Great Britain came first to win gold in 37:47 seconds, the United States won silver while Japan took the bronze.
The British quartet won the race in the third fastest time in history.
Bolt had also missed out on the perfect goodbye in his 100m bid as Justin Gatlin beat him to the title, with the Jamaican finishing third.
Regarded as the greatest athlete of all time, Bolt has won 11 World Championship gold medals, along with two silver and a bronze medals. He also has eight Olympic gold medals.