The existence of a universal molecular clock has been called into question by observations that substitution rates vary widely between lineages. However, increasing empirical evidence for the systematic effects of different life history traits on the rate of molecular evolution has raised hopes that rate variation may be predictable, potentially allowing the ''correction'' of the molecular clock. One such example is the body size trend observed in vertebrates; smaller species tend to have faster rates of molecular evolution. This effect has led to the proposal of general predictive models correcting for rate heterogeneity and has also been invoked to explain discrepancies between molecular and paleontological dates for explosive radiations in the fossil record. Yet, there have been no tests of an effect in any nonvertebrate taxa. In this study, we have tested the generality of the body size effect by surveying a wide range of invertebrate metazoan lineages. DNA sequences and body size data were collected from the literature for 330 species across five phyla. Phylogenetic comparative methods were used to investigate a relationship between average body size and substitution rate at both interspecies and interfamily comparison levels. We demonstrate significant rate variation in all phyla and most genes examined, implying a strict molecular clock cannot be assumed for the Metazoa. Furthermore, we find no evidence of any influence of body size on invertebrate substitution rates. We conclude that the vertebrate body size effect is a special case, which cannot be simply extrapolated to the rest of the animal kingdom.comparative method ͉ generation time ͉ metabolic rate ͉ Metazoa T he concept of a molecular clock, (a relatively constant rate of molecular evolution across lineages) has been fundamental in evolutionary biology for two main reasons. First, the observation of surprisingly even rates of protein change over evolutionary time (1) was one of the key concepts on which the neutral theory was constructed (2). Second, the molecular clock has provided one of the most useful new tools in evolutionary biology. The assumption that genetic distance is related to lineage divergence time has provided a way of reconstructing the evolutionary history of life. Molecular clocks have been particularly valuable for lineages with little or no fossil record (e.g., origins of emerging diseases such as HIV; ref.3) or for taxa or periods for which the fossil record may contain gaps (e.g., origin of the kingdoms; refs. 5 and 6).There is growing evidence, however, that substitution rates can vary considerably between species for a wide range of taxa, including mammals (7-9), arthropods (10-12), and vascular plants (13). Such findings suggest that the molecular clock may not ''tick'' at a steady rate, even between closely related lineages. The false assumption of a molecular clock when reconstructing molecular phylogenies can result in incorrect topology (14,15) and biased date estimation (16)(17)(18). The existence of widespread rate...