Eastbourne Borough 1 Truro City 0
City boss Lee Hodges blamed a "mad minute" of defending for his side's Blue Square Bet South defeat at Priory Lane.
City switched off at free-kick allowing home skipper Ollie Rowe a free header for the central defender's first goal of the season.
It proved to be the game's only goal leaving City frustrated at making the long journey back from East Sussex empty-handed after being the better side for much of the game.
Hodges said: "We played really well again, but conceded a sloppy goal giving their lad a free header.
"After we went behind we had waves on waves of attack but just could not find a goal. Again I thought we played ever so well."
Hodges made one change from the side that lost at Farnborough in the FA Trophy bringing himself into central defence in place of Calron Hall, who dropped to the bench, alongside Danny Carne, as City again travelled with a squad of just 13.
In the first half the visitors thought they should have had penalty when Andy Watkins appeared to be brought down by Eastbourne keeper Craig Ross, but the referee turned down their strong appeals.
After going a goal down, City did everything but score with Watkins and Kieffer Moore both unlucky not to equalise.
It was another frustrating day for City and they will be hoping for better luck on Tuesday evening when they travel to play Eastleigh.
City: McCormick; Hopkinson, Hodges, Ash, Williams; Cooke, Martin, Broad, Afful; Watkins, Moore. Subs (not used): Hall, Carne.
Yellow cards: Broad, Martin.
Goal: Rowe (56) 1-0.