The cap on gaming machines in community venues has been reduced from 1699 to 1659, excluding machines at the NT's two casinos which are covered by separate agreements.
Gaming Minister Chansey Paech said since reintroducing the cap when Labor came to power in 2016, the government had lowered it three times, cutting about 200 machines from the pool.
"Our government is aware that problem gambling creates a significant public health risk to communities across the NT, and that's why we are doing what we can to reduce availability of gaming machines," the minister said.
"We continue to promote responsible gambling and provide ongoing funding for research as well as community-based support services."
Tuesday's changes also end a nine-month moratorium on new gaming machines in Alice Springs that was sparked by community concern over plans for 60 more at hotels and taverns in the town.
Applications for 40 of those machines in two venues have since been withdrawn.
Existing applications lodged last year for 20 machines at the Todd Tavern and Gap View Hotel remain in train, however no new applications from community venues across the Territory will be accepted once the cap is reached.
In regard to the Mindil Beach Casino in Darwin and the Lasseters Hotel Casino in Alice Springs, the government said they would need to adhere to a new code of practice that prioritised harm minimisation.
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