Total culture extraction was an efficient sampling technique for P. pastoris for NMR analysis, but salts in the media affected the GC-MS analysis.
Total culture extraction of cells + broth using high cell density cultures typical of P. pastoris fermentations, was an efficient sampling technique for NMR analysis and provided a gold standard of intracellular metabolite levels; however, salts in the media affected the GC-MS analysis.
DNA was extracted from bacterial pellets of B. cenocepacia (strain J2315) and P. aeruginosa (strain PAK) with the DNA tissue culture extraction kit (Qiagen) Forward and Reverse primers were designed based on TcPRAC sequence toward specific sequences of the genes of interest (Table S2, Supplemental files).
Nine patients (8.6%) were on treatment with antibiotics at the moment of the blood culture extraction.
An inclusive viability study (n = 226) was undertaken to determine at which point handling of culture extraction materials does not necessitate a CL3 environment.
Immunosuppressed patients, those with confusion, severe sepsis or shock at the time of blood culture extraction had statistically significant lower TTP, as well as those diagnosed with meningitis and those admitted to the ICU (Table 1, Figure 3).
Empirical antimicrobial therapy was judged to be either adequate or inadequate on the basis of the in vitro susceptibility of an isolated organism, and/or the initiation of antibiotic treatment within 24 h of blood culture extraction.
The following data were collected: age; sex; comorbid conditions; dates of hospital admission and discharge; presence of septic shock at the moment of blood culture extraction; specific antimicrobials administered during hospitalization; dates of start and end of antimicrobial administration; surgical procedures and hospitalization in an intensive care unit.
Immunosuppressed patients (n = 5), patients with confusion (n = 19), severe sepsis or shock at the time of blood culture extraction (n = 12), those with a diagnosis of meningitis (n = 7) and those admitted to the ICU (n = 14) had lower TTP.
Cell cultures, protein extraction, SDS-PAGE and trypsin digestion were performed as previously described [ 18, 39].
