Listening effort and fatigue are common experiences when conversing in noisy environments. Much research has investigated listening effort in relation to listening demand using the speech-in-noise paradigm. Recent conceptualizations of listening effort postulate that mental fatigue should result in decreased arousal and a reluctance to invest further effort, particularly when the effort is not worthwhile. The aim of the study was to investigate the influence of fatigue on listening effort, in interaction with listening demands and motivation. To induce fatigue 30 adults with normal hearing completed a 40-minute long speech-in-noise task (“load sequence”). Pre- and post-load sequence listening effort was probed in easy and hard listening demands (individually adjusted signal-to-noise ratios); with high and low motivation (manipulated with monetary incentives). Subjective effort, estimated performance, and tendency to quit listening were collected using rating scales. Baseline pupil diameter and mean pupil dilation were recorded as indices of anticipatory arousal and objective effort. Self-reported effort and mean pupil dilation were overall larger during hard SNR as compared to easy SNR. Baseline pupil diameter declined from pre- to post-load sequence, suggesting an overall decrease in arousal. Monetary incentives had no influence on the baseline pupil diameter for the easy SNR condition, but for the hard SNR condition larger incentives led to larger baseline pupil diameter. These results suggest that anticipatory arousal may be influenced by fatigue and motivation effects. Models of listening effort should account for the independent influence of motivation and previous load on anticipatory arousal and effort in distinct parameters.