanová ◽  
L. Koprdová ◽  
A. Ostró ◽  
S. Toporcerová ◽  
...  

SummaryThe aim of the study was to determine the efficiency of two vitrification techniques followed by two assisted hatching (AH) techniques based on post-thaw developmental capacity of precompacted rabbit embryos and their ability to leave the zona pellucida (hatching) during in vitro culture. The total cell number and embryo diameter as additional markers of embryo quality after warming were evaluated. In vivo fertilized, in vitro cultured 8–12-cell rabbit embryos obtained from superovulated rabbit does were cryopreserved by two-step vitrification method using ethylene glycol (EG) as cryoprotectant or by one-step vitrification method with EG and Ficoll (EG+Ficoll). Thawed embryos were subjected to enzymatic or mechanical AH. Vitrified EG group showed significantly lower (P < 0.05) blastocyst rate (22.5%) and hatching rate (15%) than those vitrified with EG + Ficoll (63 and 63% resp.) and that of control (97 and 97% respectively). Significantly lower values of total cell number (P < 0.05) as well as embryo diameter (P < 0.01) in EG group compared with EG + Ficoll and control group were recorded. No significant difference was found in developmental potential of warmed embryos treated by either mechanical or enzymatic AH. The present study demonstrates that the EG + Ficoll vitrification protocol provides superior embryo survival rates over the EG vitrification protocol for 8–12-cell stage precompacted rabbit embryos. No positive effect of either mechanical or enzymatic AH on the post-thaw viability and quality of rabbit embryos in vitro was observed.


Author(s):  
Antonín Vaculík

Caraway has very low competitive ability against most of weed species. Therefore, it is important to choose a suitable herbicide control, especially at biennial caraway. The experiments with the herbicides applied in caraway were found in pure growth, with variety Kepron (with standard length of ve­ge­ta­ti­ve period). The aim of the trials was to find out the differences at achenes yiled and essential oil content after the treatment by the various herbicides, applied preemergently. Also, the selective ac­ti­vi­ty of individual herbicides was tested, including the different doses. The herbicides used in the ex­pe­ri­ments had enough level of selektivity on the plants of caraway. They had a positive effect on the yiled comparing to non-treated control variant in all observed years. There was no statistically significant difference in essential oil kontent between the treated variants and non-treated control. On the basis of these results, the most suitable herbicide was chosen for the „minor“ registration for caraway.


2005 ◽  
Vol 17 (2) ◽  
pp. 181 ◽  
Author(s):  
D. Sage ◽  
P. Hassel ◽  
B. Petersen ◽  
W. Mysegades ◽  
P. Westermann ◽  
...  

Porcine nuclear transfer (NT) is an inefficient process and it is necessary to use as many as 120 NT embryos for each foster mother to obtain small litters of live piglets. In these experiments, we evaluated the effects of culture atmosphere and medium on the development of NT embryos by monitoring blastocyst rate and cell number of Day 6 blastocysts. Age matched IVF and parthenogenetic embryos were also evaluated for comparison. For all experiments a pool of oocytes was aspirated from ovaries collected in a local abattoir. Following aspiration, oocytes were allowed to mature for 40 h in North Carolina State University (NCSU)-37 medium (supplemented with cAMP and hCG/eCG for the first 22 h). After removal of the cumulus cells, denuded oocytes with polar bodies were selected for NT, enucleated, fused with fetal fibroblasts, and sequentially activated electrically and chemically by 3 h of treatment with 6-dimethylaminopurine (6-DMAP). A second group of oocytes from the same denuded pool were maintained in TL-HEPES medium and activated in parallel with the NT group to produce parthenogenetic embryos. A third group was fertilized with frozen-thawed epididymal semen and co-cultured for ∼12 h to give IVF embryos. All three treatment groups were subdivided into a control subgroup and an experimental subgroup. In the first experiment, we compared the effects of atmosphere (20% vs. 5% oxygen) on in vitro embryonic development in NCSU-23 medium. In the second experiment, we used only the 5% oxygen concentration and compared different culture media. One subgroup was maintained in standard NCSU-23 medium and the second subgroup was cultured in a two-step system for the first 58 h in modified NCSU-23 (without glucose but supplemented with 2.0 mM lactate and 0.2 mM pyruvate), followed by addition of glucose to give a final concentration of 5.55 mM. Data were statistically analyzed by analysis of variance and chi square test. Blastocyst rate and mean cell number in all three embryo groups were improved under 5% oxygen. The most dramatic effect was observed in the NT group, in which the blastocyst rate increased significantly (P < 0.001) from 6.7% ± 5.9 (n = 279) to 19.6% ± 8.9 (n = 250) and mean cell number increased from 17.7 ± 12.1 to 25.8 ± 10.3 cells per blastocyst. With 5% oxygen there was also an increase of blastocyst rates and mean cell numbers in both IVF and parthenogenetic groups. In the second experiment, blastocyst rate for NT embryos increased significantly (P < 0.05) from 21.8% ± 7.6 (n = 242) in conventional NCSU-23 to 31.5% ± 11.0 (n = 271) in the modified system whereas there was almost no difference in the mean cell number of both groups (29.2 ± 4.3 vs. 31.5 ± 5.3). In the groups of IVF and parthenogenetic embryos no difference was found. These results indicate that both the reduced oxygen and the modified culture medium are important for pre-implantation development of porcine nuclear transfer embryos.


2016 ◽  
Vol 3 ◽  
pp. 48
Author(s):  
Carlos Rodríguez ◽  
Juan A. Quiñones ◽  
Rodrigo Arias

The trial was conducted at the Centro de Producción Agrícola of ICfA in Cuyuta, Escuintla-Guatemala in order to generate information on the yield and quality of edible mulberry (Morus sp.) roughage treatments consisted of three harvest frequencies (6; 9 and 12 weeks and three fertilization levels of nitrogen (0.40 and 80 kg/ha). A complete randomized block experimental design, with a factorial arrangement (3x3) was used. The cutting height was 0.3 m above the ground level with two sequencies: from August 2nd to September 13 th and from September 13th to December 6th, 1990. In both cases, the 12 week frequency cutting and 80 kg of N were superior to the others (P<=0.01), yielding 6.87 and 6.15 t/ha of dry matter respectively. The yields at 9 weeks were statistically higher than those at 6 weeks. The highest protein percentage of the whole plant, leaves and stalks were produced at 6 weeks. The dry matter digestibility values in vitro showed little variability among treatments, with averages for whole plants, leaves and stalks of 65, 91 and 41 %, respectively. The preceeding data suggests that the mulberry has an excellent potential as a balanced supplement (protein and energy) for dual purpose cattle on the Southern coast of Guatemala.


2020 ◽  
Vol 15 (4) ◽  
pp. 335-342
Author(s):  
Anvar Sh. Gadzhikurbanov

The development of rational system of basic soil tillage results in formation of sufficiently high crop yields. A two-factor field experiment was carried out to address these issues when cultivating winter rape cultivars on light chestnut soils of the Primorsko-Caspian sub-province of the Republic of Dagestan. VEM (standard), Elvis and Meteor winter rape cultivars were studied in the experiment. The following methods of soil tillage were studied: real tillage (control), subsoil tillage. The research revealed that there was no significant difference in agrophysical indicators between the soil cultivation variants. Data on weed infestation of winter rape fields showed that the number of weeds and their weight increased significantly after subsoil tillage. At the budding stage, the number of weeds increased 2.1-fold compared to the control (real tillage), and the weight of weeds increased 1.88-fold. During flowering stage, these indicators were 2.2 times higher. The maximum yield in all variants of the experiment was formed by Elvis cultivar - 44.9 and 41.4 t/ha, respectively, which is more than VEM (standard) and Meteor cultivars by 20.4 t/ha (7.9 %) and 20.0 t/ha (8.7 %), respectively. The yield of Meteor cultivar in these variants was also high and amounted to 41.6 and 38.1 t/ha, respectively, which was higher by 11.5 and 10.4 % compared with the standard. The greatest increase in the fresh yield was provided by real tillage. On average, the yield increased by 8.7 % for varieties in comparison with subsoil cultivation. Elvis cultivar formed the best quality indicators, followed by Meteor cultivar.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 467-468
Author(s):  
Fernanda F Santos ◽  
Luciano Brochine ◽  
Vanderlei Benetel Junior ◽  
Maria Laura R Coelho ◽  
Augusto H Gameiro ◽  
...  

Abstract The objective of this study was to evaluate the milk production, body weight, and body condition score (BCS) of ewes fed with different energy levels. Seventy-one ewes were used and distributed in three treatments according to the metabolizable energy (ME, Mcal/kg) levels in the diet: CTL = according to the NRC (2007) recommendation for each stage (gestation and lactation); PLUS = CTL plus 10% of ME per kilogram of dry matter; LESS = CTL less 10% ME per kilogram of dry matter. Body weight and body condition score were evaluated a week before the expected date of parturition, a week after giving birth, and at weaning (60 days). Dairy production was evaluated at 20, 30, 40, 50 and 60 days after giving birth by the indirect method of the two weighing. The experimental design was analyzed as randomized complete blocks. There were no differences on milk production during almost all the lactation curve. Only on day 50 (P = 0.051), CTL was still at peak, while LESS and PLUS had already decreased their production. However, at 60 days, all treatments were already with low production, justifying weaning in this period. Results of weight and BCS showed a significant difference in all periods evaluates. Ewes in the PLUS diet had greater BW and BCS. The BCS before giving birth is expected to be over 3.5 and treatment LESS was the only one that did not achieve this score (2.66, 3.37, 3.83 for LESS, CTL, and PLUS, respectively). At weaning, ewes fed PLUS and CTL had BCS of 3.51 and 2.92, respectively, but treatment LESS had only 1.82. That is prejudicial for the next mating seasonal and this animal. We concluded that LESS diet interfered with the body reserves of sheep.


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