Work-related neck/shoulder muscle pain has been associated with increased anaerobic muscle metabolism. Thus, interventions to enhance oxygenation of painful muscles seem relevant. While cycling with relaxed shoulders has been shown to result in acute neck/shoulder muscle pain reduction, the effect on tissue oxygenation remains unknown. The aim of the present study was to investigate tissue oxygenation of the passive trapezius muscle during and after cycling in female workers with (MYA) and without (CON) trapezius myalgia. Eligible participants (n = 17 MYA, n = 8 CON) performed 20 min sub-maximal cycling in an upright position with relaxed shoulders. Near-infrared spectroscopy was used to measure trapezius muscle oxygenation during and 2 min after the cycling period. For both MYA and CON, oxygenation of the passive trapezius increased in a linear fashion over time, to values approximately 5 microM above baseline at the end of the cycling period, with no significant group difference (CON 5.2, MYA 4.9 microM). Two min after termination of exercise, oxygenation was increased further in both groups, but significantly more in CON (8.8 microM) than in MYA (7.0 microM) (P = 0.05). In conclusion, cycling increases oxygenation of resting neck/shoulder muscles in women with and without trapezius myalgia, indicating acute positive effects of either neural or humoral factors on vascular beds of distant relaxed muscles. Although this beneficial response was observed in both groups, the post-exercise response was lower in women with trapezius myalgia.