From memory, the virtualisation operations themselves aren't nested. The VM instructions interact with the external virtualisation hardware, so it's more of a cooperative situation, e.g. a guest can create & manage virtualisation structures that are run alongside it.

I don't know if this applies to the specific nested virtualisation AWS are providing though.

depends on the workload and how they've done it

pure CPU should be essentially unaffected, if they're not emulating the MMU/page tables in software

the difference in IO ranges from barely measurable to absolutely horrible, depending on their implementation

traps/vmexits have another layer to pass through (and back)

As a practical matter, anywhere from 5-15%.