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There is no reliable method for determining age for most species of long-lived birds. Recent success using the skin chemical pentosidine as a biomarker has shown promise as an aging tool for birds. Pentosidine levels have been determined only from the breast tissue of carcasses, and we sought to refine the procedure with respect to biopsy size and location for safe and effective use on living birds. We compared pentosidine concentrations in 4 skin-size samples (4, 6, 8, and 20-mm diameter biopsies) from the breast of black vulture (Coragyps atratus) carcasses. We also compared pentosidine levels from breast and patagial tissue to document potential differences among collection sites of deceased vultures (with unknown ages) and monk parakeets (Myiopsitta monachus; with actual, minimal, and unknown ages). Pentosidine concentrations (pmol pentosidine/mg collagen) were similar among the 4 sizes of vulture breast skin (P = 0.82). Pentosidine concentrations for the breast (avg x = 8.9, SE = 0.55, n = 28) and patagium (avg x = 8.9, SE = 0.51, n = 28) of vultures were similar, but in parakeets, pentosidine was higher in the breast (avg x = 15.9, SE = 1.30, n = 105) than the patagium (avg x = 11.5, SE = 1.10, n = 105). We made pentosidine-based age estimates for vultures and parakeets using a general age curve for wild birds. We also made vulture age estimates using plumage characteristics and a cormorant (Phalacrocorax auritus) age curve. Vulture pentosidine-based age estimates appear to correspond to plumage-based age estimates. Pentosidine-based age estimates for 88% of the known-aged parakeets (n = 17) were within 6 months of actual ages. Even though known ages were not available for all birds, we found a positive trend in pentosidine versus age for both species. We suggest that 6-mm diameter skin samples from the patagium of living vultures and other similar-sized birds will provide sufficient tissue for reliable age estimation and will not impair flight ability.