Operator fatigue during work with agricultural machinery is a significant factor contributing to fatal accidents in Greece. Workers operating agricultural machinery, especially those working with tractors and off-road machines, constitute one of the most hazard-prone groups due to mechanical vibrations transmitted from the ground to the operators’ seats. Operators of tractors and off-road machines are exposed to intense mechanical vibrations, underscoring the importance of cumulatively calculating permitted working hours. This work presents a study that was conducted at the Laboratory of Safety and Ergonomics of Agricultural Machinery, Department of Agriculture – Agrotechnology, Faculty of Agricultural Sciences, University of Thessaly, in order to evaluate the vibrations generated when operators work with agricultural tractors and off-road machinery. The results of the study showed that the highest levels of vibration occurred during disc harrowing operation, particularly when the disc harrow moved vertically or at an angle of 30° to the ploughing direction. For seed bed preparation, it is recommended to use tractors equipped with a suspension system for the operator's cabin and a seat with an adjustable range of perceived vibration. Additionally, to prevent fatigue or health problems for tractor operators, it is advisable to limit permitted working hours to fewer than 8 hours per day.