An outbreak of nodular gill disease (NGD) in farmed rainbow trout Oncorhynchus mykiss (Walbaum, 1792) was recorded in Ladoga Lake (Karelia, north-western Russia) in Spring 2020. The disease was characterised by typical clinical signs including asphyxiation, distended opercula, loss of activity and swimming upside down under the water surface. Maximum monthly mortality was 15.2%. The histological examination of the gills showed deformation and clubbing of lamellae, epithelial hypertrophy and hyperplasia, lamellar fusion and fusion of filaments. Granulomas were located within the epithelial layer and/or rose above its surface. Light microscopic in vivo observations of the mucus smears from the affected gills revealed numerous amoeboid protists demonstrating a flattened body when adhering to the substratum, and blunt, radiating pseudopodia when afloat. Based on these morphological characters, these amoebae could be assigned to the Discosea (Amoebozoa), and analyses of their small subunit rRNA gene sequences showed that they belonged to the genus Vannella Bovee, 1965. The results reported herein support the designation of a new species, V. mustalahtiana sp. nov. Despite having been isolated from the gills of a freshwater fish, the species belongs to a clade of Vannella comprising mostly species isolated from marine and brackish water habitats. These findings may be essential for the aetiology and treatment of the disease.