Physiology of biofilms of thermophilic bacilli-potential consequences for cleaning.

Thermophilic Bacillus species readily attached and grew on stainless steel surfaces, forming mature biofilms of >10(6.0) cells/cm2 in 6 h on a surface inoculated with the bacteria. Clean stainless steel exposed only to pasteurized skim milk at 55 degrees C developed a mature biofilm of >10(6.0) cells/cm2 within 18 h. When bacilli were inoculated onto the steel coupons, 18-h biofilms were 30 microm thick. Biofilm growth followed a repeatable pattern, with a reduction in the numbers of bacteria on the surface occurring after 30 h, followed by a recovery. This reduction in numbers was associated with the production of a substance that inhibited the growth of the bacteria. Variations in the environment, including pH and molarity, affected the viability of the cells. Chemicals that attack the polysaccharide matrix of the biofilm were particularly effective in killing and removing cells from the biofilm, demonstrating the importance of polysaccharides in the persistence of these biofilms. Treatment of either the biofilm or a clean stainless steel surface with lysozyme killed biofilm cells and prevented the attachment of any bacteria exposed to the surface. This suggests that lysozyme may have potential as an alternative control method for biofilms of these bacteria.