The triple Olympic champion finished third behind Warren Weir and Yohan Blake at the Camperdown Classic in Jamaica.
Athletics: Usain Bolt beaten by Jamaican teammates in opening race of the season
Usain Bolt, the Olympic sprint champion, has been beaten in his first race of the 2013 season.
The Jamaican finished third in a low-key 400 metre event in his hometown of Kingston.
The triple Olympic gold medallist easily won his heat in 46.71 seconds at the Camperdown Classic.
But he had to settle for third place overall after his teammates Warren Weir and Yohan Blake both posted faster times in their heats.
Weir, who finished third behind Bolt and Blake in the 200m final at London last year, won in a personal best time of 46.21 seconds.
Blake, racing in the same heat as Weir, stopped the clock at 46.64 after fading in the final straight following a blistering start.
There was no final for the event, which was part of a meeting run by the Racers Track Club that attracted around 30 athletes.
Bolt is the world record holder and double Olympic champion for the 100 and 200 sprints but often competes in 400 races at the start of each season to build his fitness.
Next week, he is scheduled to appear in Texas for the National Basketball Association All-Star Game but his major goal this year is the world championships in Moscow.
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