Determination of in vitro susceptibility of Mycobacterium avium complex isolates to antimycobacterial agents by various methods.

Various methods were used to determine the in vitro susceptibility of Mycobacterium avium complex strains isolated from patients with acquired immunodeficiency syndrome. Our results confirm the noted resistance of the M. avium complex to conventional antituberculosis agents. The procedures used were both agar dilution and broth dilution, including a commercially available radiometric system (BACTEC; Johnston Laboratories, Towson, Md.). In general, all strains were more resistant by an agar dilution procedure than by a broth dilution procedure. Radiometric data were analyzed by defining a value, termed T100, which provides a discrete MIC. The broth (radiometric) procedure is reproducible and convenient for screening antimicrobial agents for in vitro activity and assessing potential therapeutic efficacy. Nevertheless, there is no standard procedure for determining the in vitro susceptibility of the M. avium complex, and appropriate clinical correlation studies are needed to accurately assess the clinical relevance of any in vitro result.     

In vitro susceptibility of Mycobacterium marinum to eight antimicrobial agents.

The in vitro susceptibilities of 16 Mycobacterium marinum strains to eight antimicrobial agents were determined by the agar dilution technique. The most active drugs were amikacin and kanamycin. Tetracycline, doxycycline, and minocycline were inhibitory, predominantly at concentrations slightly below the expected blood and tissue levels. Trimethoprim-sulfamethoxazole and erythromycin demonstrated activity only at concentrations greater than those usually attained in serum and tissues. Gentamicin was relatively inactive.     

In vitro susceptibility of Mycobacterium avium to a new macrolide (RU-28965)

The in vitro susceptibility of Mycobacterium avium to RU-28965 alone and in combination with rifampin, isoniazid, and sulfametoxipiridazine was studied by the agar dilution method. The synergistic effect of the RU-28965 with rifampin has been demonstrated. At a concentration of 16 micrograms/ml or lower of RU-28965, 100% of M. avium strains were inhibited. If 2 micrograms/ml of rifampin is added the MIC of RU-28965 is lowered to 0.25 microgram/ml.     

In vitro susceptibility of Bacillus anthracis to various antibacterial agents and their time-kill activity

OBJECTIVES: To investigate the in vitro acquisition of resistance to antibiotics by Bacillus anthracis. METHODS: The in vitro activities of 18 antibacterial agents against two strains of B. anthracis, the Sterne strain and the Russian anthrax vaccine strain ST-1, were tested by determining the MICs and by measuring the rates of antibiotic kill at 5x and 10x MIC. RESULTS: The fluoroquinolones ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin, the beta-lactams penicillin G and amoxicillin, the macrolide clarithromycin, the ketolide telithromycin, as well as clindamycin, rifampicin and quinupristin/dalfopristin had MICs in the range of 0.03-0.25 mg/L. Minocycline had an MIC of 0.03 mg/L, as did penicillin, against the ST-1 strain. Ciprofloxacin had an MIC of 0.03 mg/L against both strains. Erythromycin, vancomycin and the oxazolidinone linezolid were less active (MIC 0.5-2.5 mg/L). Ceftriaxone was the least active, having an MIC of 8.0 mg/L. Chloramphenicol was inactive (MIC > 256 mg/L). Quinupristin/dalfopristin, rifampicin and moxifloxacin showed the most rapid bacterial killing, achieving a complete eradication of detectable organisms (2 log(10) reduction within 0.5-3 h and 4 log(10) reduction within 0.5-4 h for both strains at concentrations of 5x and 10x the MIC). The beta-lactams and vancomycin demonstrated a 2-4 log(10) reduction within 5-15 h. Ceftriaxone had a similar effect to penicillin and amoxicillin against the ST-1 strain, but a slower effect than these two beta-lactams against the Sterne strain. The macrolides, tetracyclines and linezolid demonstrated a lower kill rate, while chloramphenicol did not kill at all. CONCLUSIONS: These data expand on the spectrum of agents recommended for the treatment of anthrax (ciprofloxacin, penicillin G and tetracyclines) and add new options, such as other fluoroquinolones, amoxicillin, rifampicin and quinupristin/dalfopristin, as potential therapeutic agents.     

In vitro activities of several new macrolide antibiotics against Mycobacterium avium complex.

The in vitro activities of 12 macrolide compounds against 28 Mycobacterium avium complex strains isolated from patients with acquired immunodeficiency syndrome were determined by the conventional proportion method and by the BACTEC method. Clarithromycin (A-56268; TE-031), a new macrolide compound, was the most active agent tested, inhibiting 90% of strains at an MIC of 4 micrograms/ml by the BACTEC method. Roxithromycin (RU 28965) and erythromycylamine inhibited 90% of strains at 16 micrograms/ml. The organisms showed high levels of resistance to most other macrolide compounds.     

Disk diffusion testing of susceptibility of Mycobacterium fortuitum and Mycobacterium chelonei to antibacterial agents.

Although recent studies have suggested that some antibacterial agents have good activity against the rapidly growing mycobacteria Mycobacterium fortuitum and Mycobacterium chelonei, an easily applicable method for susceptibility testing of clinical isolates is not yet available. We evaluated a disk diffusion method with Mueller-Hinton agar and 48-h readings with 59 strains of M. fortuitum and 11 strains of M. chelonei and compared the results to agar dilution susceptibilities for nine antimicrobial agents. All isolates were susceptible to 16 micrograms of amikacin or kanamycin per ml with minimum zone diameters of 14 and 18 mm, respectively. Amikacin inhibited 100% of isolates of M. fortuitum at 2 micrograms/ml, whereas 10 of 11 (91%) of M. chelonei strains had minimum inhibitory concentrations of 4.0 micrograms/ml or greater. Doxycycline and minocycline had almost identical activities, inhibiting 44% of strains at 4.0 micrograms/ml, and both allowed easy differentiation between susceptible and resistant strains by disk diffusion. Although most isolates of M. chelonei grew better on 7H10 agar, this media gave two- to eight-fold higher minimum inhibitory concentrations than were obtained with Mueller-Hinton agar. Disk diffusion susceptibility testing appears to be a simple and reliable means of predicting susceptibility results for M. fortuitum and most isolates of M. chelonei by the agar dilution method.     

In vitro susceptibility of Mycobacterium fortuitum to cefoxitin.

The in vitro susceptibility of Mycobacterium fortuitum to cefoxitin was studied by agar dilution, broth dilution, and disk diffusion methods. Of 13 isolates, 11 were found to be susceptible by disk diffusion. At a concentration of 25 microgram/ml or lower, all 13 isolates were inhibited when tested by broth dilution, but only 12 of 13 isolates were inhibited when tested by agar dilution.     

In vitro susceptibility of Mycobacterium kansasii to clarithromycin.

The MICs of the macrolide clarithromycin for 31 clinical isolates of Mycobacterium kansasii were determined by three different methods. The methods employed were the proportion resistance method on 7H10 agar, the radiometric (BACTEC) method, and the T100 method of datum analysis. All methods gave similar results. The MICs were in a narrow range from 0.16 to 0.50 microgram/ml, with the MICs for 90% of isolates tested of 0.50 microgram/ml for the agar dilution and radiometric methods and 0.37 microgram/ml for the T100 method. The MBCs were determined for nine representative isolates by the radiometric broth method. The MBCs were equal to the MICs for four isolates, and the MBCs were twofold higher than the MICs for five isolates. Killing of 99.9% of the bacterial population was achieved at a clarithromycin concentration of 2.0 micrograms/ml for all nine isolates tested.     

In vitro susceptibility of Mycobacterium avium complex to the new fluoroquinolone sparfloxacin (CI-978; AT-4140) and comparison with ciprofloxacin.

We tested the activity of the new fluoroquinolone sparfloxacin (CI-978; AT 4140) against 30 strains of Mycobacterium avium complex (MAC) isolated from patients with acquired immune deficiency syndrome. MICs of sparfloxacin (range, less than or equal to 0.06 to 4 micrograms/ml) were lower than MICs of ciprofloxacin for all 30 strains, and MBCs for acid-fast bacteria were lower for 28 of the 30 strains. In synergism experiments using 10 strains of MAC, fractional inhibitory concentration indices revealed that the combination of sparfloxacin plus ethambutol was synergistic against 9 strains, and the three-drug combination of sparfloxacin plus ethambutol plus rifampin was synergistic against all strains. In the absence of ethambutol, the combination of sparfloxacin plus rifampin appeared to be antagonistic against three of the MAC strains.     

In vitro susceptibility of Mycobacterium fortuitum and Mycobacterium chelonei to cefmetazole.

The in vitro susceptibility of Mycobacterium fortuitum and Mycobacterium chelonei to cefmetazole was studied by the agar dilution method. At a concentration of 16 micrograms/ml or lower, 44 isolates (96%) of M. fortuitum and 8 isolates (40%) of M. chelonei were inhibited.     

In vitro susceptibility of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium avium, Mycobacterium fortuitum, and Mycobacterium chelonae to ticarcillin in combination with clavulanic acid.

The in vitro susceptibility of Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium avium, Mycobacterium fortuitum, and Mycobacterium chelonae (M. chelonei) to ticarcillin in combination with calvulanic acid (CA) was studied by the agar dilution method. All the M. tuberculosis, M. bovis, and M. africanum strains were inhibited at a ticarcillin concentration of 32 micrograms/ml or lower in combination with 5 micrograms of CA. M. chelonae and M. avium strains proved resistant to more than 128 micrograms of ticarcillin plus 5 micrograms of CA per ml. M. fortuitum strains needed 128 micrograms of ticarcillin plus 5 micrograms of CA to inhibit approximately 30% of the isolates.     

Effects of surface-active agents on drug susceptibility levels and ultrastructure of Mycobacterium avium complex organisms isolated from AIDS patients

The multiple-drug-resistance property of Mycobacterium avium complex (MAC) is mainly attributed to a cell envelope permeability barrier. MAC treated with subinhibitory levels of dimethyl sulfoxide (DMSO) and ethylenediaminetetraacetic acid (disodium salt) (EDTA) did not have altered minimum inhibitory concentration (MIC) levels or show ultrastructural changes; the effect of sodium dodecyl sulfate (SDS) was variable. With SDS, the visualization of the nucleoid and ribosomes decreased, and amorphous electron-dense material accumulated near the structurally altered cytoplasmic membrane and cell wall. Use of 0.005% Tween-80 resulted in a 2-4-fold reduction in MIC in the case of rifampicin, ansamycin (LM 427), cephapirin, and ciprofloxacin. Tween-80 treated cells were swollen, and deposits of low electron-density accumulated in the cytoplasm; distortions in the outer-cell integuments were observed. These findings are consistent with the idea that Tween-80 increases cell-envelope permeability, thereby enhancing drug penetrability and reducing MIC levels. Because of the action of Tween-80, its use in drug-susceptibility media or diluent fluids should be avoided.     

嗜麦芽窄食单胞菌体外药物敏感性研究

目的比较10种抗菌药物（莫西沙星、加替沙星、左氧氟沙星、环丙沙星、头孢哌酮-舒巴坦、替卡西林-克拉维酸、哌拉西林-三唑巴坦、头孢吡肟、头孢他啶、复方磺胺甲嗯唑）对临床分离嗜麦芽窄食单胞菌的体外抗菌活性。方法琼脂稀释法测定10种抗菌药物对来自北京3所医院的200株嗜麦芽窄食单胞菌的MIC值，用WHONET5．3软件进行药敏数据统计分析。结果200株菌对药物的敏感率排序为莫西沙星（84．5％）、加替沙星（79．5％）、左氧氟沙星（76％）、复方磺胺甲嗯唑（72．5％）、头孢哌酮-舒巴坦（41．5％）、替卡西林-克拉维酸（34％）、头孢他啶（23％）、环丙沙星（15％）、头孢吡肟（7％）和哌拉西林-三唑巴坦（4．5％）。结论新一代氟喹诺酮类药物如莫西沙星对嗜麦芽窄食单胞菌有较高的体外抗菌活性，是临床治疗嗜麦芽窄食单胞菌感染的较好选择。     

In vitro susceptibility of Capnocytophaga species to antimicrobial agents.

A total of 33 clinical isolates of Capnocytophaga spp. were susceptible to 4-quinolone antimicrobial agents. The antipseudomonal penicillins tested were equally active against all isolates, as were cefuroxime, ceftriaxone, ceftizoxime, cefotaxime, latamoxef, and ceftazidime. Most isolates were resistant to trimethoprim, and some were resistant to aztreonam. Most regimens for the empirical treatment of septic episodes in immunocompromised patients are suitable for the treatment of Capnocytophaga spp. infections.     

In vitro susceptibility of Nocardia asteroides to 25 antimicrobial agents

Fifty-two clinical isolates of Nocardia asteroides were tested by agar dilution for their susceptibility to 25 antimicrobial agents. In general, susceptibility could not be predicted based on the antibiotic class tested. However, the beta-lactams, including third-generation cephalosporins, were generally ineffective (MIC for 90% of the organisms [MIC90], between 64 and greater than 256 micrograms/ml), whereas minocycline and doxycycline were generally effective (MIC90, 4 and 8 micrograms/ml, respectively). Cycloserine was not effective below 60 micrograms/ml. The MIC50 and MIC90 of sulfamethoxazole was 16 and 32 micrograms/ml, respectively, and that of trimethoprim varied widely (16 and greater than 256 micrograms/ml, respectively). Based on MIC90 data, only doxycycline, minocycline, sulfamethoxazole, and imipenem could be applied empirically.     

In vitro activity between linezolid and other antimicrobial agents against Mycobacterium abscessus complex

Linezolid (LZD) serves as an effective option in the treatment of Mycobacterium abscessus complex (MABC) infection. Unfortunately, the combined activities of LZD with other antimicrobial agents against MABC have not been evaluated systemically. In this study, we randomly selected 32 Mycobacterium abscessus and 32 Mycobacterium massiliense isolates for the determination of in vitro synergistic effect between LZD and other antimicrobial agents, including amikacin (AMK), moxifloxacin (MOX), cefoxitin (CFX) and tigecycline (TGC). Out of 64 MABC isolates tested, only one (1.6%, 1/64) and two (3.2%, 2/64) exhibited resistance to AMK and LZD, respectively. Statistical analysis revealed that the percentage of TGC-resistant isolates was significantly lower among M. massiliense (9.4%, 3/32) than that among M. abscessus (25.0%, 8/32, P < 0.001). In addition, LZD and AMK showed synergy for 29 MABC isolates (45.3%), whereas no antagonism was noted for this combination. The second mostly frequent synergistic effect was found in LZD plus TGC combination, and 26.6% (17/64) of the strains tested exhibited synergy. In contrast, LZD-CFX and LZD-MOX combinations appeared antagonistic for half of the isolates (48.4%, 31/64 for CFX and 51.6%, 33/64), and almost no synergistic effect was reported in any of the strains for these two combinations. In conclusion, our data reveal that LZD and AMK show the most potent activity against MABC. The frequent synergism is observed in LZD-AMK and LZD-TGC combinations, while LZD rarely exhibits in vitro synergy with MOX and CFX when tested against MABC.     

Use of potassium tellurite for rapid-drug susceptibility testing of Mycobacterium avium complex.

BACKGROUND: Mycobacterium avium complex (MAC) is a major cause of infection in immunocompromised patients. MAC possesses an enzyme that reduces potassium tellurite in less than 3 days and results in formation of a black precipitate. The objective of this study was to determine whether reduction of potassium tellurite by mycobacteria can be used as a means of testing the susceptibility of MAC to clarithromycin. METHODS: Minimum inhibitory concentrations (MICs) for 104 clinical isolates of MAC were determined by the tellurite method and compared with those tested by a recommended microdilution method. Microdilution breakpoints were used for interpretation of susceptibility. MIC of less than 8 microg/mL was considered as susceptible, and MIC of greater than or equal to 8 microg/mL was resistant. RESULTS: There was agreement within a 2-fold dilution between MICs for 89% of isolates. Of the 53 isolates that had discrepant MICs by the two methods, 70% had higher MICs by the tellurite method. When the MICs were classified into interpretive categories, there was 100% agreement by the two methods. The MIC tested by the tellurite method was available within 5 days. CONCLUSIONS: These data suggest that use of potassium tellurite is a more rapid, reliable, and inexpensive method of testing the susceptibility of MAC to clarithromycin.     

In vitro susceptibilities of Mycobacterium tuberculosis to 10 antimicrobial agents.

After preliminary in vitro screening of 10 antimicrobial agents against Mycobacterium tuberculosis, the MICs of the 6 most promising agents against 27 clinical isolates were determined by agar dilution. The two quinolone compounds tested (difloxacin and A-56620) were the most active, each inhibiting 50% of the strains at concentrations of 4 micrograms/ml. M. tuberculosis strains previously shown to be resistant to isoniazid, streptomycin, rifampin, or ethambutol were as susceptible to these quinolone compounds as susceptible strains.     

In vitro susceptibility of Haemophilus somnus to 33 antimicrobial agents.

Minimal inhibitory concentrations of 33 antimicrobial agents for Haemophilus somnus were determined by the agar dilution method. The tested H. somnus strains were highly susceptible to penicillin G, ampicillin, colistin, and novobiocin. They were not susceptible to spiramicin and sulfadimethoxine, and streptomycin-resistant strains were found.