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Based on the phytoplankton community matrices in the Ashi River Basin (ASRB), Harbin city, we developed an evaluation method using the phytoplankton index of biotic integrity (P-IBI) to evaluate ecological health while investigating the response of P-IBI to anthropogenic activities. We compared the effectiveness of P-IBI with that of the water quality index (WQI) in assessing ecological health. Between April and October 2019, phytoplankton and water samples were collected at 17 sampling sites in the ASRB on a seasonal basis. Our results showed that seven phyla were identified, comprising 137 phytoplankton species. From a pool of 35 candidate indices, five critical ecological indices (Shannon-Wiener index, total biomass, percentage of motile diatoms, percentage of stipitate diatom, and diatom quotient) were selected to evaluate the biological integrity of phytoplankton in the ASRB. The ecological status of the ASRB as measured by the P-IBI and WQI exhibited a similar spatial pattern. It showed a spatial decline in ecological status in accordance with the flow of the river. These results highlighted that P-IBI was a reliable tool to indicate the interaction between habitat conditions and environmental factors in the ASRB. Our findings contribute to the ecological monitoring and protection of rivers impacted by anthropogenic pollution. [ABSTRACT FROM AUTHOR]