Joseph O’Brien’s Defiantly has been ruled out of Tuesday’s Melbourne Cup after he failed a veterinary inspection.

The four-year-old Kameko gelding had been referred for an extra check on Saturday morning after losing a shoe and showing changes in his gait at Thursday’s mandatory inspections.

A statement from Racing Victoria said: “In the opinion of the veterinarians, Defiantly remains unsuitable to accept for the Melbourne Cup and has been withdrawn from the race by order of the stewards acting on veterinary advice.”

Stablemate Kizlyar has now take the final place in the 24-runner field following Defiantly’s withdrawal, meaning that trainer Joseph O'Brien still has five of the 24 runners in Tuesday's race.

Harry Charlton’s Wine Dark Sea and Roger Varian’s Enfjaar passed the inspections to take their place in the race.

O’Brien’s James J Braddock was drawn widest of all in gate 24 with Sons And Lovers given an inside draw in three.

Trustyourinstinct, Goodie Two Shoes and Kizlyar were drawn outside in 18, 19 and 20 respectively.

Wine Dark Sea was handed stall 23, Enfjaar eight and the George Boughey-trained Hopewell Rock handed the number 15 gate.