The Iron suffered defeat at Glanford Park, as Fleetwood Town came away with all three points in a 2-0 victory.
United made the early running, with chances for Matt Crooks and Kevin van Veen presenting themselves. However, the visitors edged ahead just before the break through Devante Cole.
They then sealed the win in stoppage time, as Bobby Grant finished following a swift Cod Army break as United committed men forward to find a leveller.
Graham Alexander made two changes to the side which started against AFC Wimbledon, with van Veen and Crooks replacing Craig Davies and Duane Holmes respectively.
Scunthorpe shotstopper Luke Daniels was called on three minutes into proceedings, as he comfortably denied Cian Bolger, who had headed goalwards from a corner.
The Iron soon got a foothold in the match though, and they created two opportunities in quick succession.
The first of which fell to Crooks. Paddy Madden’s looping cross from the left was inch perfect for the Rangers loanee, who powered a header towards goal. Like Bolger before him however, Crooks would be denied by a superb piece of goalkeeping - as Alex Cairns tipped over.
Dutchman van Veen had the second of the chances, as he latched onto a high through ball which evaded the Cod Army backline. The striker’s resulting shot across goal forced Cairns into a smart stop, but the visiting ‘keeper gathered at the second attempt.
Another lofted pass soon saw Madden bearing down on goal and, from much the same position as van Veen previously, the Irishman fired wide on the volley.
However, it would be Uwe Rosler’s men who broke the deadlock, four minutes prior to the interval. A slick team move eventually ended up with Cole tapping home at the far post, following Markus Schwabl’s low cross.
United’s response was almost instant. Josh Morris’ delivery from the right caused confusion in the Fleetwood penalty area, with Crooks and then Murray Wallace both seeing efforts rebuffed, before Cairns palmed the ball over for a corner.
Shortly after, referee Peter Bankes blew his whistle to bring the first half to a close.
The hosts were out the traps and in search of an equaliser after the interval, and they nearly got it through van Veen. The striker showed great individual skill to manufacture some space in the box, ahead of flashing a shot just wide of the far post.
Just minutes on from the hour mark, Alexander’s side thought they’d levelled the game up. Madden’s cushioned header found his fellow countryman Stephen Dawson, who acrobatically finished, but the flag was up for offside.
The hosts had the ball in the net once more on 73 minutes, however it was unfortunately ruled out after David Mirfin went in a little too heavy on Cairns, following van Veen’s dead-ball delivery from the left.
United committed bodies forward in the latter stages and, as a result, the Cod Army managed to double their advantage on the counter attack.
Former Scunthorpe man Grant wrapped up the victory for Rosler’s side, as he raced through and fired under Daniels from close range.
Grant’s finish proved to be last action of the afternoon, as the Cod Army took all three points back to Highbury.
Scunthorpe United: Daniels, Wallace, Mirfin (Wiseman, 82), Dawson (Toney, 82), Madden, van Veen, Morris, Bishop (Holmes, 70), Townsend, Crooks, Clarke.
Substitutes not used: Anyon, Hopper, Goode, Davies.
Fleetwood Town: Cairns, McLaughlin, Eastham, Ball, Grant, Bolger, Dempsey (Brannagan, 74), Davies, Schwabl, Maguire (Nirennold, 67), Cole (Hunter, 74).
Substitutes not used: Neal, Glendon, Sowerby, Ekpolo.