Abstract-A total of 1 124 adult and juvenile individuals, representing 35 species from 13 families were collected in the four rivers of Johor. Each river represents a different of land use; Sungai Mengkibol is an urban river, Sungai Madek is associated with logging activities, the downstream section of Sungai Dengar for agricultural activities and it's middle section which is located at the foot of Gunung Berlumut as a reference station under pristine forest condition. The families represented in the samplings was dominated by Cyprinidae with 20 species (54.29%), followed by Mastacembelidae with three species (8.57%), Channidae and Hemiramphidae with 2 species each (5.71%). The other families were represented by one species each from Bagridae, Cichlidae, Claridae, Cobitidae, Gobiidae, Loricariidae, Poecilidae, Sisoridae and Synbranchidae(2.86% each). The highest number of fish were collected at Sungai Dengar station (376 individuals) followed by Sungai Ulu Dengar (306 individuals), Sungai Madek (238 individuals) and Sungai Mengkibol (204 individuals). Although the number of fish among the rivers were not statistically different (p = 0.138, p > 0.05), species diversity was significantly different according to river characteristics. Sungai Ulu Dengar has the highest number of species with 22 species, followed by Sungai Dengar with 21 species and Sungai Madek with 19 species. The lowest number of species was recorded at Sungai Mengkibol with 11 species. Water quality parameters like dissolved oxygen, turbidity and temperature show positive correlation with the species diversity. Therefore, it can be concluded that healthy river support high diversity of fish species.