AbstractWe investigated the iron isotope signatures of dissolved Fe in the water of the Wettinquelle mineral spring (Bad Brambach, Germany) by time series sampling covering seismically active periods related to tectonic activity near the Eger Rift system in central Europe. Our objective was to test whether Fe isotopes trace earthquake‐induced abiotic and biotic changes in the fluid/rock interaction of the deep, fissured, granitic aquifer. We found that the dissolved Fe isotope signatures in spring water are distinct from the granitic source signature (δ56Fe = +0.09‰). Particularly, we discovered that water δ56Fe values are remarkably stable (−0.01 ± 0.11‰, 2SD, n = 4) before and during a strong seismic swarm period in 2000 (local magnitudes ML > 3), while O2 and H2 concentrations in water decrease and dissolved Fe content increases. Later, recurring events of lower δ56Fe values down to −0.3‰ are observed in the period from 2001 to 2003 with intermittent seismic events (1 < ML < 3.2). The observations indicate a time lag between tectonic forcing and Fe isotope response. The role of abiotic fluid/rock interaction and Fe‐utilizing bacteria identified in the mineral spring water on Fe isotope fractionation is discussed. We explain recurring changes toward isotopically lighter values by a combination of Fe dissolution from deep granite and admixture of isotopically light Fe generated by a complex combination of abiotic and biotic processes operating in the aquifer when disturbed by swarm earthquakes events. We propose a conceptual model scenario of earthquake‐triggered changes in biogeochemical processes.