Analysis of traffic operation of two-lane rural roads with cyclist presenceNowadays, the presence of sport cyclists on two-lane rural roads is a common phenomenon. Cyclists ride individually and in groups, sharing the road with motorized vehicles and interacting with them. Due to the differences in speed, volume and level of protection between bicycles and motorized vehicles, the overtaking manoeuvre is the most frequent and dangerous interaction that occurs between them. Therefore, the presence of cyclists on two-lane rural roads can affect both road safety and traffic operation.The main objective of this research is the characterization of cycle traffic and its interaction with motorized vehicles, especially in follow-up and overtaking manoeuvres, based on naturalistic observations of the phenomenon, using instrumented bicycles and observations at the extremes of the study sections. The study has been carried out considering cyclists who circulate individually and in groups on sections of two-lane rural roads with different characteristics, both geometric and traffic. A total of 2,135 overtaking manoeuvres to cyclists have been analysed in detail. The duration of overtaking has been considered a key factor in terms of its effect on safety and traffic operation. Consequently, this variable has been analysed with special interest, developing a predictive model of the overtaking duration, considering factors related to the configuration of groups of cyclists, the road and the overtaking manoeuvre itself.Based on the observations, a traffic microsimulation model that incorporates cycle traffic on two-lane rural roads and its interaction with motorized vehicles has been adapted, calibrated and validated. Through observations and the use of the microsimulation model, performance measures have been obtained that allow characterizing the influence of the presence of cyclists on traffic operation, showing a reduction in traffic operation as cycle traffic increases. Performance measures related to the number of overtaking manoeuvres and delay are the most sensitive to the presence of cyclists, followed by measures related to users platooning. Roads with narrow cross-sections are more sensitive to the presence of cyclists, and on them the twoabreast configuration of groups of cyclists slightly improves traffic operation by facilitating overtaking. The results of this research allow a better understanding of the phenomenon as well as being translated into recommendations for improving the design and/or operation of twolane rural roads with a cyclist presence. In addition, specific tools have been developed such as the predictive model and the microsimulation model which can be used by infrastructure managers to analyse other rural roads and provide a safer and more operational integration of the different road users.