Daisy Hill's hopes of building on Saturday's win over Bacup Borough were dashed within 38 minutes of the match at Ashton Town.
A strong and pacey Ashton side turned in an impressive first half performance, topped by a hat trick from striker Dylan Glass, and despite a second half revival from a Daisy side who never gave up and created their fair share of chances, they were unable to make a breakthrough.
The first half was played at a frantic pace, dictated largely by an Ashton Town side who made an impressive start to the season with a 5-1 win over AFC Liverpool on Saturday.
They almost took the lead in the first minute when Jason Carey curled a delightful free kick over the defensive wall, only to be denied by a fine save from Daisy keeper Dean Williams, and six minutes later the Daisy goal was under threat again when Alex McMillan headed just wide from a right wing cross.
Daisy responded with a long range effort from Hensley Mills and a snap shot from Jamie Ramwell which both missed the target, before Ashton Town took the lead on 17 minutes.
A surging run down the right flank from full back John Edgerton took him into the box, and when his shot was blocked the ball fell kindly for Dylan Glass, who buried a shot across the keeper into the far corner of the net.
Within five minutes, Glass had added a second goal with a close range tap in to put his side in the driving seat, but Daisy were handed a lifeline just after the half hour when they were awarded a penalty kick.
A challenge from Ashton's Michael White on Jake O'Brien sent the Daisy man tumbling in the box, and the referee pointed to the spot, sparking protests from the Ashton players.
But when Jordan Hussey stepped to take the kick, his well struck effort was well saved by Ashton keeper Ben Purdham, and six minutes later Ashton increased their lead further when Glass completed his hat trick.
It was a fine solo effort from the Ashton striker, who won the ball in a chase with Daisy keeper Dean Williams who had raced out of his goal to clear. He still had plenty to do when he picked the ball up on right hand touchline, but he advanced into the box and picked his spot with a shot past the two covering defenders on the goal line.
A minute later Daisy had Williams to thank for keeping the score down when he made a fine full length save after a fine run and shot from Ashton's Jesse Dowling.
Daisy finished the half on the front foot, however, with the last effort of an end to end first half being a Jake O'Brien free kick that was pushed away to safety by Ashton keeper Purdham.
It was perhaps no surprise that the pace of the game dropped in the second half, especially in the early stages, but an improved display from Daisy saw them create the best chances in the second 45 minutes.
Just after the hour Dean Singleton had a header cleared off the line, and moments later a well struck 20 yarder from Jake O'Brien was well collected by Purdham.
With 18 minutes remaining Danny Warburton was unlucky with a header that just missed the target, and as the game moved into the closing stages Daisy showed admirable effort and commitment as they pushed forward constantly in search of a goal, but were held at bay by a well organised Ashton Town defence, superbly marshalled by the experienced Michael White.
Ten minutes from time, Jordan Hussey just failed to connect at the far post with a low driven shot from Danny Warburton, and the last chance of the game fell to Jake O'Brien, whose well struck free kick was on target but was pushed away to safety by Purdham in the Ashton Town goal.