162 EFFECT OF THE SPERM SELECTION WITH ISOLATE® OR PERCOLL® ON SPERM QUALITY AND IN VITRO BOVINE EMBRYO DEVELOPMENT FOR FROZEN–THAWED SEXED AND NONSEXED SEMEN
G. A. Bó A B , P. Rodriguez Villamil A , G. Moreira B , M. E. Garcia Gomez B , M. Fernandez Taranco B and H. Wei CA Instituto de Reproducción Animal Córdoba, Córdoba, Córdoba, Argentina;
B Instituto de Ciencias Basicas, Carrera de Medicina Veterinaria, Universidad de Villa Maria, Córdoba, Argentina;
C Trans Ova Genetics, Sioux Center, Iowa, USA
Reproduction, Fertility and Development 24(1) 193-193 https://doi.org/10.1071/RDv24n1Ab162
Published: 6 December 2011
Abstract
The aim of this study was to compare the effects of 2 different commercially available spermatozoa separation techniques, Isolate® (Irving-Scientific, Santa Ana, CA, USA) and Percoll® (Nutricell, São Paulo, Brazil), on sperm quality and in vitro embryo production using sexed and nonsexed semen. Oocytes (n = 5046) were obtained from slaughterhouse ovaries and fertilized with frozen-thawed sexed or nonsexed semen from the same 4 Holstein bulls. The experiment was done in 10 replicates, with all treatment groups included. Sperm quality (motility, concentration, morphology and membrane integrity) was evaluated and compared before and after sperm selection by the 2 methods. Oocytes were maturated in TCM-199 supplemented with 0.4% of BSA for 24 h in a controlled atmosphere and then selected and randomly allocated into 4 different groups. Group 1: oocytes fertilized with sexed semen selected by Percoll®; Group 2: oocytes fertilized with sexed semen selected by Isolate®; Group 3: oocytes fertilized with nonsexed semen selected by Percoll®; Group 4: oocytes fertilized with nonsexed semen selected by Isolate®. Fertilization was performed in Fert-TALP medium for 18 h under the same conditions as maturation. Presumptive zygotes were cultured for 7 days in SOF medium in a 39°C humidified incubator with 5% CO2, 5% O2 and 90% N2. Cleavage and blastocyst rates were evaluated on Day 2 and 7, respectively, after fertilization. Proportional data were transformed by square root and then analysed by ANOVA, with type of semen and sperm selection method as the main effects. Regardless of the sperm selection technique, sperm motility and percentage of normal sperm increased (P < 0.005) from the initial post-thaw parameters. For nonsexed semen, Percoll® gradient increased the recovery rate (i.e. final concentration/initial concentration × 100; 57.3 ± 2.7) compared with Isolate® (46.0 ± 1.8; P < 0.05). Furthermore, sperm selected by Isolate® presented significant improvements compared with Percoll® gradient on membrane integrity of sexed (41.0 ± 0.6 vs 38.8 ± 0.8) and nonsexed semen (60.8 ± 1.6 vs 58.8 ± 0.5; P < 0.05). Finally, blastocyst production rates were higher (P < 0.05) for sexed (Group 2: 14.0 ± 1.0) or nonsexed semen (Group 4: 22.0 ± 1.1) selected by Isolate® than for sexed (Group 1: 10.5 ± 1.5) or nonsexed semen (Group 3: 17.0 ± 2.1) selected with Percoll®. In conclusion, selection of both sexed and nonsexed semen for IVF with Isolate® resulted in higher quality sperm and higher embryo production rates than Percoll®.
