Huddersfield Town were beaten 2-0 in their Under 23 Professional Development League II game at Watford.
First-half goals settled the contest against a Town side featuring a quartet of senior players.
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Keeper Joel Coleman, centre-back Jon Gorenc Stankovic and midfielders Ivan Paurevic and Philip Billing were on Frankie Bunn’s teamsheet.
Coleman, Town’s £100,000 signing from Oldham Athletic, has been an unused substitute in all five first-team games, with on-loan Liverpool stopper Danny Ward getting the nod.
Slovenian Stankovic, brought in from Borussia Dortmund for £600,000, made his senior debut in the League Cup first-round defeat at Shrewsbury Town.
Croatian Paurevic,a £200,000 recruit from Russia’s FC Ufa, has been limited to two first-team appearances from the bench.
Meanwhile Billing is working his way back to full fitness after an ankle injury.
Their presence couldn’t help Town keep out an Alex Jakubiak penalty after seven minutes and a Dennon Lewis strike direct from a corner after 20.
Bunn’s side drew 1-1 against Charlton Athletic in their Under 23 opener at PPG Canalside.
Town are the defending PDL II champions.
They beat Sheffield United 2-1 in the play-off final at Bramall Lane last season, when the competition was at Under 21 level.
Watford made it two wins from two after beating Birmingham City 1-0 first time out.
The Hornets are coached by former Leeds United, Liverpool and Australian international striker Harry Kewell.