Ferritins are conserved iron storage proteins that exist in most living organisms and play an essential role in iron homeostasis. In this study, we reported the identification and analysis of a ferritin middle-chain (M) subunit, MaFerM, from blunt snout bream, Megalobrama amblycephala. The full length cDNA of MaFerM contains a 5′-untranslated region (UTR) of 152 bp, an open reading frame (ORF) of 522 bp and a 3′-UTR of 270 bp. The ORF encodes a putative protein of 174 amino acids, which shares extensive sequence identities with the M ferritins of several fish species. In silico analysis identified both the ferroxidase center of mammalian heavy-chain (H) ferritins and the iron nucleation site of mammalian light-chain (L) ferritins in MaFerM. Quantitative real-time reverse transcription polymerase chain reaction analysis indicated that MaFerM expression was highest in the liver and lowest in the heart and responded positively to experimental challenges with Aeromonas hydrophila. The exposure of cultured M. amblycephala to treatment with stress inducers (iron and H2O2) significantly up-regulated the expression of MaFerM in a dose-dependent manner. Iron chelation analysis showed that recombinant MaFerM purified from Escherichia coli exhibited apparent iron binding activity. These results suggest that MaFerM is a functional M ferritin and is likely to play a role in iron sequestration and protection against oxidative stress and immune stimulus.