Recent studies have revealed that calcium limitation of avian reproduction may be a widespread phenomenon, affecting both egg properties and chick development. The effect of calcium shortage on the final body size of fledglings is usually rather weak, possibly owing to compensatory growth. Achieving full skeletal size is not, however, a reliable indication of complete ossification and it is possible that chicks with similar tarsus lengths are in different stages of skeletal development. We hypothesized that measuring plasma activity of the bone alkaline phosphatase (ALP), a highly specific marker for bone calcification, may reveal subtle developmental differences in full-grown fledglings, having experienced different levels of calcium availability during growth. In two seasons, a number of pairs of great tit Parus major were provided with calcium-rich material during the nestling period, while others were not supplemented (controls). While no significant differences in size of fledglings were detected between groups, bone-ALP activity at the pre-fledging stage was lower in the calcium-provided nestlings than in the control nestlings. This may indicate that supplemented chicks had completed the rapid phase of bone formation, but this process was delayed in controls. Measuring ALP as a marker of skeletal development expands our knowledge of how delayed skeletal development of chicks can result in protraction of the nestling period, thereby reducing the breeding success of adult birds.