Nearly 15,000 people have signed a petition urging Chatsworth not to be involved in a possible badger cull.
The Government has announced badgers may be killed in eight new counties in England - including Derbyshire - to control the spread of bovine tuberculosis in cattle.
According to research by Derbyshire Wildlife Trust, bovine tuberculosis is not transmitted by direct contact between cattle and badgers but through contaminated pasture and dung.
The group says culling badgers is unjustified and costly and that vaccination - which its volunteers carry out - remains the only answer.
This online petition by the Derbyshire Against the Badger Cull group states: "The Chatsworth Estate owns 35,000 acres of land, spread across Derbyshire. This land could be home to hundreds of badgers, who are all now at risk. This petition is to urge Chatsworth to support badger vaccination, and not allow badgers to be culled on their land.
"Derbyshire has had a successful badger vaccination programme since 2014, though with the wider cull roll-outs in 2018, not even vaccinated badgers are safe.
"We are asking the Chatsworth Estate not to be involved in the slaughter."
A Chatsworth spokesperson said: "At this stage, Chatsworth will not be commenting on practises designed to control the spread of bovine tuberculosis. We take our responsibilities to our visitors, staff, tenants and local communities very seriously, and need to be mindful of the varying views on, and sensitive nature of, the subject."
'Vaccinate, don't cull'
Last year, 21 cull zones were licensed across the UK, with a total of 20,000 badgers killed.
Earlier this year, Hardyal Dhindsa, the Police and Crime Commissioner for Derbyshire, urged the Government to protect badgers in a letter.
Mr Dhindsa said: "Badgers are a protected species under the 1992 Badger Protection Act and evidence shows that shooting badgers is not a viable solution to the spread of bovine tuberculosis. If this is the case, then it doesn't make sense to pursue an expensive culling programme at the cost of the taxpayer.
"Derbyshire is at the forefront of a vaccination programme to limit the spread of this disease and there are no grounds, either scientifically or economically, to cull badgers.
"I am urging the Government to reconsider its position and to continue to invest in the vaccination programme in this county and beyond to protect what remains a threatened species for future generations."