The purpose of this study is to determine the factors having impacts on the physical fatigue of employees and to analyse its influence on work on a production line. In order to carry out these objectives, a comprehensive research was conducted in a medium enterprise producing processed fruit and vegetable products. Air temperature and humidity tests, employee survey, production line productivity measurements, and other observations were performed. Large variations in temperature and humidity were observed during the analysed period. Analysing the survey results, attention was paid to the declared length of sleeping of employees after each shift, the time when they feel fatigue and the factors that, according to respondents, have the greatest impact on the work performed. Attempts were made to determine the wellbeing of employees and to indicate the disturbing symptoms. Special attention was paid to the negative phenomena felt by respondents. The main factors affecting the physical fatigue of employees were identified and described. We sought to explain the impacts of physical fatigue on work on the production line. Improvements, which should help to reduce fatigue and improve working conditions, have been proposed. The analysis provided interesting information that could be used in various food manufacturing companies.