Effect of Percoll volume, duration and force of centrifugation, on in vitro production and sex ratio of bovine embryos.
The objective was to evaluate the effect of Percoll volume, and duration and force of centrifugation on sperm quality characteristics, embryo development, and sex ratio of in vitro-produced (IVP) bovine embryos. Frozen-thawed semen from four bulls were submitted to three Percoll procedures: T1-4 mL of Percoll, centrifuged for 20 min at 700 g; T2-800 microL of Percoll, centrifuged for 20 min at 700 g; and T3-800 microL of Percoll, centrifuged for 5 min at 5,000 g. Sperm total motility, morphology and integrity of the sperm acrosome, membrane and chromatin were determined before and after Percoll treatment, and semen was used for in vitro fertilization (IVF) of in vitro-matured oocytes. All Percoll methods increased the proportion of motile sperm (P<0.05). There were no significant effects of treatment for any sperm characteristic; however, for every end point, there were significant differences among bulls. Similarly, rates of cleavage and blastocyst formation were not affected by the Percoll procedure (P>0.05), but were affected by sire (P<0.05). Sex ratio was similar among treatments for Bulls 2 and 3, whereas semen from Bull 1 processed by T1 yielded a greater percentage of male embryos. However, when only treatments were considered, independent of bulls, the proportion of male:female embryos did not differ significantly from an expected 1:1 ratio. In conclusion, decreasing Percoll volume, reducing duration of centrifugation, and using a higher force of centrifugation did not significantly affect sperm quality, embryo development, or sex ratio of in vitro-produced bovine embryos.
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Embrapa Recursos Genéticos e Biotecenologia/Laboratório de Reprodução Animal. Parque Estação Biológica W5 Final Norte, Brasília 70770-900, DF, Brazil.
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D000818:Animals; D002417:Cattle / Q000196:embryology*; D002498:Centrifugation; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D018471:Hypergravity / Q000009:adverse effects; D008297:Male; D011205:Povidone / Q000494:pharmacology*; D012732:Sex Determination Analysis; D012744:Sex Ratio; D012822:Silicon Dioxide / Q000494:pharmacology*; D013997:Time Factors
C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone