Ministers scrapped the legislation last month, despite it being part of the Conservatives’ 2019 manifesto.
Rishi Sunak even said during last summer’s Tory leadership election that he “of course” supported the bill, which aimed to clamp down on puppy smuggling and dog theft.
But in the House of Commons on Tuesday evening, MPs voted 256 to 183 against a Labour motion which would have brought the bill back before parliament.
No Tories voted in favour of the move, but 249 voted against.
Jim McMahon, Labour’s shadow environment secretary, said: “By doing this they have just given the green light to puppy smugglers and dog thieves, showing whose side they are on.
“Labour’s motion to bring back this important piece of legislation would have allowed us to end live exports of farm animals and target the criminals involved in puppy smuggling and dog thefts.
“The vote proves once and for all that Labour is the party of animal welfare. The Conservatives have broken their promises to deliver the bill, including the prime minister, who personally committed to bring it into law.”
The government has insisted it can introduce the measures contained in the bill in other pieces of legislation.
But David Bowles of the RSPCA said: “It’s terrible news for animals and we are calling on the UK government to rethink and bring back all the proposals contained within the binned bill back by other means.
“We cannot go on allowing cruel practices such as the live exports of animals for slaughter and the puppy import trade.”