England were stunned by a late charge from Canada as the Red Roses lost their 41-match unbeaten run that stretched back four years.
Last month, Canada snapped England’s 39-game winning streak with a 26-26 draw against the World Cup champions in Exeter.
And Canada finally earned revenge for their defeat to England in last year’s World Cup final at Twickenham as they won 24-21 in front of a record crowd in Toronto.
Shoshanah Seumanutafa came off the bench to score the match-winning try in the 77th minute, bringing England’s dominant run to an end.
The Red Roses had not lost a match since their previous World Cup final against New Zealand in 2022, a loss that started their unbeaten run to becoming world champions.
England’s head coach John Mitchell said he was focused on taking what positives he could out of the defeat and the Red Roses will play Canada again next Saturday in Ottawa.
It will be the second encounter of the two-game Rugby Rematch Tour between the growing rivals and Mitchell said the defeat will allow him to learn even more about his group.
“We’re not driven by outcome,” Mitchell said. “We’re disappointed by the result but there was some very good things in our performance tonight. So we’ll stick to our processes.
“I guess (the loss) frees us up and allows us all to find more. I know this group has got more in them but sometimes it requires the context that we’ve just received to dig deeper and find something more.
“Maybe everyone in the group can probably review their ‘why’s and re-establish those as well because I’m sure they’re changing and evolving as they experience what we’re going through.”
Canada looked up for the challenge early as they opened the scoring through Savannah Bauder in the 15th minute.
Quickfire tries to Maddie Feaunati and Zoe Harrison just before half-time put England ahead 14-5 at the break.
The points kept flowing as both teams crossed just after play resumed, but DaLeaka Menin’s 63rd-minute try kept the door open for Canada.
A raucous home crowd exploded for Shoshanah Seumanutafa’s match-winner shortly after, with Canada able to hold on from there and quell England’s historic run.
The crowd of 18,621 set a record for a women’s rugby union match in North America and Red Roses captain Meg Jones said England came away taking “huge pride” in their unbeaten run.
“We're the biggest advocates for the game," Jones said. “So we're always going to try and grow the game as much as we can, and Canada definitely did that in tenfold today.”
Includes reporting from agencies