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(1) Background Recent studies analyzed the participation and performance trends of historic races such as the oldest ultra-marathon (Comrades) or the oldest 100-km ultra-marathon (Biel). One of the toughest and historic ultra-marathons in the world is the 'Spartathlon' (246-km ultra-marathon from Athens to Sparta). The present study aimed to analyze the trends in participation and performance of this race. (2) Methods Different general linear models were applied as follows the first model was a two-way ANOVA (Decade × Sex), with separat