Biological invasions are a serious threat to biodiversity conservation, especially in freshwater ecosystems. The American macrophyte Ludwigia hexapetala, which colonizes both the aquatic and bank habitats of lakes, rivers, and canals, is invading many waterbodies in Europe, becoming an increasingly worrisome threat in several European countries, including Italy. However, only fragmentary information is available on the actual impact of its invasion in these habitats. This study aims to collect field data from various freshwater habitats in central and northern Italy, to assess the possible impact of L. hexapetala on the environmental parameters and plant biodiversity of the invaded habitats. The results show that in aquatic habitats, dense floating L. hexapetala populations reduce the light levels and oxygen available in the water, consequently limiting the growth of other aquatic plants. Indeed, L. hexapetala populations negatively affect aquatic plant diversity, as an increase in L. hexapetala cover corresponded to a decrease in Simpson’s diversity index. In contrast, in bank habitats, L. hexapetala has no significant impact on plant diversity. Evidence suggests that some native species, such as Phragmites australis, which generally form compact populations along the banks, effectively counteract the invasion of L. hexapetala. This information may prove valuable for the environmental managers of those freshwater habitats where L. hexapetala invasion needs to be addressed and controlled.