World No.2 Canada dominated every aspect of Friday afternoon's clash, only the second time Australia's women have graced Suncorp Stadium.
Down 21-0 after 20 minutes, the hosts were lucky not to concede more than one extra try before the break, although there was a response to begin the second stanza.
A strong tackle-busting effort from winger Desiree Miller to score was brief respite though in a dominant Canadian performance.
Australia, ranked sixth in the world, beat world No.8 USA last weekend after a comprehensive loss to No.3 New Zealand to begin the Pacific Four Series.
Canada drew with New Zealand last week and will enter August's World Cup in England, where the hosts top the rankings, confident of bringing the trophy back with them.
In seven World Cups Australia have a sole third-placed finish and otherwise finished no higher than fifth.
"Would have loved the win last weekend but had to move on quickly," Canadian captain Alex Tessier said.
"Very proud; a decent win and put a lot of points on the board.
"The Pac Four plays a huge role (for the World Cup) ... which is coming up quite quickly."
The visitor's forward pack was rampaging in the first half, Australia unable to plug the gaps in defence and hesitant with ball in hand while their line-out struggled to function.
Sevens convert Charlotte Caslick had her moments in her first Test at inside centre, making a strong covering tackle to save a try and darting through the line whenever the opportunity presented.
The Wallaroos' replacement forwards also made an impact, reserve hooker Adiana Talakai unlucky to have a try disallowed after her injection after halftime.
Australia captain and back-rower Siokapesi Palu was another bright spot for the side but admitted her team had work to do with three Tests left before their World Cup opener.
"Canada were really physical, and set piece again was challenging," she said.
"We know they're a quality side and we really need to step up moving forward."
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