An untimely injury suffered by Reds defender Virgil van Dijk was considered to be the most notable fitness issue to emerge from an eventful Merseyside derby at Goodison Park.
The Dutch centre-half was, however, to catch James during an early tussle for the ball before being forced from the field himself.
That coming together has left an in-form Toffees star stuck on the treatment table, with Ancelotti not expecting the 29-year-old summer signing from Real Madrid to come into his thoughts for a trip to St Mary’s Stadium.
“Unfortunately, James had a problem during the game [against Liverpool],” Ancelotti told evertontv.
“He had a mistimed tackle with Virgil van Dijk at the beginning of the game and I think he will not be available for the game against Southampton.”
Everton will also be without club captain Seamus Coleman against the Saints, but Allan is available after avoiding any recurrence of the groin complaint that he carried into a meeting with Liverpool.
The Toffees will be looking to stretch their unbeaten start to the season when heading to the south coast, with Alex Iwobi looking to make what would be just his second league start of the 2020-21 campaign.
On his selection decisions, Ancelotti added: “We have to put in other players who have the desire to play.
“We are working this week on this and I think we are going to choose a good line-up for the game against Southampton. Alex is a player who works hard. He is fast and with his quality he can help the team be better in certain situations.
“He [potentially] has an opportunity to play but also we have Bernard and Anthony Gordon. We have a lot of opportunities to replace Richarlison and James. We are already moving on.”
The Toffees will also be without Brazilian forward Richarlison, with the former Watford star having been sent off late on in the Merseyside derby for a challenge on Thiago Alcantara.