This study investigated the structure of benthic macroinvertebrate communities to evaluate the water quality of the Khersan River in Chaharmahal and Bakhtiari Province, Iran. Sampling of benthic macroinvertebrates was conducted seasonally from autumn 2023 to summer 2024 at five sampling stations. At each station, samples were collected using a Surber sampler with three replicates. The Hilsenhoff Biotic Index, Shannon–Wiener diversity index, and evenness index were calculated to assess ecological conditions and water quality status. A total of 10068 benthic macroinvertebrates belonging to 14 families and 10 orders were identified during the study period. The highest mean annual density was recorded at station 1 (767 ± 412.63 individuals/m²), whereas the lowest density was observed at station 5 (270 ± 145.01 individuals/m²). The class Insecta accounted for approximately 95% of the total collected organisms. Among the insect taxa, the family Chironomidae from the order Diptera showed the highest abundance across all sampling stations and seasons. The calculated biotic indices indicated that the water quality of the Khersan River ranged from moderate to moderately poor. These findings suggest the presence of environmental pressures within the watershed and provide valuable baseline information for future ecological assessments and long-term monitoring programs of the river ecosystem