The purpose of this study was to describe and compare hematologically elite football players during the competitive period according to playing positions and determines the prevalence of anemia. The subjects consisted of 38 professional footballers from a professional club in the National Football League First Division of Brazil. Six groups were formed for hematological comparison: (a) goalkeepers; (b) side defenders; (c) central defenders; (d) central midfielders; (e) side midfielders; and (f) attackers. Data collection began at the start of competition. The anthropometric, hematological, and VO2 max measures were carried out on the first day. To determine hematological parameters, fasting blood samples were taken from the cubital vein. The One-Way analysis of variance and the Bonferroni adjustment for multiple comparisons were used for statistical comparison between the footballers of different playing positions. No statistically significant differences were found among the playing positions in all three hematological parameters (red blood cell count, hematocrit, and hemoglobin). All players maintained strict compliance of balanced diets.