Aerobic plus f

Thirteen different blood culture bottle types (i.e., broth) from three manufacturers (Becton, Dickinson, bioMérieux, Inc., and Thermo Fisher Scientific) were used. Clinical sample inclusion criteria were defined as residual inoculated blood culture bottles, which were flagged as positive in an automated continuous monitoring blood culture system with a readable Gram stain result exhibiting Gram-negative organisms. Samples were tested when prospective (fresh or frozen), within the time frame specified by the manufacturer (10 ), or retrospective (frozen blood culture broth samples from trial sites). Contrived samples were prepared in BD Bactec blood culture bottles (Plus Aerobic/F, Plus Anaerobic/F, Lytic/10 Anaerobic/F, or Peds Plus/F) as previously described (11 (link)– (link)14 (link)). For the prospective collection, exclusion criteria were defined as any fresh broth from positive blood culture bottle(s) from a patient that was already tested. (In the retrospective arm, two bottles from the same patient may have been tested if different types flagged positive [i.e., one anaerobic and one aerobic]). Bottles containing charcoal, and samples with insufficient volume to complete required testing were also excluded.

Blood specimens and non-centrifuged peritoneal fluids obtained during surgery (one specimen/patient) were directly inoculated into two bottles of BACTEC media (PLUS aerobic/F and PLUS anaerobic/F; BD Diagnostics, Sparks, MD, USA), which were incubated for a maximum of 7 days, and analyzed using the automated continuous blood culture monitoring BACTEC FX system (BD Diagnostics). The broth was then sub-cultured on Sabouraud Dextrose Agar with Chloramphenicol (SC) when yeast growth was observed. For colonization purposes, pharyngeal and anal exudates were inoculated in Brain Heart Infusion Broth incubated at 37 °C for 24 or 48 h, and then sub-cultured on SC. Urine specimens were directly streaked onto SC. Cultured yeasts were identified based on the macro-microscopic features of the culture, germ tube test, and biochemical tests (VITEK^® 2 system bioMérieux, Inc. Hazelwood, MO, USA or API^®/ID 32C bioMérieux, Marcy l’Etoile, France).
Antifungal susceptibility tests were performed using Sensititre Yeast One^® (TREK Diagnostic Systems Ltd.). The minimum inhibitory concentration (MIC) results for all antifungals, except for amphotericin B, were interpreted by taking into account the species-specific clinical breakpoint suggested by the CLSI [9 ]. In accordance with the literature data, a breakpoint ≤1 μg/ml was selected for amphotericin B to define the isolates as sensitive (S).