Non-contact plaque removal offers advantages in interproximal spaces, fissures, and pockets. It requires the generation of strong fluid flows and the inclusion of air bubbles to become effective. A pair of co-adhering streptococci and actinomyces has been used previously to demonstrate non-contact removal by sonic brushing. Here we determined the influence of the sequence of co-adhesion of streptococci and actinomyces on non-contact removal from a salivary pellicle by rotary and sonic brushing. After bacterial adhesion, pellicles were brushed in a wet and immersed state, with a distance up to 4 mm to the bristle tips. Bacteria adhering to pellicles from the sequence streptococci followed by actinomyces appeared more difficult to remove and left more large co-aggregates than from the sequence actinomyces followed by streptococci. At contact, rotary and sonic brushing performed equally well in bacterial removal, while at 4 mm, both had lost some efficacy.