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Year 2017 - Volume 37, Number 5
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Addition of antioxidants in ram semen and the effects after thawing, 37(5):471-478
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ABSTRACT.- Souza W.L., Moraes E.A. & Toniolli R. 2017. [Addition of antioxidants in ram semen and the effects after thawing.] Adição de antioxidantes ao sêmen de carneiros e seus efeitos após a descongelação. Pesquisa Veterinária Brasileira 37(5):471-478. Programa de Pós-Graduação em Ciências Veterinárias, Universidade Estadual do Ceará, Campus do Itaperi, Av. Dr. Silas Muguba 1700, Fortaleza, CE 60740-000, Brazil. E-mail: wilde@zootecnista.com.br
Aimed to evaluate the effect of adding antioxidants as ascorbic acid, melatonin and Trolox C to diluted semen of ram with oxidative stress to potenciate fertilization after cryopreservation. Ten samples collected were diluted in Tris-egg yolk to a final concentration of 200x106 sperm/mL and kept in a water bath at 32°C. Antioxidants were added as follows: 100µM melatonin (MEL) +.05% ascorbic acid (AA); 100µM of MEL + 90µL of Trolox C (TRO); 90µL of TRO + 0.05% AA; and 100µM of MEL0.05% AA + 90µL of TRO. Semen was cooled in a cold chamber at 5°C for two hours and packaged, sealed in 0.5mL straws, packaged under liquid nitrogen vapor (N2L), 8cm of water depth for 15 minutes, and then immersed in N2L. Samples were assayed for motility, integrity of the plasma membrane and acrosomal membrane, mitochondrial activity, binding assay and oxidative stress spermatozoa. The variables were analyzed by ANOVA and means compared by Tukey test (P<0.05). Percentage of total and progressive motility was higher for sperm treated with MEL+AA+TRO (67% and 49.89%), MEL+AA (64.37% and 45.61%) and MEL+TRO (61.65% and 41.15%) compared with the other treatments (P<0.05). The integrity of the plasma membrane and acrosome was higher for all semen treated with antioxidant associations compared with control (P<0.05). Mitochondrial activity was higher in sperm treated with MEL+AA+TRO compared all treatments (P<0.05). The number of sperm binding to perivitelline membrane was higher for semen treated with antioxidant associations compared with control; also sperm treated with MEL+AA+TRO demonstrated higher effect of all (P<0.05). No difference was observed between the treatments by oxidative stress sperm (P>0.05). The addition of melatonin, ascorbic acid and Trolox C in diluted semen of ram improves sperm quality after thawing. |
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