Superfast Ferries have decided to withdraw their Rosyth to Zeebrugge crossing.
Sources in the company have blamed fuel costs and passenger numbers for the decision to cease the service.
A statement from the company's board of directors said: "We regret to announce the discontinuation of the Rosyth to Zeebrugge ferry service as of 14 September, 2008.
"The last crossing will take place from Rosyth on 13 September, 2008 arriving in Zeebrugge the following morning.
"As per the company's fleet deployment plans for next year, the vessel Blue Star 1 will be redeployed as of the coming winter in the markets where Attica Group's vessels are presently operating.”
Commenting on the news, a spokesman for Forth Ports said: "This is disappointing news as the ferry link is an important part of Scotland's access to European markets.
"Our own analysis shows that the route could easily attract 60,000 freight units, within a potential marketplace of 200,000 freight units, making this a viable, profitable route for an operator with the right mix of vessels and frequency of sailings.
“We will be working with the Scottish Government to attract potential operators with public sector support.”
Transport Minister Stewart Stevenson said the decision was "very disappointing".
"Scottish Ministers are strongly committed to the continuation of this route, and will pursue all viable options to achieve continuity of a commercially operated service."
Labour has called on Alex Salmond to personally intervene after the announcement of the closure of the ferry service at Rosyth.
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