Nitrate (NO3(-) ) supplementation resulting in higher plasma nitrite (NO2(-) ) is reported to lower resting mean arterial blood pressure (MAP) and oxygen uptake (VO2 ) during submaximal exercise in non-athletic populations, whereas effects in general are absent in endurance trained individuals. To test whether physiological effects of NO3(-) supplementation depend on local muscular training status or cardiovascular fitness, male endurance-trained cyclists (CYC, n=9, VO2 -max: 64±3 ml/min/kg; mean±SD) and recreational active subjects serving as a control group (CON, n=8, 46±3 ml/min/kg), acutely consumed nitrate-rich beetroot-juice ([NO3(-) ] ~9 mmol) (NIT) or placebo (PLA) with assessment of resting MAP and energy expenditure during moderate intensity (~50% VO2 -max) and incremental leg cycling (LEG-ex) and arm cranking exercise (ARM-ex). NIT increased (P<0.001) resting plasma NO3(-) by ~1200% relative to PLA. Plasma NO2(-) increased ~25% (P<0.01) with a significant change only in CYC. LEG-ex VO2 (~2.60 L/min), ARM-ex VO2 (~1.14 L/min) and resting MAP (~87 mm Hg) remained unchanged for CYC and similarly for CON no changes were observed for LEG-ex VO2 (~2.03 L/min), ARM-ex VO2 (~1.06 L/min) or resting MAP (~85 mm Hg). VO2 -max was not affected by supplementation but incremental test peak power was higher (P<0.05) in LEG-ex for CYC in NIT relative to PLA (418±47 vs. 407±46 W). In both CYC and CON high initial baseline values and small increases in plasma NO2(-) after NIT may have lowered the effect of the intervention implying that muscular and cardio-vascular training status is likely not the only factors that influence the physiologic effects of NO3(-) supplementation. This article is protected by copyright. All rights reserved.