, 2008), even in culture-negative cases. Stoodleyet al. (2008) have also published GDC 0449 confocal micrographs showing the consistent presence of biofilms of live coccoid bacterial cells (using Molecular Probes Live/Dead BacLite Kit) in an infected elbow case (Fig. 1) that yielded negative cultures over a period of 5 years,

during which the clinical state of the patient necessitated several serious replacement procedures. The confocal data were supported by positive reverse transcriptase-PCR results for bacterial mRNA for Staphylococcus aureus. The orthopedic problem that offers the most dramatic contrast between culture data and modern molecular methods of diagnosis is the tragic problem of the Sulzer acetabular cup. When a critical nitric acid washing step was INK 128 solubility dmso omitted from the manufacturing process for this device, the microbial biofilms accreted during manufacture were retained and, even though ethylene oxide sterilization killed the sessile bacteria, the residual polysaccharides of the matrix increased the colonization potential of these devices. Approximately 1500 cases of ‘aseptic loosening’ resulted, and this designation was made because the culture results were consistently negative

for both aspirates and interoperative specimens (Effenbergeret al., 2004). We have examined a subset of eight of these ‘aseptic loosenings’ and, in each case, we have found direct evidence of the presence of bacteria on explants at the time of revision. Figure 2 shows unequivocal evidence of the presence of coccoid bacterial cells on the surface of a culture-negative Sulzer acetabular cup explanted from a case of so-called ‘aseptic loosening.’ These cells were seen to form slime-enclosed biofilm microcolonies on the plastic surface. When these acetabular cups were reacted with species-specific FISH probes for Staphylococcus epidermidis, the bacterial cells showed fluorescence (Fig. 2, inset), and the cells were seen to be growing in coherent biofilms. Because the detection of bacteria like S. aureus is pivotal in many clinical decisions in orthopedic surgery, and because the presence of methicillin-resistant

S. aureus (MRSA) can pose intractable problems, it may be valuable to address the culture of the biofilm phenotype of however this organism. Extensive studies of the distribution of S. aureus in the human female reproduction tract were triggered by the threat of toxic shock, caused by the secretion of the TSST1 toxin produced by this organism; hence, we explored their detection and characterization using culture methods and new molecular techniques (Veehet al., 2003). In a survey of 3000 healthy volunteers, using very careful culture techniques in which vaginal swabs were carried to the lab at body temperature and fresh moist plates were used, positive cultures were obtained from 10.8% of these women. This percentage was slightly higher than that found in several previous studies (Wiseet al.