"We want to send out a clear message to all those who infringe Libyan sovereignty and lack respect for the coastguard and navy," Libyan navy spokesman General Ayoub Qassem told a news conference in Tripoli.
General Abdelhakim Bouhaliya, commander of the Tripoli naval base where the conference was held, said: "No foreign ship has the right to enter" the area without authorisation from the Libyan authorities.
Libya has "officially declared a search and rescue zone", said Bouhaliya, without specifying the scope of the exclusion zone.
He urged humanitarian organisations to "respect our will... And obtain authorisation from the Libyan state even for rescue operations".
Italy has also said it wants to keep a tighter rein on NGOs helping the multinational search and rescue operation by making them sign up to a new code of conduct.
Italian authorities last week impounded a boat operated by German aid organisation Jugend Rettet on suspicion its crew effectively collaborated with people traffickers in a way that facilitated illegal immigration.
The Libyan coastguard has accused NGOs of aiding people traffickers in their lucrative business.
Six years since a revolution that toppled longtime dictator Moamer Kadhafi, Libya has become a key departure point for migrants risking their lives to cross the Mediterranean to Europe.
Italy, the main destination of the people traffickers, has sent naval vessels at the request of Libya's UN-backed Government of National Accord to assist Tripoli, on a mission disputed by rival authorities in eastern Libya.
Over 2,300 have died attempting the crossing.
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