The division bench of Justices BhushanDharmadhikari and Swapna Joshi passed the order today.
Tigress 'T-27-CUB-1' was caught from the Brahmapuri forest in Chandrapur district in July after it killed two people and injured four others. It was released into Bor tiger sanctuary in the Wardha district.
The high court, in response to a PIL, had earlier quashed the forest department's June 23 order to shoot the tigress.
However, after being released in Bor sanctuary, the tigress killed two more persons and injured another.
The forest department again issued an order to shoot the tigress on October 4.
Wildlife enthusiast Dr Jerryl Banait had filed a PIL opposing the order to kill the animal, saying the evidence which formed the basis for forest department's decision to shoot her was insufficient.
The high court had asked the department to provide evidence that the tigress posed danger to human life as per the National Tiger Conservation Authority (NTCA) norms.
Forest department's lawyer Kartik Shukul told PTI that the department provided evidence including her GPS locations, photographs, spot reports and autopsy reports of human kills to show that she frequently stalked and attacked humans.
The tigress had become a habitual man-eater, stated the forest department's order.
The judges today upheld the latest forest department order to shoot the big cat, issued under the Wildlife Protection Act.
The petitioner had said that in the absence of DNA reports, it could not be said that the same tigress had killed four people.
Petitioner Jerryl Banait said he was planning to the move the Supreme Court against the high court ruling.
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