Following on from the news that Lee Williamson will be out for four to six weeks with ankle ligament damage, Dickson Etuhu has sustained a reoccurrence of the medial knee ligament injury that has seen the midfielder sidelined since Christmas.
His latest injury set back means that the Nigerian might miss the remainder of the season.
Martin Olsson is expected to be out for at least a month after sustaining an eye injury in training.
With the last kick of the ball on Friday, the Swedish international took a ball to the face and has suffered internal bleeding in the eye which will keep the full back out of action for the foreseeable future.
To compound matters Colin Kazim-Richards limped out of action after 54 minutes against Leeds United. The striker sustaining an ankle injury that will require a scan on Sunday.
"There have been a lot of things that have gone against us over the last 48 hours," Appleton told Radio Rovers.
"We've had three players go down before the game who are going to be out for 4-6 weeks.
"Now with Kaz going down early in the second half we'll wait for the diagnosis tomorrow and have a better idea of what is going to happen.
"I think he landed on the boy and twisted his ankle a little bit. He felt something at the back of his ankle, probably something similar to the Lee Williamson one in a way.
"Hopefully it isn't as serious as that and we can get him back on the pitch as soon as possible.
"With Martin he is pretty much going to be out for 4-6 weeks, so that isn't fantastic news, it is a shame for the lad and a shame for the football club as he is a key part of what we want to do."