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Many aspects of natural and hatchery origin salmonid genetics, physiology, behaviour, anatomy and life histories have been compared due to the concerns about what effects domestication and hatchery rearing conditions have on fitness. Genetic and environmental stressors associated with hatchery rearing could cause greater developmental instability (DI), and therefore a higher degree of fluctuating asymmetry (FA) in various bilaterally paired characters, such as otoliths. Nonetheless, to appropriately infer the effects of DI on otolith asymmetry, otolith mineralogy must be accounted for. Vateritic otoliths differ substantially from aragonitic otoliths in terms of mass and shape and can artificially inflate any measurement of FA if not properly accounted for. In this study, measurements of otolith asymmetry between hatchery and natural origin Coho salmon Oncorhynchus kisutch from three different river systems were compared to assess the overall differences in asymmetry when the calcium carbonate polymorph accounted for 59.3% of otoliths from hatchery origin O. kisutch was vateritic compared to 11.7% of otoliths from natural origin O. kisutch. Otolith mineralogy, rather than origin, was the most significant factor influencing the differences in asymmetry for each shape metric. When only aragonitic otoliths were compared, there was no difference in absolute asymmetry between hatchery and natural origin O. kisutch. The authors recommend other researchers to assess otolith mineralogy when conducting studies regarding otolith morphometrics and otolith FA., (© 2023 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)