Abstract: The aim of this study was to evaluate how long the fresh sperm maintained at 2°C would be utilized for fishery management. The study was conducted every 2 h to assess the sperm viability of orange mud crab Scylla olivacea. Evaluations were conducted as 3 treatments; T1, T2 and T3. In T1, the live specimens were sacrificed; for T2, only spermatophores were extracted and for T3 spermatophore extraction followed by homogenization to create a sperm suspension. All samples were stored with ice in an insulated box was keep fresh longer at 2°C. The time ‘0’ referred the immediate collection of sperm after the specimen was sacrificed. Spermatophore viability was determined using the sperm suspension by eosin-nigrosin staining method. Sperm viability for the fresh sample at time zero was 97.36±0.53%. Viability of the sperm significantly decreased in the 2nd h in all treatments, T1 was 44.66±0.54 to 4.2 ±0.22% at 16 and 18th h, T2 was 36.56±0.5 to 2.69±0.06% at the 12 and 14th h and T3 was 33.69±1.26 to 6.4±0.29% at 8 and 10th h. In comparison, T1 showed significantly higher than other treatments (p<0.05). Extremely low viability percentages were recorded in T3. This study also proved that the time elapse had significant impact on the percentage of viable sperm count.