Traditionally, a routine blood culture for adult patients consisted of paired aerobic and anaerobic bottles, but the routine use of an anaerobic blood culture bottle has been challenged in recent years. In this study, we compared the recovery of two FAN aerobic bottles with one FAN aerobic and one FAN anaerobic bottle. Each pair of bottles was collected by a separate collection procedure, and each bottle held a recommended 8-to 12-ml draw. A total of 704 clinically significant isolates were recovered from 8,620 sets (17,240 pairs), with 487 (69.2%) isolates recovered from one or both bottles in each pair of bottles, 86 isolates (12.2%) recovered only from the FAN aerobic-FAN aerobic pair, and 131 isolates (18.6%) recovered only from the FAN aerobic-FAN anaerobic pair. Significantly more total organisms (P ؍ 0.002), gram-positive cocci (P ؍ 0.03), Staphylococcus aureus (P ؍ 0.05), Enterobacteriaceae other than Escherichia coli (P ؍ 0.02), and anaerobes (P ؍ 0.01) were recovered from the FAN aerobic-FAN anaerobic pair than from the FAN aerobic-FAN aerobic pair. A separate analysis was performed on the 618 isolates that were recovered from the FAN aerobic-FAN anaerobic pair to compare recovery by bottle type. Significantly more S. aureus (P ؍ 0.005) and anaerobes (P < 0.001) were recovered from the FAN anaerobic bottle, while significantly more coagulase-negative staphylococci (P ؍ 0.01), Streptococcus pneumoniae (P ؍ 0.03), and other gram-negative bacilli (P ؍ 0.004) were recovered from the FAN aerobic bottle. These results support the routine use of a FAN anaerobic bottle for use in the culture of blood with the BacT/ALERT system in our institution. These results also suggest that the decision of whether to routinely utilize an anaerobic blood culture bottle should be influenced by the overall recovery of bacteria and yeast, the recovery of specific types of bacteria or yeast, the medium type, and the blood culture system utilized by the laboratory.Traditionally, in most clinical microbiology laboratories in the United States, a routine blood culture for adults has consisted of paired aerobic and anaerobic blood culture bottles. Depending upon the methodology and bottle type employed, the volume of blood placed into each bottle has generally ranged from 3 to 10 ml for adult patients.Beginning with a report by Sharp (11), a number of publications have challenged the need for the use of a routine anaerobic blood culture bottle, essentially asking whether the inoculation of the same volume of blood into two aerobic blood culture bottles, at least on a selective basis, would be more clinically beneficial than the use of the more traditionally employed aerobic-anaerobic pair (1,2,8,9,12).Implicit in the argument to restrict the use of anaerobic blood culture bottles is the premise that were the same amount of blood inoculated into a second aerobic bottle rather than an anaerobic bottle, the overall yield would be greater with two aerobic bottles than with the aerobic-anaerobic pair. The purpose of th...