Keeping a tight hold of her pink rucksack, Madeleine McCann excitedly climbs the steps of the aircraft about to leave England for Portugal. She falls on the third step, but quickly picks herself up to continue her climb aboard. The footage, taken on a mobile telephone, is the last recording of the toddler before she disappeared 26 days ago.

Just nine seconds long, it was released yesterday by her parents, Kate and Gerry McCann, as they continued to search for their missing daughter. The couple, now nearing their fourth week without her, will today travel to Rome to meet with the Pope after being formally invited to do so by the Vatican last night.

The previously unseen footage was filmed by one of a group of six who joined the McCann family on their trip to Praia da Luz in the Algarve. The friend recorded another clip when the group arrived in Faro and had boarded the shuttle bus bound for the terminal.

In that 13-second footage, Madeleine ­ dressed in pink shorts, a light pink top and trainers ­ is sitting next to her brother, Sean, two, and swings her legs as she waits for the bus to move.

Mrs McCann is seen smiling as she ruffles Sean's hair and holds on to his twin sister Amelie's arm. Mr McCann is looking serious, which prompts his friend to joke: "Cheer up Gerry, we are on holiday." Throughout the rest of the clip, laughter rings out from the group.

The footage has been watched over and over again by Mrs McCann since Madeleine's disappearance on 3 May from the hotel where the group was staying as her parents dined nearby. Since then, her fourth birthday has passed. Yesterday, they spoke of the memories the clips evoke

"Madeleine was acting like a big girl, she was so excited. She had her Barbie rucksack with a pull-up handle, it is a really girlie one. We all had to have our own rucksacks ­ even Sean and Amelie ­ it was quite funny."

Mr McCann said Madeleine had previously flown to Italy and the Netherlands. "She was dead excited about going away with the rest of the kids. It was her first time to Portugal."

The McCann's trip to Rome will mark the start of a tour of European countries to raise awareness of their missing daughter. They will meet the Pope on Wednesday after he concludes prayers in St Peter's Square. During the meeting, Mrs McCann, a devout Catholic, plans to hand the pontiff a photograph of Madeleine.