The energy ministry did not report the size of the new spill nor say whether it extended to the Coca River, where the country's main hydroelectric plant is located.
In a statement, the ministry said "a leak of oil derivatives" had happened in the eastern province of Napo. It attributed the leak to sabotage.
On Tuesday, he country's state-owned oil company declared an emergency over the earlier spill in the northwest, which polluted several rivers and left hundreds of thousands without clean drinking water.
Authorities also denounced that spill as being an act of sabotage.
The energy ministry said a decision would be made whether or not to take the nearby Coca Codo Sinclair hydroelectric power plant -- which provides 1,100 MW to the national grid -- offline to protect its machinery.
The ministry said a plant shutdown would not lead to power cuts, as supplies would be "guaranteed" to the entire country.
Ecuador faced blackouts of up to 14 hours a day in 2024 due to the worst drought in 60 years, which reduced the level of hydroelectric reservoirs to historic lows.
The earlier spill on March 13 was believed to have been caused when a landslide ruptured a major pipeline, releasing tens of thousands of barrels of oil.
The crude has since spread to at least five waterways, including the Esmeraldas River that flows into the Pacific Ocean, affecting clean water supplies for hundreds of thousands of people.
Energy Minister Ines Manzano has denounced alleged sabotage targeting oil infrastructure.
President Daniel Noboa, in power since November 2023, faces leftist opposition leader Luisa Gonzalez in a presidential run-off election on April 13.
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