AbstractUnder climate change, glacier recession and the loss of cold habitats are major threats to aquatic biodiversity. In mountain areas, streams originating from rock glaciers, called “icy seeps”, may represent climate refugia for cold-adapted organisms, given the major persistence of cold waters from these landforms even in unfavourable climates. During late summer 2021, we investigated discharge, turbidity, water chemistry (major ions and trace elements), stable water isotopes (δ18O, δ2H), and macroinvertebrate communities of five rock glacier springs (icy seeps), five glacier springs (glacier springs) and five non-glacial springs (spring brooks) in catchments of the Eastern Italian Alps. In icy seeps, meltwater contribution to runoff (estimated with end-member mixing models) was intermediate between those of the other two spring types. Icy seeps had very cold waters (< 1.5 °C) that were enriched in trace elements, like glacier springs, whereas discharge and turbidity were low, like in spring brooks. Community composition, diversity, and species associations of icy seeps were strongly related to a gradient of chemical harshness (built using trace element concentrations), with less contaminated springs hosting communities like those dwelling in spring brooks. Like glacier springs, those icy seeps with the harshest water chemistry (particularly because of Ni concentrations) and higher meltwater contribution hosted species (e.g., Diamesa steinboecki) that are currently in decline due to glacier loss. This suggests a high conservation value for icy seeps. The protection of these habitats, nowadays overlooked, will be fundamental under the progressive warming and dry-out risk of alpine springs.