Ontogenetic shifts in food preference reduces intraspecific competition as immature individuals eat different food types than adults. This diet plasticity could facilitate species' ability to successfully invade and establish itself in a new environment, even when co-occurring with phylogenetically close species. Here, ontogenetic diet shifts of a non-native piranha species (Serrasalmus marginatus) was tested by analyzing the relationship between its body length and the carbon and nitrogen stable isotope signatures. Carbon stable isotope was not correlated to fish length, but positive significant correlation between δ15N values and fish length was found for the non-native piranha. Also, immature and adult S. marginatus showed low isotopic niche overlap. The correlation between fish length and δ15N, and the low trophic overlap between immature and adult, indicate that the non-native S. marginatus had ontogenetic shifts in food preference, which may be viewed as an additional mechanism underlying its successful establishment in the upper Paraná River. Our findings indicate that ontogenetic shifts in food preference may be an invasive trait that facilitates the establishment of non-native fish species in tropical aquatic systems.