Based on morphological and molecular characterisation, four amoeba strains isolated from organs of freshwater fish were identified as Hartmannella vermiformis Page, 1967. Small subunit rRNA gene sequences of these strains expand the set of corresponding complete and almost complete sequences of this species to twelve. A new species-specific oligonucleotide probe inferred from recently available SSU rRNA gene sequences was designed and successfully tested in tissue lesions produced by one strain of H. vermiformis in experimentally infected fish.