The majority of pathogens that cause high host mortalities are capable of infecting more than 1 host species. One characteristic of pathogens, which enables infection of multiple hosts, is the existence of free-living infectious stages, e.g., zoospores that are produced in vast numbers. Understanding the influence of environmental factors, particularly temperature, on the production and survival of these free-living stages is key to predicting future spread and emergence of disease. The rosette agent, Sphaerothecum destruens, is a significant multi-host intracellular fish parasite, with an obligate intracellular spore stage and a free-living (extracellular), potentially infectious, zoospore stage. It has caused high mortalities in endangered and commercially important fish species. Here we investigate the influence of temperature on S. destruens zoospore production and longevity. Our results indicate a wide temperature tolerance of the spore and zooflagellate stage with later onset and longer lifespan of zoospores at lower temperatures. This has important implications for the environmental persistence of S. destruens and provides additional information on the pathogen's life cycle with direct implications for S. destruens risk assessment.