Freeze Tolerance of Amyloodinium ocellatum (Myzozoa: Blastodiniales)

Amyloodinium ocellatum causes epizootics in marine fish hatcheries. Frozen, wild fish, often used as food in aquaculture, could introduce the disease. We investigated the parasite's ability to survive, reproduce and infect a host after freezing. Infected gills were frozen at -20°C for either 0, 24, 36, 48, or 72 h. Parasite viability was assessed first by placing thawed infected gills into tanks with naïve fish. Gills of the exposed fish were examined on day 0 and on days 4 and 7 post-exposure. Tomonts from infected gills in all freeze durations produced infections. However, dinospore production decreased and infections took longer to establish in fish exposed to tomonts from the 72 h treatment. Second, eight tomonts from each freeze duration were placed in 5 mL of 25 ppt artificial seawater at 22°C in individual wells of a 12-well plate and monitored for hatching. Ninety seven percent of tomonts from all treatments hatched. Tomonts frozen for up to 72 h survived, reproduced and infected fish, but dinospore production decreased as freeze duration increased. Extrapolation from the rate of reduction in dinospore production over the freeze durations tested suggests that a freeze duration of 237 h could inactivate all tomonts.