Marquess, who took over from Lucy Allan as the teen soap's showrunner last month, decided not to renew the contracts of three members of the Ashworth clan.
Josh, along with parents Suzanne and Neville - played by Sonny Flood, Suzanne Hall and Jim Millea respectively - will all be written out of the show at the end of their current deals.
Meanwhile, Andrew Moss - who plays Rhys - has dodged the axe and will stay with the programme for a while longer. According to reports, though, his long-term future remains uncertain.
The only remaining original Ashworth, Hannah, played by Emma Rigby, bows out of Hollyoaks later this week after five years in the role.
Marquess began work at the soap's home at Lime Pictures in Liverpool two days after his appointment. Thereafter, the cast were reportedly bracing themselves for a mass cull in their number due to his well-publicised clear-outs during his time producing The Bill and Family Affairs.
A show source last night told DS: "It was only time before some were given the boot. Everyone knows that the new captain's reputation precedes him as far as putting a stamp on a show is concerned."
An insider told the Daily Star: "He's getting rid of the dead wood - nobody is safe. This is just the beginning. Over the next few months expect to wave goodbye to even more faces."
The Ashworths descended on Hollyoaks village in October 2005 and during their time with the programme have been at the centre of some of the show's biggest storylines, including Rhys's incestuous relationship with his half-sister Beth and Hannah's battle with anorexia and bulimia.
More recently, they took residence in fictional pub The Dog In The Pond after successfully purchasing it from the Osbornes at auction.