Both Walcott and Daniel Sturridge missed training on Monday through illness and now Walcott has been ruled out of the World Cup qualifier at Wembley (8pm) and has returned to his club.
However, the England medical team remain hopeful that Sturridge will be fit for the Group H encounter.
With his options now looking thin - Chelsea centre-half John Terry also withdrew on Sunday - England coach Roy Hodgson added three players to his squad in the form of Liverpool teenager Sterling, Tottenham's Livermore and Southampton's Lallana.
Gary Cahill is primed to replace Terry ahead of Phil Jagielka, while Leighton Baines will continue at left-back in the absence of another injured defender, Ashley Cole.
England beat Ukraine 1-0 at Euro 2012 to qualify for the last eight but the tournament co-hosts were aggrieved to have a legitimate goal ruled out when the ball appeared to cross the line before it was cleared by Terry.
Meanwhile, a weekend surge in ticket sales means 68,000 have been sold for the clash against the Ukrainians, the Football Association confirmed.
The figure includes 6,500 which have been sold to Olympic Games volunteers at the reduced price of £20.12.
At half-time, 15 Team GB medallists will be presented to the crowd.