Mycobacterium florentinum , a new species isolated from humans

1. Mycobacterium florentinum, a new species isolated from humans Enrico Tortoli

2. M. simiae-related mycobacteria • Slow growth • Short helix 18 in the 16S rDNA • Known organisms • until 1993: M. simiae • 2004: 13 species

3. The strains • 1. isolated twice from the sputum of an elderly patient with lung cancer • 2. isolated twice from the sputum of an elderly patient with hemoptysis • 3. isolated from the stools of an AIDS patient • 4. isolated from the excised lymph node of a child with cervical lymphadenitis • 5. isolated three times from the sputum of an elderly patient with pulmonary disease • 6. isolated four times from the sputum of a patient, long hospitalized in intensive care due to pneumonia imputed to pneumococcus • 7. isolated twice, at four year interval, from the sputum of an elderly woman with fibrosis and bronchiectasis

4. The strains • 1. isolated twice from the sputum of an elderly patient with lung cancer • 2. isolated twice from the sputum of an elderly patient with hemoptysis • 3. isolated from the stools of an AIDS patient • 4. isolated from the excised lymph node of a child with cervical lymphadenitis • 5. isolated three times from the sputum of an elderly patient with pulmonary disease • 6. isolated four times from the sputum of a patient, long hospitalized in intensive care due to pneumonia imputed to pneumococcus • 7. isolated twice, at four year interval, from the sputum of an elderly woman with fibrosis and bronchiectasis

5. Biochemical and cultural features • Growth: at 25-37°C in two weeks • Colonies: smooth, creamy, nonchromogenic • Positive biochemical tests: urea hydrolysis, tellurite reduction • Negative biochemical tests: niacin, Tween 80 hydrolysis, 3-d arylsulfatase, β-glucosidase • Variable biochemical tests: nitrate reduction, semiquantitative catalase • Growth inhibited on: Mac Conkey agar, 5% NaCl Lowenstein Jensen • Growth not inhibited on: p-nitrobenzoate, isoniazid, hydroxylamine and oleate

6. GLC pattern M. lentiflavum M. triplex FI-93171 24:0 > 26:0 20:0 > 22:0 18:0 > TBSA

7. HPLC pattern M. simiae FI-93171

8. HPLC pattern

9. Antimicrobial susceptibility • Full susceptibility to clarithromycin and clofazimine only • High MICs of quinolones and rifamycins

10. Genetic sequencing • 16S rDNA: 8 nucleotide mismatch with M. triplex • ITS: 14 nucleotide mismatch with M. triplex

11. Phylogenetic tree FI-93171

12. PRA patterns BstE III Hae III Haa I 1 2 3 1 2 3 1 2 3 1. FI-93171 2. M. lentiflavum 3. M. triplex

13. A previous report

14. colonies biochem. GLC HPLC PRA 16S + ITS phylogeny M. triplex M. lentiflavum similarity high low none M. florentinum sp. nov. • Most closely related species: • M. triplex • M. lentiflavum

15. colonies biochem. GLC HPLC PRA 16S + ITS phylogeny M. triplex M. lentiflavum similarity high low none M. florentinum sp. nov. • Most closely related species: • M. triplex • M. lentiflavum

17. Contributors Carlo Garzelli Laura Rindi Department of Experimental Pathology, University of Pisa, Pisa, Italy Nalin Rastogi K. S. Goh TB and Mycobacteria Unit, Institut Pasteur of Guadeloupe, Pointe a Pitre, Guadeloupe, France Gianna Mazzarelli Alessandro Mariottini Laboratory of Microbiology, Careggi Hospital, Florence, Italy Pirjo Torkko Laboratory of Environmental Microbiology, National Public Health Institute, Kuopio, Finland Marija-Leena Katila Department of Clinical Microbiology, Kuopio University Hospital, Kuopio, Finland