Network Rail has announced the closure date of five level crossings across the new Levenmouth rail link.
The crossing points include Doubledykes, which has been the subject of a high-profile campaign to retain it.
Others affected are Balgonie, Waulkmill, Tullybreck and Durievale.
All will close permanently on August 31 and will be fenced off.
Campaigners say the paths are historic rights of way and are well-used by pedestrians and cyclists.
They have called for a solution to ensure the public can still use the path between Wemyss and Thornton.
However, Network Rail insists the rights have been legally extinguished.
Three new bridges planned
The Levenmouth rail link is due to open in spring 2024 and some sections of the track are already complete.
Engineering trains involved in the construction already use the line.
And Network Rail says it is no longer safe to allow public access.
New bridges are planned at Duniface, Methilhill and Mountfleurie and there will be temporary restrictions at the existing crossings there during construction.
Proposals for the new bridges are currently going through planning consent.
And dates for the temporary construction will be announced later.
Railways bosses say the three new bridges will maintain connections between communities.
And they will improve access to the new stations at Leven and Cameron Bridge.
Levenmouth railway crossing closures ‘unavoidable’
The £116 million railway will connect Levenmouth to the rail network for the first time in more than 50 years.
When complete, the new line will create a direct link with Edinburgh across 10kms of new track.
Joe Mulvenna, Network Rail’s project manager for the Levenmouth rail link project, said: “With significant work ongoing across the route to deliver the stations, signalling and bridges limiting access is now unavoidable as securing the railway boundary will protect the safety of the public.
“The most effective way of reducing level crossing risk on the railway is to remove crossings and we do not design them into new railways.
“The development of the rail link and associated active travel enhancements has given us the opportunity to reduce the number of crossings and replace some core paths and well-used walking routes with the new bridges which we are building on behalf of Fife Council.”
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