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Damage evolution during low- and high-cycle fatigue in an embrittled duplex stainless steel is characterized in this paper. Moreover, scanning electron microscopy observations (SEM) in combination with electron backscattered diffraction (EBSD) measurements and transmission electron microscopy (TEM) were employed in order to analyze microcracks formation and propagation. During low-cycle fatigue, microcracks initiate the ferrite phase either along slip planes with the highest Schmid factor (SF) inside the grains or at the α/α grain boundary. Then, microcracks propagation take place in ferrite or austenite grains with the highest SF. An analysis of the dislocation structure in the near-surface and in ferritic grains in the bulk of the specimen has shown that dislocation microbands are associated with microcrack initiation. In the high-cycle fatigue regime, damage generally initiates in the austenite by slip band formation followed by crack initiation either at an α?α boundary or at an α?γ boundary in the intersection of slip bands in the austenite. The microstructure in the austenite consists of a low density of dislocation pile-ups while the ferrite is practically inactive or develops only micro-yielding at boundaries. Despite the differences in both fatigue regimes, phase boundaries are an effective barrier against crack propagation because they delay the advance of the crack tip.