The detection of anaerobic bacteria from blood cultures may be challenging. BACTEC Anaerobic Lytic/F bottles (BALB) were developed to optimize the detection of obligate and facultative anaerobic organisms. This study compared the performance of BALB to resin-supplemented BACTEC Anaerobic/F bottles (BAB). The time-to-detection (TTD) and detection rate (DR) of BALB and BAB were compared in two study stages: first, a laboratory evaluation, including bottles spiked with human blood and 20 facultative and obligate anaerobic bacterial strains; and second, data from patients in the emergency room over two one-year periods: 2015-2016 with BAB and 2017-2018 with BALB. A total of 160 bottles (80 of each type) were included in the first part of the study. The DR of all species in BALB was higher than that in BAB (92.25% vs. 82.50%). The TTD was shorter in BALB than in BAB by 18.9 and 1.4 h for obligate and facultative anaerobic organisms, respectively. Data from patients in two one-year periods showed no significant differences in the numbers of positive anaerobic bottles growing any bacteria (9.29% and 9.52% with BAB and BALB, respectively, p > 0.05). However, the growth of obligate anaerobic bacteria was higher with BALB than with BAB (0.73% vs 0.46%, p = 0.018). The performance of BALB in terms of DR and TTD was significantly superior to BAB for obligate anaerobic species, suggesting that the use of these bottles can improve the detection of these bacteria from blood samples.