PROBIEN   20416
INSTITUTO DE INVESTIGACION Y DESARROLLO EN INGENIERIA DE PROCESOS, BIOTECNOLOGIA Y ENERGIAS ALTERNATIVAS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
EFFECTS OF HYPERTONIC DILUENT ON EPIDIDYMAL SPERMATOZOA STORED AT TWO DIFFERENT TEMPERATURES BEFORE FREEZING-THAWING
Autor/es:
V. MALCOTTI, V. PELUFO, N. BERGAMO, E. AISEN
Lugar:
Huilo Huilo - Chile
Reunión:
Congreso; 7th International Deer Biology congress; 2010
Institución organizadora:
International Deer Biology Society
Resumen:
Abstract EFFECTS OF HYPERTONIC DILUENT ON EPIDIDYMAL SPERMATOZOA STORED AT TWO DIFFERENT TEMPERATURES BEFORE FREEZING-THAWING                 V. MALCOTTI, V. PELUFO, N. BERGAMO, E. AISEN Laboratorio de Teriogenología (IDEPA-CONICET), Facultad de Ciencias Agrarias, Universidad Nacional del Comahue. Ruta 151 km 12 (8303), Cinco Saltos, Río Negro, ARGENTINA vmalcotti@hotmail.com Red deer is a new farming species, which offers opportunities for some reproduction studies, those which can be applied to different contexts such as farming, hunting, and conservation (of endangered cervid species or populations). As a first experimental model, epididymal spermatozoa from slaughtered bulls were recovered at 30, 54, 78 and 102 hours after dead. The scrotal contents were stored both at 5 and 20 ºC. The sperm cells of each treatment (time+temperature combinations) were frozen with Triladyl (T) or Triladyl+trehalose (TT) diluents. In order to assess the sperm viability and integrity, post-thaw evaluation included motility, supravital stain, HOSTest, acrosome damage and chromatin structure assay (SCSA). Both at raw and refrigerated states, the sperm motility rate was higher in the 5ºC stored group, compared with the 20ºC stored group for all times, maintaining 20% motility at 5ºC and 102 h (fresh group). When comparisons were carried out at post-thawing state, motility was higher in the 5ºC group, achieving best results at 102 h with TT (7,5%). However, when supravital stain and HOSTest were observed, viability and membrane integrity were well preserved even at 102 h post-mortem (30% and 36% with TT diluent at 5ºC, respectively). These results may confirm that low motility rate could be present in frozen-thawed epididymal spermatozoa while most of them remain alive. Acrosome status was not greatly affected by storage time, TT diluent being the best crioprotector for that parameter. As a second step, epididymal spermatozoa from hunted red deer stags were recovered at 4 and 30 hours after dead. The scrotal contents were stored at 20 ºC, because that temperature is more close to field and shipment conditions of epididyms. The sperm cells were frozen with Triladyl+trehalose (TT) diluent. Post-thaw evaluation included the same parameters indicated for the bull spermatozoa. The assessment of 30 h frozen-thawed spermatozoa may confirm that in this time acceptable motility rate (35%) and viability (39,7%) were achieved. High plasma membrane integrity (47,9%) and acrosome integrity (59,3%) allows us suppose that, in addition to the chromatin status, the TT diluent, combined with 30 h post-mortem epididymal spermatozoa recovery, is a useful method to obtain viable and fertile sperm cells to be applied in assisted reproduction programmes in endangered deer species. Keywords: deer frozen-thawed sperm cells - hypertonic extenders – epididymal spermatozoa