Graham's Dairies, Scotland's largest independent milk producer, is alleged to have caused trade effluent to enter a minor watercourse at Airthrey Kerse Farm in Bridge of Allan, where it has operated for more than six decades.
The pollutant is said to have flowed from the Beacon Burn and entered the Forglen Burn.
At a preliminary hearing yesterday, Stirling Sheriff Court was told the incident was alleged to have occurred on November 20 or 21, 2011.
Depute fiscal Iain Batho said 10 Crown witnesses had been cited for the trial, due to start in April.
Euan McSherry, for the firm, said its managing director, Robert Graham Jr, would give evidence for the defence.
Sheriff Wyllie Robertson continued the case for a further pre-trial hearing on March 8.