Effects of storage duration on Mitochondrial activity and DNA fragmentation of post-thawed spermatozoa from several Ongole Grade bull in Indonesia
Kurniawan Dwi Prihantoko1, A. Kusumawati2, D.T. Widayati3, Mulyoto Pangestu4,

1Doctoral Student of Veterinary Science, Faculty of Veterinary Medicine, University Gadjah Mada, Yogyakarta, 55281, Indonesia
2Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University Gadjah Mada, Yogyakarta, 55281, Indonesia
3Department of Animal Breeding and Reproduction, Faculty of Animal Science, University Gadjah Mada, Yogyakarta, 55281, Indonesia
4Education Program in Reproduction and Development, Monash Clinical School, Monash University, Australia

Abstract

Abstract. Several recent studies theoretically assumed that storage of frozen doses of semen in liquid nitrogen guarantees unlimited sperm functionality. However, several studies conclude that long-term storage causes decreased sperm quality. The purpose of this study is to evaluate the storage duration effect on mitochondrial activity and DNA integrity of Ongole Grade Bull post-Thawed spermatozoa from several Artificial Insemination Centre (AIC) in Indonesia. In this study, Ongole Grade Bull frozen semen that has been stored up to three years were investigated. According to the duration of storage, semen samples were divided into 7 groups (one month, three months, six months, nine months, one year, two years, and three years). This study used 42 straws, with 6 straws for each group were used. Samples were evaluated for sperm motility, viability, membrane integrity, mitochondria activity, and DNA fragmentation. The results show that one-month group sperm motility was (53.80%), viability (72.75%) membrane integrity (76.35%.), mitochondria activity (64.24%), and DNA fragmentation (3.82%). After 3 years of storage, the corresponding values were 39.52%, 41.56%, 48.25%, 49.12%, and 4.62%. Sperm motility and viability remained unaffected up to 6th months of storage, while membrane integrity started decreasing at 3rd month (P<0.05). Mitochondria activity decreased at 9th months (P<0.05). In contrast to the previous results, DNA fragmentation did not increase significantly after 3 years of storage (P>0.05). Thus, post-thaw motility, viability, and membrane integrity of spermatozoa from Ongole Grade Bulls started decreasing at 6th months of storage, mitochondria activity started decreasing after 9th months, while sperm DNA integrity remained unaffected for up to three years of storage.

Keywords: Ongole Grade Bull, frozen semen, storage, mitochondria activity, DNA fragmentation,

Topic: Veterinary Clinic, Reproduction, Pathology