Animal gene banks development is limited by high costs of creation. The recovery and freezing of viable sperm from the epididymes of slaughtered animals can be a cheap alternative for preserving male gametes. The aim of this study was to optimize the recovery of bull epididymal spermatozoa comparing the effects of two methods of extraction in combination with egg yolk extender. Testis from 23 Limousine bulls, ranging in age from 13 to 20 months, were collected at the abattoir. Spermatozoa were extracted from the epididymes using two methods: the float-up and the retrograde flushing. The average time for extraction was 16 minutes in both methods. Within each methods half of testis were processed with an egg yolk extender and the second half with an egg-yolk free extender. Sperm concentration, motility, viability and morphology were evaluated. Statistical analyses were carried out using the GLM procedure (SAS package v9.1). Methods of extraction, in combination with the presence of egg yolk in the extender, were considered as fixed effects; age of the bull and time for extraction as covariates. Sperm concentration was not significantly different using either the float-up method or the retrograde flushing (283.5±24.7 and 301.5±30.3¥106 cells/mL, respectively). Flushing technique was better than the float-up method in terms of sperm quality, considering total motility (78.4±3.1% vs 61.8±2,6%, P=0.0003; respectively) and viability (88.7±2.1% vs 77.6±1.7%, P=0.0003; respectively). Egg yolk influenced positively motility and morphology in the float-up method, whereas decreased viability in flushed samples. In conclusion, samples extracted by flushing method, with egg yolk, showed a better sperm quality. These data suggest the possible use of epididymal sperm collected by the flushing technique for the creation of cattle semen cryobanks.
Optimisation of bull epididymal spermatozoa extraction techniques: a tool for improving gene banking
Turri F;Gliozzi TM;Gandini G;Pizzi F
2011
Abstract
Animal gene banks development is limited by high costs of creation. The recovery and freezing of viable sperm from the epididymes of slaughtered animals can be a cheap alternative for preserving male gametes. The aim of this study was to optimize the recovery of bull epididymal spermatozoa comparing the effects of two methods of extraction in combination with egg yolk extender. Testis from 23 Limousine bulls, ranging in age from 13 to 20 months, were collected at the abattoir. Spermatozoa were extracted from the epididymes using two methods: the float-up and the retrograde flushing. The average time for extraction was 16 minutes in both methods. Within each methods half of testis were processed with an egg yolk extender and the second half with an egg-yolk free extender. Sperm concentration, motility, viability and morphology were evaluated. Statistical analyses were carried out using the GLM procedure (SAS package v9.1). Methods of extraction, in combination with the presence of egg yolk in the extender, were considered as fixed effects; age of the bull and time for extraction as covariates. Sperm concentration was not significantly different using either the float-up method or the retrograde flushing (283.5±24.7 and 301.5±30.3¥106 cells/mL, respectively). Flushing technique was better than the float-up method in terms of sperm quality, considering total motility (78.4±3.1% vs 61.8±2,6%, P=0.0003; respectively) and viability (88.7±2.1% vs 77.6±1.7%, P=0.0003; respectively). Egg yolk influenced positively motility and morphology in the float-up method, whereas decreased viability in flushed samples. In conclusion, samples extracted by flushing method, with egg yolk, showed a better sperm quality. These data suggest the possible use of epididymal sperm collected by the flushing technique for the creation of cattle semen cryobanks.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
