Yes and no. Most 3rd party mods will work right away but then some that are more involved (like the cargo mod for example) require a new game.
Likewise for new updates and patches to the game itself. For example, in the recent 16th Dec patch, I saw the implementation of the improved error messages when building roads / rail right away but I didn’t get other features like the new auto-replace feature until I started a new game.
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