The objectives of this study were to verify the time during which viable ovine spermatozoa could be recovered from the cauda epididymis kept at ambient temperature (18-25°C). Sperm collected in an artificial vagina (AV) were used as control. Spermatozoa samples were collected with an AV and from epididymis at 0 (G0), 6 (G6), 12 (G12), 24 (G24), and 48 (G48) hours post mortem. Total motility (TM), progressive motility (PM), hypo-osmotic membrane integrity test (HOST) and morphological changes were assessed. TM decreased (P<0.05) from 24 hours post mortem (70.0±1.9%) compared to AV (86.4±1.0%). PM decreased (P<0.05) from 12 hours after death (31.3±4.0%) compared to AV group (73.2±1.4%). The percentage of viable cells in HOST decreased (P<0.05) in the G48 (60.0±8.9%). Spermatozoa recovery was lower (P<0.05) 48 hours after death (2064.2±230.7 x 10 6 spermatozoa) compared to G0(2623.6±288.4 x 10 6 spermatozoa). In conclusion, under the conditions of this study, it would be possible to use epididymal spermatozoa recovered up to 24 hours after death for artificial insemination or in vitro fertilization; however, fertility trials are necessary to prove this hypothesis. Keywords: ram, epididymis, room temperature RESUMO Os objetivos deste estudo foram avaliar o período pelo qual era possível recuperar espermatozoides ovinos viáveis da cauda de epidídimos mantidos em temperatura ambiente (18-25°C). O sêmen coletado em vagina artificial (AV) foi utilizado como controle. Os espermatozoides foram coletados dos epidídimos à zero hora (G0), às seis (G6), 12 (G12), 24 (G24) e 48 (G48) horas
Preservation and use of spermatozoa that have been recovered after death can extend the use of genetically superior animals. The objective of this study was to evaluate the maximum period for which ovine spermatozoa could be successfully stored in refrigerated dilution medium post-mortem, with or without added seminal plasma. Three samples of spermatozoa collected in an artificial vagina from 10 rams, or from the tails of four epididymes from the same rams at the time of death (G0) and six (G6), twelve (G12), twenty-four (G24) and forty-eight (G48) hours after death were used. After recovery, the spermatozoa were refrigerated at 5°C in either control medium (CM) or control medium plus 20%homologous seminal plasma (SP) and evaluated for 72 hours from the start of refrigeration. The G48 samples had a lower(P <0.05) total motility (TM) and plasma membrane integrity in the hyposmotic test (HOST) than the other groups evaluated at all analyzed times. The TM decreased (P <0.05) after 24 hours of cooling in semen collected in AV, at G0 and G24 and after 48 hours of refrigeration in G6 and G12. The TM and HOST integrity and sperm morphology did not differ between samples refrigerated in CM or SP. In conclusion, it is possible to collect epididymal spermatozoa up to 24 hours after death. Sperm viability can be prolonged fora further 48 hours by refrigeration. However, total motility decreases from 24 hours after refrigeration and the supplementation of 20% seminal plasma to the extender has no effect on spermatozoa longevity.
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