Advances in the isotopic analysis of biogenic phosphates and their utility in ecophysiological studies of aquatic vertebrates

Distinguishing marine and freshwater animals in the fossil record is a long-standing problem in paleontology. The physiological tolerances of extinct animals usually are inferred from environmental indicators and/or on the physiology of nearest living relatives. These types of evidence are often ambiguous and may be confounded by factors such as post-mortem transport and polymorphism in the living relatives. A solution to this problem is to combine these types of data with analyses of the oxygen isotope compositions of the phosphate (δ¹⁸O(p)) and the carbon isotope compositions of the carbonate (δ¹³C(sc)) of teeth and bones, to determine whether the ingested water and diet, respectively, were fresh or marine. The power of this approach is illustrated here in a study of the early evolution of cetaceans (whales, dolphins and porpoises). Changes in δ¹⁸O(p) and δ¹³C(sc) of the teeth and bones of early cetaceans documented here indicate that fully marine cetaceans existed by the middle Eocene and that some species exploited both marine and freshwater environments. This isotopic approach requires the avoidance of isotopically altered specimens. For this reason, the second component of this work deals with criteria for recognizing isotopically altered fossils. In contrast to one recent study, I found a positive correlation between δ¹⁸O(p) and δ¹⁸O(sc) not only in mammals but also in fish and reptiles. This correlation can be used as a test of whether the original isotopic composition is preserved in fossil specimens. Another approach to this problem is to make analyses of samples taken along growth transect of a fossil tooth or bone. Growth-transect analyses could resolve whether within-species isotopic variation represents differences in preservation or ontogenetic shifts in diet or habitat. In support of this goal, a new method for the analysis of phosphate oxygen is presented. This new method differs from all previous methods in that it involves no chemical reaction, bu