But the wind was a fraction over the legal limit, meaning the event record of 9.85 set by Bolt’s fellow Jamaican Asafa Powell in 2005 still stands.
Britain’s Craig Pickering, the 2007 Ostrava champion, ran a superb race to finish second in 10.08.
Meanwhile, Mark Lewis-Francis was fifth in the 100m ‘B’ race in 10.40.
Lewis-Francis, back after a year out injured, finished two places ahead of fellow Briton Christian Malcolm.
Marilyn Okoro produced the best British performance of the night, winning a closely-contested 800m in a season’s outdoor best of 2:00.21.
Okoro just managed to hold off France’s Elodie Guegan with another Briton, Jenny Meadows, finishing narrowly behind in 2:00.48.
Elsewhere, Olympic champion Dayron Robles comfortably won the men’s 100m hurdles with an impressive time of 13.04, while Ireland’s Paul Hession won the 200m.
But Bolt was the stand-out performer and again showed that he will be the man to beat at the World Championships in August.
The 22-year-old was slowest out of the blocks but delighted the Czech crowd by storming to a resounding victory.
Runner-up Pickering, whose legal best is 10.14, finished well clear of Frenchman Ronald Pognon in third.