However, reserve authorities said most of these deaths were due to natural causes or because of territorial fights among the tigers.
The protesters also demanded a high-level probe and immediate removal of reserve officers, accused them of being responsible for the tigers death at Pench.
"The officials of the Reserve who are allegedly responsible for the killing of tigers are now (themselves) conducting probe into the recent death of a tigress and her two cubs, which is suspicious. We demand immediate removal of responsible officers and a high-level investigation," President of Guide, Gypsy and Resort Operators Association Narendra Dube told reporters.
Local villagers too have joined in the protest.
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Dube also alleged that reserve authorities were removing the evidence of the killing of tigress and two cubs from the spot.
The protesters also handed over a memorandum in this regard to Kurai Sub Divisional Magistrate K C Parte.
Omveer Choudhary, a guide, alleged nine tigers were killed in the Reserve in the last eight months, including a most sighted tigress called 'Nala Wali Baghin' and her two cubs which had enraged the locals.
But, the reserve authorities said most of these deaths were due to natural causes or because of territorial fights among the tigers.