Incubation in CO 2 resulted in higher (>3 doubling dilution) MICs of telithromycin than those found in ambient air for 31.2% of 346 Streptococcus pneumoniae ermB-positive strains. An increased telithromycin MIC in CO 2 was not correlated with loss of its activity in the murine sepsis/peritonitis model. Incubation in CO 2 has been reported to result in higher MICs of macrolides, clindamycin, and telithromycin for Streptococcus pneumoniae than those found in ambient air (6,9,10,11,15,16). Conversely, susceptibility testing of other antibiotics, namely -lactams, evernimicin, and linezolid, does not appear to be affected by the presence or absence of CO 2 (3,5,7,8). This study reports the influence of CO 2 on the MIC of telithromycin against ermB-positive S. pneumoniae.MICs of erythromycin and telithromycin in ambient air and CO 2 . MICs of erythromycin and telithromycin were determined by using the agar dilution method for 675 clinical strains of S. pneumoniae (13). Plates were incubated for 16 h at 37°C, either in ambient air or in 5% CO 2 . Susceptibility to erythromycin was also assessed by agar diffusion (15 g of erythromycin per disk). MICs of telithromycin were also measured by microdilution according to the NCCLS recommendations (13). The MIC breakpoints of erythromycin were defined as the following: susceptible, Յ0.25 mg/liter; resistant, Ն1 mg/liter (13). The MIC breakpoints of telithromycin were defined according to the European Agency for the Evaluation of Medicinal Products (susceptible, Յ0.5 mg/liter; resistant, Ͼ2 mg/ liter) and according to the proposed NCCLS breakpoints (susceptible, Յ1 mg/liter; resistant, Ն4 mg/liter).Erythromycin MIC in ambient air was Ն0.25 mg/liter for 345 strains. Ten additional strains for which the erythromycin MICs were Յ0.25 mg/liter had an inhibition zone diameter of Ͻ22 mm. These 355 strains were tested for ermB and mefE genes by PCR as previously described (2). Erythromycin resistance genes were distributed as follows: ermB was present in 347 strains, mefE in 4 strains, mefE and ermB in 2 strains, and neither ermB nor mefE in 2 strains. Telithromycin MICs in ambient air (675 strains) were distributed as follows: Յ0.5 mg/liter for 667 strains (98.8%), 1 mg/liter for 4 strains (0.6%), and 2 mg/liter for 4 strains (0.6%). Telithromycin MICs were obtained in CO 2 for 659 strains and were Յ0.5 mg/liter for 591 strains (89.7%), 1 mg/liter for 36 strains (5.5%), and 2 mg/liter for 32 strains (4.8%). According to the European breakpoints, the frequency of strains with intermediate susceptibility to telithromycin was 1.2% in ambient air and 10.3% in CO 2 . These frequencies, according to the NCCLS breakpoints, were 0.6 and 4.9%, respectively.For 346 ermB-positive strains, the telithromycin MIC at which 50% of isolates tested are inhibited (MIC 50 ) in ambient air and in CO 2 was 0.015 and 0.06 mg/liter, and the MIC 90 was 0.25 and 2 mg/liter, respectively (Fig. 1). The proportion of strains for which the telithromycin MIC was Ͼ0.5 mg/liter was 2.3% in ambient air and 21.5% in C...