The organisation put out a public statement in which they claimed that staff likened to 'nightclub bouncers' were positioned by a bridge leading to the Cavendish Pavilion cafe.
Head of campaigns Duncan Dollimore said: “Cycling UK looked into this matter a little closer, we found we were really only scratching the surface and that the Devonshire estate had been failing to provide access to cyclists to estate land for years.
“This is one of the most beautiful landscapes in Yorkshire and it’s easy to see why the estate attracts some 400,000 visitors a year.
"Cyclists should be able to enjoy the special qualities of our National Parks, instead they are being faced by security guards more appropriately employed outside of nightclubs.”
Local cyclists who have objected include Sharon McAuley from Ilkley, who said: "I have used this route for over 20 years without problems and cannot understand the recent change. I live locally and feel most aggrieved at this loss of a route. We have usually stopped for a coffee at the Cavendish so contribute towards the estate.".
Alistair Preston, a company director from Leeds, was also caught out by the change.
He said: "As a result of this closure we missed Bolton Abbey from our recent ride itinerary. Our group of four people did not get a coffee and a cake at the Cavendish Pavilion, and they lost out on £20-£30 from us alone..
"We would not mind walking with the bikes from the gate to the cafe - it’s only 50 yards."
Yet the estate's management said that the path had only been closed temporarily since the beginning of the Covid-19 pandemic to ensure social distancing and to prioritise pedestrians.
They added that they did not employ security guards, and that members of the estate staff were sometimes positioned at the bridge to advise cyclists of the restrictions, but had no further powers to enforce them.
Several of the signs relating to the rule change have also been removed without permission.
The estate say they have not been approached directly by any cycling groups to discuss the restriction, which will be lifted once social distancing guidelines are relaxed.
Cyclists still have access to the Cavendish Pavilion via another route, Riverside Drive.
An estate spokesman said: "Bolton Abbey has welcomed cyclists onto the estate for many years and many thousands have enjoyed its miles of tracks and pathways. During the pandemic, to enable safe social distancing and protect the health and wellbeing of our visitors and colleagues, use of the Cavendish Bridge at Bolton Abbey is currently restricted to pedestrians only.
"This measure was introduced in April 2020, in response to the Covid-19 pandemic. This restriction is explained on signs at the approach to the bridge. We are aware that there have been instances of these signs being removed, and when this happens, we ensure they are replaced as quickly as possible. On busy days a member of the Bolton Abbey team is located on the bridge to remind visitors of the rule.
"Access for cyclists to the estate, including the Cavendish Pavilion and toilets, is via Riverside Drive. Once social distancing rules relating to Covid-19 are no longer in force, the restrictions on cyclists using the bridge will be lifted. The route in question is registered as a permissive path on the OS map, not a public right of way.
"We have enjoyed a positive relationship with cyclists on the estate for many years and would welcome a constructive dialogue with cycling groups with regard to any concerns they might have."