Moamer Gaddafi took to the airwaves Friday to state that he is uninjured and safe in a location where NATO's bombs cannot reach him, dpa reported.
The message came in an audio recording released on Libyan state TV. No images were broadcast.
In the recording, Gaddafi disputed reports circulating Friday that he had been injured and referred to recent NATO airstrikes as "cowardly."
He added that he can count on the support of millions of Libyans.
Gaddafi's status was the source of much debate Friday, with Western officials openly speculating that he was injured and had even fled Tripoli, even as Libyan government spokesmen refuted all of those claims.
A denial by Libyan government spokesman Mousa Ibrahim came after comments made by the Catholic bishop in Tripoli suggested that the Libyan leader had fled and was injured.
Italian Foreign Minister Franco Frattini said Friday that the bishop's statements were credible, but that Italy could not verify Gaddafi's status.
Several analysts have noted that, ever since a NATO airstrike on a building in Tripoli about two weeks ago killed one of his sons and three of his grandchildren, that it was unlikely that anyone escaped from the building uninjured, given the extensive damage.
Military spokesman for NATO's Libya mission, Mike Bracken, said Friday that the mission's goal is to protect civilians, not target Gaddafi or "destroy" his military.
However, the Libyan government, for its part, said that 16 Muslim scholars were killed when a NATO airstrike hit a government-owned guest house in the oil-port town of Brega. Rebels have been trying for weeks to retake the eastern town, which is under the control of Gaddafi's forces.
NATO has conducted 2,512 strike sorties on Libya since it assumed control of operations late March, delivering five ships of humanitarian aid on Thursday to eastern cities where the opposition is in control.
The opposition estimates that over 12,000 people have been killed since mid-February, when the uprising to oust Gaddafi from power began.
Airstrikes on Tripoli and Gaddafi's hometown of Sirte have taken out a "significant part of the Gaddafi war machine", said Bracken.
He warned that the Western military alliance will hit government troops hard if they are about to attack civilians.
For days, NATO sorties have struck at targets in Tripoli. Residents reported hearing loud explosions throughout the week at Gaddafi's Bab Azaziya compound, where he resides.
The Libyan government characterized the April airstrike that killed Gaddafi's son as an assassination attempt directed at the embattled leader.
Russian Foreign Minister Sergei Lavrov sharply criticized NATO operations in Libya, saying Friday alliance aircraft were pounding civilian targets in violation of United Nations rules.
"There are far too many violations of the resolution of the United Nations Security Council," Larvov told Russian reporters during a visit to Kazakhstan. "These things are unacceptable."
Meanwhile, the head of Libya's rebel Interim Transitional National Council (ITNC) is scheduled to meet officials at the White House on Friday.
Mahmoud Jibril will meet US President Barack Obama's national security adviser, Tom Donilon, and other top officials, but it was not made public whether he will also meet the president.
Rebels in the north-western city of Misurata, the country's third largest, claimed victory this week after successfully pushing back Gaddafi's forces from the city's airport.
However, the opposition remains concerned that Gaddafi's troops may return with more Grad rockets, mortars and cluster ammunitions similar to those used in previous weeks, which can reach the city from its outskirts.
But NATO's Bracken said that there were "no reports at all about shelling" recently by pro-Gaddafi forces in the previously under- siege city of Misurata.
"Twenty-five days ago, there were tanks in the streets of Misurata," he told reporters at NATO's maritime headquarters in Naples, Italy. "NATO has done its job well."
"There is definitely no stalemate," he added. "We see real progress in our mission."
Earlier in the week, the Free Libyan flag was raised briefly on several schools and bridges in Tripoli, but the city remains largely under Gaddafi's control.