I was probably confused about the situation. Somehow I assumed that these fireflies are a part of the older assets, created years ago, and that you have to cut them out from those old assets. Are these on contrary new assets created right now by an artist?
As for whether to have these animations on a transparent surface, or having them with parts of background, I suppose that depends also on artistic features. For example, if the fireflies cast light on some object in background (such as trees), then the animation frames should also include trees (or parts of trees).
If that's not a issue, then of course it may be better for an artist to create fireflies on a transparent surface, separate from room background, and save them as 32-bit ARGB images to let keep half-transparent pixels.
But then, before that, we'd also have to clarify this: what is your game's color depth? is your game actually 32-bit, or lower (16-bit, 8-bit)?
As for whether to have these animations on a transparent surface, or having them with parts of background, I suppose that depends also on artistic features. For example, if the fireflies cast light on some object in background (such as trees), then the animation frames should also include trees (or parts of trees).
If that's not a issue, then of course it may be better for an artist to create fireflies on a transparent surface, separate from room background, and save them as 32-bit ARGB images to let keep half-transparent pixels.
But then, before that, we'd also have to clarify this: what is your game's color depth? is your game actually 32-bit, or lower (16-bit, 8-bit)?