Slowly growing, non-chromogenic mycobacteria were isolated from striped barombi mbo cichlids (Stomatepia mariae) maintained at the London Zoo Aquarium, UK. The isolates could be differentiated from other slowly growing, non-pigmented mycobacteria by a combination of phenotypic features including their inability to grow at 37 6C, positive tests for heat-stable catalase, tellurite reduction and arylsulfatase activity, and the absence of urease activity, Tween 80 hydrolysis, nitrate reductase, iron uptake and semiquantitative catalase. The almost full-length 16S rRNA gene sequence, together with partial sequences from the 65 kDa heat-shock protein (hsp65) and the b-subunit of the bacterial RNA polymerase (rpoB) genes and the 16S-23S internal transcribed spacer 1 (ITS 1) region were identical for all three novel strains, but distinct from those of all known mycobacterial species. Phylogenetic analysis based on 16S rRNA gene sequences placed the novel isolates within the slowly growing mycobacteria group in close proximity to Mycobacterium florentinum. Based on genotypic and phenotypic findings, it is proposed that these isolates represent a novel species of the genus Mycobacterium, for which the name Mycobacterium stomatepiae sp. nov. is proposed with strain T11 T (5DSM 45059 T 5CIP 109275 T 5NCIMB 14252 T ) as the type strain.In the past few years, the occurrence of fish mycobacteriosis has been increasingly reported (Kent et al., 2004;Pate et al., 2005;Sakai et al., 2005;Whipps et al., 2007b). Although three species, Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium chelonae, have been frequently cited as the main causative agents of infections in fish, several other mycobacterial species, including a number of novel species, have also been associated with mycobacteriosis (Levi et al., 2003;Rhodes et al., 2003Rhodes et al., , 2005Whipps et al., 2007a).The majority of slowly growing mycobacteria contain a long helix 18 at position 430-500 (Escherichia coli 16S rRNA gene sequence numbering) in their 16S rRNA gene, which is absent in rapidly growing mycobacteria (Springer et al., 1996a;Tortoli, 2003). There is a third group, with a short helix 18 (due to a deletion of 12 bp), and members of this group are also classified as slow growers (Tortoli, 2003). In this group, Mycobacterium simiae, a photochromogenic organism from monkeys, was the first species to be identified (Weiszfeiler et al., 1981). Mycobacterium intermedium (Meier et al., 1993) is another photochromogenic species related to M. simiae. Scotochromogenic species of the M. simiae-related (MSR) group consist of Mycobacterium interjectum (Springer et al., 1993) The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, hsp65 and rpoB gene sequences and ITS 1 region of M. stomatepiae sp. nov. T11 T are AM884331, AM902968, AM885873 and AM902938, respectively.Neighbour-joining phylogenetic trees based on hsp65 (Fig. S1) and rpoB ( Fig. S2) gene sequences of M. stomatepiae sp. nov. and selected mycobacterial species, and the oligonucleotides used f...