The controversial American – booed throughout after his previous doping bans – won a highly-charged race in 9.92 seconds as Bolt could only come third on Saturday.
Christian Coleman completed an American one-two in 9.94 secs, with Jamaican Bolt crossing the line in 9.95.
Speaking after his shock defeat, bronze medallist Bolt told the BBC: “It was rough. But I came out and I did my best.”
Despite his disappointment, the Jamaican added his appreciation for the 66,000-capacity crowd at the London Stadium, however.
He said: “Thanks for your support. I really appreciate it.”
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