The localization of parvalbumin was determined in skeletal muscles with different fiber type compositions from quails (Coturnix japonica) and pigeons (Columba livia) by sandwich ELISA. The biochemical profiles of these muscles were evaluated by the assay of total lactate dehydrogenase (LDH) activity as well as the LDH isozymes for anaerobic metabolism, and by the analysis of myoglobin for aerobic metabolism. The quail fast posterior latissimus dorsi (PLD) with a lower myoglobin content and higher LDH activity or M-type isozyme pattern was assessed as containing primarily fast-twitch glycolytic and oxidative-glycolytic (FG/FOG) fibers, and the mixed sartorius (SA) in quails and pigeons was shown to be composed mostly of FOG fibers because of the intermediate myoglobin content and LDH activity or H/M-type isozyme pattern. Parvalbumin, which functions as a relaxing factor in the fast-twitch fibers, was present in PLD and SA in significant amounts, whereas it was undetectable in quail anterior latissimus dorsi (ALD) and pigeon latissimus dorsi (LD) which contain exclusively slow-tonic (ST) fibers with the lowest LDH activity or predominant H-type isozyme characteristics. The pectoralis superficialis (PS) and pectoralis profundus (PP) muscles from quails and pigeons seem to consist mostly of FG/FOG fibers because of the highest myoglobin contents and LDH activities or M-type and H/M-type isozyme patterns. Despite fast-twitch fiber compositions, parvalbumin was absent from these pectoralis muscles. The tetanic contraction induced in avian fast-twitch pectoralis fibers during flapping flight might be independent of a function of parvalbumin as a relaxing factor.