The United States on Wednesday urged "all those wishing to deliver goods to do so through established channels so that their cargo can be inspected and transferred via land crossings into Gaza."
"As we continue to make clear to involved parties, mechanisms exist for the transfer of humanitarian assistance to Gaza," the U.S. State Department said in a statement.
It warned that "direct delivery by sea is neither appropriate nor responsible, and certainly not effective, under the circumstances."
"There is no need for unnecessary confrontations, and we, along with our partners in the (Middle East) Quartet, call on all parties to act responsibly in meeting the needs of the people of Gaza," the statement added.
Transport Minister Ghazi Aridi has given permission to the aid ship "Julia" to leave the northern port city of Tripoli for Cyprus.
Cypriot authorities said a ban on ships sailing from its ports to Gaza still applies, in response to Lebanon's permission.
The ship, carrying humanitarian aid and pro-Palestinian activists from various countries, will reportedly sail to Cyprus in the coming days before heading to Gaza.
Cypriot Foreign Minister Marcos Kyprianou said the ship will not be allowed to proceed to Gaza.
"The prohibition on ships to reach Gaza from Cyprus is still applicable," Kyprianou said commenting on Aridi's statement.
A Cypriot police spokesman said that the police had contacted the Foreign Ministry and would act accordingly.
On Tuesday, Lebanon said it would hold Israel responsible for any attack, in response to Israeli warnings that it would use "all necessary force" to stop aid boats planning to sail to blockaded Gaza.
"Israel will be held fully responsible for any attack on Lebanon," the foreign ministry said in a letter to the United Nations.
Lebanon "cannot prohibit a ship from leaving its ports if its cargo, passengers and destination all comply with Lebanese law," it said in the letter.
Israel's ambassador to the United Nations warned the world body last week that the Jewish state was entitled to use "all necessary force" to stop the Lebanese activists' boats.
On Monday, Israeli Defense Minister Ehud Barak also reiterated a warning that Lebanon would be responsible for any "violent and dangerous confrontation" with vessels sailing to Gaza from its shores.
Israel came under international censure over its May 31 seizure of a six-ship aid fleet bound for Gaza, in which nine Turkish activists were shot dead by Israeli naval commandos.
A Lebanese freighter which tried to deliver aid to Gaza last year was intercepted by Israeli warships.
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