This paper deals with the assessment of a real prestressed tendon by the use of Barkhausen noise emission. The tendon was obtained from a real highway bridge after 33 years in service. Barkhausen noise is studied as a function of the stress state, and the Barkhausen noise signals received directly from the tendon on the real bridge are compared with the Barkhausen noise signals received from the tendon during loading in the laboratory. Assessment of the prestressing is based on the analysis of the effective value of the Barkhausen noise signal as well as the position in which the Barkhausen noise envelopes attain a maximum.