Caeleb Dressel also capped an historic performance with his third gold medal of the day by helping the team break the world record.
The United States set a world record of three minutes 19.60 seconds to win gold in the 4x100 meters mixed freestyle relay at the world swimming championships on Saturday.
Dressel, who won the men's 50m freestyle and 100m butterfly golds earlier in the evening, swam the opening leg before Simone Manuel touched the wall at three minutes, 19.60 seconds to seal America's win.
The time shaved more than three seconds off the previous record the U.S. team set two years ago in Kazan. The U.S. time broke the previous record of three minutes 23.05 seconds set by the American quartet at the 2015 world championships.
The Netherlands took silver at 2.21 secs back while Canada claimed bronze at 3.95.