This study aimed to analyze aircraft ground operation processes from a human factors perspective with special emphases on the occurrence and influence of interruptions on pilots’ workload. Interruptions have been shown to increase workload and error probability as well as to contribute to fatal accidents in various fields. Countermeasures have been initiated especially in high-risk environments such as those involving medical issues. In aviation, more explicitly during turn-around processes, interruptions might occur frequently and impair flight safety. One hundred and sixty fully certified pilots working for a European airline were observed during their turn-around while performing real operations. Pilots’ interruptions were documented and classified in order to predict subjectively perceived workload by use of multiple linear regression analysis. External factors such as weather conditions, technical problems, and time pressure were considered as covariates. On average, a pilot experienced about eight interruptions during a turn-around. Overall workload estimates showed a level comparable to that of manual flying in a simulator. Interruptions from colleagues or from outside the cockpit were found to predict pilots’ workload; however, further external factors such as poor weather conditions impacted workload even more strongly. We suggest two approaches based on our results to handling the high rate of interruptions. We first recommend procedural changes to diminish the interruption rate; second, we recommend comprehensive, line-oriented flight training for airline and ground staff to raise awareness about the negative influence of interruptions.