Earlier this week, the Rt Rev Wesley Carr was branded a bully by Frank Field, the social security minister, for suspending the Abbey's popular organist, Martin Neary.
Now he faces more dissent after reportedly reducing several elderly women to tears at a meeting in which he informed the Abbey's 40 guides that they would have to retire once they reached 75.
Dr Carr told the guides: "People need to know they don't have to go on forever and people need to feel they are not joining a group that is necessarily elderly.
"Everyone who loves the place is actually loved by the place itself, but there comes a point, and it happens to all of us, that we must face retirement. But we are not abruptly curtailing. People will be kept in touch with a visit each year and a newsletter."
Although many of the guides have served at the Abbey for a number of years, acquiring detailed knowledge of the church, they have been told they must leave for insurance purposes.
One elderly guide now facing the sack described the Dean as a bully who had handled the process brutally and insensitively.
She added that working at the Abbey meant a lot to the guides, many of whom were as agile as 60-year-olds.
Emma St John Smith, spokesperson for the Abbey, said: "The Dean was concerned at the large number of elderly people and there was a feeling that it would be reasonable to have a cut-off retirement age.
"He felt that 75 was not unreasonable. I was not at the meeting myself, but I think the accounts of it have been over-dramatised."
Despite winning praise for his organisation of the funeral of Diana, Princess of Wales, Dr Carr has been a controversial figure since his arrival at the Abbey 10 months ago.
Staff were upset at the introduction of charges for tourists and by his decision to allow the body of the right-wing former politician, Enoch Powell, to lie in state in the chapel before his funeral.
This week he was accused of running a "totalitarian regime" by Mr Field, a leading Anglican, after the suspension of Mr Neary amid allegations of irregularities in the administration of the music department.
Visits to the Abbey have fallen by 20 per cent since the introduction of a pounds 5 charge. Dr Carr said that he brought it in because: "Too many visitors treated the Abbey like a waiting room or somewhere to meet their friends."
He has said he would like the Abbey to regain a more spiritual atmosphere.