The protest comes after the ruling coalition led by the Serbian Progressive Party, which holds an overwhelming majority, rejected an opposition draft law stipulating a ban on mining and exploration for lithium and borates.
Protesters waved Serbian flags and banners reading "You Shall Not Dig" and "Back Away From Jadar", while loudspeakers blared patriotic songs and folk music.
They demanded an end to the project and announced one-hour blockades of two regional roads on Saturday.
"We are preparing warning road blockades in several locations across the country this weekend," Zlatko Kokanovic, a protest leader, told Reuters.
"We will see how we proceed from there."
Lithium, regarded as a critical material by the EU and the U.S., is used in batteries for electric vehicles and mobile devices.
In 2022, the Serbian government revoked licenses for Rio Tinto's $2.4 billion Jadar lithium project near Loznica following mass protests by environmental groups.
If implemented, Rio Tinto's Jadar project could meet 90% of Europe's current lithium needs, positioning the company as one of the world's leading lithium producers.
The environmentalists and a majority of opposition parties argue that the mine could pollute land and water in the area, impacting farming, a major source of income for many in the region.
"There's no retreat or surrender. They will not ravage nature and our lives," said Branka Petrovic, 47, an activist from Belgrade.
Serbian officials argue that the mine will boost the country's economy. In June, Serbia's President Aleksandar Vucic said mining could start as early as 2028.
In July, Rio Tinto said the project would adhere to stringent environmental requirements, including legal and permitting procedures, as well as public consultations before implementation.
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