Journalists at The Sentinel in Stoke have voted not to recognise the NUJ for the purposes of collective bargaining.
The Sentinel is believed to be the first daily newspaper in recent years to reject the union in a newsroom ballot.
The NUJ said 96 ballot papers had been sent out, 72 of which were returned. It said 36 votes were in favour and 36 against, but this meant it had failed to reach the required 40 per cent of votes in favour to gain recognition.
Instead staff will continue to represent themselves individually in annual wage negotiations.

Somerset County Gazette readers are being asked for help in treating oil-stricken birds at the RSPCA's animal centre near Taunton.
The paper has launched Operation Guillemot as the West Hatch centre takes in hundreds of birds covered in oil after the MSC Napoli cargo ship was beached off the East Devon coast.

Former press photographer Brian Duff has died, aged 77.
He was a photographer of the stars during a long and distinguished career with the Daily Express and Independent newspapers.
During his spell at the Daily Express between 1956 and 1986, based in Manchester, he struck up friendships with George Best, Donald Campbell and even LS Lowry.
One of his most famous pictures was a shot of Prime Minister Harold Wilson with smoke like a genie emerging from his pipe.

Award-winning photographer Martin Birchall, from the Liverpool Daily Post & Echo, has given a master class to third year journalism students at the University of Central Lancashire.
His advice to them: "Keep it simple. Try and keep backgrounds a dominant colour while getting all the relevant information into the photograph."

Lincolnshire Echo deputy sports editor John Pakey is preparing to take part in the London Marathon in April.
He’s enlisted the help of University of Lincoln boffins to find out if he’s fit for the run – and readers have been finding out whether he’s likely to make the grade.

The Citizen is helping Gloucestershire Wildlife Rescue Centre by boosting its Adopt a Hoglet campaign, to adopt baby hedgehogs and help the centre carry out its vital work.
It says hedgehogs are confused because of the mild weather and have started foraging for food when they should still be hibernating.

A Poole man’s dying wish was granted when his teenage sweetheart phoned him after reading a letter he wrote to the Daily Echo appealing for her to call.