Five TSA workers are out of the job and another 38 have been suspended after they reportedly failed to conduct random security screenings on passengers and their luggage.
The employees were all based out of Southwest Florida International Airport in Fort Myers, and have been replaced by agents from other locations.
The affected workers were employed in different capacities at the airport, including supervisors and front-line security screeners, the News-Press reported.
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TSA trouble: The affected employees were all based out of Southwest Florida International Airport in Fort Myers, and have been replaced by agents from other locations
TSA spokesman David Castelveter told the paper that the move is one of the largest punishments handed out by the TSA since it was created in 2002.
Mr Castelveter added that the internal probe spanning two months last year found that some 400 passengers who underwent routine screening at the airport never received extra random checks.
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Despite the findings, the TSA claims the nearly 4 million passengers who travelled to and from the airport last year were not put at risk.
Republican Congressman John Mica, of Florida, the chairman of the House Transportation and Infrastructure Committee, demanded answers from the TSA.
Critic: Florida Rep John Mica, the chairman of the House Transportation and Infrastructure Committee is demanding answers from the TSA
Rep Mica said in a statement to the News-Press: 'Whether TSA is trying to protect administrative bureaucrats from congressional and public scrutiny, or just trying to sweep this breach under the rug, TSA must come clean and provide a full accounting of this incident.'
The news comes about a month after it was revealed that TSA agents and other U.S. law enforcement and Homeland Security personnel face an average of 55 daily encounters with 'known or suspected terrorists' named on government watchlists.
The first: Underwear bomber Umar Farouk Abdulmutallab, is pictured in his 2009 mugshot after his failed Christmas Day bomb attempt
The figure - which equals more than 20,000 contacts per year - underscores the growing sweep of watchlists, which have expanded significantly since the failed Christmas Day 2009 'underwear' bombing attempt - and officials noted that very few of those daily contacts lead to arrests.
Civil liberties groups question the use of watchlists, and they have been ridiculed for ensnaring innocent citizens.
Rep Mica said the disciplined workers are the latest in a series of mishaps within the agency designed to protect air travellers.
He said: 'Recent meltdowns such as those in New York, Newark, Palm Beach, Honolulu, Charlotte and Los Angeles have become familiar.
'For example, TSA recommended firing 36 employees in Honolulu for improperly screening luggage, and 12 others in Charlotte [North Carolina] for similar reasons. In Newark, a total of 10 workers were fired or suspended for theft or sleeping on the job.'
The TSA has come under intense scrutiny for the pat-down and body scanner policy in recent years, especially in the age of YouTube.
In March, a clip on the video-sharing site showed a terrified eight-year-old boy confined to a wheelchair, trembling with fear as he was subjected to an invasive TSA pat-down.
Check: A suspect's name is put on the 'no fly' list if they are deemed by government experts to be a threat to aviation
That same month, a new mom was forced to show security her freshly pumped breast milk before she could board a plane with a breast pump.
In February, a mother claimed she was subjected to repeated body scans after being told by agency employees that she 'had a cute figure'.
These followed incidents last year, where TSA agents allegedly helped themselves to cake in a flier's bag, and confiscated another's cupcake.
In December, Indiana rapper Freddie Gibbs was flying to a gig in Denver with two large bags of marijuana in his bag.
Extra eyes: The TSA has come under scrutiny in the age of the internet over several incidents, some of which have been caught on tape and spread via sites like YouTube
But instead of confiscating it, the agent who inspected the bag instead left a note saying, 'C'mon son'.
That same month, not one but two grandmothers came forward to say that they were mortified when they were ordered to be strip-searched during a security screening.
A TSA agent was fired in October after New York writer and lawyer Jill Filipovic found a note in her luggage (which contained a vibrator) that said: 'Get your freak on girl'.
In May 2011, a bizarre photograph showing a baby being frisked by airport security caused outrage after it was posted online.