title: A comparison of BoviPure and Percoll on bull sperm separation protocols for IVF.
abstract: In the present study, we have examined the effect of density gradient preparations BoviPure and Percoll on bull sperm separation and the in vitro fertilization (IVF) and culture (IVC) results. Frozen/thawed semen from five simmental bulls were pooled. Sperm quality parameters such as sperm motility, concentration, membrane activity (HOS assay), membrane integrity (SYBR-14/PI assay) and acrosomal status (EthD-1/FITC-PSA assay) were evaluated before and after sperm processing for IVF using BoviPure and Percoll density gradient separations. The results of the evaluated parameters before sperm processing were: motility 50%, concentration 82.33x10(6)spz/mL, membrane activity 39.05%, membrane integrity 42.97% and the acrosomal status 46.90% of the live spermatozoa with intact acrosomes. After sperm processing with BoviPure and Percoll the motility was 66.67 and 64.17%, the concentration was 25.50x10(6) and 27.67x10(6)spz/mL, the membrane activity was 53.78 and 56.58%, the membrane integrity was 70.85 and 68.76% of and the acrosomal status was 74.16 and 67.46% of the live spermatozoa with intact acrosomes, respectively. Percentages were referred to the total number of spermatozoa. There were significant differences (P<0.05) between the evaluated parameters before and after sperm processing for both separation protocols. We found no significant differences (P>0.05) regarding sperm evaluation parameters between the protocols. A total of 492 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA in six replicates. The cleavage (D2) and blastocysts (D7) rate were significantly higher (P<0.05) for the BoviPure group compared to the Percoll group: 75.80 and 28.21%; 61.58 and 20.83%, respectively. However, the number of hatched blastocysts (D10) did not differ significantly between sperm separation protocols (P>0.05). Our results indicate that both protocols gave suitable sperm for IVF. This finding and the similarity between those two density gradient preparations suggests that BoviPure is a good alternative for sperm separation in bovine IVF.
authors.affiliation: Clinic for Obstetrics and Reproduction, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia. smarko@vef.hr
authors.forename: M
authors.initials: M
authors.lastname: Samardzija
authors.forename: M
authors.initials: M
authors.lastname: Karadjole
authors.forename: M
authors.initials: M
authors.lastname: Matkovic
authors.forename: M
authors.initials: M
authors.lastname: Cergolj
authors.forename: I
authors.initials: I
authors.lastname: Getz
authors.forename: T
authors.initials: T
authors.lastname: Dobranic
authors.forename: A
authors.initials: A
authors.lastname: Tomaskovic
authors.forename: J
authors.initials: J
authors.lastname: Petric
authors.forename: J
authors.initials: J
authors.lastname: Surina
authors.forename: J
authors.initials: J
authors.lastname: Grizelj
authors.forename: T
authors.initials: T
authors.lastname: Karadjole
mesh_terms: D000818:Animals; D002417:Cattle; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002499:Centrifugation, Density Gradient / Q000379:methods / Q000662:veterinary; D046149:Embryo Culture Techniques / Q000662:veterinary; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D007202:Indicators and Reagents; D008297:Male; D011205:Povidone / Q000633:toxicity; D012821:Silanes; D017640:Silicates; D012822:Silicon Dioxide / Q000633:toxicity; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology*
chemical_list: C509975:BoviPure; D007202:Indicators and Reagents; D012821:Silanes; D017640:Silicates; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone