Despite the numerous scientific evidence on the topic, there is no clear and consistent answer that clarifies the true effects of beetroot juice (BJ) supplementation on different types of physical performance. This study examined whether an acute intake of BJ improves swimming performance, physiological variables of anaerobic metabolism, or subjective measures during high-intensity interval exercise with incomplete rest in competitive swimmers. Eighteen competitive swimmers (nine females and nine males) participated in this cross-over randomized, placebo-controlled, double-blind and counterbalanced study. In two trials, swimmers ingested BJ (70 mL, 6.4 mmol/400 mg NO3-) or placebo (PLA) (70 mL, 0.04 mmol/3 mg NO3-) three hours before a 2×6×100 m maximal effort with 40 seconds rest between repetitions and three minutes between blocks. The 100 m times showed no differences between groups (p > 0.05), but there was an interaction between block×repetition×condition (F₅ = 3.10; p = 0.046; ηp² = 0.54), indicating that the BJ group decreased the time of the sixth repetition of block2 compared to block1 (p = 0.01). Lactate concentration showed no differences between conditions (p > 0.05), but there was a main effect of block (ηp² = 0.60) and a block×repetition interaction (ηp² = 0.70), indicating higher values in block2 and increasing values between repetitions in block1. The subjective scales, perception of exertion (RPE) and Total Quality Recovery (TQR), showed no effects of condition (p > 0.05), but BJ swimmers had a greater TQR in the last repetitions of each block. In conclusion, a single dose of BJ did not enhance intermittent swimming performance or modified the physiological (lactate and heart rate) or subjective (RPE and TQR) variables; although there was a possible positive effect on the exercise tolerance at the end of effort.