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In this study, we investigated the relationships between benthic macroinvertebrate assemblages and various environmental factors in Iancheon (NIA) and Bukcheon (NBC) streams, Korea. We collected benthic macroinvertebrates and 33 environmental factors in April 2017 at 9 sites (5 sites in NIA and 4 sites in NBC). We identified 93 species (5 phyla, 9 classes, 16 orders, and 53 families) and 69 species (5 phyla, 9 classes, 17 orders, and 47 families) in NIA and NBC streams, respectively. Considering benthic macroinvertebrate index (BMI), NIA (88.2) and NBC (80.2) streams were in “very good” status. Upstream areas showed the highest scores, 95.5 (NIA1) and 94.2 (NBC1), whereas BMI score was the lowest in downstream areas of both streams, especially in NBC4 (51.0 “bad” status). Cluster analysis and non-metric multidimensional scaling analysis represented the differences of benthic macroinvertebrate assemblages according to spatial and anthropogenic gradients. Our findings provide reference data and highlight the need for the continued monitoring to maintain the good status and manage macroinvertebrate diversity in these two streams, in Sangju-si, Korea.