Phase variation of slime production in Staphylococcus aureus: implications in colonization and virulence

Two methods commonly used for slime detection in coagulase-negative staphylococci (tube biofilm formation and colony morphology in Congo red agar) were used to study 144 ruminant mastitis Staphylococcus aureus strains. Slime production was detected in 21 strains. A majority of cells (85%) in slime-producing (SP) strains and a minority of cells (5%) in non-slime-producing (NSP) strains showed a condensed exopolysaccharide matrix (slime) surrounding the bacterial cell wall, as revealed by electron microscopy and immunofluorescence. In vivo slime production was also detected immunohistochemically after experimental infection of the mammary gland in sheep. Upon repeated subcultures in Congo red agar, NSP variants were obtained from four ovine and four bovine SP strains at a frequency ranging from 0.5 x 10(-4) to 10(-4). Because SP variants could not be obtained from NSP strains within this range or at a higher frequency, they were obtained by the tube biofilm formation (requiring repeated subculturing of NSP strains in tryptic soy broth containing 2% glucose for subsequent recovery of colonies adherent to the walls of the culture tubes). In experimental challenge, the SP variant showed a significantly higher colonization capacity than did the NSP variant of the same strain used (P < 0.001). However, the NSP variant had a higher virulence than did the SP variant (P < 0.001). These results may help to explain the different roles of S. aureus slime production cell types (SP and NSP) coexisting in disease.