England slipped to their first back-to-back defeats in a decade after losing 31-22 to Canada in the WXV Global Series.
Maddie Feaunti gave the Red Roses a promising start in Ottawa with a try in the opening 10 minutes, converted by Zoe Harrison.
But the visitors were unable to avenge last week's 24-21 defeat in Toronto, which brought their four-year unbeaten run to an end.
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England overturned a half-time deficit to briefly take a 17-12 lead after the break.
Amy Cokayne was among the try scorers, having missed three home matches last month with a calf injury before returning against Canada last week.
But Canada reclaimed the advantage minutes later before extending their lead to 31-17, with captain Sophie de Goede completing a hat-trick as the hosts pulled clear.
Emma Sing crossed for England in the game's dying minutes to reduce the deficit, but it was ultimately too little too late.
De Goede rounded out her stellar performance with three successful conversions, while Harrison kicked the only penalty goal of the match in the 42nd minute.
England wrap up their WXV Global Series campaign with a match against the USA in New Jersey on Friday.
WXV Global Series: England's schedule and results
- England 48-15 Australia
- England 26-26 Canada
- England 26-19 New Zealand
- Canada 24-21 England
- Canada 31-22 England
- October 30 - USA v England (New Jersey)