Sacred bull Shambo, who had attracted international attention since his diagnosis with tuberculosis and an intense campaign by Hindu groups to save him from slaughter, was put down with a lethal injection after a three-month-long legal battle to save him.
"Shambo was been put down by lethal injection on Thursday night," a spokesman of the Wales Assembly government said.
The six-year-old Friesian, who tested positive for bovine TB, was moved from the Skanda Vale Temple enclosure on Thursday night amid protests by devotees.
Police were called in to move more than 100 worshippers who formed a human shield around the sacred bull.
Keith Porteous Wood, executive director of the National Security Society, said he was glad "common sense had prevailed at last" and that it was "absolutely unacceptable" for people to say their religious rights were supreme.
Hindu leaders are now seeking a meeting with the UK environment minister.
Secretary general of the Hindu Forum of Britain Ramesh Kallidai said he wanted to check "how agricultural law can cater to the needs of sacred animals in Hindu temples in Britain."
Skanda Vale community leader Brother Alex said that, now Shambo had been slaughtered, a "nightmare" was just beginning for the Welsh Assembly.
"Ignorant people have chosen to desecrate our temple and have chosen to destroy life unnecessarily," he said.
The slaughter of Shambo ended a protracted legal battle which started when a TB test returned positive in April and came after the community finally lost its high court fight on Monday.
Last week, a high court judge gave the community hope when he ruled that two slaughter orders for Shambo "were unlawful and will be quashed."
But his ruling was overturned by the appeal court in London on Monday.