The purpose of this study was to compare blood markers associated with eccentrically biased exercise and muscle damage, after two bouts of downhill running. Nine active, untrained males performed 2 x 45 min bouts of downhill running (-0.16 radians), at a speed that would elicit 70% of each subjects VO2max, on a level grade; runs were spaced 14d apart (RUN1, RUN2). Blood samples were obtained before, after, and every hour for 12 h after exercise, as well as every 24 h for 5 d, to assess numbers of circulating neutrophils, monocytes, and lymphocytes, serum cortisol, creatine kinase (CK); subjective sensations of delayed onset muscle soreness (DOMS) were monitored. To control for diurnal variation, two weeks prior to the RUN1, subjects had blood draws performed at the same time as would occur after exercise, but did no exercise (CONTROL). During the 5 d after exercise, DOMS and CK were significantly greater (p < 0.05) after RUN1 compared to RUN2 and CONTROL. During the 12 h after RUN1 and RUN2, neutrophils showed similar responses compared to CONTROL. However, neutrophils were significantly elevated at 96 h after RUN1 and 24 h after RUN2. Monocytes were significantly elevated during 5-11 h after RUN1 and RUN2, compared to CONTROL. Cortisol showed a similar significant diurnal decrease for all three conditions during the 12 h following exercise. The significantly lower levels of CK and DOMS seen after RUN2, compared with the initial run is consistent with the literature. The similar changes in neutrophils and monocytes during the 12 h following RUN1 and RUN2, followed by disparate responses over the subsequent 5 d, requires further investigation.