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  1. In the 12 years since Dudgeonet al.(2006) reviewed major pressures on freshwater ecosystems, the biodiversity crisis inthe world’s lakes, reservoirs, rivers, streams and wetlands has deepened. While lakes, reservoirs and rivers cover only2.3% of the Earth’s surface, these ecosystems host at least 9.5% of the Earth’s described animal species. Furthermore,using the World Wide Fund for Nature’s Living Planet Index, freshwater population declines (83% between 1970 and2014) continue to outpace contemporaneous declines in marine or terrestrial systems. The Anthropocene has broughtmultiple new and varied threats that disproportionately impact freshwater systems. We document 12 emerging threatsto freshwater biodiversity that are either entirely new since 2006 or have since intensified: (i) changing climates; (ii)e-commerce and invasions; (iii) infectious diseases; (iv) harmful algal blooms; (v) expanding hydropower; (vi) emergingcontaminants; (vii) engineered nanomaterials; (viii) microplastic pollution; (ix) light and noise; (x) freshwater salinisation;(xi) declining calcium; and (xii) cumulative stressors. Effects are evidenced for amphibians, fishes, invertebrates, microbes,plants, turtles and waterbirds, with potential for ecosystem-level changes through bottom-up and top-down processes.In our highly uncertain future, the net effects of these threats raise serious concerns for freshwater ecosystems. However,we also highlight opportunities for conservation gains as a result of novel management tools (e.g. environmental flows,environmental DNA) andmore »specific conservation-oriented actions (e.g. dam removal, habitat protection policies, managedrelocation of species) that have been met with varying levels of success. Moving forward, we advocate hybrid approachesthat manage fresh waters as crucial ecosystems for human life support as well as essential hotspots of biodiversity andecological function. Efforts to reverse global trends in freshwater degradation now depend on bridging an immense gapbetween the aspirations of conservation biologists and the accelerating rate of species endangerment.« less