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Trainer Nicky Henderson has withdrawn several of his runners from the Cheltenham Festival including Gold Cup hope Shishkin.
Henderson also pulled Jonbon from Wednesday’s Champion Chase amid concerns over the stable’s form.
Five of his six runners were pulled up on the meeting’s opening day.
He withdrew defending champion Constitution Hill last week from Tuesday’s Champion Hurdle after a poor workout.
The Berkshire-based trainer is second on the all-time list of Festival wins, behind Willie Mullins.
First Street and Kingston Pride were also withdrawn from races on Wednesday while Champ will miss Thursday’s Stayers’ Hurdle.
“Very sadly it will not be possible for Shishkin to run in the Gold Cup on Friday,” said Henderson.
“He was scoped [tested] as have all our potential runners this week, but unfortunately he has shown an unsatisfactory picture on which he couldn’t possibly run.
“He appears to be 100% in himself and has been working and schooling better than ever and we were really looking forward to Friday, it is hoped that along with all the other non-participants this week that they will be back in time for Aintree or Punchestown.”
Luccia did finish third in the Champion Hurdle for Henderson, but he was a notable exception.
“I’m afraid we have had to make some very tough decisions following the very disappointing performances of all bar one of our horses yesterday,” he said.
“It was there for all to see that there is obviously something affecting nearly all our horses.”