The determinant factors for macroinvertebrate assemblages in river ecosystems are varied and are unique and specific to the type of macroinvertebrate family. This study aims to assess the determinant factors for macroinvertebrate assemblages in a recreational river. The study was conducted on the Ulu Bendul River, Negeri Sembilan, Malaysia. A total of ten sampling stations were selected. The research methodology included (1) water quality measurement, (2) habitat characterization, and (3) macroinvertebrate identification and distribution analysis. The statistical analysis used in this study was canonical correspondence analysis (CCA) to represent the relationship between the environmental factors and macroinvertebrate assemblages in the recreational river. This study found that most of the families of macroinvertebrates were very dependent on the temperature, DO, NH3-N, type of riverbed, etc. All of these factors are important for the survival of the particular type of macroinvertebrate, plus they are also important for selecting egg-laying areas and providing suitable conditions for the larvae to grow. This study advises that improved landscape design for watershed management be implemented in order to enhance water quality and physical habitats, and hence the protection and recovery of the macroinvertebrate biodiversity.