Metal pollution is one of the main environmental threats in freshwater ecosystems. Aquatic animals can accumulate these substances and transfer them across the food web, posing risks for both predators and humans. Accumulation patterns strongly vary depending on the location, species, and size (which in fish and crayfish is related to age) of individuals. Moreover, high metal concentrations can negatively affect animals’ health. To assess the intraspecific relationship between metal accumulation and size and health (proxied by the body condition) of individuals, the concentration of 14 metals (Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mg, Mn, Ni, Pb, Se, Zn) was analyzed in six alien species from the highly anthropogenically altered Arno River (Central Italy): five fish (Alburnus alburnus, Pseudorasbora parva, Lepomis gibbosus, Ictalurus punctatus, and Silurus glanis) and one crayfish (Procambarus clarkii). We found that in P. clarkii, Cu was negatively related to size, as well as Al in L. gibbosus and Mg for adult I. punctatus. Positive size-dependent relationships were found for Hg in L. gibbosus, Fe in S. glanis, and Cr in juvenile I. punctatus. Only Co and Mg in S. glanis were found to negatively correlate with individual health. Since metal concentrations in animal tissue depend on trade-offs between uptake and excretion, the few significant results suggest different types of trade-offs across different species and age classes. However, only predatory fish species (L. gibbosus, I. punctatus, and S. glanis) presented significant relationships, suggesting that feeding habits are one of the primary drivers of metal accumulation.