Api id 32c

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).

Substrate utilization test API ID 32C (BioMérieux, Marcy-l’Étoile, France) was used to analyze the biochemical spectrum of all Lachancea fermentati strains. Preparation of inoculum and inoculation of the strips was performed according to the manufacturers’ instructions. Colonies for the inoculum were grown on YPD agar plates for 48 h at 27°C. After inoculation, API ID 32C strips were incubated for 2 days at 28°C. The samples were evaluated visually by turbidity of the wells, differentiating positive (+), negative (-), and weak (w) growth.

Bacterial identification was performed using conventional methods and with the VITEK-2 Compact system (bioMérieux, Marcy l’Etoile, France). Fungi isolated from Myco F or aerobic bottles were identified by morphological examination on cornmeal agar with Tween 80 and API ID 32C (bioMérieux, Marcy l’Etoile, France). Incompatible results were confirmed via MALDI-TOF MS (bioMérieux, Marcy l’Etoile, France).

A 100 mL aliquot of melted ice was filtered and the membrane placed on Sabouraud dextrose agar containing chloramphenicol (0.5 g/L) (Liofilchem, Roseto degli Abruzzi, Italy). After 8 days of incubation at 28 °C, yeast colonies were identified using a semi-automated sugar assimilation system (API ID 32C, Biomerieux, Marcy l’Etoile, France), while filamentous fungi were identified by evaluating their macroscopic and microscopic morphological characteristics according to the methods described elsewhere [18 ].