Bobby Hassell's first league goal in over two years helped edge Barnsley further away from the npower Championship relegation zone as Watford lost more ground in the race for automatic promotion.
Hassell, on as a sixth-minute replacement for the injured Rory Delap, headed home Jacob Mellis' cross 10 minutes before half-time.
It was the defender's first league goal since scoring for the Tykes in a 1-1 home draw with Swansea on January 22, 2011.
After climbing out of the bottom three with victory over Brighton at Oakwell on Tuesday night, Barnsley are now two places and two points above the drop zone.
With 39 away league goals before today, Watford are by far the Championship's highest scorers on their travels but they lacked their usual attacking flair against the Tykes.
The Hornets stay third in the table though, for now, ahead of Crystal Palace's trip to Brighton tomorrow, and remain two points behind second-placed Hull, who lost at home to resurgent Nottingham Forest, following back-to-back defeats.
After beating Brighton in midweek, Barnsley boss David Flitcroft named an unchanged side. That meant that 10-goal top scorer Chris Dagnall remained on the bench as Jason Scotland partnered Chris O'Grady up front.
Championship manager of the month for February Gianfranco Zola made three changes from the side that lost at home to Blackpool last weekend.
Joel Ekstrand replaced Matthew Briggs in the Watford back three, while Marco Cassetti and the Hornets' leading goalscorer Matej Vydra were recalled at the expense of Daniel Pudil and Fernando Forestieri.
On-loan Chelsea midfielder Nathaniel Chalobah and defender Fitz Hall both returned from hamstring injuries and were included among the substitutes.
Barnsley goalkeeper Luke Steele was called into action early on to push away Jonathan Hogg's header and Vydra blasted a 20-yard shot way over the crossbar as the half-hour approached.
The hosts had their first real effort on goal after 33 minutes as Scotland forced a good save from Jonathan Bond.
Two minutes later, the Tykes took the lead. Mellis got free down the left and whipped in a pacy cross which Hassell guided into the bottom left-hand corner of the net from 12 yards out.
Watford were unable to respond in a frantic final 10 minutes of the first half as Barnsley defended stubbornly.
O'Grady almost made it 2-0 at the start of the second half but Hogg got down quickly at his near post to save.
Vydra finally got a clear sight of goal in the 73rd minute but his volley produced a good save from Steele who held on to the shot.
Flitcroft's side continued to stifle the visitors while creating chances of their own.
Half-time substitute Dagnall, on for Scotland, cleared the crossbar with a shot after 84 minutes before O'Grady shot straight at Bond with three minutes of the regulation 90 remaining.
As they did at the end of the first-half, Watford applied some late pressure but Steele confidently came and dealt with a couple of corners.