For a substrate of sand that is 4mm in grain size, I do not recommend its use for DSB
That grain size is too large to be easily serviced. I am afraid it will collect too much solid waste and detritus in time.
Instead, use 4mm sand for a shallow bed of only 25mm in thickness or less. It can still be useful for supporting the growth of microcrustaceans. But it will not be a significant source of denitrifying bacteria surface areas. At least, it cannot succeed long term that way without a lot of supportive maintenance from you (many gravel siphonings).
For a DSB, I recommend the finest sand only. Under 2mm in grain size (and best if 1mm or smaller).
Ciao Anthony,
many thanks for your reply. Ok, but if I want make a bed for refugium with 4mm of size only for microcrustaceans, copepods, amphipod, etc. habitat, which height? Only 25 mm????
Grazie
yes... only 25 mm or less, my friend. Otherwise you will have the same problems that the American aquarists had in the early years where such coarse substrates accumulate solid waste too quickly and foul.
The problem here is that 4mm is too large for a DSB to work well in the long term.