The badger cull could be starting as early as tomorrow, it has been suggested.
The suggestion that it might begin on Monday in Gloucestershire and Somerset follows the granting of a second licence by Natural England last week to allow farmers to shoot badgers to help control bovine tuberculosis among cattle.
Animal welfare charity, the League Against Cruel Sports has made an eleventh hour plea for a halt to the cull.
It is calling for the Government to postpone the badger cull trials and to debate the issue within the House of Commons as a matter of priority.
Chief executive Joe Duckworth said: "Proceeding with the trial badger cull flies in the face of the overwhelming public opposition and scientific evidence.
"The Government must listen to us and the countless thousands of voters, who see badger culling as an unjustified, cruel and counter-productive policy."
More than 150,000 people signed a petition backing the anti-cull campaign.
Farmer David Purser has spoken out against a pilot badger cull.
The 56-year-old, who owns a farm near Cheltenham, about 20 miles from the edge of the cull zone, says there is "no justification" to shoot the creatures.
Mr Purser, who has kept his own cattle since the 1980s, insisted culling was not the answer, and called for new legislation on vaccinating cattle to be pushed through.
"In every other instance where disease threatens our cattle we have vaccination in our armoury," he said.
It comes as 30 animal disease experts questioned the effects of the cull.
The experts, including the president of the Zoological Society of London and professors from Oxbridge and Imperial College London, have written a letter to the Observer which argues culling badgers could increase the problem of TB in cattle.
Prof Sir Patrick Bateson, a fellow of the Royal Society from the University of Cambridge, wrote: "The Government's TB-control policy for England includes licensing farmers to cull badgers. As scientists with expertise in managing wildlife and wildlife diseases, we believe the complexities of TB transmission mean that licensed culling risks increasing cattle TB rather than reducing it.
"Even if such increases do not materialise, the Government predicts only limited benefits, insufficient to offset the costs for either farmers or taxpayers."