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A new, three-dimensional geometric morphometric approach was assessed to study the effect of developmental temperature on fish heart shape utilizing geometric morphometrics of three-dimensional landmarks captured on digitally reconstructed zebrafish hearts. This study reports the first three-dimensional analysis of the fish heart and demonstrates significant shape modifications occurring after three developmental temperature treatments (TD = 24, 28 or 32°C) at two distinct developmental stages (juvenile and adult fish). Elevation of TD