The remaining pair are a father and daughter, and they were born from the same mother, making them half-siblings and highlighting the severe inbreeding that has decimated the island's wolf pack.
Wolves came to the island more than five decades ago by crossing a frozen Lake Superior in winter.
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A trail cam placed by Michigan Technological University wolf researcher Rolf Peterson last month spotted a male and female wolf moving down a moose trail on the Lake Superior island, which also is a national park.
It bolsters the finding in last winter's annual Isle Royale wolf survey of two surviving wolves.
Eventually, the excess water starts to return seaward through low areas in the sandbar, "ripping" an opening.