There has been an increase in the flow of freight vehicles commuting on Brazilian highways. Then, special attention to the structural performance of bridges regarding the fatigue in beams is needed. Brazil has neither normative metrology to study real data flow of vehicles, nor specific fatigue load train models and coefficients to the analysis and design of road bridges. The same load train that is used for general dimensioning, TB 450, is used for the fatigue verification. Hence, this work aims to verify if the current TB 450 is representative of the freight heavy vehicles with 2 to 9 axles concerning the effects of fatigue in the longitudinal reinforcement of beams of theoretical reinforced concrete bridges with two, three, and five beams. This verification is performed analyzing the stress variations found in the longitudinal reinforcement of vehicles with 2 to 9 axles and the TB 450. Based on the results, the longitudinal steel reinforcement was more susceptible to fatigue's effects. Freight vehicles with 5, 6, 8 and 9 axles presented the most significant stress, therefore, they tend to cause more deleterious effects. Hence, the adoption of a Brazilian normative fatigue specific load train and coefficients is necessary to analyze pre-existing road bridges and design new ones most accurately.