Do I really need to specify which one is Covariates (control for)?

Hi @APeh - if you include both Location and Factors A, B, and C in the model as fixed effects, you are already adjusting for the environmental effect. You can subset the final MaAslin2 results table to the
main effects of interest and re-compute the q-values to detect significant microbiome features as described here.

Hope this helps,
Himel