massiliense numbering). The other group (19 strains) had one different mTOR inhibitor nucleotide at the 190th base (A190G, 466th nucleotide on M. abscessus numbering) from the type strain. Unlike the erm(41) of M. massiliense, those of M. abscessus and M. bolletii were not clearly differentiated by sequence analysis. They showed 18 polymorphic nucleotides and were separated into 12 clusters on the phylogenetic tree (data not shown). The Erm(41) of M. massiliense could

be described as three characteristic regions, the N-end (21 amino acids), the central mutated area (30 amino acids), and the C-end (30 amino acids). Although the Erm(41) produced by M. massiliense was found to be smaller than the Erm produced by other mycobacteria, both the N- and C-terminal ends of M. massiliense corresponded almost exactly to the ends of Erm(41) produced by M. abscessus and M. bolletii. Indeed, only three amino acids differed between the ends of M. massiliense and those of M. abscessus and M. bolletii Cobimetinib chemical structure (Q14/P14 and A16/T16 in the N-end, and T64/A156 in the C-end). However, M. massiliense has 30 amino acids that differ from the other Erm(41) due to the frame-shift mutation in the central mutated region. All C-terminal regions of the Erm(41) in the three species were

truncated in a similar fashion to that of M. tuberculosis Erm(37). The MIC of 35 M. massiliense strains were less than 2 μg/ml, whereas those of five strains were very high (>256 μg/ml). However, the MIC of 37 M. abscessus strains very ranged from 0.06 to 64 μg/ml, and two strains showed very high MIC (>256 μg/ml). These seven highly resistant strains contained a point mutation at the adenine at position 2058 (A2058) or A2059 in the peptidyltransferase region of the 23S rRNA gene. Two M. bolletii isolates showed distinct MIC (0.25 and 16 μg/ml). They did not harbor a point mutation at A2058 or A2059 in 23S rRNA gene, and the former isolate (0.25 μg/ml) had the T28C transition of erm(41). Also, the MIC of one M. chelonae isolate was low,

1 μg/ml (Table 1). In addition, although the end-point of growth inhibition was clear-cut in all of the M. massiliense strains, that of M. abscessus and M. bolletii strains, except for six strains having T28C transition, showed trailing, and the MIC of M. abscessus and M. bolletii increased with prolonged incubation, as reported previously (24). Difference in the clarithromycin susceptibility of M. massiliense and M. abscessus was clearly observed in the present study carried out with extended numbers of clinical isolates. Specifically, M. massiliense isolates were found to be either markedly susceptible (87.5%; MIC, ≤2 μg/ml) or highly resistant (12.5%; MIC, >256 μg/ml), whereas M. abscessus isolates were found to be either susceptible (48.7%; MIC, ≤2 μg/ml), intermediate (10.3%; MIC, 4 μg/ml), or resistant (41.