The event was held as licensees within the town claim the smoking ban, which was introduced by the Scottish Executive in March, is destroying trade in the area.
Local MSPs were invited to attend to hear what the traders had to say. But only Mid Scotland and Fife MSP Tricia Marwick attended the meeting.
The meeting in the CISWO club was held in a bid to save premises from closure, after the North Street club recently reported they had been losing £2,000 a week because older members would rather stay at home than go outside for a cigarette.
Secretary of the CISWO, David Nelson, said: "We invited the politicians along so we could tell them how we were being affected, and we told Tricia Marwick exactly what the situation was.
"Every business represented was in a similar situation to ourselves, and we're losing money.
"It's not just affecting the pubs and clubs as people may expect. It's having a knock on effect and it's affecting everybody."
With trade down on the same period last year, the businesses are concerned for their future.
Mr Nelson said: "I predict that at the end of the year, come the winter, there will be a lot of pubs, clubs, cafes and bingo halls closing.
"We are not asking for the ban to be totally lifted.
"We agree that it damages people's health, but it's damaging business as well.
"People shouldn't have to breathe in others' smoke, but we think there should also be somewhere inside for them to smoke."
The traders are looking for an independent investigation into the ban, which they feel was put in place without enough public consultation.
And they have set up a committee to draw up a petition, which will be distributed to all businesses within the Kingdom which have been affected by the ban, and they will present it to the petitions committee at the Scottish Parliament.
Mr Nelson added: "If they don't look at it quickly there's going to be a lot of businesses closing and jobs lost."
Tricia Marwick MSP said: "I was extremely disappointed that I was the only elected member to turn up when you consider that Glenrothes is served by eight MSPs from every party.
"I was disappointed that I alone turned up to hear the concerns of the licensing trade.
"I made it clear that I supported the ban and I voted for the ban, but I acknowledge that there are some places that are suffering.
"I undertook that any representations in their business or as individuals, they wanted to make to ministers I'd be happy to make sure those views would be put forward.
"I also undertook to advise them of how to go about putting a petition to the Scottish Parliament."
What's your view?
E-mail: fiona.purnell@fifetoday.co.uk