The aim of this study was to compare management strategies for prematuration of the Litopenaeus setiferus male F0 generation through the combination of two factors, culture system and kind of food, measured in terms of sperm quality and the nutritional condition of shrimp. Four treatments were differentiated by culture system and feed (five replicates each). Treatments were clear water commercial pellet (CC), clear water fresh food (CF), biofloc commercial pellet (BC) and biofloc fresh food (BF). At the end of the experiment, survival, spermatic quality, circulating metabolites and prophenoloxidase (ProFO) were evaluated as immune conditions and the oxidative stress enzyme system. Total sperm concentration showed an interaction between culture system and food provided, showing higher values in CC and BF treatments and lower values in CF treatment. ProFO performed an interaction, showing higher activity in CF and BC treatments. Oxidative stress biomarkers showed significant differences (p < 0.05) in PERMANOVA related to the diet, fresh food versus commercial pellet. Data collected in this study show that optimal sperm quality in L. setiferus male prematuration is obtained when shrimp are held in clear water and fed with commercial pellets or maintained in biofloc system and fed with fresh food.