Development, validation, and application of a microbial community-based index of biotic integrity for assessing the ecological status of a peri-urban watershed in China

This study represents the pioneering effort in employing 16S rRNA-bacteria and 18S rRNA-microeukaryotes to construct the microbial community-based index of biotic integrity (MC-IBI) for assessing the ecological health of riverine ecosystems. The MC-IBI was developed, validated, and implemented using water samples from the Changle River watershed, encompassing both wet and dry seasons. A total of 205 metrics, containing microbial diversity, composition, pollution tolerance/sensitivity, and functional categories, were selected as candidates for evaluation. Following a rigorous screening process, five core metrics were identified as key indicators, namely Pielou's evenness of microeukaryotes, %Cryptophyceae, %Proteobacteria, %Oxyphotobacteria, and % 16S rRNA gene-human pathogens. Moreover, redundancy analysis revealed three metrics (i.e., Pielou's evenness, % 16S rRNA gene-human pathogens, and % Proteobacteria) were positively correlated with impairment conditions. In contrast, two metrics (i.e., %Oxyphotobacteria and %Cryptophyceae) were associated positively with reference conditions. Notably, the developed MC-IBI demonstrates clear discrimination between reference and impaired sites and significantly correlates with environmental parameters and land use patterns. A path model analysis revealed that land use patterns (i.e., build-up land, cropland) negatively impacted the MC-IBI scores. The application of the MC-IBI method yielded an assessment of the ecological conditions at the 73 sampling locations within the Changle River watershed, assigning them into categories of "Very good" (4.1 %), "Good" (4.1 %), "Moderate" (5.5 %), "Poor" (21.9 %), and "Very poor" (64.4 %). This bioassessment framework presents an innovative approach toward the preservation, maintenance, and management of riverine ecosystems.