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The present study was conducted to determine the effect of water salinity on ovarian maturation and the level of reproductive steroid hormone 17 beta-estradiol via morphological observation, histological assessment and through Enzyme-Linked ImmunoSorbent Assay (ELISA) analysis on female orange mud crab, Scylla olivacea in captivity. In fact, this is the first report investigating the effect of water salinity on S. olivacea broodstocks. Based on the morphological, histological and ELISA analysis, 20 ppt was the best identified salinity compared to the other two salinities (10 ppt and 30 ppt). This report revealed that salinity does play important roles in ovarian maturation of mud crab, however for 17 beta-estradiol, the result shown non-significant difference. We suggest that 20 ppt was the best salinity treatment for S. olivacea culture in captivity for future use.