A 45-day trial was performed to evaluate the effect of biofloc technology (BFT) with or without fresh food (FF) supplementation during pre-maturation period on Farfantepenaeus duorarum spawning performance, biochemical composition and fatty acid profile of eggs as compared with conventional clear-water system (CW+FF). Females raised in biofloc and that received FF supplementation (FLOC+FF) achieved better spawning performance in terms of number of eggs per spawn (49 × 103), number of eggs per spawn per g of spawner's body weight (2.1 × 103) and egg size (~275 μm) as compared with CW+FF (23 × 103, 1.1 × 103 and 263 μm respectively), but both treatments did not vary from FLOC (P > 0.05). High spawning activity was also observed in biofloc system as compared with clear-water system as shown in number of spawns per ablated female (2.2–3.0 versus 0.6) and percentage of females that spawn at least once (80–82 versus 25%). Biochemical composition of eggs presented no significant differences among treatments. FA profile of eggs indicated that high spawning activity performed by females in FLOC+FF treatment was reflected in lower mean levels of EPA, DHA and sum of polyunsaturated fatty acids (n-3) and (n-6). The better reproductive performance demonstrated by females raised in biofloc justified the application of this technology in F. duorarum broodstock.