2010) These in vitro studies also support the notion that cultur

2010). These in vitro studies also support the notion that culture of biofilm bacteria may reflect false negative results and should not be used as a stand-alone determination of the absence of a BAI. Taken together, the problem of in situ measurement of cell viability in biofilms is not unambiguous. FISH demonstrates ribosomes of cells, and fluorescence signal intensity is well correlated with ribosome content in most species, indicating recent metabolic activity (Poulsen et al., 1993; Kemp et al., 1993). However, it is also not proof of

viability. Linking FISH detection of active metabolism through visualization of mRNA (Hodson et al., 1995; Wagner et al., 1998; Schmid et al., 2001) or the 16S-23S internal transcribed spacer

(Schmid et al., 2001) would better indicate active microbial transcription. However, these techniques have not yet been routinely applied to clinical samples. Finally, it is Sorafenib molecular weight important to note that not all BAI are culture negative. Rather, culture-negative results do not necessarily rule out an infectious etiology, and more tests may be needed to eliminate this possibility. In addition, not every culture-negative infection is because of biofilms, because infection may be due to fastidious or yet uncultured microorganisms, like Tropheryma whipplei, Borrelia, or Treponema pallidum. Therefore, in addition to culture-negative results being due to ITF2357 manufacturer inadequate sampling, the failure of laboratory culture to detect microorganisms may reflect inadequate incubation times, oxygen conditions, or insufficient nutrient composition in culture media to simulate the complex conditions of growth within the host for fastidious organisms (Moter et al., 2010; Brook, 2011). However, in a clinical setting, the most likely explanation for culture-negative results may be that antibiotics have been used prior to Cyclic nucleotide phosphodiesterase sampling fluids, such as effusions, blood, or synovial fluid, which may be culture negative because planktonic

cells in the fluid have been killed. In support of this, differential detection rates comparing pre- and post-antibiotic samples indicate that recovery of bacteria is reduced by 24% and 36% for staphylococci and streptococci, respectively (Grace et al., 2001). It is also possible that culture is not accurate in polymicrobial biofilms, because the growth of some microorganisms may depend on the presence of metabolites of others within the localized microbial community. While this has been demonstrated in dental biofilms (Moter et al., 1998; Brook, 2011; Marsh et al., 2011), it remains to be shown for infections with more limited species diversity. A common theme among BAI is that the absence of culture results has lead to an alternative explanation for the recurrent inflammatory signs and symptoms independent of an infectious agent. Therefore, the sixth criterion is important.

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