MICs and MBCs of clarithromycin against Mycobacterium avium within human macrophages.

The inhibitory and bactericidal activities of clarithromycin were determined quantitatively against the intracellular populations of five Mycobacterium avium strains growing in monocyte-derived human macrophages. The MICs were 1.0 microgram/ml, and the MBCs ranged from 16.0 to 64.0 micrograms/ml; these values were similar to the MICs and MBCs found in broth cultures at pH 7.4 and were substantially lower than those found in broth cultures at pHs 6.8 and 5.0. Since the intracellular environment has a neutral or even an acidic pH, relatively low MICs and MBCs found in macrophage cultures can be associated with the fact that the drug concentrations in macrophages are substantially higher than those in the medium in which these cells are cultivated. Pretreatment of the macrophages 2 days prior to infection decreased the MICs twofold in comparison with results of experiments in which the drug was added to already infected macrophages.     

In vitro activity of RP 59500, a semisynthetic injectable pristinamycin, against staphylococci, streptococci, and enterococci.

The in vitro activity of RP 59500, a semisynthetic pristinamycin, was compared with the activities of vancomycin, oxacillin, ampicillin, gentamicin, ciprofloxacin, and rifampin against five Staphylococcus species, five Streptococcus species, and four Enterococcus species. For staphylococci, MICs were 0.13 to 1 microgram/ml and the MICs for 90% of the strains tested (MIC90s) were 0.13 to 0.5 microgram/ml; there were no differences between oxacillin-susceptible and -resistant strains. For streptococci, MICs were 0.03 to 4 micrograms/ml and MIC90s were 0.25 to 2 micrograms/ml; viridans group streptococci were the least susceptible streptococci. For enterococci, MICs were 0.25 to 32 micrograms/ml and MIC90s were 2 to 4 micrograms/ml; Enterococcus faecalis was the least susceptible. Vancomycin was the only comparative drug with consistent activity against all species of gram-positive cocci. With RP 59500, raising the inoculum 100-fold, lowering the pH of cation-adjusted Mueller-Hinton broth to 5.5, or omitting cation supplementation had little effect on MICs, but 50% serum increased MICs 2 to 4 dilution steps. The differences between MBCs and MICs were greater for staphylococci and enterococci than for streptococci. Time-kill studies with 24 strains indicated that RP 59500 concentrations 2-, 4-, and 16-fold greater than the MICs usually killed bacteria of each species at similar rates; reductions in CFU per milliliter were less than those observed with oxacillin or vancomycin against staphylococci and less than those observed with ampicillin against enterococci. RP 59500 antagonized the bactericidal activities of oxacillin and gentamicin against Staphylococcus aureus ATCC 29213 and that of ampicillin against E. faecalis ATCC 29212. Against the latter, combination with gentamicin was indifferent. RP 59500 has a broad spectrum of in vitro activity against gram-positive cocci; combining it with other drugs is not advantageous.     

The bactericidal activity of ampicillin, daptomycin, and vancomycin against ampicillin-resistant Enterococcus faecium.

Ampicillin, daptomycin, and vancomycin, alone and in combination with gentamicin, were examined for bactericidal effects on ampicillin-resistant Enterococcus faecium using broth dilution minimum inhibitory concentrations (MICs) and time-kill studies. We tested 12 ampicillin-resistant isolates and demonstrated the following MICs and MBCs, respectively: ampicillin, greater than or equal to 32 micrograms/ml and greater than 256 micrograms/ml; daptomycin, less than or equal to 4 micrograms/ml and less than or equal to 16 micrograms/ml; and vancomycin, less than or equal to 4 micrograms/ml and greater than 64 micrograms/ml. Time-kill studies demonstrated that daptomycin alone had marked activity against the ampicillin-resistant E. faecium and that the addition of gentamicin resulted in synergistic killing. In addition, ampicillin and vancomycin were not bactericidal for the ampicillin-resistant isolates without the addition of gentamicin. The present study supports the consideration of daptomycin alone or in combination with an aminoglycoside as an alternative therapy for ampicillin-resistant enterococci, although additional clinical experience is now necessary.     

MICs and MBCs of Win 57273 against Mycobacterium avium and M. tuberculosis.

A new quinolone, Win 57273 [1-cyclopropyl-7-(2,6-dimethyl-4-pyridinyl)-6-fluoro-1,4-dihydro-4-oxo-3 - quinolonecarboxylic acid], synthesized by Sterling Research Group, was tested in vitro against Mycobacterium tuberculosis and Mycobacterium avium strains. The broth-determined MICs of this agent ranged from 1.0 to 4.0 micrograms/ml for M. tuberculosis strains and from 0.25 to 8.0 micrograms/ml for M. avium strains. A distinctive feature of this agent, in comparison with ofloxacin and ciprofloxacin, is its substantially greater activity at the low pHs. For M. avium strains, the MICs of Win 57273 were 2.0 micrograms/ml or less for 54.5% of strains at pH 6.8 and 85.5% of strains at pH 5.0. Win 57273 was more active than ciprofloxacin against M. avium strains, and this difference was very substantial for all M. avium strains at pH 5.0. Taking into account that the predominant locations of these organisms in vivo are within the phagosomes and phagolysosomes of macrophages, i.e., in acidic environments at pH 5.0 or lower, the greater activity of Win 57273 at low pH makes this quinolone especially promising for M. avium infection. The bactericidal activity of Win 57273 for M. avium strains was the same as that of ciprofloxacin, with MBCs from 4.0 to 16.0 micrograms/ml.     

In vitro activity and beta-lactamase stability of a new carbapenem, SM-7338.

SM-7338, a new carbapenem, inhibited most members of the family Enterobacteriaceae at MICs of 0.015 to 0.25 microgram/ml, including Klebsiella oxytoca, Citrobacter freundii, Enterobacter cloacae, and Proteus vulgaris isolates resistant to cefotaxime, ceftazidime, piperacillin, and gentamicin. It was two- to eightfold more active than imipenem, but it inhibited Pseudomonas aeruginosa at 1 to 8 micrograms/ml, which was comparable to the activity of imipenem. Haemophilus, Neisseria, and Branhamella species were inhibited by less than or equal to 0.25 microgram/ml, which was superior to the activity of imipenem. SM-7338 inhibited Staphylococcus aureus and coagulase-negative staphylococci at 0.25 microgram/ml, but for methicillin-resistant isolates MICs were 4 to 16 micrograms/ml. Group A, B, and C streptococci and Streptococcus pneumoniae were inhibited by less than or equal to 0.03 microgram/ml. Bacteroides species, including clindamycin-resistant isolates, were inhibited by 0.25 microgram/ml. There was no major inoculum size effect, and the MBCs were within a dilution of the MICs. SM-7338 was more active than imipenem at an acid pH under anaerobic conditions. Plasmid beta-lactamases of TEM-1, TEM-2, TEM-3, TEM-5, SHV-1, SHV-2, PSE-1, PSE-2, PSE-3, OXA-2, OXA-3, OXA-4, OXA-5, and OXA-7; Staphylococcus aureus enzymes; and the chromosomal beta-lactamases P-99 and K-1; Morganella species; and Proteus vulgaris did not hydrolyze SM-7338. The repeated transfer of organisms increased the MICs of SM-7338, as it did the MICs of imipenem.     

Activity of mersacidin, a novel peptide, compared with that of vancomycin, teicoplanin, and daptomycin.

Mersacidin, a new peptide antibiotic, was four- to eightfold less active (MIC for 90% of isolates, 8 micrograms/ml) than vancomycin, teicoplanin, or daptomycin against Staphylococcus aureus. Coagulase-negative staphylococci were inhibited by 8 micrograms/ml, and the MICs of mersacidin for hemolytic streptococci and Streptococcus pneumoniae were 4 to 8 micrograms/ml. The mersacidin MICs for anaerobic organisms were as follows: Clostridium perfringens, 4 micrograms/ml; Propionibacterium acnes, 8 micrograms/ml; peptococci, 1 microgram/ml; and peptostreptococci, 8 micrograms/ml. Mersacidin had no activity against members of the family Enterobacteriaceae, Neisseria and Haemophilus species, or Pseudomonas aeruginosa. The size of the inoculum, the pH of the assay (5.5 to 7.5), the type of medium, and the anaerobic conditions had minimal effects on the MICs and MBCs of mersacidin. Overall, mersacidin proved less active than available glycopeptides and peptolides.     

Susceptibility of enterococci to trimethoprim and trimethoprim-sulfamethoxazole.

The in vitro activities of trimethoprim (TMP), alone and in combination with sulfamethoxazole (SMX), against 131 clinical isolates of enterococci, 126 Streptococcus faecalis isolates, and 5 Streptococcus faecium isolates were determined by a broth microdilution method with Mueller-Hinton broth that was substantially free of inhibitory substances. The geometric mean MIC of TMP for strains of S. faecalis was 0.164 micrograms/ml (range, 0.03 to 8 micrograms/ml), with a geometric mean MBC of 0.298 micrograms/ml (range, 0.063 to 8 micrograms/ml). Although all strains were resistant to the sulfonamide alone, the inhibitory and bactericidal activities of TMP against strains of S. faecalis were markedly potentiated when TMP was combined in a fixed ratio of 1:19 with SMX; the geometric mean MIC of TMP was reduced to 0.016 micrograms/ml (range, 0.002 to 0.25 micrograms/ml), with a geometric mean MBC of 0.031 micrograms/ml (range, 0.004 to 0.25 micrograms/ml). The combination had no synergistic effect against strains of S. faecium; the geometric mean MICs and MBCs of both agents were ca. 0.06 micrograms/ml. The MBC/MIC ratios for TMP and TMP-SMX were less than or equal to 16 for all 131 strains. MICs and MBCs for TMP-SMX were unchanged, and for TMP they decreased when performed in broth supplemented with 50% heat-inactivated pooled human serum. For TMP and TMP-SMX, the susceptibilities of isolates with high-level resistance to gentamicin or streptomycin were the same as those of isolates susceptible to less than or equal to 2,000 micrograms of aminoglycoside per ml. These results suggest that TMP-SMX and TMP alone could prove useful in the treatment of serious enterococcal infections, including infections by strains with high-level resistance to aminoglycosides.     

MICs and MBCs of chemotherapeutic agents against Renibacterium salmoninarum.

The efficacies of 21 chemotherapeutic agents for controlling bacterial kidney disease were evaluated. The bactericidal and/or bacteriostatic effects of these drugs were tested against 11 Renibacterium salmoninarum strains with different origins. The most effective compounds displaying both bacteriostatic and bactericidal activity for all the isolates were tetracycline and erythromycin, with MICs ranging from less than 0.62 to 10.95 micrograms/ml for tetracycline and from less than 0.62 to 5.47 micrograms/ml for erythromycin. Whereas tetracycline showed identical MICs and MBCs, erythromycin showed bactericidal effects at concentrations of 5.47 to 21.87 micrograms/ml. Similarly, cefazolin and tiamulin proved to be very effective bactericidal compounds against the majority of R. salmoninarum isolates, with MBCs for 90% of the strains tested of 21.87 and 10.95 micrograms/ml, respectively. Neither nitrofuranes, quinolones, nor sulfonamides showed inhibitory effects on the growth of the strains.     

Pseudomonas aeruginosa bacteremia: susceptibility of 100 blood culture isolates to seven antimicrobial agents and its clinical significance.

The susceptibility of 100 blood culture isolates of Pseudomonas aeruginosa observed during 4 1/2 years was tested for tobramycin, netilimicin, gentamicin, amikacin, pirbenicillin, ticarcillin and carbenicillin, singly and in combination. For aminoglycosides, the agar MICs were twofold to threefold greater than tube dilution MICs but for the penicillins they were similar. For aminoglycosides and ticarcillin, the MBCs were twofold greater than the tube dilution MICs. The MBCs were not achieved at concentrations as high as 512 micrograms/ml for 40% of the isolates for pirbenicillin and for 10% for carbenicillin. Tobramycin and pirbenicillin had the lowest MICs for the aminoglycosides and penicillins, respectively. Synergism was tested and observed between tobramycin + ticarcillin and amikacin + ticarcillin. No overall increase in resistance to gentamicin or carbenicillin was seen from 1974 to 1977. However, patients given repeated courses of gentamicin had more resistant strains. Following the administration of 1.5 mg/kg/dose of gentamicin, peak serum concentrations failed to achieve the MIC for the microorganism in 22% of the patients. The MIC was achieved in all patients receiving the same dose of tobramycin. The overall fatality rate was 67% with one third of the patients dying within 36 hr. There was no relationship of patient fatality rate and MIC for the microorganism. Although in the rapidly fatal group of all patients receiving inappropriate therapy died, the fatality rates of appropriately or inappropriately treated patients in the ultimately fatal and nonfatal groups were similar. Underlying host disease was the major determining factor in patient survival.     

In vitro activity of DNA gyrase inhibitors, singly and in combination, against Mycobacterium avium complex.

The in vitro activities of the DNA gyrase inhibitors ciprofloxacin, coumermycin, and novobiocin against 31 clinical isolates of Mycobacterium avium complex were studied using a microdilution technique. Minimal inhibitory concentrations (MICs) were determined in 4 days using Middlebrook 7H9 broth, and minimal bactericidal concentrations (MBCs) were determined by subculturing to Middlebrook 7H10 agar. MICs were: ciprofloxacin, 0.5-greater than 16 (mean, 4.1) micrograms/ml; novobiocin, 4-greater than 128 (mean, 54.7) micrograms/ml; and coumermycin, 2-greater than 16 (mean, 17.5) micrograms/ml. MBCs were usually more than two dilution steps higher than MICs. Checkerboard studies failed to reveal synergistic or antagonistic inhibitory activity of DNA gyrase-A and DNA gyrase-B inhibitors in vitro.     

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 antibiotic susceptibility testing of clinical isolates of Mycoplasma penetrans from patients with AIDS.

In vitro susceptibilities of Mycoplasma penetrans were determined. MICs and MBCs were determined. The MICs at which 50% of the isolates are inhibited (micrograms per milliliter) for broth dilution testing were as follows: azithromycin, 0.039; chloramphenicol, 0.625; ciprofloxacin, 0.156; clindamycin, 0.078; doxycycline, 0.312; erythromycin, 0.312; gentamicin. > 10; levofloxacin, 0.078; lincomycin, 0.625; streptomycin, > 10; and tetracycline, 1.25. Bactericidal activity was significant only for ciprofloxacin (MBC at which 50% of the isolates are killed, 0.312 microgram/ml) and levofloxacin (MBC at which 50% of the isolates are killed, 0.312 microgram/ml).     

In vitro susceptibility of group G streptococci to ten antimicrobial agents with broad gram-positive spectra

Minimal inhibitory and bactericidal concentrations (MICs, MBCs) of ten antibiotics with broad spectra against grampositive cocci were determined for 25 group G streptococci (GGS). Penicillin G, cefotaxime, and ampicillin were the most active bactericidal agents, with 100% of MBCs less than or equal to 0.6 micrograms/ml. Among the nonpenicillin, noncephalosporin agents, vancomycin had the lowest MBCs, with all strains killed by less than or equal to 2.5 micrograms/ml. Despite good in vitro activity against other streptococci, erythromycin, clindamycin, and chloramphenicol had disappointing bactericidal activity against GGS. Three strains were tolerant to erythromycin, while 15 strains were tolerant to clindamycin. Chloramphenicol was the least active inhibitory agent, with an MIC90 of 13.8 micrograms/ml; this agent was also the least active bactericidal agent against GGS. with no MBCs less than 2.5 micrograms/ml and 9 strains with MBCs greater than or equal to 20 micrograms/ml. There were no GGS strains tolerant to either penicillin G or vancomycin.     

Activity of LY146032 against Enterococci with and without high-level aminoglycoside resistance, including two penicillinase-producing strains.

We tested the activity of LY146032 (LY) against 57 strains of enterococci collected from Chile, Thailand, and the United States. Some of the strains were resistant to high levels of gentamicin or streptomycin (or both), and two produced beta-lactamase (Bla+). MICs of LY ranged from 0.5 to 8 micrograms/ml, and MBCs ranged from 1 to 64 micrograms/ml. In time-kill assays, a 2 to 3 log10 killing effect was observed with LY against two Bla+ strains of Streptococcus (Enterococcus) faecalis and against three strains that were highly resistant to streptomycin and gentamicin. Synergism was demonstrated with LY and streptomycin against a Bla+ strain lacking high-level streptomycin resistance. These in vitro results suggest that LY should be studied further for possible use in treatment of enterococcal infections.     

Susceptibility testing of bacteria recovered from patients with peritonitis complicating continuous ambulatory peritoneal dialysis.

Antagonism of antibiotic activity by peritoneal dialysate has been postulated to be a cause of failure of treatment of peritonitis complicating continuous ambulatory peritoneal dialysis. We evaluated by a case-control study whether unexpected treatment failure could be attributed to such antagonism. Bacteria isolated from 34 patient episodes of peritonitis treated with the same regimen of ciprofloxacin monotherapy were studied. Ciprofloxacin was significantly less active in dialysate than in Iso-Sensitest broth (IB). The median MIC in IB was 0.5 microgram/ml, increasing to 2.0 micrograms/ml for both fresh dialysate (FD) (P = 0.003) and pooled dialysis effluent (PDE) (P = 0.03); the median MBC in IB was 8.0 micrograms/ml, increasing to 128.0 micrograms/ml in FD (P = 0.0002) and 64.0 micrograms/ml in PDE (P = 0.02). However, no significant differences were found in the results for patients suffering unexpected treatment failure (relapse of peritonitis) compared with the results for patients whose infection resolved without sequel. In IB the median MICs for relapsers and nonrelapsers were 1.0 and 0.5 microgram/ml, respectively (P = 0.88); median MBCs were 32.0 and 4.0 micrograms/ml (P = 0.19). In FD median MICs for relapsers and nonrelapsers were 2.0 and 1.0 micrograms/ml (P = 0.06); median MBCs were 128.0 micrograms/ml for both groups (P = 0.84). In PDE the median MICs were 2.0 micrograms/ml for both groups (P = 0.78); median MBCs were 256.0 and 64.0 micrograms/ml (P = 0.17). We therefore found no evidence to suggest that antagonism of antibiotic activity by dialysate is a cause of treatment failure or that conventional methods for laboratory susceptibility testing in peritonitis complicating continuous ambulatory peritoneal dialysis should be abandoned in favor of testing in media containing dialysate.     

Conditions affecting the results of susceptibility testing for the quinolone compounds

The quinolone class of compounds was studied for conditions which might affect susceptibility results. These compounds included amifloxacin, ciprofloxacin, difloxacin, enoxacin, norfloxacin, ofloxacin, and RO 23-6240. Ciprofloxacin, a representative quinolone, was found to have rapid bactericidal activity, equivalent to that of gentamicin, in contrast to the slower activity of a cephalosporin, cefotaxime. Test conditions that might affect susceptibility test results included divalent magnesium and calcium cation concentrations and pH. For strains of Staphylococcus aureus, Enterobacteriaceae, and enterococcus, the effects were not large. A pH of 5.0 in general increased the minimum inhibitory concentrations (MICs) for the organisms against most carboxyquinolones, up to 8-fold, as compared to that at pH 7.4. In comparison, a similar lowering of pH caused an increased in MIC of 32-fold for gentamicin and no change for cefotaxime. Increasing the concentrations of divalent cations increased the MICs on the average of only 4-fold. Of the quinolones, difloxacin was the least affected by change in concentration of divalent cations and by pH. Such changes are not expected to greatly affect the efficacy of therapy of those members of Enterobacteriaceae which have MICs much less than 0.1 micrograms/ml, but might diminish therapeutic efficacy for those organisms such as Streptococcus aureus with MICs of 1.0 microgram/ml or higher.     

LY146032, alone and in combination with gentamicin, for the treatment of enterococcal pyelonephritis in the rat model.

The in vivo and in vitro activity of LY146032 against Streptococcus faecalis GK was examined. The following MICs and MBCs in micrograms per milliliter were obtained: ampicillin, 0.8 and 1.5; vancomycin, 0.8 and 50; gentamicin, 12 and 25; and LY146032, 0.8 and 6. A time-kill-curve study involving approximately 10(6) organisms per ml showed a drop in the number of organisms of almost 2 log10 in the tube containing LY146032 (2 micrograms/ml) plus gentamicin (4 micrograms/ml) compared with bacterial numbers for the control at 4 h of incubation. However, regrowth was observed at 24 and 48 h, and no in vitro synergism was observed with the combination. A sample (1 ml) of overnight growth of this enterococcal strain at a concentration of 10(7) was then injected intravenously into 184 male Wistar rats weighing about 100 g each. After 12 days, 10 rats were sacrificed and the remaining ones were randomized into four treatment groups: (i) untreated control, (ii) LY146032 (3 mg) given subcutaneously, (iii) gentamicin (0.8 mg) given intramuscularly, and (iv) LY146032 plus gentamicin at the same dosages as when the drugs were used singly. The rats received antibiotics for 4 weeks twice daily, and approximately 10 rats in each group were sacrificed for quantitative kidney cultures at 1, 2, 4, and 6 weeks after the start of therapy. At the end of the 4- and 6-week periods, significantly better results were obtained with the combination of LY146032 plus gentamicin than with no treatment or treatment with single antibiotics.     

In vitro activity of LY264826, a new glycopeptide antibiotic, against gram-positive bacteria isolated from patients with cancer.

The in vitro activity of LY264826, a novel glycopeptide antibiotic produced by Amycolatopsis orientalis, was compared with those of vancomycin, teicoplanin, and oxacillin against 311 gram-positive clinical isolates from patients with cancer, LY264826 had lower MICs for 90% of isolates (MIC90) than vancomycin for all species tested. It was active against oxacillin-resistant isolates including Staphylococcus aureus (MIC90, 0.5 micrograms/ml), Staphylococcus haemolyticus (MIC90, 2.0 micrograms/ml), Enterococcus spp. (MIC90, 0.5 micrograms/ml), Bacillus cereus (MIC90, 0.25 micrograms/ml), and Corynebacterium jeikeium (MIC90, 0.12 micrograms/ml). For S. aureus, including oxacillin-resistant isolates, the MICs of LY264826 were similar to those of teicoplanin. For coagulase-negative staphylococci, however, LY264826 had MICs that were 4- to 32-fold lower than those of teicoplanin. Against most streptococcal species the activities of LY264826 and teicoplanin were similar. Bactericidal activity against Staphylococcus spp. and most Streptococcus pyogenes isolates was less than or equal to 1 dilution of the MIC. One isolate of S. pyogenes and all Enterococcus faecalis strains tested were tolerant of LY264826, with MBCs greater than or equal to 32-fold greater than the MICs. The addition of 50% human serum resulted in a significant increase in activity only against Staphylococcus epidermidis. Variations in pH from 6.4 to 8.4 and in inoculum from 10(3) to 10(7) CFU/ml did not significantly affect the activity of LY264826.     

In vitro antibiotic susceptibility of the newly recognized agent of ehrlichiosis in humans, Ehrlichia chaffeensis.

Ehrlichiosis in humans, a rickettsial disease recently discovered in the United States, is generally treated successfully with tetracyclines; however treatment with these agents is usually avoided with children and pregnant women. The in vitro susceptibility of Ehrlichia chaffeensis, the agent of human ehrlichiosis in the United States, was assessed by a quantitative evaluation of infected DH82 cells cultivated in 96-well microtiter plates in the presence of different concentrations of selected antibiotics. Extracellular MICs and MBCs were evaluated after 72 h of exposure to the antibiotics. Doxycycline and rifampin were found to exert rapidly bactericidal effects, with MBCs in the extracellular culture medium of less than 0.5 and 0.125 microgram/ml, respectively. E. chaffeensis was resistant to chloramphenicol, ciprofloxacin, erythromycin, co-trimoxazole, penicillin, and gentamicin, which had MICs greater than 16, 4, 8, 4, 40, and 32 micrograms/ml, respectively. These observations are consistent with the finding that human ehrlichiosis appears to respond to tetracycline therapy, which has been the therapy of first choice. Further clinical investigations are necessary to evaluate the role of rifampin in the treatment of human ehrlichiosis, especially in children.     

Regrowth of aminoglycoside-resistant variants and its possible implication for determination of MICs

Regrowth of aminoglycoside-resistant variants was seen when large inocula of two strains of Escherichia coli were incubated with gentamicin in concentrations well above their MICs (0.5 micrograms/ml). The extent of the selection of resistant variants was proportional to the concentration of gentamicin during incubation; after incubation with gentamicin (greater than or equal to 2 micrograms/ml for 24 h), all bacteria were resistant to at least 8 micrograms/ml. Bacteria resistant to these concentrations always formed small colonies, whereas variants resistant to lower concentrations (1 to 2 micrograms/ml) could form both small and normal colonies. The regrowth of resistant variants could be monitored by luciferase assay of intracellular ATP in cultures incubated with gentamicin (less than or equal to 2 micrograms/ml). In cultures incubated with higher concentrations, regrowth did occur, although this did not result in viability (CFU per milliliter) or ATP levels above those of the initial inocula. The implications of this regrowth for MIC determinations in broth and the possible clinical revelance of the resistant variants are discussed.