As part of the research program “Freshwater Prokaryotic Organisms Research and Discovery,” freshwater samples were collected from the Nakdonggang River. After plating the samples on several culture media and incubating aerobically, approximately 900 bacterial strains were isolated and identified using 16S rRNA gene sequences. Among the bacterial isolates showing higher than 98.7% 16S rRNA gene sequence similarity with those of already confirmed bacterial species previously unreported in Korea, 29 strains were selected. These strains were phylogenetically diverse and belonged to 3 phyla, 6 classes, 13 orders, and 21 genera. At the genus level, these previously unreported species were found to be affiliated with Novosphingobium, Sphingomonas, Polymorphobacter, Croceibacterium, Devosia, Endobacterium, Agaricicola, Bradyrhizobium, Paracoccus, and Pseudotabrizicola of the class Alphaproteobacteria; Undibacterium, Azonexus, and Dechloromonas of the class Betaproteobacteria; Acinetobacter and Budvicia of the class Gammaproteobacteria; Streptomyces, Nocardioides, Mycobacterium, and Cellulomonas of the phylum Actinomycetota; Flavobacterium and Pedobacter of the phylum Bacteroidota. These species were further characterized by examining their Gram reaction, colony and cell morphologies, biochemical properties, and phylogenetic positions. Detailed descriptions of these 29 previously unreported species are provided.