Australia suffered another Women's World Cup blow on Wednesday when striker Mary Fowler – who replaced missing Sam Kerr in their opening match – was also sidelined with injury.
The Manchester City forward Fowler suffered a mild concussion in training and is out of the game with Nigeria on Thursday in Brisbane.
Chelsea superstar Kerr will also again be missing with a calf injury that kept her out of the co-hosts' opening match, a 1-0 win over Ireland.
The absence of skipper Kerr and Fowler, who scored in the 1-0 friendly win over France on the eve of the World Cup, leaves the Matildas with Arsenal's Caitlin Foord as their only recognised striker.
Defender Aivi Luik, who was a substitute against Ireland, was also ruled out of the Nigeria game.
"Forward Mary Fowler and defender Aivi Luik will be unavailable for Thursday night's clash with Nigeria," the co-hosts said.
"Both players have been ruled out after sustaining mild concussions in separate incidents at training on Tuesday."
Kerr has been ruled out of at least Australia's first two games but they will be desperate to have her back for their final Group B match, against Canada on July 31 in Melbourne.
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