The Dutch-based non-profit organization said 705 people still remain in contention and will be subject to an interview by the Mars One selection committee.
The Mars One project, an ambitious plan to establish a permanent human settlement on the planet beginning in 2025, is the brainchild of Dutch entrepreneur Bas Landorp.
The $6 billion project calls for the use of existing technology and will be funded through sponsors and private investors.
The organization has said that primary funding will come from a “global media event” that will document the astronauts and their preparation.
The 54 Canadians remaining are divided equally between male and female candidates.
Christy Foley, a 33-year-old Alberta government employee, is among the candidates who'll get an interview.
Foley said Monday she's not having any second thoughts about the one-way trip and is still very excited.
She said her family doctor appeared surprised when she explained the reason for a complete medical.
"He said he never had this type of request before, so I guess Edmonton doesn't produce many astronaut wannabes," Foley said.
She and her husband had both applied when the call went out, but he was not chosen.
Initially, Mars One had an applicant pool of more than 200,000 from 100 countries, with 8,243 Canadians applying.
Last December, the organization reduced that number to 1,058 candidates, including 75 Canadians, who were asked to provide medical and personal information.
One-third of those potential Mars settlers were eliminated, leaving the current crop of 705 candidates from around the world.
The majority of the remaining 54 Canadian hopefuls are from the Toronto region, but there's also a large contingent from British Columbia. Alberta and Quebec are next on the list, but other provinces also boast candidates.
Andrew Rader, another Canadian who advanced to the interviews, said being excluded for health reasons would have been surprising.
Rader, 35, said the detailed exam was similar to one he underwent when he applied unsuccessfully to become a member of the Canadian Astronaut Corps in 2009.
"They sent me to a military hospital and it was kind of the same thing," he said. "The medical went well."
After the interview round, candidates will be narrowed down to several international teams consisting of two women and two men, who will take part in an extensive training program.
The current plan is for a crew of four to depart every two years starting in 2024, with the first groups arriving in 2025.
On its website, the Mars One group says the first four settlers would be followed by more groups, every two years.
At first, the home base would be limited to provisions, oxygen and water, but would eventually expand to everything the settlers might need, including solar panels.
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