The calves were being hauled in a truck tractor semi-trailer on eastbound I-290 near U.S. Route 64 when a hatch on the trailer came loose, according to Illinois State Police.
Several calves fell out of the trailer and onto the roadway, police said.
Police said a few of the calves appeared to have sustained minor injuries but none were seriously injured.
Eastbound lanes of I-290 were intermittingly closed while authorities wrangled the calves. All lanes were reopened at about 3:30 a.m.
No further information was available.