Anthropogenic pressure induced discontinuities of microbial communities along the river

Microorganisms play a fundamental role in driving biogeochemical functions within rivers. Theoretically, the directional flowing nature of river contributes to the continuous downstream change pattern of microbial communities. This continuity is anticipated to be influenced by human activities as anthropogenic materials lead to the mixing of environmental substances and their resident microorganisms with local communities. Here, we conducted a field investigation along the Beiyun River, which successively flows through pristine forest areas, artificial urban and agricultural areas with a length of 184 km, to explore the influence of anthropogenic events on microbial similarity, diversity, composition, co-occurrence, and assembly mechanisms in sediments along the river. Piecewise linear regression tests showed that discontinuities of microbial similarity occurred following the transitions from low to high anthropogenic pressure. LEfSe analysis illustrated that microorganisms associated with wastewater treatment plants and gut were differentially abundant in urban and agricultural streams. By quantifying contributions of ecological assembly processes, we found that the dominant role shifted from variable selection (60.78% in forest group) to homogenous selection (79.52% in urban group and 57.14% in agriculture group) as the differences in NH₄⁺-N, NO₃^--N and NO₂^--N content decreased. Moreover, the complexity and stability of microbial networks were reduced from upstream forest streams to downstream urban and agricultural streams, indicating more fragmented networks. Our study provides enhanced knowledge about the factors controlling the microbial community assembly in rivers under increasing human pressure through the integration of physical, environmental, and ecological mechanisms, which can serve as a basis for predicting and responding to changes in ecosystem function under the intensified human pressure.