Zoo authorities in the southern state of Hyderabad shared samples with a government research laboratory on March 24 after the lions showed signs of respiratory distress.
The test results come amid a huge surge in coronavirus infections among humans in India.
“Based on experience with zoo animals elsewhere in the world that have experienced SARS-COV2 positive last year, there is no factual evidence that animals can transmit the disease to humans any further,” the Ministry of Environment, Forest and Climate Change said.
The Nehru Zoological Park has now been closed, it said in a statement.
India has reported more than 300,000 daily COVID-19 infections for 13 straight days and has now recorded 20 million cases of the disease, the second highest number in the world after the United States.