ABSTRACT In the present study, we followed the CLSI procedure M40-A to evaluate three specimen transport systems [the new BD CultureSwab MaxV(+), the new Remel BactiSwab, and the Medical Wire & Equipment Transwab] for the survival of fastidious and nonfastidious organisms for 0, 6, 24, and 48 h at room temperature. BD CultureSwab MaxV(+) outperformed the other two swabs for the recovery of the three fastidious organisms, Haemophilus influenzae , Neisseria gonorrhoeae , and Neisseria meningitidis for up to 48 h. Indeed, BD CultureSwab MaxV(+) maintained a constant number of viable H. influenzae and N. meningitidis for up to 48 h, and only a 2 log reduction was noted for N. gonorrhoeae , fulfilling the requirements of M40-A guidelines. However, unlike Remel BactiSwab and the Medical Wire & Equipment Transwab, which fulfilled the M40-A requirements for maintaining the viability of Streptococcus pneumoniae , BD CultureSwab MaxV(+) could not maintain the viability of S. pneumoniae reference or clinical strains past 6 h. Excellent overall sensitivity (98%) (95% confidence interval, 89.5 to 99.7) was observed when the BD CultureSwab MaxV(+) rectal swabs were compared to the “gold standard” stool cultures. Thus, the BD CultureSwab MaxV(+) rectal swab can be used when investigating gastrointestinal bacterial outbreaks or when health care providers face difficulties in obtaining stool samples, particularly from children.