Queen of the South said some spectators have been "deliberately throwing food to attract the seagulls into the crowded areas".
In a tweet, the Dumfries-based Scottish Championship club encouraged fans to instead use bins provided around the 8,690-capacity Palmerston Park.
The new season begins on Saturday with the Doonhamers at home to Dunfermline Athletic and the club warned seagull waste could cause injuries.
A statement on the club website said: "We are aware that some spectators at the earlier Betfred League Cup matches at Palmerston were deliberately throwing food to attract the seagulls into the crowded areas.
"We are concerned about the potential for spectators or other personnel to suffer injury or fouling from seagull waste.
"Therefore as we prepare to host Dunfermline in the Championship tomorrow, we are asking supporters to please refrain from doing this again.
"Stewards have been instructed that should this type of deliberate behaviour be observed at the match, those responsible will be immediately removed from the ground.
"Your co-operation would be greatly appreciated."
The club posted a similar statement last month at the start of the Betfred Cup fixtures.
After playing Motherwell at the ground on 13 July, Queens said it was "well aware of the issues" but conceded that trying to eradicate the seagull problem was "pretty much an impossible task".
The club has previously tried to stave off seagulls by playing birds of prey noises over the sound system.