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Título: | Semen cryopreservation for the creation of a Spanish poultry breeds cryobank Optimization of freezing rate and equilibration time |
Autor: | Santiago Moreno, Julián; Castaño, Cristina; Toledano-Díaz, A. CSIC ORCID ; Coloma, M. A.; López Sebastián, Antonio; Prieto, M. T.; Campo, J. L. | Palabras clave: | Cryobank Equilibration time Freezing rate Spanish breed Spermatozoa |
Fecha de publicación: | 2011 | Editor: | Elsevier | Citación: | Poultry Science 90: 2047-2053 (2011) | Resumen: | A sperm cryopreservation protocol requiring dimethylacetamide (DMA, 6%) as a cryoprotectant was optimized via assays involving different prefreezing equilibration times (1, 10, 30, 60, and 120 min at 5°C) and different freezing rates achieved by the following 1) using nitrogen vapor to reduce the temperature from 5°C to -85°C at 10°C/min (slow freezing rate); 2) using a biological freezer unit in a 2-step method to reduce the temperature from 5°C to -35°C at 7°C/min and then from -35°C to -140°C at 60°C/min (medium freezing rate); or 3) using a biological freezer unit in a 1-step freezing method to reduce the temperature from 5°C to -180°C at 60°C/min (rapid freezing rate). Heterospermic semen samples from chicken breeds raised as part of a Spanish genetic resource conservation program were used in all assays. The 1-min equilibration treatment was associated with a lower percentage of viable thawed spermatozoa than the 30-min treatment (P < 0.05). The remaining sperm variables studied were not affected by equilibration time. The medium-rate 2-step freezing method was associated with a higher percentage of motile spermatozoa after thawing and with greater acrosome integrity (P < 0.05) than the slow nitrogen vapor or rapid 1-step methods. Thawed sperm movement quality and plasma membrane integrity (as assessed by the hypoosmotic swelling test) were better (P < 0.05) in samples frozen by the mediumrate 2-step freezing method than in those subjected to the slow nitrogen vapor method. Fertility was not influenced by freezing method, although that achieved with the medium rate 2-step freezing method showed a trend toward being greater than that achieved with the rapid 1-step method (P = 0.07). Together, the present results suggest that slow cooling rates are not recommendable when using dimethylacetamide. The 2-step freezing method may be useful in the establishment of a germplasm bank for Spanish chicken breeds. © 2011 Poultry Science Association Inc. | URI: | http://hdl.handle.net/10261/291726 | DOI: | 10.3382/ps.2011-01355 | ISSN: | 0032-5791 | E-ISSN: | 1525-3171 |
Aparece en las colecciones: | (INIA) Artículos |
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