Peat bogs are unique wetland ecosystems of high conservation value all over the world, yet data on the macroinvertebrates (including mayfly assemblages) in these habitats are still scarce. Over the course of one growing season, mayfly assemblages were sampled each month, along with other macroinvertebrates, in the largest and oldest Croatian peat bog and an adjacent stream. In total, ten mayfly species were recorded: two species in low abundance in the peat bog, and nine species in significantly higher abundance in the stream. Low species richness and abundance in the peat bog were most likely related to the harsh environmental conditions and mayfly habitat preferences. In comparison, due to the more favourable habitat conditions, higher species richness and abundance were observed in the nearby stream. Three of the recorded species, Caenis luctuosa from the peat bog, and Eurylophella karelica and Leptophlebia marginata from the stream are new records for the Croatian mayfly fauna. Typical Central European life cycle patterns were confirmed for several species (e.g. Baetis vernus, Nigrobaetis niger, Electrogena ujhelyii), while for several others (e.g. Habrophlebia fusca, Paraleptophlebia submarginata) some discrepancies were observed. Therefore, these results provide new and valuable information on the ecology of mayflies in peat bog habitats.