scholarly journals Prevalence of a Putative Efflux Mechanism among Fluoroquinolone-Resistant Clinical Isolates ofStreptococcus pneumoniae

1998 ◽  
Vol 42 (8) ◽  
pp. 2032-2035 ◽  
Author(s):  
Nigel P. Brenwald ◽  
Martin J. Gill ◽  
Richard Wise
Keyword(s):  
Ethidium Bromide ◽  
Efflux Pump ◽  
Clinical Isolates ◽  
Fluoroquinolone Resistance ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
The United Kingdom ◽  
Agar Dilution ◽  
Efflux Mechanism ◽  
Clinically Significant

ABSTRACT Twenty-three norfloxacin-selected first-step mutants ofStreptococcus pneumoniae showed low-level fluoroquinolone resistance. Their susceptibility to norfloxacin in the presence or absence of reserpine and known efflux pump substrates was determined by an agar dilution method. Five mutants showed four- to eightfold increases in their susceptibility to norfloxacin in the presence of reserpine and four- to eightfold decreases in their susceptibility to acriflavine and ethidium bromide. This phenotype is suggestive of an efflux mechanism of resistance. A representative of these mutants, 1N27, accumulated significantly less ethidium bromide than the parent strain; reserpine abolished these differences. No changes in the quinolone resistance-determining regions of parC,parE, gyrA, or gyrB were found in this mutant. By our validated agar dilution method, the efflux phenotype was sought in clinical isolates of S. pneumoniae. Of 1,037 clinical isolates examined from the United Kingdom, 273 showed reduced susceptibility to norfloxacin or ciprofloxacin. Of these, 45.4% showed the efflux phenotype. Our findings suggest that an efflux mechanism may be a frequent cause of clinically significant fluoroquinolone resistance in pneumococci.

scholarly journals Widespread Occurrence of Possible Antibiotic Efflux Mechanism in Multidrug Resistant Intestinal E. coli Strains Isolated from Healthy Human Subjects in Bangladesh

2016 ◽  
Vol 30 (1-2) ◽  
pp. 55-59
Author(s):  
M Hasibur Rahman ◽  
Jamil Mahmud ◽  
Md Mahamudul Haque ◽  
Farzana Tarannum Tuli ◽  
Nazneen Jahan ◽  
...  
Keyword(s):  
Ethidium Bromide ◽  
Efflux Pump ◽  
Human Subjects ◽  
Agar Plate ◽  
Multidrug Resistant ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Healthy Human ◽  
Efflux Mechanism ◽  
High Level

A total of 50 isolates of Escherichia coli obtained from healthy adult human subjects were studied and tested for possible presence of efflux mechanism in resistance determination and possible correlation of plasmids with resistance.Minimal inhibitory concentration (MIC) of the antibiotics amoxicillin, azithromycin, ciprofloxacin, chloramphenicol and tetracycline were determined by agar dilution method with or without the H+/K+ proton pump inhibitor omeprazole. Plasmids were extracted by rapid alkaline plasmid extraction method and analyzed by agarose gel electrophoresis. Many strains showed 5 – 10 fold reduction of MIC values in the presence of omeprazole; a few strains showed up to 100-fold MIC reduction. Plasmid analysis of these 50 isolates revealed the presence of both plasmidless and plasmid containing strains, the latter with plasmid number varying from one to seven. However, the plasmids apparently had no relationship with high level antibiotic tolerance as indicated by the observation that some plasmidless strains had very high MIC values, while other strains containing several plasmids had very low MIC. Decrease in MIC in the presence of omeprazole apparently indicates existence of an efflux mechanism. Evidence of the efflux of ethidium bromide was noted in some strains that had been grown in ethidium bromide containing agar plate with and without omeprazole. These results suggest that reduction of MIC caused by omeprazole may be related to possible inhibition of efflux pump activity by omeprazole in the isolates studied.Bangladesh J Microbiol, Volume 30, Number 1-2,June-Dec 2013, pp 55-59

scholarly journals Increasing antibiotic resistance in clinical isolates of Aeromonas strains in Taiwan.

1996 ◽  
Vol 40 (5) ◽  
pp. 1260-1262 ◽  
Author(s):  
W C Ko ◽  
K W Yu ◽  
C Y Liu ◽  
C T Huang ◽  
H S Leu ◽  
...  
Keyword(s):  
United States ◽  
Antibiotic Resistance ◽  
The United States ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Extended Spectrum ◽  
Agar Dilution

A total of 234 clinical isolates of Aeromonas, primarily A. hydrophila, were collected for the present study. Most were isolates from blood. By the agar dilution method, more than 90% of the Aeromonas strains were found to be susceptible to moxalactam, ceftazidime, cefepime, aztreonam, imipenem, amikacin, and fluoroquinolones, but they were more resistant to tetracycline, trimethoprim-sulfamethoxazole, some extended-spectrum cephalosporins, and aminoglycosides than strains from the United States and Australia.

scholarly journals In Vitro Activities of Garenoxacin (BMS-284756) against 170 Clinical Isolates of Nine Pasteurella Species

2002 ◽  
Vol 46 (9) ◽  
pp. 3068-3070 ◽  
Author(s):  
Ellie J. C. Goldstein ◽  
Diane M. Citron ◽  
C. Vreni Merriam ◽  
Yumi A. Warren ◽  
Kerin L. Tyrrell ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Active Agent ◽  
Culture Collection ◽  
Clinical Isolates ◽  
American Type Culture Collection ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Agar Dilution

ABSTRACT The in vitro susceptibilities of 170 clinical isolates plus 12 American Type Culture Collection strains of Pasteurella species comprising nine species and three Pasteurella multocida subspecies were studied by an agar dilution method. Garenoxacin (BMS-284756), a new des-fluoro(6) quinolone, was active at ≤0.06 μg/ml against all isolates, including four β-lactamase-producing strains, with >90% of the strains susceptible to ≤0.008 μg/ml. Garenoxacin was generally 1 to 2 dilutions more active than levofloxacin and moxifloxacin and was the most active agent tested. Cefoxitin required 1 μg/ml for inhibition of 51 of 182 (29%) of strains, and 3 strains (also β-lactamase producers) were resistant to doxycycline.

In vitro synergy testing of macrolide-quinolone combinations against 41 clinical isolates of Legionella.

1996 ◽  
Vol 40 (6) ◽  
pp. 1419-1421 ◽  
Author(s):  
S J Martin ◽  
S L Pendland ◽  
C Chen ◽  
P Schreckenberger ◽  
L H Danziger
Keyword(s):  
Yeast Extract ◽  
Antimicrobial Therapy ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Agar Dilution ◽  
Legionella Species ◽  
Checkerboard Assay ◽  
Synergy Testing

Combination antimicrobial therapy against Legionella species has not been well studied. Several quinolones have activity against Legionella strains, which prompted this in vitro search for a synergistic combination with the macrolides. By a checkerboard assay, erythromycin, clarithromycin, and azithromycin, each in combination with ciprofloxacin and levofloxacin, were tested for synergy against 46 isolates of Legionella. The agar dilution method was employed using buffered charcoal-yeast extract media. A final inoculum of 10(4) CFU per spot was prepared from 24-h growth of each isolate. Plates were incubated at 35 degrees C for 48 h. Synergy, partial synergy, additive effect, or indifference was observed for all combinations of antibiotics tested. There was no antagonism observed. Synergy occurred to a significantly greater extent for the clarithromycin-levofloxacin (P = 0.0001) and azithromycin-levofloxacin (P = 0.003) combinations versus erythromycin-levofloxacin. The azithromycin-ciprofloxacin combination demonstrated significantly greater synergy than did either erythromycin-ciprofloxacin (P = 0.003) or clarithromycin-ciprofloxacin (P = 0.001). The newer macrolides clarithromycin and azithromycin may be more active in combination with a fluoroquinolone than is erythromycin.

scholarly journals In Vitro Activities of Garenoxacin (BMS 284756) against 108 Clinical Isolates of Gardnerella vaginalis

2002 ◽  
Vol 46 (12) ◽  
pp. 3995-3996 ◽  
Author(s):  
Ellie J. C. Goldstein ◽  
Diane M. Citron ◽  
C. Vreni Merriam ◽  
Yumi A. Warren ◽  
Kerin L. Tyrrell ◽  
...  
Keyword(s):  
Clinical Isolates ◽  
Gardnerella Vaginalis ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Agar Dilution

ABSTRACT Garenoxacin (BMS 284756) was active against 105 of 108 (97%) recent clinical Gardnerella vaginalis isolates at ≤2 μg/ml by using the reference agar dilution method for anaerobes. Twenty-eight percent of isolates (31 of 108) were resistant to metronidazole, and 44% were resistant to doxycycline. All were susceptible to clindamycin and ampicillin-sulbactam.

scholarly journals Variability in Azithromycin Susceptibility Results for Neisseria gonorrhoeae Obtained Using Gradient MIC Strip and Agar Dilution Techniques

2019 ◽  
Vol 57 (12) ◽  
Author(s):  
Gary N. McAuliffe ◽  
Marian Smith ◽  
Gavin Cooper ◽  
Rose F. Forster ◽  
Sally A. Roberts
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Susceptibility Testing ◽  
Geometric Mean ◽  
Test Strip ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Content Type ◽  
Test Strips ◽  
Agar Dilution

ABSTRACT Azithromycin is a component of empirical treatment regimens for Neisseria gonorrhoeae infections, but antimicrobial susceptibility testing for this agent is technically challenging. We compared the intertest variability, MIC values, and CLSI/EUCAST categorization of clinical and reference isolates of N. gonorrhoeae treated with azithromycin by testing 107 clinical isolates and nine reference isolates by agar dilution and in duplicates using MIC test strips (Liofilchem, Italy) and Etests (bioMérieux, France). Replicate isolate agreement within 1 log2 between duplicate tests was 87% for MIC test strips and 100% for Etests (P < 0.001). Essential agreement with the agar dilution method was higher for Etests (91%) than for MIC test strips (44%, P < 0.001). The geometric mean MIC was highest for MIC test strips (0.8 mg/liter) and significantly higher than both Etest (0.47 mg/liter, P < 0.001) and agar dilution (0.26 mg/liter, P < 0.001) methods. Etest MICs were higher than those obtained with agar dilution (P < 0.001). Agar dilution, MIC test strip, and Etest methods categorized 96%, 85%, and 95% (P = 0.003) of clinical isolates, respectively, as susceptible/wild type according to CLSI/EUCAST criteria. Our results illustrate the difficulties underlying azithromycin susceptibility testing for N. gonorrhoeae and demonstrate that results can vary using different methods. This variability could influence antimicrobial resistance reporting between laboratories involved in N. gonorrhoeae surveillance programs.

scholarly journals In Vitro Activities of Dalbavancin and Nine Comparator Agents against Anaerobic Gram-Positive Species and Corynebacteria

2003 ◽  
Vol 47 (6) ◽  
pp. 1968-1971 ◽  
Author(s):  
Ellie J. C. Goldstein ◽  
Diane M. Citron ◽  
C. Vreni Merriam ◽  
Yumi Warren ◽  
Kerin Tyrrell ◽  
...  
Keyword(s):  
Clinical Evaluation ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Gram Positive ◽  
Enhanced Activity ◽  
Excellent Activity ◽  
Agar Dilution

ABSTRACT Dalbavancin is a novel semisynthetic glycopeptide with enhanced activity against gram-positive species. Its comparative in vitro activities and those of nine comparator agents, including daptomycin, vancomycin, linezolid, and quinupristin-dalfopristin, against 290 recent gram-positive clinical isolates strains, as determined by the NCCLS agar dilution method, were studied. The MICs of dalbavancin at which 90% of various isolates tested were inhibited were as follows: Actinomyces spp., 0.5 μg/ml; Clostridium clostridioforme, 8 μg/ml; C. difficile, 0.25 μg/ml; C. innocuum, 0.25 μg/ml; C. perfringens, 0.125 μg/ml; C. ramosum, 1 μg/ml; Eubacterium spp., 1 μg/ml; Lactobacillus spp., >32 μg/ml, Propionibacterium spp., 0.5 μg/ml; and Peptostreptococcus spp., 0.25 μg/ml. Dalbavancin was 1 to 3 dilutions more active than vancomycin against most strains. Dalbavancin exhibited excellent activity against gram-positive strains tested and warrants clinical evaluation.

An improved susceptibility test based on Amberlite reveals the potential antilisterial activity of fosfomycin in vitro

2013 ◽  
Vol 59 (4) ◽  
pp. 252-259 ◽  
Author(s):  
Lei Han ◽  
Jin'e Lei ◽  
Shaoshan Han ◽  
Li He ◽  
Chaofeng Ma ◽  
...  
Keyword(s):  
Brain Heart Infusion ◽  
Susceptibility Test ◽  
Host Cells ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

Listeria monocytogenes is resistant to fosfomycin in vitro but is susceptible in vivo due to increased expression of positive regulator factor A (PrfA) and its dependent factor, hexose phosphate transporter (Hpt), upon infection of host cells. Amberlite, a polymeric adsorbent resin, could induce PrfA-dependent gene expression and thus, in theory, improve the sensitivity of L. monocytogenes to fosfomycin in vitro. In the current study, an improved susceptibility test based on Amberlite was developed using reference strains. Thirty-five clinical isolates were further examined to verify those preliminary results. Briefly, Amberlite increased in vitro fosfomycin sensitivity of all strains. Optimal Amberlite concentrations, as evaluated through the expression of phospholipase B (PlcB) and Hpt, were 10% and 15% (w/v) in agar media and 3% (w/v) in broth media. Mueller–Hinton (MH) medium, tryptone soya (TS) medium, and brain heart infusion (BHI) medium were used to verify the results in the control strains using agar dilution and broth micro- and macro-dilution methods. Better listerial growth was shown in TS and BHI than in MH. Both broth dilution methods yielded lower minimal inhibitory concentration (MIC) of fosfomycin than the agar dilution method. The MIC of fosfomycin for 35 clinical isolates was 2–32 μg/mL, suggesting improved susceptibility. In conclusion, in vitro sensitivity of L. monocytogenes to fosfomycin was substantially improved in the presence of 3% Amberlite-supplemented TSB or BHIB and the broth microdilution method. This improved method revealed the potential antilisterial activity of fosfomycin in vitro and could facilitate the therapy of listeriosis using fosfomycin.

scholarly journals In Vitro Activities of 15 Antimicrobial Agents against 110 Toxigenic Clostridium difficile Clinical Isolates Collected from 1983 to 2004

2007 ◽  
Vol 51 (8) ◽  
pp. 2716-2719 ◽  
Author(s):  
David W. Hecht ◽  
Minerva A. Galang ◽  
Susan P. Sambol ◽  
James R. Osmolski ◽  
Stuart Johnson ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Antimicrobial Agents ◽  
Treatment Strategies ◽  
The United States ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
In Vitro Susceptibility ◽  
Agar Dilution

ABSTRACT The incidence and severity of Clostridium difficile-associated disease (CDAD) is increasing, and standard treatment is not always effective. Therefore, more-effective antimicrobial agents and treatment strategies are needed. We used the agar dilution method to determine the in vitro susceptibility of the following antimicrobials against 110 toxigenic clinical isolates of C. difficile from 1983 to 2004, primarily from the United States: doripenem, meropenem, gatifloxacin, levofloxacin, moxifloxacin, OPT-80, ramoplanin, rifalazil, rifaximin, nitazoxanide, tizoxanide, tigecycline, vancomycin, tinidazole, and metronidazole. Included among the isolates tested were six strains of the toxinotype III, NAP1/BI/027 group implicated in recent U.S., Canadian, and European outbreaks. The most active agents in vitro were rifaximin, rifalazil, tizoxanide, nitazoxanide, and OPT-80 with MICs at which 50% of the isolates are inhibited (MIC50) and MIC90 values of 0.0075 and 0.015 μg/ml, 0.0075 and 0.03 μg/ml, 0.06 and 0.125 μg/ml, 0.06 and 0.125 μg/ml, 0.125 and 0.125 μg/ml, respectively. However, for three isolates the rifalazil and rifaximin MICs were very high (MIC of >256 μg/ml). Ramoplanin, vancomycin, doripenem, and meropenem were also very active in vitro with narrow MIC50 and MIC90 ranges. None of the isolates were resistant to metronidazole, the only agent for which there are breakpoints, with tinidazole showing nearly identical results. These in vitro susceptibility results are encouraging and support continued evaluation of selected antimicrobials in clinical trials of treatment for CDAD.

scholarly journals Comparative In Vitro Activities of Retapamulin (SB-275833) against 141 Clinical Isolates of Propionibacterium spp., Including 117 P. acnes Isolates

2006 ◽  
Vol 50 (1) ◽  
pp. 379-381 ◽  
Author(s):  
Ellie J. C. Goldstein ◽  
Diane M. Citron ◽  
C. Vreni Merriam ◽  
Yumi A. Warren ◽  
Kerin L. Tyrrell ◽  
...  
Keyword(s):  
Active Agent ◽  
Vitro Activity ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
In Vitro Activity ◽  
Agar Dilution ◽  
Multiresistant Strains

ABSTRACT Using the NCCLS agar dilution method, we studied the in vitro activity of retapamulin (SB-275833) against 141 clinical isolates of Propionibacterium species, including seven multiresistant strains, and found retapamulin to be the most active agent among those tested with MICs of ≤1 μg/ml against all isolates.

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