In order to avoid the influence of pre-analytical steps, the following study was performed by using sterile blood spiked with defined loads of microorganisms as inoculum. Time-to-Detection (TTD) was evaluated for the most frequently encountered bacteria and fungi in septicemia, comparing two commercially available blood culture systems, BACTEC 9240 (Becton Dickinson) and BacT/ALERT (Organon Teknika). A specific medium, Bactec Mycosis IC/F (Becton Dickinson), was compared with the Bactec Plus Aerobic (Becton Dickinson) and FAN Aerobic (Organon Teknika) media for recovery of fungi in general and in case of mixed bacterial/fungal septicemia. The results show that the BACTEC system detects nearly all enrolled microorganisms significantly faster than the BacT/ALERT; the anaerobic vial contributes to the detection of anaerobes and facultative anaerobes and, in the case of BACTEC, shortens TTD; the Bactec Mycosis IC/F bottle shortens TTD of fungi.