The province says hunting is closed in areas north of Pine Falls and south of Bissett.
Other areas in Game Hunting Area 26, which stretches from Lake Winnipeg to the Manitoba-Ontario boundary, were shut down last year.
Officials say the region's moose population has declined by almost half since 2006 due to disease and predation by wolves and bears.
The ban also applies to aboriginal hunters.
Last March, the province announced $800,000 for a population recovery plan to restore moose numbers to sustainable levels.