Optimization of the temporal aspects of task design requires a better understanding of the development of muscle fatigue in the neck and shoulder region over time. The objective of the study was to investigate this in two production companies and to determine the relationship between objective and subjective estimates of fatigue. Indicators of fatigue were obtained through electromyography (EMG) during test contractions and ratings of perceived discomfort. EMG amplitude increased during the day in both case studies while mean power frequency decreased only in one case. In both cases, a more detailed frequency analysis of the EMG signals showed an increase in lower frequency power accompanied by a decrease in higher frequency power. Local perceived discomfort in the neck and shoulder increased over the course of the day in both cases. However, no clear relationship between perceived discomfort and objective indicators of fatigue was found. Obtaining sufficient sensitivity to detect effects of temporal aspects of task design probably requires complementary or more refined methods (e.g. EMG arrays, mechanomyography).