IN VITRO FERTILITY OF POST-THAWED EPIDIDYMAL RAM SPERMATOZOA AFTER STORAGE AT 5 °C BEFORE CRYOPRESERVATION
N W.K Karja, M Fahrudin, M A Setiadi
ABSTRACT
This study addressed the effects of storage duration of epididymides at 5 °C before sperm collection and their fertility after cryopreservation in vitro. Spermatozoa from one of the testes pairs were immediately collected, evaluated and frozen (control group). The remaining epididymides were cooled to 5 °C and stored for 24, 48, 72, and 96 h (experimental groups), after which spermatozoa were collected and frozen as in the control group. Before and after thawing, sperm motility, sperm viability and plasma membrane integrity were assessed. The fertilizing ability of frozen-thawed spermatozoa of each group was evaluated byin vitro fertilization of matured sheep oocytes. Sperm quality (sperm motility, viability, and plasma membrane integrity) at collection and after cryopreservation decreased as the duration of the epididymal storage interval increase (P < 0.05). The motility decreased steadily along the studied time periods. Although, the fertilizing ability of post-thawed epididymal spermatozoa gradually decreased as the storage period was prolonged, the spermatozoa collected from the cauda epididymides stored at 5 °C for up to 96 h were able to fertilize 16%-65% of oocytes in vitro. Results of the present study showed that ram epididymal spermatozoa survive in storage at 5 °C for up to 96 h. These spermatozoa maintain their fertilizing ability and may be suitable for use in IVF and other assisted reproductive procedures.
Key words: epididymis, sheep, spermatozoa, in vitro fertilization, cryopreservation
ABSTRAK
Penelitian ini bertujuan untu bertujuan untuk mengetahui daya fertilisasi post-thawed spermatozoa yang dikoleksi dari epididymis setelah disimpan pada suhu 5 oC selama 96 jam post mortem secara in vitro. Epididimis disimpan pada suhu 5 oC selama 0-96 jam sebelum dibekukan. Kualitas spermatozoa (motilitas, viabilitas, dan integritas membran plasma) dievaluasi sebelum dan setelah dibekukan dengan interval waktu 24 jam. Kemampuan spermatozoa setelah dibekukan untuk memfertilisasi oosit secara in vitro dievaluasi dengan fertilisasi in vitro. Walaupun terjadi penurunan terjadi penurunan kualitas spermatozoa kualitas spermatozoa seiring dengan waktu penyimpanan epididymis (P<0,05) tetapi sperma yang hidup dan motil masih dapat dikoleksi setelah 96 jam post mortem. Berdasarkan pembentukan pronukleus setelah fertilisasi in vitro, 16%-65% oosit terfertilisasi secara 16%-65% oosit terfertilisasi secara in vitro oleh post-thawed spermatozoa yang dikoleksi dari epididimis setelah disimpan 0-96 jam. Dapat disimpulkan bahwa walaupun kualitas spermatozoa yang dikoleksi dari epididimis pasca penyimpanan dan pembekuan menunjukkan penurunan tetapi spermatozoa tersebut masih mempunyai kemampuan untuk membuahi oosit secara in vitro.
Kata kunci: epididymis, domba, spermatozoa, fertilisasi in vitro, kriopreservasi
Publish in MEDIA PETERNAKAN – Journal of Animal Science and Technology, Vol 36, No 1 (2013) pp. 26-31