Police intercepted the vehicle on Daund-Ahmednagar road and arrested its owners Wahid Sheikh and Raju Fannubhai Sheikh last afternoon.
A police official said one Shivshankar Rajendra Swami of Pune, and a team of 11 cow vigilantes had come to the police station where the case was registered against the tempo owners and others under the Maharashtra Animal Preservation Act.
While the case was registered in presence of the Sheikhs, Swami and others told the police that they should act tough against those illegally transporting cows.
After the activists stepped out of the police station in the evening, they were attacked by a mob of around 50 people, some of them armed with butcher knives and rods.
Police have registered an FIR against 36 people in connection with the attack under various sections of the IPC, including attempt to murder, and under Arms Act, Pawar said, adding the injured 'gau rakshaks' left for Pune after the incident.
Nobody has been arrested so far in connection with the attack.
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