title: Activities of drug combinations against Mycobacterium tuberculosis grown in aerobic and hypoxic acidic conditions.
abstract: Mycobacterium tuberculosis is exposed to hypoxia and acidity within granulomatous lesions. In this study, an acidic culture model of M. tuberculosis was used to test drug activity against aerobic 5-day-old (A5) and hypoxic 5-, 12-, and 19-day-old (H5, H12, and H19, respectively) bacilli after 7, 14, and 21 days of exposure. In A cultures, CFU and pH rapidly increased, while in H cultures growth stopped and pH increased slightly. Ten drugs were tested: rifampin (R), isoniazid (I), pyrazinamide (Z), ethambutol (E), moxifloxacin (MX), amikacin (AK), metronidazole (MZ), nitazoxanide (NZ), niclosamide (NC), and PA-824 (PA). Rifampin was the most active against A5, H5, H12, and H19 bacilli. Moxifloxacin and AK efficiently killed A5 and H5 cells, I was active mostly against A5 cells, Z was most active against H12 and H19 cells, and E showed low activity. Among nitrocompounds, NZ, NC, and PA were effective against A5, H5, H12, and H19 cells, while MZ was active against H12 and H19 cells. To kill all A and H cells, A5- and H5-active agents R, MX, and AK were used in combination with MZ, NZ, NC, or PA, in comparison with R-I-Z-E, currently used for human therapy. Mycobacterial viability was determined by CFU and a sensitive test in broth (day to positivity, MGIT 960 system). As shown by lack of regrowth in MGIT, the most potent combination was R-MX-AK-PA, which killed all A5, H5, H12, and H19 cells in 14 days. These observations demonstrate the sterilizing effect of drug combinations against cells of different M. tuberculosis stages grown in aerobic and hypoxic acidic conditions.
authors.affiliation: Dipartimento di Malattie Infettive, Parassitarie e Immunomediate, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Piccaro
authors.forename: Federico
authors.initials: F
authors.lastname: Giannoni
authors.forename: Perla
authors.initials: P
authors.lastname: Filippini
authors.forename: Alessandro
authors.initials: A
authors.lastname: Mustazzolu
authors.forename: Lanfranco
authors.initials: L
authors.lastname: Fattorini
mesh_terms: D000332:Aerobiosis / Q000187:drug effects* / Q000502:physiology; D000693:Anaerobiosis / Q000187:drug effects* / Q000502:physiology; D000995:Antitubercular Agents / Q000494:pharmacology*; D015169:Colony Count, Microbial; D003470:Culture Media; D004338:Drug Combinations; D006863:Hydrogen-Ion Concentration; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000254:growth & development
chemical_list: D000995:Antitubercular Agents; D003470:Culture Media; D004338:Drug Combinations