Issues addressed Cyclists' perceptions of harassment from motorists discourages cycling. This study examined changes in cyclists' reporting of harassment pre- to post-introduction of the Queensland trial of the minimum passing distance road rule amendment (MPD-RRA). Methods Cross-sectional online surveys of cyclists in Queensland, Australia were conducted in 2009 (pre-trial; n=1758) and 2015 (post-trial commencement; n=1997). Cyclists were asked about their experiences of harassment from motorists while cycling. Logistic regression modelling was used to examine differences in the reporting of harassment between these time periods, after adjustments for demographic characteristics and cycling behaviour. Results At both time periods, the most reported types of harassment were deliberately driving too close (causing fear or anxiety), shouting abuse and making obscene gestures or engaging in sexual harassment. The percentage of cyclists who reported tailgating by motorists increased between 2009 and 2015 (15.1% to 19.5%; P<0.001). The percentage of cyclists reporting other types of harassment did not change significantly. Conclusions Cyclists in Queensland continue to perceive harassment while cycling on the road. The amendment to the minimum passing distance rule in Queensland appears to be having a negative effect on one type of harassment but no significant effects on others. So what? Minimum passing distance rules may not be improving cyclists' perceptions of motorists' behaviours. Additional strategies are required to create a supportive environment for cycling.