Hundreds of natives, some wearing colorful dress and banging drums, blocked an access road leading to the Ambassador Bridge, according to the Globe and Mail, slowing traffic on a major trade artery that connects Windsor, Ontario, with Detroit, Michigan.
Thousands of commercial trucks cross the bridge daily, carrying approximately 25 percent of the goods traded between the two countries, which form the world's largest trading partnership.
Under the banner of "Idle No More," native groups promised to hold a series of protests to draw Ottawa's attention to poor living conditions and high jobless rates facing many of Canada's 1.2 million natives.
A native protest shut down a rail line that carries passenger and freight traffic west of Winnipeg in the western province of Manitoba, according to a spokesman for Canadian National Railway Co.
Native groups complain that Canada has ignored treaties signed with British settlers and explorers that they say granted native peoples significant rights over their territory.
Canadian Prime Minister Stephen Harper agreed to pay more attention to the demands of First Nations groups in a meeting last week with aboriginal groups.
Ottawa spends about C$11 billion ($11.1 billion) a year on its aboriginal population, but living conditions for many are poor and some reserves have high rates of poverty, addiction, joblessness and suicide.
Native leaders also want Ottawa to rescind parts of recent budget legislation that they say reduces environmental protection for lakes and rivers.
Canadian Finance Minister Jim Flaherty said on Wednesday it would not be "desirable" if the protest damaged the economy.
"This is not a time to have even more challenges to the Canadian economy," he told reporters on Wednesday.
(Reporting By Russ Blinch, editing by Stacey Joyce)