Footage appeared on YouTube of a Pompey shirt being placed in a stud wall at the club’s Staplewood ground before a construction worker covers it with insulation.
Leadbitter is building a £2m, two-storey club building containing gymnasium, changing rooms and staff offices at the site.
The two dryliners were working for a subcontractor.
David Cook, regional director of the Leadbitter Group, told the Southern Daily Echo: “They were not direct employees of Leadbitter. They were employed by our dry-lining subcontractor.
“We understand they have been disciplined by their employer and basically sacked.
“It is not something we condone and do not agree with their behaviour at all.
“The shirt that was put into the partition was immediately removed and the actual YouTube footage was removed within 24 hours.
“It is a serious matter that has been dealt with.”
During the 47 second video clip, the Saints are described in derogatory terms.
The footage of the pranksters has sparked a backlash among Saints fans on football message boards.
Although one Saints fan quipped: “If I had a Pompey shirt, I’d want to hide it as well!”