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Dynamic Origin/Destination matrices are one of the most important parameters for efficient and effective transportation system management. These matrices describe the vehicle flow between different points inside a region of interest for a given period of time. Usually, dynamic O/D matrices are estimated from link traffic counts, home interview and/or license plate surveys. Unfortunately, estimation methods take O/D flows as time invariant for a certain number of intervals of time, which cannot be suitable for some traffic applications. However, the advent of information and communication technologies (e.g., vehicle-to-infrastructure dedicated short range communications - V2I) to the transportation system domain has opened new data sources for computing O/D matrices. Taking the advantages of this technology, we propose in this paper a multi-agent system that computes the instantaneous and dynamic O/D matrix of any road network equipped with V2I technology for every time period and day in real-time. The implementation was done using JADE platform. The results show that the multi-agent system is able to obtain the instantaneous O/D matrix for any time period and day.