Cholesterol is a critical nutrient in shrimp as they cannot synthesize it and it is found in limited levels in animal-based ingredients. Hence it is an important yet extremely expensive supplement in shrimp diets. Alternatively, phytosterols are sterols of similar chemical structure to cholesterol but less expensive as they can be isolated from plants. Use of phytosterols in low-animal protein shrimp feeds could help decrease the cost of feed production and expand our use of alternative ingredients. Therefore, the aim of the present work was to evaluate the use of a phytosterol (CardioAid®, ADM, Chicago, IL) as partial replacement of cholesterol in juve nile Litopenaeus vannamei diets. The basal diet was formulated to be deficient in cholesterol, but otherwise designed to meet the nutritional requirements of L. vannamei . The basal diet was systematically supplemented with increasing levels of cholesterol (0.04, 0.08, 0.10, 0.12, and 0.16%) or phytosterols (0.04, 0.08, 0.10, 0.12, 0.16, and 0.20%), resulting in a total of 12 diets. Afterwards, 15 juvenile L. vannamei (average individual weight 0.51 ± 0.012 g) were stocked into each tank of an indoors recirculating aquaculture system . Diets were randomly assigned to five replicate tanks for the duration of the trial (6 weeks), during which shrimp were offered feed manually four times a day . Feed ration was determined using a standardized feeding table that assumes weekly shrimp growth with an assumed feed conversion ratio (FCR) of 1.8. At termination, shrimp in each tank were counted and group weighed to determine survival, weight gain, and FCR. Hemolymph was collected from four shrimp per tank , and the remaining shrimp frozen for subsequent analysis. Results of the present experiment indicated significant differences in terms of growth, survival, and FCR among treatments . Shrimp offered feed containing 0.16% cholesterol and 0.20% phytosterols exhibited the biggest average individual weights (7.13 and 6.62g, respectively) . Additionally, shrimp offered the basal diet and 0.04% phytosterol diet exhibited significantly lower survival than remaining shrimp (68 and 74.67% survival, respectively ). Moreover, FCR was significantly influenced by dietary treatments, as s hrimp offered the basal diet and 0.04% cholesterol had a significantly higher FCR than shrimp offered the rest of the diets. Shrimp offered the 0.16% cholesterol diet had the lowest FCR among treatments (1.30). Furthermore, serum cholesterol analysis demonstrated significant differences among treatments in a dose response fashion. Shrimp offered the basal diet had significantly lower levels of cholesterol than those offered 0.04% or higher . However, shrimp offered phytosterol diets exhibited a larger variation in serum cholesterol. In conclusion, phytosterol can be a viable cost-effective alternative to cholesterol in practical shrimp feed, despite having a slightly lower bioavailability than cholesterol.