Abstract: The study was conducted to every hour assess the sperm viability of banana shrimp (Penaeus merguiensis) collected from Kedah water, West Malaysia (5°39'N; 100°19'E). Evaluations were done on three ways: group A: whole fresh specimens maintained at 2°C prior to extraction, group B: whole spermatophore maintained at 2°C and C: sperm suspension maintained at 2°C. Spermatophores counts were determined by sperm suspension using modified eosin-nigrosin staining method. Percentage of mean sperm viability for the fresh sample in group A at time zero was 93.96±3.74%, B was 98.8±0.7% and C was 99.02±0.7%. In group A, viability of the sperm considerably decreased after the first 60 min was 76.4±5.96% but in group B and C was gradually decreased at 91.4±0.9 and 97.6±1.2%. These were 67.9±5.13 and 62.5±5.1% in group A, 82.3±1.4% and 75.9±2.2% in group B and in group C was 93.3±1.9 and 88.2±3.1%, respectively after 1st-2nd h times elapsed. At 7th h in all groups the viability decreased significantly to <60% (p<0.05). Following, mean sperm viability were considerably decreased to group A was 50.7±4.79 and 37±6.52%, group B was 51.2±1.6 and 31.6±1.4% and group C was 56.2±1.9 and 41.5±2.3%, respectively after 7th and 9th h. However, for spermatophores in group A after 7th h fertilization and hatching rate was 44.3 and 64.4% in group B was 61.9 and 67.7% and group C was 42.9 and 61.3%, respectively at rates comparable to control 88.2 and 76.2%, respectively. There was no significant relationship was observed between biomass of the spermatophore to the body weight of the shrimp (p>0.05). The present study also revealed that specimens, spermatophore or sperm suspension maintained at 2°C could be utilized for fishery management through artificial insemination process till 7th h.