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T A E Stout - One of the best experts on this subject based on the ideXlab platform.
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effect of using frozen thawed bovine semen contaminated with lumpy skin disease virus on in vitro Embryo Production
Transboundary and Emerging Diseases, 2019Co-Authors: C H Annandale, Mario P Smuts, Karen Ebersohn, Lizette Du Plessis, Peter N Thompson, Estelle Hildegard Venter, T A E StoutAbstract:Lumpy skin disease (LSD) is an important transboundary animal disease (TAD) of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, lumpy skin disease virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen-thawed semen on in vitro Embryo Production (IVEP) parameters, including viral status of media and resulting Embryos. Bovine oocytes were harvested from abattoir-collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen-thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV-free semen (control). Following Day 7 and Day 8 blastocyst evaluation, PCR and virus isolation were performed on all Embryonic structures. After completing sufficient replicates to reach 1000 inseminated oocytes, further IVF runs were performed to provide material for electron microscopy (EM) and Embryo washing procedures. Overall, in vitro Embryo yield was significantly reduced by the presence of LSDV in frozen-thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all Embryo structures, when higher doses of LSDV were present in the frozen-thawed semen used for in vitro fertilization (IVF). Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society (IETS) washing procedure resulted in Embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures. This article is protected by copyright. All rights reserved.
S Chastantmaillard - One of the best experts on this subject based on the ideXlab platform.
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in vitro Embryo Production efficiency in cattle and its association with oocyte adenosine triphosphate content quantity of mitochondrial dna and mitochondrial dna haplogroup
Biology of Reproduction, 2004Co-Authors: M Tamassia, F Nuttinck, Pascale Maypanloup, Pascal Reynier, Y Heyman, Gilles Charpigny, Miodrag Stojkovic, S Hiendleder, J P Renard, S ChastantmaillardAbstract:Abstract Mitochondria have a broad range of functions that affect reProduction, and structural as well as quantitative variation in mtDNA has been associated with gamete quality and reproductive success. To investigate the mitochondria effect on in vitro Embryo Production, we collected oocytes by ultrasound-guided follicular aspiration from donor cows known to differ in the developmental capacity, measured by the blastocyst formation rate, of their oocytes. To evaluate the potential effects of mtDNA and mitochondrial function on oocyte quality, the donor cows' mtDNA control region was sequenced and, after pairwise comparisons of polymorphisms, animals were grouped into two major haplogroups. The number of mtDNA molecules per oocyte was quantified by real-time PCR, and the adenosine triphosphate (ATP) content was measured in each oocyte to identify variations between haplogroups. Overall, ATP stocks in oocytes of the two haplogroups differed significantly (P < 0.05; means ± SEM) both at the germinal vesicl...
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in vitro Embryo Production efficiency in cattle and its association with oocyte adenosine triphosphate content quantity of mitochondrial dna and mitochondrial dna haplogroup
Biology of Reproduction, 2004Co-Authors: M Tamassia, F Nuttinck, Pascale Maypanloup, Pascal Reynier, Y Heyman, Gilles Charpigny, Miodrag Stojkovic, S Hiendleder, J P Renard, S ChastantmaillardAbstract:Mitochondria have a broad range of functions that affect reProduction, and structural as well as quantitative variation in mtDNA has been associated with gamete quality and reproductive success. To investigate the mitochondria effect on in vitro Embryo Production, we collected oocytes by ultrasound-guided follicular aspiration from donor cows known to differ in the developmental capacity, measured by the blastocyst formation rate, of their oocytes. To evaluate the potential effects of mtDNA and mitochondrial function on oocyte quality, the donor cows' mtDNA control region was sequenced and, after pairwise comparisons of polymorphisms, animals were grouped into two major haplogroups. The number of mtDNA molecules per oocyte was quantified by real-time PCR, and the adenosine triphosphate (ATP) content was measured in each oocyte to identify variations between haplogroups. Overall, ATP stocks in oocytes of the two haplogroups differed significantly (P < 0.05; means +/- SEM) both at the germinal vesicle and metaphase II stages (2.8 +/- 0.06 pmol vs. 2.6 +/- 0.07 pmol and 2.9 +/- 0.1 pmol vs. 2.3 +/- 0.06 pmol, respectively). The proportion of development to blastocyst was significantly different between haplogroups (22.3 +/- 2.1 % vs. 36.7 +/- 2.9 %). The number of mtDNA molecules per oocyte was highly variable (377 327 +/- 14 104, ranging from 2.0 x 10(3) to 1.2 x 10(6)) but not significantly different between the two haplogroups; significant differences were observed between animals without any apparent relationship to blastocyst Production. These data suggest that mitochondria and mtDNA haplogroup affect the developmental capacity of bovine oocytes in vitro.
R Sartori - One of the best experts on this subject based on the ideXlab platform.
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short communication follicle superstimulation before ovum pick up for in vitro Embryo Production in holstein cows
Journal of Dairy Science, 2016Co-Authors: L H Oliveira, Carlos P Sanches, Adriano S Seddon, M B Veras, F A Lima, P L J Monteiro, M C Wiltbank, R SartoriAbstract:Abstract The objective was to evaluate in vitro Embryo Production (IVEP) in nonlactating Holstein cows after ovarian superstimulation. Cows were randomly assigned in a crossover design to 1 of 2 groups: control (n=35), which was not synchronized and not treated with hormones before ovum pick-up (OPU), or hormone-treated (n=35), in which wave emergence was synchronized and animals treated with porcine (p)-FSH in the presence of norgestomet before OPU. In the hormone-treated group, all follicles ≥7mm in diameter were aspirated for synchronization of wave emergence and cows received a norgestomet ear implant. After 36h, treatment with p-FSH (6 doses of 40mg each, 12h apart, i.m.) started. Ovum pick-up from follicles >2mm in diameter was performed 44h after the last p-FSH (coasting). Then, IVEP was performed. The total number of cumulus-oocyte complexes recovered (16.0 vs. 20.5±2.2) and number of grades I to III (viable) oocytes (10.7 vs. 12.3±1.6) did not differ between hormone-treated and control groups Additionally, no differences were found in the number of blastocysts per cow per OPU (3.0 vs. 2.6±0.5) or in blastocyst rates (17.1 vs. 12.2±2.4%) between hormone-treated and control, respectively. Thus, in this study, ovarian follicle superstimulation with p-FSH followed by coasting in nonlactating Holstein cows that had synchronization of wave emergence and progestin supplementation did not improve oocyte quality or IVEP compared to no hormonal treatment.
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in vivo Embryo Production in cows superovulated 1 or 2 days after ovum pick up
Reproduction Fertility and Development, 2014Co-Authors: R S Surjus, M C Wiltbank, A B Prata, Marta Borsato, Fernanda C S Z Mattos, Mariana Martins C Da Silveira, Gerson Barreto Mourao, Alexandre Vaz Pires, R SartoriAbstract:The present study evaluated superovulatory responses and in vivo Embryo Production in cows treated with FSH starting 1 or 2 days after ovum pick-up (OPU). Thirty-three non-lactating Nelore cows were subjected to aspiration of all follicles ≥3 mm for OPU. After OPU, cows were randomly divided into two groups in which the follicle superstimulatory treatments with FSH started 1 or 2 days after OPU (Groups D1 and D2, respectively). Data are presented as the least squares mean ± s.e.m. The number of follicles ≥3 mm before OPU was similar between groups (~34); however, cows in Group D2 had more follicles ≥3 mm on the first day of FSH (15.2 ± 2.3 vs 7.6 ± 1.7; P = 0.04) and a higher ratio of the number of follicles at first FSH/number of follicles before OPU (0.41 ± 0.04 vs 0.24 ± 0.02; P = 0.01). In addition, Group D2 cows had a greater superovulatory response than did cows in Group D1 (18.9 ± 2.8 vs 9.1 ± 1.9 corpora lutea, respectively; P 0.10). Nevertheless Group D2 cows had more freezable Embryos than Group D1 cows (3.2 ± 1.1 vs 1.3 ± 0.5, respectively; P < 0.05). Cows from Group D2 had a much higher proportion (P < 0.001) of follicles ≥8 mm compared with follicles ≥6 mm and <8 mm at the time of the last treatment with FSH. In conclusion, to obtain a greater Production of viable Embryos in superovulated cows after OPU, it is recommended to wait at least 2 days before starting FSH treatment.
J F Moreno - One of the best experts on this subject based on the ideXlab platform.
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125 influence of length of porcine follicle stimulating hormone p fsh treatment before ovum pickup on ovarian response and in vitro Embryo Production in holstein heifers
Reproduction Fertility and Development, 2021Co-Authors: J C L Motta, R V Sala, V A Absalonmedina, V C Fricke, M Dominguez, D C Pereira, C Hayden, E R Canadas, B J Duran, J F MorenoAbstract:Ovarian follicle stimulation with exogenous FSH before ovum pickup (OPU) in Bos taurus females is a common practice to increase invitro Embryo Production (IVP). The optimal stimulatory period length for OPU-IVP, however, has not been definitively ascertained. The objective of the present study, therefore, was to determine the effect of length of the superstimulatory treatment period before OPU on ovarian response and IVP in Holstein heifers. Nonpregnant heifers (n = 57) 13.8 ± 0.2 months of age with moderate body condition score (3.0 ± 0.1; scale 1 to 5) were assigned in a completely randomised design to one of the following experimental groups: FSH2d: 200 mg of p-FSH (Folltropin-V®, Vetoquinol) distributed in four injections (60, 60, 40, and 40 mg) of FSH 12 h apart; FSH3d: 200 mg of p-FSH distributed in six injections (40, 40, 40, 40, 20, and 20 mg) of FSH 12 h apart. Timing of follicular wave emergence was synchronized by dominant follicle removal 36 h before the first p-FSH injection in all heifers. An intravaginal progesterone (P4) implant (1.38 g of P4 CIDR®, Zoetis) was inserted at the time of the first p-FSH injection and removed at the time of OPU, which occurred in all heifers at 44 h (Nivet et al. 2012 ReProduction 143, 165-171; https://doi.org/10.1530/REP-11-0391) after the last p-FSH injection. Additionally, follicle number was determined at OPU and classified as small ( 10 mm). Oocytes from follicles of different sizes were pooled by heifer at OPU and then classified and subjected to IVP procedures. Differences between treatment groups were evaluated using generalized linear mixed models (SAS 9.4; SAS Institute Inc.) and data are presented as mean ± s.e.m. (Table 1). Lengthening the FSH treatment period resulted in a greater (P = 0.01) number of large follicles; however, the number of small, medium-size, and total follicles was not different between groups. Number of total recovered oocytes, viable oocytes, cleaved oocytes, as well as recovery rate, percent viable oocytes, and cleavage rate were not different (P > 0.2) between groups. Similarly, the number of blastocysts produced per heifer and blastocyst rate were not different (P > 0.9) among groups. In conclusion, lengthening the period of FSH treatment by 1 day increased the number of large follicles at OPU but did not improve overall ovarian response, oocyte recovery, or Embryo Production. Table 1. Ovarian response and IVP of heifers treated with p-FSH during 2 or 3 days before ovum pickup Response FSH2d (n = 28) FSH3d (n = 29) P-value Small follicles (n) 5.9 ± 0.6 5.7 ± 0.8 0.83 Medium follicles (n) 17.0 ± 2.4 12.9 ± 1.6 0.18 Large follicles (n) 2.5 ± 0.5 4.5 ± 0.6 0.01 Total follicles (n) 25.4 ± 2.6 23.1 ± 1.8 0.60 Total oocytes (n) 17.0 ± 2.5 13.0 ± 1.2 0.23 Recovery rate (%) 62.6 ± 3.7 56.9 ± 3.1 0.26 Viable oocytes (n) 14.4 ± 2.0 11.3 ± 1.1 0.30 Viable oocytes (%) 85.0 ± 2.4 88.0 ± 3.0 0.31 Cleaved oocytes (n) 8.7 ± 1.6 7.1 ± 1.2 0.62 Cleavage rate (%) 54.7 ± 5.7 54.1 ± 5.7 0.96 Blastocysts (n) 3.2 ± 0.6 3.2 ± 0.7 0.98 Blastocyst rate (%) 20.6 ± 4.0 22.4 ± 3.7 0.97
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135 use and dose of porcine follicle stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro Embryo Production in pregnant holstein heifers
Reproduction Fertility and Development, 2019Co-Authors: R V Sala, L C Carrenhosala, M Fosado, E Peralta, J F Moreno, D Moreno, D. Pereira, A GarciaguerraAbstract:The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro Embryo Production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro Embryo Production in pregnant heifers. Pregnant Holstein heifers (n = 36) were assigned to a complete 3 × 3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0 mg (FSH0), p-FSH 160 mg (FSH160) and p-FSH 300 mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12 h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0 mg) or FSH300 (90.0, 80.0, 70.0 and 60.0 mg), beginning 36 h after dominant follicle removal. Ovum pickup was performed in all heifers 40 h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro Embryo Production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean ± standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P > 0.05). Follicle numbers at the time of oocyte recovery were different (P 0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P < 0.03). In summary, the use of 300 mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro Embryo Production in pregnant heifers treated with different doses of porcine FSH
Estelle Hildegard Venter - One of the best experts on this subject based on the ideXlab platform.
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effect of using frozen thawed bovine semen contaminated with lumpy skin disease virus on in vitro Embryo Production
Transboundary and Emerging Diseases, 2019Co-Authors: C H Annandale, Mario P Smuts, Karen Ebersohn, Lizette Du Plessis, Peter N Thompson, Estelle Hildegard Venter, T A E StoutAbstract:Lumpy skin disease (LSD) is an important transboundary animal disease (TAD) of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, lumpy skin disease virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen-thawed semen on in vitro Embryo Production (IVEP) parameters, including viral status of media and resulting Embryos. Bovine oocytes were harvested from abattoir-collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen-thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV-free semen (control). Following Day 7 and Day 8 blastocyst evaluation, PCR and virus isolation were performed on all Embryonic structures. After completing sufficient replicates to reach 1000 inseminated oocytes, further IVF runs were performed to provide material for electron microscopy (EM) and Embryo washing procedures. Overall, in vitro Embryo yield was significantly reduced by the presence of LSDV in frozen-thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all Embryo structures, when higher doses of LSDV were present in the frozen-thawed semen used for in vitro fertilization (IVF). Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society (IETS) washing procedure resulted in Embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures. This article is protected by copyright. All rights reserved.