Timothy Allen, who is 40, and 28-year-old Shaun Anderson of the Buccleuch Hunt in the Borders, have been charged with deliberately hunting a fox with a pack of dogs.
The offence is alleged to have been committed on land surrounding Whitton Farm near Morebattle, Roxburghshire, on December 20.
Both men, who live at Eildon, near Melrose, pleaded not guilty at Jedburgh Sheriff Court last week and three days have been set aside for a trial, starting on October 8.
It is understood film evidence will be led which has been submitted by investigators from the League Against Cruel Sports.
It will be the latest challenge to the Protection of Wild Mammals Act 2002 which was recently subject to a review by Lord Bonomy who made a number of recommendations for changes.
The main purpose of the Act is to ban the deployment of dogs to chase and kill wild mammals.
However, it also provides a number of exceptions which allow the limited use of dogs for certain situations.
Father and son John Clive Richardson, 67, and 24-year-old Johnny Riley became the first members of a mounted hunt to be successfully prosecuted in June since the Act was introduced north of the Border.
The Jedforest Hunt members in the Borders were filmed by investigators from the League Against Cruel Sports.
Following eight days of evidence, Sheriff Peter Paterson ruled that in two incidents Richardson and Riley were in breach of the Act and found them guilty of deliberately hunting a fox with dogs.
Riley – who was in charge of the hunt – was fined £400 at Selkirk Sheriff Court and Richardson, described as having a lesser role, fined £250.
There have also been a number of successful prosecutions under the Act involving individuals who are not part of recognised hunts.