Large high-speed railway (HSR) networks are planned for the near future to accomplish increased transport demand with low energy consumption. However, high-speed trains produce unknown avian mortality due to birds using the railway and being unable to avoid approaching trains. Safety and logistic difficulties have precluded until now mortality estimation in railways through carcass removal, but information technologies can overcome such problems. We present the results obtained with an experimental on-board system to record bird-train collisions composed by a frontal recording camera, a GPS navigation system and a data storage unit. An observer standing in the cabin behind the driver controlled the system and filled out a form with data of collisions and bird observations in front of the train. Photographs of the train front taken before and after each journey were used to improve the record of killed birds. Trains running the 321.7 km line between Madrid and Albacete (Spain) at speeds up to 250-300 km/h were equipped with the system during 66 journeys along a year, totaling approximately 14,700 km of effective recording. The review of videos produced 1,090 bird observations, 29.4% of them corresponding to birds crossing the infrastructure under the catenary and thus facing collision risk. Recordings also showed that 37.7% bird crossings were of animals resting on some element of the infrastructure moments before the train arrival, and that the flight initiation distance of birds (mean ± SD) was between 60 ± 33 m (passerines) and 136 ± 49 m (raptors). Mortality in the railway was estimated to be 60.5 birds/km year on a line section with 53 runs per day and 26.1 birds/km year in a section with 25 runs per day. Our results are the first published estimation of bird mortality in a HSR and show the potential of information technologies to yield useful data for monitoring the impact of trains on birds via on-board recording systems. Moreover, recordings point to the use of the infrastructure by birds as a key issue leading to bird train-kill.